From SJones <@t> cvm.tamu.edu Sun Aug 1 14:11:12 2004 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] transferring paraffin ribbon Message-ID: Hi Jackie, I use a wooden applicator stick that I angle cut, at the tip, with a single edge razor blade, making a fine point. I dip the tip of the stick into my waterbath before touching the ribbon and the water makes the ribbon adhere to the stick. I use them for both the beginning and the end of the ribbon. When doing serial sections, it's difficult to save every section if you need more than one ribbon. At the best of times, I'll usually lose one section at the beginning and one at the end. If I have trouble getting the ribbon started, I may lose even more than that. If researchers are doing 3D reconstruction, I just keep a record of where sections were lost and how many sections were lost. I'm doing that right now with some laser induced retinal lesions in the eye of the rat. If you need thin sections (3-5 microns), the blocks need to be cold. If the block isn't cold, the ribbon will be compressed. I always cut thin sections off of an ice tray. I use the dissecting pans (without wax) available from Fisher for my ice trays. If you are doing thicker sections, you can cut at room temperature. I once had a disposable knife holder where the sections would stick to the metal. I ended up putting a piece of wide, clear, plastic packaging tape over it and that kept the ribbon from sticking to it. You might try Paraguard too, a spray used for keeping paraffin from sticking to surfaces. It has a nasty smell that makes me nauseous so I don't use it myself. Also, make sure there isn't any water on the face of the block or on the knife before you start a ribbon. Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Jacqueline Miller" 7/30/2004 2:16:00 PM >>> Hi everyone, I would like to know about the techniques that histotechs use to cut paraffin sections--especially how the ribbon is transferred from microtome to water bath while preserving the first and last sections. I've been sectioning paraffin blocks for almost 4 years now. However, I've never needed to cut serial sections and the samples were always large. So, I never worried about losing the first section of each ribbon (and sometimes the last). I would like to know what techniques the histologists out there use to transfer the ribbon from the microtome to the water bath. I've used metal forceps and my fingers (gloved and not gloved) to grab the first section (but the section sticks to the forceps), and I'm using the wooden part of a cotton swab to lift off the last section, which is working fairly well. I'm also using a new microtome, Leica RM2235, and having some trouble with the first section coming off the knife just wrinkling up. And, the surface below the knife is such that even if I grab the edge of the section with a paintbrush, it won't budge. Is there anything I can do to correct these problems? Also, which is preferred, to cut the blocks chilled or to leave them RT. I used to cut blocks chilled all the time, but here it's preferred that I cut them RT unless I have a problem. Thanks, Jackie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jengirl1014 <@t> yahoo.com Mon Aug 2 06:47:33 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] MAC-387 Message-ID: <20040802114733.31059.qmail@web60605.mail.yahoo.com> A guy in my lab has asked me to do some MAC-387 staining. Has anyone heard of it? Could someone please tell me what is used and what the protocol is? It would be greatly appreciated. Thanks in advanced! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. From MAUGER <@t> email.chop.edu Mon Aug 2 07:05:49 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] MAC-387 Message-ID: Hi Jennifer, We use this marker for macrophages,monocytes. We get it from Dako #M0747. The dilution that's best for us is 1:200, and we use pepsin digestion for Ag retrieval. It is a mouse monoclonal Ab. Jo >>> "Jennifer Sipes" 08/02/04 07:47AM >>> A guy in my lab has asked me to do some MAC-387 staining. Has anyone heard of it? Could someone please tell me what is used and what the protocol is? It would be greatly appreciated. Thanks in advanced! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From settembr <@t> umdnj.edu Mon Aug 2 07:58:51 2004 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Negative Control Reagents Message-ID: Is there anyone out there who is running negative controls that are: 1. Species specific? What are you using? 2. Isotype specific? What are you using? 3. Running the negative control reagents at the same protein concentration as each and every antibody? Any response from anyone and especially CAP guideline administrators, would be appreciated. Dana Settembre University Hospital Newark, NJ USA From akwilliams75 <@t> hotmail.com Mon Aug 2 10:10:24 2004 From: akwilliams75 <@t> hotmail.com (kevin williams) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Resubscribeing Message-ID: I am trying to resubscribe, I was thrown out for bouncing, sorry, In the future I guess all I have to do is unsubscribe to stop that happening when I am away? Please put me back on. Yours faithfully, A Kevin Williams _________________________________________________________________ [1]Dont just search. Find. Check out the new MSN Search! References 1. http://g.msn.com/8HMBENUS/2731??PS=47575 From cfavara <@t> niaid.nih.gov Mon Aug 2 10:21:53 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] bleaching Message-ID: I need to bleach the retinal pigmented epithelial layer of some B6 mice. I have searched the archives and the literature have some interesting prospects. Would like to know if anyone has any hands on experience. Most interesting was from some one working at the AFIOP that bleached section in ribbon form. Any guidance would be greatly appreciated! Cynthia Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From ftulenko06 <@t> jcu.edu Mon Aug 2 10:32:45 2004 From: ftulenko06 <@t> jcu.edu (ftulenko06@jcu.edu) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] thanks Message-ID: <80def67b.699ce62b.8275200@mirapoint.jcu.edu> Thanks to everybody who responded to my question. Your advice is appreciated. Frank From Dixon.Leslie <@t> mayo.edu Mon Aug 2 11:37:55 2004 From: Dixon.Leslie <@t> mayo.edu (Dixon, Leslie E.) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] blocking charges Message-ID: <115ED45FBA6D3D4B89E7F40A9EB5339D90B9D9@excsrv60.mayo.edu> Good morning, We are currently looking at our charges in comparison with others. I was hoping that those in the animal research field could help me out. I know some of this is confidential so a personal e-mail would be fine and I would delete your name from the information for my research. I am looking for the charges to take a wet tissue through processing to an H&E. Also, the charges for pathologic interpretation if requested. Thank you in advance for your help. Thanks, Leslie Dixon From asmith <@t> mail.barry.edu Mon Aug 2 12:04:30 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] DAB fading/pH range Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3BA9@exchsrv01.barrynet.barry.edu> Spend $12 on a little bottle of "Permount" (resinous mounting medium)! HSR is just as good, but harder to find. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Melissa P Wu Sent: Friday, July 30, 2004 1:06 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] DAB fading/pH range Hi, I know this has been addressed in previous posts but I haven't been able to find an answer to this question specifically. What is the pH range that the DAB stain is stable in? Supporting info: I am using DAB (sigma, tablet form) on Drosophila ovaries (enhanced with Nickel). After about 8 minutes of the reaction, the tissues will stain black. I mount in 70% glycerol/PBS (pH 7.2), and after a day the stain fades or disappears. I also tried 90% glycerol/PBS, but the stain faded here as well. I read somewhere that the nailpolish could affect the staining, so I stored one sample in 70% glycerol/PBS, but the color faded in these samples after a day. I called Sigma and they suggested using a resin mount, but I don't have access to that so I used Vectashield mounting media, the sample's color faded with this one also. Also, a side question. This is my lab's first time using HRP reactions, I tried one on a control sample and let it incubate for about 20 minutes in the DAB reaction. I was surprised to see the tissue stain black. I am pretty sure it is not reacting to endogenous HRP as I used methanol earlier to destroy the activity. I am also pretty sure it is not due to the primary antibody non-specifically binding, as its antigen is BrdU. Is it just background staining that I am seeing, or perhaps my sample was contaminated? Any help with this situation would be appreciated. Thanks, Melissa Wu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet /listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From laurie.colbert <@t> huntingtonhospital.com Mon Aug 2 13:09:25 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Lymph Node Revealing Solution Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BFCE@EXCHANGE1.huntingtonhospital.com> Someone told one of my pathologists about a lymph node revealing solution that is made by Sigma than can be disposed of down the drain. I called Sigma, and no one there knows what I'm talking about. Has anyone out there heard of this product? Laurie Colbert Huntington Memorial Hospital Pasadena, CA From cleger <@t> seattlecca.org Mon Aug 2 13:13:25 2004 From: cleger <@t> seattlecca.org (Leger, Carolyn A) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] PAS Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B94841BAB9@wala01.seattlecca.org> I am comparing our made in-home Schiffs stain with the commercially made Schiffs. Ours is clearly more vibrant then the Richard-Allen Schiffs. Is there another brand that anyone is using that has good quality especially on bone marrows? Also is there anything that can be added to the commerically bought Schiffs that would make it more vibrant. Thanks Carolyn A. Leger Pathology Department 206/288-2232 206/288-1355 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please notify me by telephone at (206) 288-2232 or by electronic reply, and delete this message. Thank you This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From mari.ann.mailhiot <@t> leica-microsystems.com Mon Aug 2 13:37:27 2004 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] PAS Message-ID: Carolyn Try ordering Schiff's from Surgipath. When I used it in the lab I had great results! Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com "Leger, Carolyn A" To: "'histonet@lists.utsouthwestern.edu'" Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] PAS western.edu 08/02/2004 01:13 PM I am comparing our made in-home Schiffs stain with the commercially made Schiffs. Ours is clearly more vibrant then the Richard-Allen Schiffs. Is there another brand that anyone is using that has good quality especially on bone marrows? Also is there anything that can be added to the commerically bought Schiffs that would make it more vibrant. Thanks Carolyn A. Leger Pathology Department 206/288-2232 206/288-1355 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please notify me by telephone at (206) 288-2232 or by electronic reply, and delete this message. Thank you This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From JQB7 <@t> CDC.GOV Mon Aug 2 13:42:49 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] PAS Message-ID: Newcomer Supply has a great Schiff's. It's very strong and has a long, room temperature shelf life. Jeanine Bartlett CDC, Atlanta -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Leger, Carolyn A Sent: Mon 8/2/2004 2:13 PM To: 'histonet@lists.utsouthwestern.edu' Cc: Subject: [Histonet] PAS I am comparing our made in-home Schiffs stain with the commercially made Schiffs. Ours is clearly more vibrant then the Richard-Allen Schiffs. Is there another brand that anyone is using that has good quality especially on bone marrows? Also is there anything that can be added to the commerically bought Schiffs that would make it more vibrant. Thanks Carolyn A. Leger Pathology Department 206/288-2232 206/288-1355 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please notify me by telephone at (206) 288-2232 or by electronic reply, and delete this message. Thank you This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Mon Aug 2 13:47:06 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] PAS In-Reply-To: <5E6BFDF4F0AB2C4DA69CF4473FC7B94841BAB9@wala01.seattlecca.org> Message-ID: Carolyn, In my experience, homemade always is far better than any commercially prepared solution. At least you know how long the solution has been sitting around and what's in it. We use Anapath's Schiff's from StatLab Medical products 1-800-442-3573. In my opinion (and you know how everyone feels about opinions) this is the closest to homemade that I saw. Once again, the views of this author do not reflect the views of the employer. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Leger, Carolyn A Sent: Monday, August 02, 2004 1:13 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] PAS I am comparing our made in-home Schiffs stain with the commercially made Schiffs. Ours is clearly more vibrant then the Richard-Allen Schiffs. Is there another brand that anyone is using that has good quality especially on bone marrows? Also is there anything that can be added to the commerically bought Schiffs that would make it more vibrant. Thanks Carolyn A. Leger Pathology Department 206/288-2232 206/288-1355 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please notify me by telephone at (206) 288-2232 or by electronic reply, and delete this message. Thank you This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From algranth <@t> u.arizona.edu Mon Aug 2 14:58:55 2004 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] PAS In-Reply-To: References: <5E6BFDF4F0AB2C4DA69CF4473FC7B94841BAB9@wala01.seattlecca.org> Message-ID: <4.3.2.7.2.20040802125549.00c7bba8@algranth.inbox.email.arizona.edu> I've tried several commercial Schiff's and I'd like to second the vote for the Newcomer Supply Schiff's. Haven't tried the StatLab solution but I may have to check into that one. Andi Grantham At 01:47 PM 8/2/2004 -0500, Joe Nocito wrote: >Carolyn, > In my experience, homemade always is far better than any commercially >prepared solution. At least you know how long the solution has been sitting >around and what's in it. We use Anapath's Schiff's from StatLab Medical >products 1-800-442-3573. In my opinion (and you know how everyone feels >about opinions) this is the closest to homemade that I saw. Once again, the >views of this author do not reflect the views of the employer. > >Joe Nocito, BS, HT(ASCP) QIHC >Histology Manager >Pathology Reference Lab >San Antonio, TX > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Leger, >Carolyn A >Sent: Monday, August 02, 2004 1:13 PM >To: 'histonet@lists.utsouthwestern.edu' >Subject: [Histonet] PAS > >I am comparing our made in-home Schiffs stain with the commercially made >Schiffs. Ours is clearly more vibrant then the Richard-Allen Schiffs. Is >there another brand that anyone is using that has good quality especially on >bone marrows? Also is there anything that can be added to the commerically >bought Schiffs that would make it more vibrant. Thanks > >Carolyn A. Leger >Pathology Department >206/288-2232 >206/288-1355 >This electronic message transmission contains information which may be >confidential or privileged. The information is intended to be for the use of >the individual or entity named above. If you are not the intended recipient, >be aware that any disclosure, copying, distribution or use of the contents >of this information is prohibited. If you have received this electronic >transmission in error, please notify me by telephone at (206) 288-2232 or by >electronic reply, and delete this message. Thank you > > > > > This electronic message transmission contains information which may be >confidential or privileged. The information is intended to be for the >use of the individual or entity named above. If you are not the >intended recipient, be aware that any disclosure, copying, distribution >or use of the contents of this information is prohibited. If you have >received this electronic transmission in error, please leave a message >via telephone at (206) 288-6266, notify me by electronic reply, and >delete this message. Opinions and ideas in this message that do not >relate to official business are understood as neither given nor >endorsed by the Seattle Cancer Care Alliance. To view our complete >Notice of Privacy Practices, visit our web site at www.seattlecca.org. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From gcallis <@t> montana.edu Mon Aug 2 14:59:15 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] PAS and how you do the staining Message-ID: <6.0.0.22.1.20040802131137.01b2a338@gemini.msu.montana.edu> We tested commercial against in house preparation and found NO difference in staining quality. We do not use kits since buying Schiffs and making up Periodic acid is very minimal for cost and time. In general, Scjhiffs is stable but it MUST be tested. We purchase our Schiffs from Fisher, and store it in the refrigerator, although mfr say it is stable at Room temperature. We stopped making it in house to eliminate carcinogenic dye weighing plus having to filter the final solution - in our estimation time consuming and messy and without any staining difference. Have used Surgipath, Sigma, Fisher's - they all worked well. Vibrancy of staining could be linked to more than just the Schiff's reagent. After a lecture from the famous Culling, a true expert in PAS staining, how we did performed the PAS stain changed forever. 1) Periodic acid is NOT stable forever, Culling advised make this oxidizing reagent fresh each time for PAS protocol. His premise, this solution was cheap and one must insure good, consistent oxidation. 1% periodic acid is common, time 5 - 10 min. Some use 0.5% for 5 - 10 min. Overoxidation will affect results, not a bright, pink-red. Periodic acid freshness issue is also addressed in Hrapchak and Sheehans Theory and Practice of Histotechnology, plus they also advised weekly changes of Schiffs. 2) Do you reuse the Schiffs reagent without testing it? Test is 10 ml formaldehyde,(37%) add few drops of Schiffs, watch it turn red-purple. If you have used Schiffs, it should be stored in a separate bottle labeled accordingly. Do not pour it back into unused stock. Change it regularly (weekly per Hrapchak and Sheehan). If it has a hint of pink to it, discard. Do not allow to freeze. 3) Thin 1 - 3 um sections need longer time in both oxidizer and Schiffs. 4) Culling did not bother with sulfurous acid rinses, he just rinsed 10 min in running tap water. If you use sulfurous acid rinses, running tap water rinse afterwards will intensify color. 5) time in Schiffs reagent can vary from 10 to 30 min, we often do 15 and/or 20 min. This time can vary with strength and age of Schiffs reagent. We made Schiff using pararosaninline hydrochloride - the pure form of basic fucshin mixture). Strength could be linked to dye content of Basic fuchsin or pararosaniline hydrochloride or if mfr has changed concentration of these dyes when making up their product. It is a good idea to go back and look at how Schiffs is made, including dye concentration versus what you are using commercially. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From JQB7 <@t> CDC.GOV Mon Aug 2 15:15:36 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] PAS and how you do the staining Message-ID: Gayle makes some good points: I never reuse the Schiff's and I make fresh periodic acid every 6 months. I also test the Schiff's as I approach the month before the expiration date. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Gayle Callis Sent: Mon 8/2/2004 3:59 PM To: Histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] PAS and how you do the staining We tested commercial against in house preparation and found NO difference in staining quality. We do not use kits since buying Schiffs and making up Periodic acid is very minimal for cost and time. In general, Scjhiffs is stable but it MUST be tested. We purchase our Schiffs from Fisher, and store it in the refrigerator, although mfr say it is stable at Room temperature. We stopped making it in house to eliminate carcinogenic dye weighing plus having to filter the final solution - in our estimation time consuming and messy and without any staining difference. Have used Surgipath, Sigma, Fisher's - they all worked well. Vibrancy of staining could be linked to more than just the Schiff's reagent. After a lecture from the famous Culling, a true expert in PAS staining, how we did performed the PAS stain changed forever. 1) Periodic acid is NOT stable forever, Culling advised make this oxidizing reagent fresh each time for PAS protocol. His premise, this solution was cheap and one must insure good, consistent oxidation. 1% periodic acid is common, time 5 - 10 min. Some use 0.5% for 5 - 10 min. Overoxidation will affect results, not a bright, pink-red. Periodic acid freshness issue is also addressed in Hrapchak and Sheehans Theory and Practice of Histotechnology, plus they also advised weekly changes of Schiffs. 2) Do you reuse the Schiffs reagent without testing it? Test is 10 ml formaldehyde,(37%) add few drops of Schiffs, watch it turn red-purple. If you have used Schiffs, it should be stored in a separate bottle labeled accordingly. Do not pour it back into unused stock. Change it regularly (weekly per Hrapchak and Sheehan). If it has a hint of pink to it, discard. Do not allow to freeze. 3) Thin 1 - 3 um sections need longer time in both oxidizer and Schiffs. 4) Culling did not bother with sulfurous acid rinses, he just rinsed 10 min in running tap water. If you use sulfurous acid rinses, running tap water rinse afterwards will intensify color. 5) time in Schiffs reagent can vary from 10 to 30 min, we often do 15 and/or 20 min. This time can vary with strength and age of Schiffs reagent. We made Schiff using pararosaninline hydrochloride - the pure form of basic fucshin mixture). Strength could be linked to dye content of Basic fuchsin or pararosaniline hydrochloride or if mfr has changed concentration of these dyes when making up their product. It is a good idea to go back and look at how Schiffs is made, including dye concentration versus what you are using commercially. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rockbeki <@t> ufl.edu Mon Aug 2 15:41:48 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] sheep placenta histology Message-ID: <412562421.1091479308597.JavaMail.osg@spnode33> Hey all! I'm sorry to be bugging y'all again, but I've actually finished my sheep placenta eNOS slides (with much thanks to y'all for your advice)and now I'm trying to learn more about the histology of sheep placenta so I can describe what I'm looking at in the slides more completely. Anybody know of a book or anything that has good pictures and descriptions of slides of sheep placenta? Thanks, Rebekah Smith -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From RFail <@t> Charleston.net Mon Aug 2 18:09:53 2004 From: RFail <@t> Charleston.net (Rena) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Schiff's Message-ID: <000001c478e5$d2f8a150$da11a6a5@renad4yk9b8abe> Dr. McManus worked here at the medical University of South Carolina, when I started as a tech. He said the commercial was every bit as good as the homemade. I prefer the 4% from Sigma Rena Fail From histolog <@t> fcv.unl.edu.ar Mon Aug 2 18:38:06 2004 From: histolog <@t> fcv.unl.edu.ar (=?iso-8859-1?Q?Laboratorio_de_Histolog=EDa?=) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Secondary Antibodies Message-ID: <000e01c478e9$c381e700$0100a8c0@casa> I wanted to know if some of you uses the following dogs, and to that dilution uses them in immunohistochemistry. Biotin-Goat anti-Rabbit IgG (Zymed 65-6140 Biotin-Goat anti-Mouse (Chemicon AP181B) Thanks Dr. Hugo H. Ortega (DMV, PhD) Departament of Cellular Biology Faculty of Veterinary Sciences Universidad Nacional del Litoral R.P. Kreder 2805 - Esperanza (3080) Santa Fe - ARGENTINA Tel. (54)3496-420639 Fax. (54)3496-426304 http://fcv.unl.edu.ar/histolog/ http://fcv.unl.edu.ar/bioterio/ From histolog <@t> fcv.unl.edu.ar Mon Aug 2 18:40:53 2004 From: histolog <@t> fcv.unl.edu.ar (=?iso-8859-1?Q?Laboratorio_de_Histolog=EDa?=) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] =?iso-8859-1?q?Secondary_Antibodies_=28Sorry_by_the_?= =?iso-8859-1?q?=A8Dogs=29=2E=2E?= Message-ID: <001301c478ea$2963c020$0100a8c0@casa> I wanted to know if some of you uses the following Antibodies , and to that dilution uses them in immunohistochemistry. Biotin-Goat anti-Rabbit IgG (Zymed 65-6140 Biotin-Goat anti-Mouse (Chemicon AP181B) Thanks Dr. Hugo H. Ortega (DMV, PhD) Departament of Cellular Biology Faculty of Veterinary Sciences Universidad Nacional del Litoral R.P. Kreder 2805 - Esperanza (3080) Santa Fe - ARGENTINA Tel. (54)3496-420639 Fax. (54)3496-426304 http://fcv.unl.edu.ar/histolog/ http://fcv.unl.edu.ar/bioterio/ From histolog <@t> fcv.unl.edu.ar Mon Aug 2 18:41:31 2004 From: histolog <@t> fcv.unl.edu.ar (=?iso-8859-1?Q?Laboratorio_de_Histolog=EDa?=) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Secondary Antibodies (Sorry by the "Dogs" error in translation).. Message-ID: <001801c478ea$3d96f440$0100a8c0@casa> I wanted to know if some of you uses the following Antibodies , and to that dilution uses them in immunohistochemistry. Biotin-Goat anti-Rabbit IgG (Zymed 65-6140 Biotin-Goat anti-Mouse (Chemicon AP181B) Thanks Dr. Hugo H. Ortega (DMV, PhD) Departament of Cellular Biology Faculty of Veterinary Sciences Universidad Nacional del Litoral R.P. Kreder 2805 - Esperanza (3080) Santa Fe - ARGENTINA Tel. (54)3496-420639 Fax. (54)3496-426304 http://fcv.unl.edu.ar/histolog/ http://fcv.unl.edu.ar/bioterio/ From WWmn916 <@t> aol.com Mon Aug 2 21:12:49 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Recycle alcohol, xylene, americlear Message-ID: <7e.54d4fe25.2e404ea1@aol.com> Greetings everyone, I'm looking for a unit to recycle used alcohol, xylene, americlear. I can't seem to find info on line.....perhaps I'm not using a good keyword. Anyone know of a place I can find infomation, cost, specs....etc.? Thanks! Deb King, HT(ASCP) Sacramento,CA From WWmn916 <@t> aol.com Mon Aug 2 21:15:11 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] ShurmarkPlus slide cassette labeler Message-ID: <199.2cb42461.2e404f2f@aol.com> Hello everyone, Does anyone have experience with the slide/cassette labeling unit from Shurmark? I'm interested in real world experience/comments/opinions. Thanks, Deb King, HT(ASCP) Sacramento, CA From WWmn916 <@t> aol.com Mon Aug 2 21:24:08 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Fume hoods Message-ID: <1ec.269dbb1f.2e405148@aol.com> Hello again! Can anyone lead me in a direction in finding information about fume hoods (or type of ventilation systems that can cover a sink area and draw-up fumes? I'm not exactly sure how to describe what I need. (I don't think I need a fume hood that is a countertop model. I need to include a sink area.) Most of the fume hoods I see in supply catalogs are countertop in nature. The exhaust system I'm thinking of would drop down from a ceiling and cover a sink area. Any help is appreciated. Thanks, Deb King, HT(ASCP) From mira <@t> sctimst.ker.nic.in Mon Aug 2 21:30:13 2004 From: mira <@t> sctimst.ker.nic.in (Dr Mira Mohanty) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Removal of polymer particles from tissue Message-ID: Is there a method to remove polymer paticles less than 5 micron size from cryostat sections of tissue. Tried using a laser dissection microscope, but the laser burnt off the polymer. The particle has to be removed very carefully for further analysis. Hence, tissue cannot be digested and particles sieved. Mira From int09018 <@t> alphahunt.com Tue Aug 3 00:08:46 2004 From: int09018 <@t> alphahunt.com (HCS) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] looking for reference for a PAS/Mucicarmine stain Message-ID: <000701c47917$f51af5d0$6501a8c0@hp> Does someone have a procedure for a PAS/Mucicarmine stain? This is for a paraffin section. Thanks, LeRoy Brown HCS Everson, WA 98247 360-966-7300 From mira <@t> sctimst.ker.nic.in Tue Aug 3 01:25:47 2004 From: mira <@t> sctimst.ker.nic.in (Dr Mira Mohanty) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] removal of polymer particles from tissue Message-ID: Is there a method to remove polymer paticles less than 5 micron size from cryostat sections of tissue. Tried using a laser dissection microscope, but the laser burnt off the polymer. The particle has to be removed very carefully for further analysis. Hence, tissue cannot be digested and particles sieved. Mira From rentonlf <@t> bru.wits.ac.za Tue Aug 3 02:13:18 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] D-limonene Message-ID: <1091517198.9401c420rentonlf@bru.wits.ac.za> Hi all, I'd like to get an update on how many labs out there are currently using a xylene substitute, particularly limonene, for processing. It may be better to reply to me personally so as not to clog up the histonet, then I'll collate the info and post it later. Many, many thanks Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From pengbw <@t> sjtu.edu.cn Tue Aug 3 02:26:22 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] difference between wright-giemsa &giemsa staining Message-ID: <20040803072622.E1BC21105B4F@sjtu.edu.cn> Hi, all Would anyone please tell me the difference between wright-giemsa & giemsa staining. I want to use one of the method to study the morphology of monocytes, which should I choose? And hopefully, tell me the protocol. Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai,200030 China From carl.hobbs <@t> kcl.ac.uk Mon Aug 2 13:34:47 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] re Cryosectioning, fixing and H&E staining Message-ID: <001301c478bf$6400fed0$cf949a51@home> Completely agree with Gayle..... folow Gayle's advice. In my experience, omit Glutaraldhehyde ; this really messes up achieving a good H&E easily. You don't need it if you are only doing H&Es, anyway. If you want a "crisper" result, use Bouin or Carnoy, for example. . --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.726 / Virus Database: 481 - Release Date: 22/07/2004 From gcallis <@t> montana.edu Tue Aug 3 08:59:31 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Secondary Antibodies dilutions In-Reply-To: <001801c478ea$3d96f440$0100a8c0@casa> References: <001801c478ea$3d96f440$0100a8c0@casa> Message-ID: <6.0.0.22.1.20040803075317.01b29d40@gemini.msu.montana.edu> A rule of thumb is to use 2 - 10 ug/ml, do the calculation from concentration of antibody found as mg/ml in the specification sheets. Some people even use a lesser concentration - we have done dilution panels on our secondaries at times to determine the optimal concentration needed in ug/ml. We tend to average around 1:250 dilution on 0.5mg/ml concentration. Specification sheets usually give the recommended dilution for immunohistochemistry usage. At 05:41 PM 8/2/2004, you wrote: >I wanted to know if some of you uses the following Antibodies , and to >that dilution uses them in immunohistochemistry. > >Biotin-Goat anti-Rabbit IgG (Zymed 65-6140 > >Biotin-Goat anti-Mouse (Chemicon AP181B) > >Thanks > >Dr. Hugo H. Ortega (DMV, PhD) > >Departament of Cellular Biology > >Faculty of Veterinary Sciences > >Universidad Nacional del Litoral > >R.P. Kreder 2805 - Esperanza (3080) > >Santa Fe - ARGENTINA > >Tel. (54)3496-420639 > >Fax. (54)3496-426304 > > http://fcv.unl.edu.ar/histolog/ > > http://fcv.unl.edu.ar/bioterio/ > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From mbryhan <@t> NORTHERNHEALTH.ORG Tue Aug 3 09:30:09 2004 From: mbryhan <@t> NORTHERNHEALTH.ORG (Mary Bryhan) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] testing Message-ID: testing From GAshton <@t> PICR.man.ac.uk Tue Aug 3 10:27:30 2004 From: GAshton <@t> PICR.man.ac.uk (Garry Ashton) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] XIAP Message-ID: Dear Histonetters, I'm having problems using the above antibody from various sources. I'm getting non-specific staining on my knock out mice specimens, with no true positive staining seen in any of the WT mice. Firstly has anybody out there used a XIAP antibody on FFPE sections, and if so which one of the several available, (or even on frozens). Secondly what is the best method of preventing non specific staining of the antibody. In the past I've made the antibody in 1% casein or normal serum from which the secondary is made, after applying a 10% solution to the sections prior to the antibody incubation. I've also tried various retrieval methods. Many thanks in advance for any much needed help. Garry PICR UK -------------------------------------------------------- This email is confidential and intended solely for the use of the person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute for Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From histo007 <@t> hotmail.com Tue Aug 3 11:37:05 2004 From: histo007 <@t> hotmail.com (Jim Ball) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Digest Message-ID: _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfeeŽ Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From lteves <@t> uhnres.utoronto.ca Tue Aug 3 15:17:07 2004 From: lteves <@t> uhnres.utoronto.ca (Lucy Teves) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] scanners Message-ID: <410FF2C3.EF127EFA@uhnres.utoronto.ca> Hello, I'm looking for a scanner that will scan whole mounted histological samples on regular slides. I would like to show whole rat brain. Which product do you recommend? Thanks in advance -- Lucy Teves Lab Manager Neuroprotection Lab. From lteves <@t> uhnres.utoronto.ca Tue Aug 3 15:17:19 2004 From: lteves <@t> uhnres.utoronto.ca (Lucy Teves) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] scanners Message-ID: <410FF2CF.AC2D99A8@uhnres.utoronto.ca> Hello, I'm looking for a scanner that will scan whole mounted histological samples on regular slides. I would like to show whole rat brain. Which product do you recommend? Thanks in advance -- Lucy Teves Lab Manager Neuroprotection Lab. From lteves <@t> uhnres.utoronto.ca Tue Aug 3 15:17:24 2004 From: lteves <@t> uhnres.utoronto.ca (Lucy Teves) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] scanners Message-ID: <410FF2D4.1FB22ECC@uhnres.utoronto.ca> Hello, I'm looking for a scanner that will scan whole mounted histological samples on regular slides. I would like to show whole rat brain. Which product do you recommend? Thanks in advance -- Lucy Teves Lab Manager Neuroprotection Lab. From pengbw <@t> sjtu.edu.cn Tue Aug 3 20:34:03 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Thanks Message-ID: <20040804013403.6F80C1113A9D@sjtu.edu.cn> Dear Robert Krug Thank you detailed explaintation of difference between the several Giemsa stain. I appreciate it very much. Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From kennedya <@t> email.cs.nsw.gov.au Tue Aug 3 22:05:05 2004 From: kennedya <@t> email.cs.nsw.gov.au (Andrew Kennedy) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Recycle alcohol, xylene, americlear Message-ID: <200408041301667.SM01312@crgcsls813> Hi Deb, For info on recyclers, try these web sites: http://www.brinstrument.com/ http://www.cbgbiotech.com Andrew Kennedy Senior Science Officer Anatomical Pathology Concord Repatriation General Hospital Hospital Road Concord NSW 2139 ph: +612 9767 6115 Fax +612 9767 8427 "corpora non agunt nisi fixata" From AnthonyH <@t> chw.edu.au Tue Aug 3 22:58:23 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Thanks Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E22D@simba.kids> Unfortunately, no one else on the Server saw it. Just a friendly reminder to reply to all when you answer a posting so that everyone else can become involved in the discussion if they wish. Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] Sent: Wednesday, 4 August 2004 11:34 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Thanks Dear Robert Krug Thank you detailed explaintation of difference between the several Giemsa stain. I appreciate it very much. Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From David.Edmondson <@t> christie-tr.nwest.nhs.uk Wed Aug 4 06:08:55 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] paraffin blocks Message-ID: Hi, Did you have any replies? Our files go back sixty or seventy years, so the issue has not arisen. I imagine incineration to be the "normal" course of events. That is how the left overs from the macro are handled. Recording the blocks/samples disposed of would be important along with someones signature. A decision made that the patients are deceased and never to need the material reviewed. etc David Histology Christie Hosp Manchester -----Original Message----- From: Kim.Kolman@med.va.gov [mailto:Kim.Kolman@med.va.gov] Sent: 29 July 2004 20:25 To: convmcm@cc.usu.edu; ernestinemiddleton@yahoo.ca; histonet@pathology.swmed.edu Subject: RE: [Histonet] paraffin blocks Did I miss all the replies on this subject? I was looking forward to reading all the reponses; my lab is currently in a dilemma about how to handle disposal of these items. Afraid to share/admit? Kim Kolman, HT (ASCP) VA Eastern Kansas Health Care System Eisenhower VA Medical Center Leavenworth, Ks. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, July 27, 2004 9:37 AM To: 'Ernestine Middleton'; histonet@pathology.swmed.edu Subject: RE: [Histonet] paraffin blocks We have our own incinerator. That's where all this stuff goes. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ernestine Middleton Sent: Monday, July 26, 2004 6:03 PM To: histonet@pathology.swmed.edu Subject: [Histonet] paraffin blocks Hi; I like to know how do other Histology Lab. get rid of paraffin blocks that are 20+years. Do you have a medical waste company destroy them or do you do it your self? Thank you for answer me as quickly as possible. Ernestine Middleton, Manager Montefiore Med. Ct. Bronx, NY 914-920-4157 --------------------------------- Post your free ad now! Yahoo! Canada Personals _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pam <@t> ategra.com Wed Aug 4 08:06:49 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Job Opportunities in Histology Latest Update 08/04/04 Message-ID: Hi Histonetters, Here is the latest update on opportunities with some of my best clients throughout the US who are seeking Histology Supervisors, Histo Technologists and Histo Technicians. These are positions as direct employees of our client. There are fulltime 40 hour per week positions.. As a direct employee of one of our clients you will be provided with full benefits including Health Insurance, Vacation, Sick Pay, Relocation money and a lucrative sign-on bonus. I have part time and temporary positions as well. I have supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are some of my HOTTEST Histology Supervisor positions: 1. Alabama - Histology Supervisor 2. Colorado - Histology Manager/PA (if you know anyone I also need a cytology manager/supervisor for this client) 3. Virginia - Histology Supervisor Here are some of my HOTTEST Histo Tech and Histology Supervisor positions: 1. Georgia - Histo Tech 2. Texas - Histo Tech temp to perm 3. Maine - Histo Tech 4. Maine - IHC Tech 5. N. California - Histo Tech 6. North Carolina - Histo Tech 7. NYC - Lead Histo Tech - evenings 2p-11p 8. NYC - IHC Tech - nights 10p-7a 9. Virginia - Histo Tech 10. Michigan - Histo Tech 11. Nebraska - Histo Tech If you are interested in these jobs, please CALL ME ASAP at 800 466 9919 x234. To speed things up, please also send me a copy of your resume, (if you haven't already done so). If you are interested in jobs outside the above-mentioned areas, please send me your resume as well. I have clients throughout the US. I will keep your resume confidential and will not release it to anyone without your permission (This is Ategra policy as well as my own). My services are at no charge to you. Of course, you may be happy in your present job, but it never hurts to to keep an eye open. Also, if you have friends/peers who might be interested as well, if you could pass my query & name on to them I'd be very grateful. However, if you are interested in any of the jobs above, please call me. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Pam Barker Senior Lab Recruiter Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd. Winter Park, FL 32792 VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com To Learn More About Ategra: http://www.ategra.com From jengirl1014 <@t> yahoo.com Wed Aug 4 09:28:56 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:23:49 2005 Subject: [Histonet] Staining for Macrophages Message-ID: <20040804142856.20519.qmail@web60608.mail.yahoo.com> I had previously asked about MAC-387 staining. Unfortunately, we can't use the staining because we are staining mouse kidney and it doesn't look right if you stain with a mouse anti-body. Does anyone know of a way to stain for macrophages with out an antibody??? Or perhaps a polyclonal antibody???? Thanks a bunch! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! From JWEEMS <@t> sjha.org Wed Aug 4 09:45:19 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Color-Coding Biopsies Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A622A@sjhaexc02.sjha.org> Hello Group, Is anyone color coding prostate and/or breast biopsies to help with identification of these little critters? The color would be dictated into the gross description and would aid in being certain multiple biopsies of like tissue did not get mixed up. We're thinking of doing this, and I would like to get your ideas. Thanks in advance. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From joseph-galbraith <@t> uiowa.edu Wed Aug 4 09:50:06 2004 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Staining for Macrophages Message-ID: <5D03ED7B9391D4119D9B0008C76B7B2403008618@uihc-mail1.uihc.uiowa.edu> Jennifer: Have you tried any mouse on mouse systems for this? Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jennifer Sipes Sent: Wednesday, August 04, 2004 9:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Staining for Macrophages I had previously asked about MAC-387 staining. Unfortunately, we can't use the staining because we are staining mouse kidney and it doesn't look right if you stain with a mouse anti-body. Does anyone know of a way to stain for macrophages with out an antibody??? Or perhaps a polyclonal antibody???? Thanks a bunch! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joseph-galbraith <@t> uiowa.edu Wed Aug 4 09:55:18 2004 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Color-Coding Biopsies Message-ID: <5D03ED7B9391D4119D9B0008C76B7B2403008619@uihc-mail1.uihc.uiowa.edu> Joyce: We tried eosin and it usually washed off during processing as you might expect (though it was good for making the small biopsies more visible during handling). Also our regular marking dyes and India ink were too messy and the dyes actually made the tiniest specimens harder to find in a pool of thick dye. I'd be interested in hearing what experiences others have had. Best wishes, Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Weems, Joyce Sent: Wednesday, August 04, 2004 9:45 AM To: Histonet Subject: [Histonet] Color-Coding Biopsies Hello Group, Is anyone color coding prostate and/or breast biopsies to help with identification of these little critters? The color would be dictated into the gross description and would aid in being certain multiple biopsies of like tissue did not get mixed up. We're thinking of doing this, and I would like to get your ideas. Thanks in advance. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From tpmorken <@t> labvision.com Wed Aug 4 10:24:02 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:50 2005 Subject: "...No one... saw it..." RE: [Histonet] Thanks Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2D94B15@usca0082k08.labvision.apogent.com> Tony, I have a feeling that more reponses are private these days. Maybe people are leery having their responses flamed. Whatever the reason, I miss the variety of reponses we used to see here and encourage people to take the risk. Just be judicial, professional and non-judgemental in how you respond and no one is going to bother you. I've posted to Histonet for about 8 years now and have never had an ill word against me. I hope others can see the value in responding publically . That's what Histonet was designed for - the free flow of information among a very diverse group. Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Tuesday, August 03, 2004 8:58 PM To: 'pengbw@sjtu.edu.cn'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks Unfortunately, no one else on the Server saw it. Just a friendly reminder to reply to all when you answer a posting so that everyone else can become involved in the discussion if they wish. Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] Sent: Wednesday, 4 August 2004 11:34 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Thanks Dear Robert Krug Thank you detailed explaintation of difference between the several Giemsa stain. I appreciate it very much. Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cgfields <@t> lexhealth.org Wed Aug 4 10:27:11 2004 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Color-Coding Biopsies Message-ID: We have used mercurochrome and Cobalt Blue to color biopsies. However the mercurochrome has mercury in it so we are going to switch to something else also...not sure what. Cobalt Chloride works great but I'm not sure this group will go for blue tissue. You can put it in your last alcohol on the processor and you cannot see it on the finished slide. Worked great in Texas. You can also make it as dark or light as you want. We use agar for prostate biopsies. www.palpath.com/agar.htm after that click on more agar tips.. It will explain how we process biopsies. My pathologist Dr. Shaw came up with the procedures. Good luck. Carole Fields Lex Med Ctn W.Columbia, SC -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Wednesday, August 04, 2004 10:45 AM To: Histonet Subject: [Histonet] Color-Coding Biopsies Hello Group, Is anyone color coding prostate and/or breast biopsies to help with identification of these little critters? The color would be dictated into the gross description and would aid in being certain multiple biopsies of like tissue did not get mixed up. We're thinking of doing this, and I would like to get your ideas. Thanks in advance. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From Jackie.O'Connor <@t> abbott.com Wed Aug 4 10:27:43 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] scanners Message-ID: We have a computer system on our Zeiss microscope which enables us to take photo-micrographs of an entire section, such as a rodent brain, or whole xenograft in our case, and put the photo on a CD to use in presentations, posters, etc. It's called a Matrix system, and works similar to Chromavision, but is cheaper. The stage is motorized, and is programmed to move to take sequential photographs of a section on a slide, and then put them all together for an intact photo. I love it - you can probably contact Zeiss for more info. Jackie O'Connor Abbott Labs Lucy Teves Sent by: histonet-bounces@lists.utsouthwestern.edu 08/03/2004 03:17 PM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] scanners Hello, I'm looking for a scanner that will scan whole mounted histological samples on regular slides. I would like to show whole rat brain. Which product do you recommend? Thanks in advance -- Lucy Teves Lab Manager Neuroprotection Lab. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Aug 4 10:36:05 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Staining for Macrophages In-Reply-To: <20040804142856.20519.qmail@web60608.mail.yahoo.com> References: <20040804142856.20519.qmail@web60608.mail.yahoo.com> Message-ID: <6.0.0.22.1.20040804093213.01b16cf8@gemini.msu.montana.edu> There are other monoclonal antibodies for macrophages in the mouse, generally rat antimouse, try BD Pharmingen. F4/80 (mature macrophages) is another available from Serotec. You can use a mouse monoclonal on mouse tissue, but should use the DAkO ARK kit or Chemicon mouse on mouse kit, there are others on the market. Hopefully, you have a positive control expressing the macrophage population you wish to see, this way you can optimize your antibody before going to experimental animal. At 08:28 AM 8/4/2004, you wrote: >I had previously asked about MAC-387 staining. Unfortunately, we can't >use the staining because we are staining mouse kidney and it doesn't look >right if you stain with a mouse anti-body. Does anyone know of a way to >stain for macrophages with out an antibody??? Or perhaps a polyclonal >antibody???? Thanks a bunch! > > >Jennifer K. Sipes, RALAT >Sr. Laboratory Technician >Johns Hopkins University >Ross 929 >720 Rutland Avenue >Baltimore, MD 21205 >phone: 410-614-0131 >cell: 443-413-0853 >e-mail: jengirl1014@yahoo.com > > > > > > > > > > > >--------------------------------- >Do you Yahoo!? >Yahoo! Mail - 50x more storage than other providers! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Rcartun <@t> harthosp.org Wed Aug 4 10:41:04 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Androgen receptor IHC Message-ID: Is anyone doing IHC for androgen receptors in prostate CA? If so, whose antibody (or clone) are you using? Thank you. Richard Cartun From jwatson <@t> gnf.org Wed Aug 4 10:54:50 2004 From: jwatson <@t> gnf.org (James Watson) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Part time position in San Diego Message-ID: <7BA50F61CB491B4692B8BCFBB656B5F924ABAA@EXCHCLUSTER01.lj.gnf.org> I have a part time position, 20 hrs per week, available in San Diego at the Genomics Institute of the Novartis Research Foundation. Hours are flexible. The position requires a HT with experience with Frozen sectioning, Paraffin sectioning, routine and special stains, and Immunohistochemistry. Experience with animal tissue is preferred. James Watson HT, ASCP Facilities Manager of Histology GNF, C015 858-332-4647 jwatson@gnf.org From ynwang <@t> u.washington.edu Wed Aug 4 11:33:41 2004 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Activated macrophages in porcine tissue Message-ID: Does anyone know of an antibody that can be used to label (IHC) activated macrophages in porcine tissue? Thank you Yak-Nam Wang From Dorothy.L.Webb <@t> HealthPartners.Com Wed Aug 4 12:22:40 2004 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] inking needle biopsies Message-ID: At the facility I used to work at, they did exactly as you worded it! Different colored inks were used for different patient specimens. We kept track of the last color used on a logsheet with a histotech verifying what the PA or pathologist used when grossing in, even inasmuch as the number of pieces submitted! It may seem like overkill, but there were 2 breast specimens interchanged by a pathologist when reading the slides out and it was bad! They use a marking eosin that is made up in the lab and sticks well to the tissue as well as other colors they purchase from a local art supply place. The inks are called "Pennco". I have introduced these at the facility I now work at and they love them! They do not get "gunked" up and adhere nicely with a mordant like Davidson's. I would be happy to share more if you EMail me! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From jluis.palazon <@t> icman.csic.es Wed Aug 4 13:10:21 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Hellerstrom and Hellman alcoholic silver nitrate Message-ID: <20040804181021.BE2D9278575@perceval.uca.es> Dear histonet fellows Can any of you tell me how to prepare the working solution of alcoholic silver nitrate for the Hellerstrom and Hellman protocol for argyrophil cells? I have two books that describe it but they are not coincident in the silver nitrate concentration. thanks in advance Jos? Luis From HornHV <@t> archildrens.org Wed Aug 4 13:30:43 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Color-Coding Biopsies Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B91A@EMAIL.archildrens.org> While we don't color the biopsies themselves we color code the cassettes. All GI's on one case will be for example pink, the next green.......We also use the same color slides to coordinate with the biopsy cassette. We put together a poster on this and one of our residents presented it at one the their meetings. It has helped us. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Wednesday, August 04, 2004 9:45 AM To: Histonet Subject: [Histonet] Color-Coding Biopsies Hello Group, Is anyone color coding prostate and/or breast biopsies to help with identification of these little critters? The color would be dictated into the gross description and would aid in being certain multiple biopsies of like tissue did not get mixed up. We're thinking of doing this, and I would like to get your ideas. Thanks in advance. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From Janet.Bonner <@t> FLHOSP.ORG Wed Aug 4 14:05:49 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:23:50 2005 Subject: "...No one... saw it..." RE: [Histonet] Thanks Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB3FFB@fh2k093.fhmis.net> Tim, I've answered alot of the questions privately because my initial "opinions" were rapidly flamed - a sad state of affairs. I've been in the field 32 years and know some things about what works. People who disagree really should just express their opinions and exchange ideas - not try to prove their worth by exacting Histo Web Wars. I can be aggressive but have come to prefer not showing my backside to the world on a consistent basis because I might fancy myself wonderful with dilutions of grandeur! Even skewed answers make people think!!! -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Wednesday, August 04, 2004 11:24 AM To: 'Histonet@lists.utsouthwestern.edu' Subject: "...No one... saw it..." RE: [Histonet] Thanks Tony, I have a feeling that more reponses are private these days. Maybe people are leery having their responses flamed. Whatever the reason, I miss the variety of reponses we used to see here and encourage people to take the risk. Just be judicial, professional and non-judgemental in how you respond and no one is going to bother you. I've posted to Histonet for about 8 years now and have never had an ill word against me. I hope others can see the value in responding publically . That's what Histonet was designed for - the free flow of information among a very diverse group. Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Tuesday, August 03, 2004 8:58 PM To: 'pengbw@sjtu.edu.cn'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks Unfortunately, no one else on the Server saw it. Just a friendly reminder to reply to all when you answer a posting so that everyone else can become involved in the discussion if they wish. Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] Sent: Wednesday, 4 August 2004 11:34 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Thanks Dear Robert Krug Thank you detailed explaintation of difference between the several Giemsa stain. I appreciate it very much. Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From tpschaer <@t> vet.upenn.edu Wed Aug 4 14:15:24 2004 From: tpschaer <@t> vet.upenn.edu (Tom Schaer) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] open position References: Message-ID: <004301c47a57$65190b20$38595b82@vet.upenn.edu> Greetings list: The University of Pennsylvania School of Veterinary Medicine New Bolton Center Campus has an immediate opening for a full or part time Histopathology Technician in the Comparative Orthopaedic Research Laboratory. Responsibilities include but are not limited to histology (and IHC) involving a variety of different orthopaedic tissue engineering projects (bone, cartilage, spine). Expanding contracted histology services is desired and financial reward can be expected. Interaction with and teaching of residents is possible if desired. We are a small group of enthusiastic people working in the field of equine orthopaedics, orthopaedic tissue engineering and intervertebral disc research, collaborating with a variety of industrial and academic partners in the Tri-State area. The large animal hospital of the University of Pennsylvania School of Veterinary Medicine is about 50 miles south of Philadelphia in the beautiful Chester County horse country. Salary commensurate with education and experience. Position offers full benefits package including vacation, sick-leave, free parking, retirement plan, health and life insurance. For further details about the position please contact the lab at 610-925-6261 and ask for Tom or email me directly at tpschaer@vet.upenn.edu. Thank you, ----------------------------------------- Thomas P. Sch?r, VMD Asst Prof School of Biomedical Engineering, Drexel University & Comparative Orthopaedic Research & Tissue Engineering Department of Clinical Studies University of Pennsylvania New Bolton Center 382 West Street Road Kennett Square, PA 19348 tel. 610-444-5800 (x6261 office) tel. 610-444-5800 (x6131 lab) fax. 610-925-8100 tpschaer@mail.vet.upenn.edu http://www2.vet.upenn.edu/labs/corl/drtps.html From thallada <@t> noch.org Wed Aug 4 14:16:01 2004 From: thallada <@t> noch.org (Hallada, Teri) Date: Fri Sep 16 15:23:50 2005 Subject: "...No one... saw it..." RE: [Histonet] Thanks Message-ID: "Ditto." I have experienced the same problem from the same parties on the Cytopath.net. Teri Hallada BS MT CT (ASCP) thallada@noch.org > -----Original Message----- > From: Bonner, Janet [SMTP:Janet.Bonner@FLHOSP.ORG] > Sent: Wednesday, August 04, 2004 15:06 > To: 'Morken, Tim - Labvision'; 'Histonet@lists.utsouthwestern.edu' > Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks > > Tim, > I've answered alot of the questions privately because my initial "opinions" > were rapidly flamed - a sad state of affairs. I've been in the field 32 > years and know some things about what works. People who disagree really > should just express their opinions and exchange ideas - not try to prove > their worth by exacting Histo Web Wars. I can be aggressive but have come > to prefer not showing my backside to the world on a consistent basis because > I might fancy myself wonderful with dilutions of grandeur! Even skewed > answers make people think!!! > > -----Original Message----- > From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] > Sent: Wednesday, August 04, 2004 11:24 AM > To: 'Histonet@lists.utsouthwestern.edu' > Subject: "...No one... saw it..." RE: [Histonet] Thanks > > > Tony, > I have a feeling that more reponses are private these days. Maybe people are > leery having their responses flamed. Whatever the reason, I miss the variety > of reponses we used to see here and encourage people to take the risk. Just > be judicial, professional and non-judgemental in how you respond and no one > is going to bother you. I've posted to Histonet for about 8 years now and > have never had an ill word against me. I hope others can see the value in > responding publically . That's what Histonet was designed for - the free > flow of information among a very diverse group. > > Tim Morken > Lab Vision - Neomarkers > www.labvision.com > > Free webhosting for US State Histotechnology Societies: > http://www.labvisioncorp.com/demowebsite/index.cfm > > > > -----Original Message----- > From: Tony Henwood [mailto:AnthonyH@chw.edu.au] > Sent: Tuesday, August 03, 2004 8:58 PM > To: 'pengbw@sjtu.edu.cn'; Histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Thanks > > > Unfortunately, no one else on the Server saw it. > Just a friendly reminder to reply to all when you answer a posting so that > everyone else can become involved in the discussion if they wish. > > Regards, > > > Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager > The Children's Hospital at Westmead, > Locked Bag 4001, Westmead, 2145, AUSTRALIA. > Tel: 612 9845 3306 > Fax: 612 9845 3318 > > > -----Original Message----- > From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] > Sent: Wednesday, 4 August 2004 11:34 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Thanks > > > Dear Robert Krug > > Thank you detailed explaintation of difference between the several Giemsa > stain. I appreciate it very much. > > > > Baowei Peng > Pharmacy School > Shanghai Jiaotong University > Shanghai, 200030 > China > > > ********************************************************************** > This email and any files transmitted with it are confidential and intended > solely for the use of the individual or entity to whom they are addressed. > If you are not the intended recipient, please > delete it and notify the sender. > > Views expressed in this message and any attachments are those > of the individual sender, and are not necessarily the views of The > Children's Hospital at Westmead > > This footnote also confirms that this email message has been > virus scanned and although no computer viruses were detected, the Childrens > Hospital at Westmead accepts no liability for any > consequential damage resulting from email containing computer > viruses. > ********************************************************************** > > > _______________________________________________> > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > The information contained in this message may be privileged and/or > confidential and protected from disclosure. If the reader of this message > is not the intended recipient or agent responsible for delivering this > message to the intended recipient, you are hereby notified that any > dissemination, distribution or copying of this communication is strictly > prohibited. If you have received this communication in error, please notify > the sender immediately by replying to this message and deleting the material > from any computer. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Statement: This message is intended only for the individual or entity to which it is addressed. It may contain privileged, confidential information which is exempt from disclosure under applicable laws. If you are not the intended recipient, please note that you are strictly prohibited from disseminating or distributing this information (other than to the intended recipient) or copying this information. If you have received this communication in error, please notify us immediately by e-mail or by telephone at (616) 842-3600. Thank you. From subratab <@t> bdonline.com Wed Aug 4 14:25:37 2004 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] difference between the several Giemsa stain. Message-ID: <200408041930.i74JUlCa002977@mailout.proshikanet.com> Dear Baowei Peng Could you please send the replys to your question on Giemsa staining to histonet? I think many persons (including me) are interested to read the discusion. Sincerely Dr Subrata Biswas University of Campinas SP, Brazil From scoop <@t> mail.nih.gov Wed Aug 4 15:02:32 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Spleen autofluorescence In-Reply-To: <1085476466.40b30e723e67f@staffmail.ed.ac.uk> References: <1085476466.40b30e723e67f@staffmail.ed.ac.uk> Message-ID: I have had pretty good luck quenching autofluorescence in FFPE spleen with sodium borohydride 0.5 mg/ml in PBS pH 8.0 x 10 minutes x 3 changes. It works better than H2O2 for me. Sharon >Hi histonetters, > >Apologies if my query is really basic but I am wanting to do immunofluorescent >antibody staining on mouse spleen sections (paraffin-embedded) and am seeing a >lot of autofluorescence, which I presume is due to platelets. Is there any way >around this problem? > >Helen Newbery, >Medical Genetics Section, >Molecular Medicine Centre, >Western General Hospital, >Edinburgh, >EH4 2XU. >0131 651 1047 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From nena.dimaano <@t> stryker.com Wed Aug 4 15:03:45 2004 From: nena.dimaano <@t> stryker.com (Dimaano, Nena) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Tissue Processor Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD06210D87@HOS2KEXCHCL.howost.strykercorp.com> We are in the process of buying a new tissue processor. Can anyone recommend a good and reliable processor good for R&D purposes? thanks, Nena Dimaano Advanced Technology Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com From gcallis <@t> montana.edu Wed Aug 4 16:52:37 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] mouse spleen autofluorescence quenching and other considerations In-Reply-To: References: <1085476466.40b30e723e67f@staffmail.ed.ac.uk> Message-ID: <6.0.0.22.1.20040804150129.01b005a8@gemini.msu.montana.edu> I didn't think that hydrogen peroxide is meant to quench autofluorescence (but I could be misinformed) but rather it quenches endogenous peroxidase for immunohistochemistry peroxidase method. Sodium borohydride (blocks free aldehydes and not without possible problems on antigenicity loss per discussion by Jules Elias) has been used to quench autofluorescence. Some people have a terrible time getting rid of autofluorescence and often never completely succeed. Autofluorescence can be worse aldehyde fixation. As offered before, I have a review of autofluorescence in tissues with GFP work in mind, but the discussion is very thorough, and pertains to nonGFP work also, it includes fixation and what in tissue autofluorescences. I will send it to anyone via personal email. A good way to avoid autofluorescence in mouse spleen (an any other tissue) is cut frozen sections and use either acetone or acetone/alcohol fixative (whichever fixative gives the best immunfluorescent results for your antibody) avoid NBF or PFA fixation. Be sure to have one unstained section to see degree of autofluorescence and and if you use oil immersion, the oil must be PCB free and not contributing to autofluorescence. If you insist on FFPE with spleen, you can cleverly use the autofluorescence as a "counterstain", so to speak, have your fluorophore be in contrast to the slight greenish-yellow, say an Alexa 546 or 555 - red or Rhodamine or even Texas Red. Jackson has secondary antibodies conjugated to a rhodamine RRX that is bright, and more resistant to photobleaching versus standard TRITC or Texas Red. The joy of Alexa dyes - very bright and resistant to photobleaching. A side note: Molecular Probes has "ready to use" Prolong Gold antifade mounting media (although pricey and requires sealing with diluted permanent mounting media, not nail polish) - it sets up hard and is excellent for keeping your fluorophores glowing longer. We are delighted with this media, even with eGFP. At 02:02 PM 8/4/2004, you wrote: I have had pretty good luck quenching autofluorescence in FFPE spleen with sodium borohydride 0.5 mg/ml in PBS pH 8.0 x 10 minutes x 3 changes. It works better than H2O2 for me. Sharon Hi histonetters, Apologies if my query is really basic but I am wanting to do immunofluorescent antibody staining on mouse spleen sections (paraffin-embedded) and am seeing a lot of autofluorescence, which I presume is due to platelets. Is there any way around this problem? Helen Newbery, Medical Genetics Section, Molecular Medicine Centre, Western General Hospital, Edinburgh, EH4 2XU. 0131 651 1047 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu [1]http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu [2]http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) References 1. http://lists.utsouthwestern.edu/mailman/listinfo/histonet 2. http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ihc <@t> unipathllc.com Wed Aug 4 16:57:51 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Microwave Processing Message-ID: <000001c47a6e$16e95c40$4500a8c0@unipath02> I don't mean to beat a dead horse, but I have some questions about microwave processing and what I found in the archives didn't exactly answer them. Does anyone use the microwave to process ALL of their tissues? If so do you have different runs for large vs. small bxs, prostates, GIs, etc.? Do you find the quality to be as good as or better than conventional processing, or just quicker? Thank you, Brianna Jackson UniPath, LLC Denver, CO 303-512-2220 From bills <@t> icpmr.wsahs.nsw.gov.au Wed Aug 4 17:13:48 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Spleen autofluorescence In-Reply-To: Message-ID: <000101c47a70$510d2bc0$e1ce080a@wsahs.nsw.gov.au> Many many years ago I had a similar problem in gut sections. The autofluorescence after formaldehyde fixation was very strong in the argentaffin cells. Our Histochemist at that time suggested I do a haematoxylin stain (very briefly Mayer's 30sec - 1min no differentiation) and I was very surprised when it quenched most of the autofluoresence. Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sharon Cooperman Sent: Thursday, 05 August 2004 6:03 AM To: Helen Newbery Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Spleen autofluorescence I have had pretty good luck quenching autofluorescence in FFPE spleen with sodium borohydride 0.5 mg/ml in PBS pH 8.0 x 10 minutes x 3 changes. It works better than H2O2 for me. Sharon >Hi histonetters, > >Apologies if my query is really basic but I am wanting to do immunofluorescent >antibody staining on mouse spleen sections (paraffin-embedded) and am seeing a >lot of autofluorescence, which I presume is due to platelets. Is there any way >around this problem? > >Helen Newbery, >Medical Genetics Section, >Molecular Medicine Centre, >Western General Hospital, >Edinburgh, >EH4 2XU. >0131 651 1047 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From JWEEMS <@t> sjha.org Wed Aug 4 17:35:34 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Microwave Processing Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A6258@sjhaexc02.sjha.org> Have you checked into the one Sakura has? It will be the wave of the future, I'm sure. Check it out on their web site http://www.sakuraus.com/ . Good luck! j:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of UniPath IHC Sent: Wednesday, August 04, 2004 5:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave Processing I don't mean to beat a dead horse, but I have some questions about microwave processing and what I found in the archives didn't exactly answer them. Does anyone use the microwave to process ALL of their tissues? If so do you have different runs for large vs. small bxs, prostates, GIs, etc.? Do you find the quality to be as good as or better than conventional processing, or just quicker? Thank you, Brianna Jackson UniPath, LLC Denver, CO 303-512-2220 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From JNocito <@t> Pathreflab.com Wed Aug 4 18:46:13 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:50 2005 Subject: "...No one... saw it..." RE: [Histonet] Thanks In-Reply-To: <07AB60D5D7B9754EBF56F360F98D083DEB3FFB@fh2k093.fhmis.net> Message-ID: Hey y'all it's me again. Sometimes, I'll answer privately only because a couple of times, I've had vendors call my employers telling me to back off on my comments (yeah, that worked well now, didn't it). Sometimes I say things tongue in cheek, but that always doesn't come across that way either. Then, I get the phone calls (this just happened last month) Many times though, I'll get a private response thanking me for helping out. It's those special times that keep me coming back to the Histonet. Sometimes I, myself need a reality check because I have this big fear that I start taking myself too seriously (okay, it can happen you know). I've been around the block once or twice, but I'm still learning too. Just the other day I printed out the responses about PAS staining and made it our monthly inservice. It's been a while since I've looked to see how long periodic acid stays fresh. Without the public postings, it would have been another item that I forgot about. Yeah, I've been hooked and skinned to. Just have to add another layer of skin back on. As always, the opinions of this author do not reflect the opinions of my employer. Have a good one. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonner, Janet Sent: Wednesday, August 04, 2004 2:06 PM To: 'Morken, Tim - Labvision'; 'Histonet@lists.utsouthwestern.edu' Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks Tim, I've answered alot of the questions privately because my initial "opinions" were rapidly flamed - a sad state of affairs. I've been in the field 32 years and know some things about what works. People who disagree really should just express their opinions and exchange ideas - not try to prove their worth by exacting Histo Web Wars. I can be aggressive but have come to prefer not showing my backside to the world on a consistent basis because I might fancy myself wonderful with dilutions of grandeur! Even skewed answers make people think!!! -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Wednesday, August 04, 2004 11:24 AM To: 'Histonet@lists.utsouthwestern.edu' Subject: "...No one... saw it..." RE: [Histonet] Thanks Tony, I have a feeling that more reponses are private these days. Maybe people are leery having their responses flamed. Whatever the reason, I miss the variety of reponses we used to see here and encourage people to take the risk. Just be judicial, professional and non-judgemental in how you respond and no one is going to bother you. I've posted to Histonet for about 8 years now and have never had an ill word against me. I hope others can see the value in responding publically . That's what Histonet was designed for - the free flow of information among a very diverse group. Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Tuesday, August 03, 2004 8:58 PM To: 'pengbw@sjtu.edu.cn'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks Unfortunately, no one else on the Server saw it. Just a friendly reminder to reply to all when you answer a posting so that everyone else can become involved in the discussion if they wish. Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] Sent: Wednesday, 4 August 2004 11:34 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Thanks Dear Robert Krug Thank you detailed explaintation of difference between the several Giemsa stain. I appreciate it very much. Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From scoop <@t> mail.nih.gov Wed Aug 4 19:58:37 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Staining for Macrophages In-Reply-To: <20040804142856.20519.qmail@web60608.mail.yahoo.com> References: <20040804142856.20519.qmail@web60608.mail.yahoo.com> Message-ID: You can use ED-1 from Serotec. It works on FFPE tissue with pH 6.0 citrate antigen retrieval and on frozen tissue without AR. You can also use F4/80 on frozen tissue. Both are rat monoclonals and you can use an anti-rat secondary Ab cross adsorbed against mouse IgG. You could also also use ricin 1 from Vector or E-Y Laboratories which binds mouse macrophages. Ricin 1 is not the dangerous ricin. I don't know details of how to use it (I did once, but I don't remember). There are books available or the manufacturers may know or maybe some other Histonet folks. I have used ED1 and F4/80 on mouse tissues and I think it's your best bet. Sharon >I had previously asked about MAC-387 staining. Unfortunately, we >can't use the staining because we are staining mouse kidney and it >doesn't look right if you stain with a mouse anti-body. Does anyone >know of a way to stain for macrophages with out an antibody??? Or >perhaps a polyclonal antibody???? Thanks a bunch! > > >Jennifer K. Sipes, RALAT >Sr. Laboratory Technician >Johns Hopkins University >Ross 929 >720 Rutland Avenue >Baltimore, MD 21205 >phone: 410-614-0131 >cell: 443-413-0853 >e-mail: jengirl1014@yahoo.com > > > > > > > > > > > >--------------------------------- >Do you Yahoo!? >Yahoo! Mail - 50x more storage than other providers! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From pengbw <@t> sjtu.edu.cn Wed Aug 4 20:01:28 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:23:50 2005 Subject: "...No one... saw it..." RE: [Histonet] Thanks Message-ID: <20040805010128.0D0861105C66@sjtu.edu.cn> En~~~, sorry to bother all in the histonet. But you know, I did not realize it was a private reply from Mr Rober Krug at first. I\'m not intend to answer private mails in public. ----- Original Message ----- From: "Joe Nocito" To: "Bonner, Janet" , "\'Morken, Tim - Labvision\'" , Cc: Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks Hey y\'all it\'s me again. Sometimes, I\'ll answer privately only because a couple of times, I\'ve had vendors call my employers telling me to back off on my comments (yeah, that worked well now, didn\'t it). Sometimes I say things tongue in cheek, but that always doesn\'t come across that way either. Then, I get the phone calls (this just happened last month) Many times though, I\'ll get a private response thanking me for helping out. It\'s those special times that keep me coming back to the Histonet. Sometimes I, myself need a reality check because I have this big fear that I start taking myself too seriously (okay, it can happen you know). I\'ve been around the block once or twice, but I\'m still learning too. Just the other day I printed out the responses about PAS staining and made it our monthly inservice. It\'s been a while since I\'ve looked to see how long periodic acid stays fresh. Without the public postings, it would have been another item that I forgot about. Yeah, I\'ve been hooked and skinned to. Just have to add another layer of skin back on. As always, the opinions of this author do not reflect the opinions of my employer. Have a good one. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonner, Janet Sent: Wednesday, August 04, 2004 2:06 PM To: \'Morken, Tim - Labvision\'; \'Histonet@lists.utsouthwestern.edu\' Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks Tim, I\'ve answered alot of the questions privately because my initial "opinions" were rapidly flamed - a sad state of affairs. I\'ve been in the field 32 years and know some things about what works. People who disagree really should just express their opinions and exchange ideas - not try to prove their worth by exacting Histo Web Wars. I can be aggressive but have come to prefer not showing my backside to the world on a consistent basis because I might fancy myself wonderful with dilutions of grandeur! Even skewed answers make people think!!! -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Wednesday, August 04, 2004 11:24 AM To: \'Histonet@lists.utsouthwestern.edu\' Subject: "...No one... saw it..." RE: [Histonet] Thanks Tony, I have a feeling that more reponses are private these days. Maybe people are leery having their responses flamed. Whatever the reason, I miss the variety of reponses we used to see here and encourage people to take the risk. Just be judicial, professional and non-judgemental in how you respond and no one is going to bother you. I\'ve posted to Histonet for about 8 years now and have never had an ill word against me. I hope others can see the value in responding publically . That\'s what Histonet was designed for - the free flow of information among a very diverse group. Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Tuesday, August 03, 2004 8:58 PM To: \'pengbw@sjtu.edu.cn\'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks Unfortunately, no one else on the Server saw it. Just a friendly reminder to reply to all when you answer a posting so that everyone else can become involved in the discussion if they wish. Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children\'s Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] Sent: Wednesday, 4 August 2004 11:34 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Thanks Dear Robert Krug Thank you detailed explaintation of difference between the several Giemsa stain. I appreciate it very much. Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children\'s Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From AnthonyH <@t> chw.edu.au Wed Aug 4 20:39:37 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:50 2005 Subject: "...No one... saw it..." RE: [Histonet] Thanks Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E22E@simba.kids> Joe, Unfortunately if no one else sees the replies then our in-house sessions will lose a valuable resource. If you are worried about being flamed, as Joe says, we all have thick skins. I personally feel embarrassed when I see someone else being flamed, so I then disregard that message and any further threads that appear. The DELETE button is invaluable. If I'm being flamed, then I ignore it. I don't bother responding. Few people know where Australia is anyway!! Please respond to all. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Thursday, 5 August 2004 9:46 AM To: Bonner, Janet; 'Morken, Tim - Labvision'; Histonet@lists.utsouthwestern.edu Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks Hey y'all it's me again. Sometimes, I'll answer privately only because a couple of times, I've had vendors call my employers telling me to back off on my comments (yeah, that worked well now, didn't it). Sometimes I say things tongue in cheek, but that always doesn't come across that way either. Then, I get the phone calls (this just happened last month) Many times though, I'll get a private response thanking me for helping out. It's those special times that keep me coming back to the Histonet. Sometimes I, myself need a reality check because I have this big fear that I start taking myself too seriously (okay, it can happen you know). I've been around the block once or twice, but I'm still learning too. Just the other day I printed out the responses about PAS staining and made it our monthly inservice. It's been a while since I've looked to see how long periodic acid stays fresh. Without the public postings, it would have been another item that I forgot about. Yeah, I've been hooked and skinned to. Just have to add another layer of skin back on. As always, the opinions of this author do not reflect the opinions of my employer. Have a good one. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonner, Janet Sent: Wednesday, August 04, 2004 2:06 PM To: 'Morken, Tim - Labvision'; 'Histonet@lists.utsouthwestern.edu' Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks Tim, I've answered alot of the questions privately because my initial "opinions" were rapidly flamed - a sad state of affairs. I've been in the field 32 years and know some things about what works. People who disagree really should just express their opinions and exchange ideas - not try to prove their worth by exacting Histo Web Wars. I can be aggressive but have come to prefer not showing my backside to the world on a consistent basis because I might fancy myself wonderful with dilutions of grandeur! Even skewed answers make people think!!! -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Wednesday, August 04, 2004 11:24 AM To: 'Histonet@lists.utsouthwestern.edu' Subject: "...No one... saw it..." RE: [Histonet] Thanks Tony, I have a feeling that more reponses are private these days. Maybe people are leery having their responses flamed. Whatever the reason, I miss the variety of reponses we used to see here and encourage people to take the risk. Just be judicial, professional and non-judgemental in how you respond and no one is going to bother you. I've posted to Histonet for about 8 years now and have never had an ill word against me. I hope others can see the value in responding publically . That's what Histonet was designed for - the free flow of information among a very diverse group. Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Tuesday, August 03, 2004 8:58 PM To: 'pengbw@sjtu.edu.cn'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks Unfortunately, no one else on the Server saw it. Just a friendly reminder to reply to all when you answer a posting so that everyone else can become involved in the discussion if they wish. Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] Sent: Wednesday, 4 August 2004 11:34 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Thanks Dear Robert Krug Thank you detailed explaintation of difference between the several Giemsa stain. I appreciate it very much. Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Wed Aug 4 20:42:37 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:50 2005 Subject: "...No one... saw it..." RE: [Histonet] Thank s Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E22F@simba.kids> Baowei, Do not be sorry. This was not your fault. You reponded the way everyone on Histonet should do (or try to remember to do). Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] Sent: Thursday, 5 August 2004 11:01 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: RE: "...No one... saw it..." RE: [Histonet] Thanks En~~~, sorry to bother all in the histonet. But you know, I did not realize it was a private reply from Mr Rober Krug at first. I\'m not intend to answer private mails in public. ----- Original Message ----- From: "Joe Nocito" To: "Bonner, Janet" , "\'Morken, Tim - Labvision\'" , Cc: Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks Hey y\'all it\'s me again. Sometimes, I\'ll answer privately only because a couple of times, I\'ve had vendors call my employers telling me to back off on my comments (yeah, that worked well now, didn\'t it). Sometimes I say things tongue in cheek, but that always doesn\'t come across that way either. Then, I get the phone calls (this just happened last month) Many times though, I\'ll get a private response thanking me for helping out. It\'s those special times that keep me coming back to the Histonet. Sometimes I, myself need a reality check because I have this big fear that I start taking myself too seriously (okay, it can happen you know). I\'ve been around the block once or twice, but I\'m still learning too. Just the other day I printed out the responses about PAS staining and made it our monthly inservice. It\'s been a while since I\'ve looked to see how long periodic acid stays fresh. Without the public postings, it would have been another item that I forgot about. Yeah, I\'ve been hooked and skinned to. Just have to add another layer of skin back on. As always, the opinions of this author do not reflect the opinions of my employer. Have a good one. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonner, Janet Sent: Wednesday, August 04, 2004 2:06 PM To: \'Morken, Tim - Labvision\'; \'Histonet@lists.utsouthwestern.edu\' Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks Tim, I\'ve answered alot of the questions privately because my initial "opinions" were rapidly flamed - a sad state of affairs. I\'ve been in the field 32 years and know some things about what works. People who disagree really should just express their opinions and exchange ideas - not try to prove their worth by exacting Histo Web Wars. I can be aggressive but have come to prefer not showing my backside to the world on a consistent basis because I might fancy myself wonderful with dilutions of grandeur! Even skewed answers make people think!!! -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Wednesday, August 04, 2004 11:24 AM To: \'Histonet@lists.utsouthwestern.edu\' Subject: "...No one... saw it..." RE: [Histonet] Thanks Tony, I have a feeling that more reponses are private these days. Maybe people are leery having their responses flamed. Whatever the reason, I miss the variety of reponses we used to see here and encourage people to take the risk. Just be judicial, professional and non-judgemental in how you respond and no one is going to bother you. I\'ve posted to Histonet for about 8 years now and have never had an ill word against me. I hope others can see the value in responding publically . That\'s what Histonet was designed for - the free flow of information among a very diverse group. Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Tuesday, August 03, 2004 8:58 PM To: \'pengbw@sjtu.edu.cn\'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks Unfortunately, no one else on the Server saw it. Just a friendly reminder to reply to all when you answer a posting so that everyone else can become involved in the discussion if they wish. Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children\'s Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] Sent: Wednesday, 4 August 2004 11:34 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Thanks Dear Robert Krug Thank you detailed explaintation of difference between the several Giemsa stain. I appreciate it very much. Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children\'s Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From jnocito <@t> satx.rr.com Wed Aug 4 21:07:14 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:23:50 2005 Subject: "...No one... saw it..." RE: [Histonet] Thanks References: <1CF2E2E5BB36D5119E7A0008C791F3740800E22E@simba.kids> Message-ID: <008201c47a90$ef811530$5d6bce44@yourxhtr8hvc4p> Tony, I also love my delete button. How's it go? Gaday mate! Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Tony Henwood" To: "'Joe Nocito'" ; "Bonner, Janet" ; "'Morken, Tim - Labvision'" ; Sent: Wednesday, August 04, 2004 8:39 PM Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks > Joe, > > Unfortunately if no one else sees the replies then our in-house sessions > will lose a valuable resource. If you are worried about being flamed, as Joe > says, we all have thick skins. > > I personally feel embarrassed when I see someone else being flamed, so I > then disregard that message and any further threads that appear. The DELETE > button is invaluable. > > If I'm being flamed, then I ignore it. I don't bother responding. Few people > know where Australia is anyway!! > > Please respond to all. > > > Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager > The Children's Hospital at Westmead, > Locked Bag 4001, Westmead, 2145, AUSTRALIA. > Tel: 612 9845 3306 > Fax: 612 9845 3318 > > > > -----Original Message----- > From: Joe Nocito [mailto:JNocito@Pathreflab.com] > Sent: Thursday, 5 August 2004 9:46 AM > To: Bonner, Janet; 'Morken, Tim - Labvision'; > Histonet@lists.utsouthwestern.edu > Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks > > > Hey y'all it's me again. > Sometimes, I'll answer privately only because a couple of times, > I've had > vendors call my employers telling me to back off on my comments (yeah, that > worked well now, didn't it). > Sometimes I say things tongue in cheek, but that always doesn't come > across > that way either. Then, I get the phone calls (this just happened last month) > Many times though, I'll get a private response thanking me for > helping out. > It's those special times that keep me coming back to the Histonet. > Sometimes I, myself need a reality check because I have this big > fear that > I start taking myself too seriously (okay, it can happen you know). > I've been around the block once or twice, but I'm still learning > too. Just > the other day I printed out the responses about PAS staining and made it our > monthly inservice. It's been a while since I've looked to see how long > periodic acid stays fresh. Without the public postings, it would have been > another item that I forgot about. > Yeah, I've been hooked and skinned to. Just have to add another > layer of > skin back on. > As always, the opinions of this author do not reflect the opinions of my > employer. Have a good one. > > Joe Nocito, BS, HT(ASCP) QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonner, Janet > Sent: Wednesday, August 04, 2004 2:06 PM > To: 'Morken, Tim - Labvision'; 'Histonet@lists.utsouthwestern.edu' > Subject: RE: "...No one... saw it..." RE: [Histonet] Thanks > > Tim, > I've answered alot of the questions privately because my initial "opinions" > were rapidly flamed - a sad state of affairs. I've been in the field 32 > years and know some things about what works. People who disagree really > should just express their opinions and exchange ideas - not try to prove > their worth by exacting Histo Web Wars. I can be aggressive but have come > to prefer not showing my backside to the world on a consistent basis because > I might fancy myself wonderful with dilutions of grandeur! Even skewed > answers make people think!!! > > -----Original Message----- > From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] > Sent: Wednesday, August 04, 2004 11:24 AM > To: 'Histonet@lists.utsouthwestern.edu' > Subject: "...No one... saw it..." RE: [Histonet] Thanks > > > Tony, > I have a feeling that more reponses are private these days. Maybe people are > leery having their responses flamed. Whatever the reason, I miss the variety > of reponses we used to see here and encourage people to take the risk. Just > be judicial, professional and non-judgemental in how you respond and no one > is going to bother you. I've posted to Histonet for about 8 years now and > have never had an ill word against me. I hope others can see the value in > responding publically . That's what Histonet was designed for - the free > flow of information among a very diverse group. > > Tim Morken > Lab Vision - Neomarkers > www.labvision.com > > Free webhosting for US State Histotechnology Societies: > http://www.labvisioncorp.com/demowebsite/index.cfm > > > > -----Original Message----- > From: Tony Henwood [mailto:AnthonyH@chw.edu.au] > Sent: Tuesday, August 03, 2004 8:58 PM > To: 'pengbw@sjtu.edu.cn'; Histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Thanks > > > Unfortunately, no one else on the Server saw it. > Just a friendly reminder to reply to all when you answer a posting so that > everyone else can become involved in the discussion if they wish. > > Regards, > > > Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager > The Children's Hospital at Westmead, > Locked Bag 4001, Westmead, 2145, AUSTRALIA. > Tel: 612 9845 3306 > Fax: 612 9845 3318 > > > -----Original Message----- > From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] > Sent: Wednesday, 4 August 2004 11:34 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Thanks > > > Dear Robert Krug > > Thank you detailed explaintation of difference between the several Giemsa > stain. I appreciate it very much. > > > > Baowei Peng > Pharmacy School > Shanghai Jiaotong University > Shanghai, 200030 > China > > > ********************************************************************** > This email and any files transmitted with it are confidential and intended > solely for the use of the individual or entity to whom they are addressed. > If you are not the intended recipient, please > delete it and notify the sender. > > Views expressed in this message and any attachments are those > of the individual sender, and are not necessarily the views of The > Children's Hospital at Westmead > > This footnote also confirms that this email message has been > virus scanned and although no computer viruses were detected, the Childrens > Hospital at Westmead accepts no liability for any > consequential damage resulting from email containing computer > viruses. > ********************************************************************** > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > The information contained in this message may be privileged and/or > confidential and protected from disclosure. If the reader of this message > is not the intended recipient or agent responsible for delivering this > message to the intended recipient, you are hereby notified that any > dissemination, distribution or copying of this communication is strictly > prohibited. If you have received this communication in error, please notify > the sender immediately by replying to this message and deleting the material > from any computer. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Ianbernard <@t> netzero.com Wed Aug 4 21:30:48 2004 From: Ianbernard <@t> netzero.com (Ian) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Tissue Processor In-Reply-To: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD06210D87@HOS2KEXCHCL.howost.strykercorp.com> Message-ID: The new VIP's -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dimaano, Nena Sent: Wednesday, August 04, 2004 1:04 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue Processor We are in the process of buying a new tissue processor. Can anyone recommend a good and reliable processor good for R&D purposes? thanks, Nena Dimaano Advanced Technology Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com From WWmn916 <@t> aol.com Wed Aug 4 22:04:47 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers Message-ID: <15c.3afd8299.2e42fdcf@aol.com> Hello again, I'm looking for opinions on the subject of glass coverslippers versus tape coverslipping. I have the opportunity to decide on a system. My only experience has been with tape coverslipping. I understand machines that glass coverslip are slower than tape systems. Is the refractive index better with glass coverslips under the microscope? Opinions pros/cons are appreciated. Deb King, HT(ASCP) Sacramento, CA From lfaucette <@t> niaid.nih.gov Thu Aug 5 05:38:03 2004 From: lfaucette <@t> niaid.nih.gov (Faucette, Lawrence (NIH/NIAID)) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Tissue Processor Message-ID: <78F91CA4B8F3504E8FE5F13AAF122ED804A02CA6@nihexchange27.nih.gov> I have always been a strong fan of the Sakura tissue processors. But I have to admit, the Shandon unit, with its on-board QA/QC features combined with their system of rotating reagents have caught my eye and would have be be seriously evaluated Lawrence J Faucette Contractor HT ASCP http://www.niaid.nih.gov/dir/services/animalcare/VetPathology/VetPathology-i ndex.html Infectious Disease Pathogenesis Section Comparative Medicine Branch Division of Intramural Research, NIAID, NIH Twinbrook III, Room 2W-01A, MSC 8135 12735 Twinbrook Parkway Bethesda, MD 20892-8135 Telephone 301-451-1056 Fax 301-480-2343 SoBran, Inc. Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. The National Institute of Allergy and Infectious Diseases (NIAID) shall not accept liability for any statement made that are the sender's own and not expressly made on behalf of the NIAID by one of its representatives. -----Original Message----- From: Ian [mailto:Ianbernard@netzero.com] Sent: Wednesday, August 04, 2004 10:31 PM To: 'Dimaano, Nena'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Tissue Processor The new VIP's -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dimaano, Nena Sent: Wednesday, August 04, 2004 1:04 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue Processor We are in the process of buying a new tissue processor. Can anyone recommend a good and reliable processor good for R&D purposes? thanks, Nena Dimaano Advanced Technology Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Thu Aug 5 06:34:58 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers Message-ID: I have used both. Yes, the film coverslippers are faster, but after using one for five years, this institution is considering going to glass. My former employer also chose glass over tape. Glass just holds up better over the long run. I think some stains fade over time with tape. Hope I was of help. >>> 08/04/04 11:04PM >>> Hello again, I'm looking for opinions on the subject of glass coverslippers versus tape coverslipping. I have the opportunity to decide on a system. My only experience has been with tape coverslipping. I understand machines that glass coverslip are slower than tape systems. Is the refractive index better with glass coverslips under the microscope? Opinions pros/cons are appreciated. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From akbitting <@t> geisinger.edu Thu Aug 5 06:37:40 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Tissue Processor Message-ID: My vote is for the VIP. I have a Shandon Pathcenter and never again. I think every part has been repalced on it, some twice. If you decide to go with a Shandon, get a good service contract and have back up. I don't know how their new one is. I'd be interested to hear comments. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Laboratories (570)214-9634 >>> "Faucette, Lawrence (NIH/NIAID)" 08/05/04 06:38AM >>> I have always been a strong fan of the Sakura tissue processors. But I have to admit, the Shandon unit, with its on-board QA/QC features combined with their system of rotating reagents have caught my eye and would have be be seriously evaluated Lawrence J Faucette Contractor HT ASCP http://www.niaid.nih.gov/dir/services/animalcare/VetPathology/VetPathology-i ndex.html Infectious Disease Pathogenesis Section Comparative Medicine Branch Division of Intramural Research, NIAID, NIH Twinbrook III, Room 2W-01A, MSC 8135 12735 Twinbrook Parkway Bethesda, MD 20892-8135 Telephone 301-451-1056 Fax 301-480-2343 SoBran, Inc. Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. The National Institute of Allergy and Infectious Diseases (NIAID) shall not accept liability for any statement made that are the sender's own and not expressly made on behalf of the NIAID by one of its representatives. -----Original Message----- From: Ian [mailto:Ianbernard@netzero.com] Sent: Wednesday, August 04, 2004 10:31 PM To: 'Dimaano, Nena'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Tissue Processor The new VIP's -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dimaano, Nena Sent: Wednesday, August 04, 2004 1:04 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue Processor We are in the process of buying a new tissue processor. Can anyone recommend a good and reliable processor good for R&D purposes? thanks, Nena Dimaano Advanced Technology Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From Barbara_Lentz <@t> dahlchase.com Thu Aug 5 07:18:23 2004 From: Barbara_Lentz <@t> dahlchase.com (Barbara Lentz) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Tissue Processor Message-ID: We have 6 VIP's and have just recently purchased 2 Shandon Electrons's. I think it is too early to judge their worth. The cost made it worthwhile to purchase. Barb Barbara Lentz, HTL/MT (ASCP) Dahl-Chase Diagnostic Services Webber West, Suite 545 417 State St Bangor, ME 04401 phone (207) 941-8261 fax (207) 990-2987 Barbara_Lentz@dahlchase.com From Barbara_Lentz <@t> dahlchase.com Thu Aug 5 07:40:35 2004 From: Barbara_Lentz <@t> dahlchase.com (Barbara Lentz) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Conundrum Message-ID: I do not understand why formalin keeps the tissue from falling off, but I swear by it. It's especially useful when there is no other tissue available and we cannot take the chance of the tissue falling off. Barb >>> Jennifer MacDonald 07/25/04 10:12PM >>> I am curious about the treatment of the slides in formalin. I have heard of a few places that do this but no one seems to know why. Does anyone have a theory on why this helps the tissue stay on the slide? Jennifer MacDonald We use the Ventana XT and place some of our slides in 10% NBF for 30 minutes. This helps the tissue stay on even when the slides haven't been rehydrated. Barb _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nena.dimaano <@t> stryker.com Thu Aug 5 07:49:30 2004 From: nena.dimaano <@t> stryker.com (Dimaano, Nena) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Tissue Processor Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD13F2DE@HOS2KEXCHCL.howost.strykercorp.com> To all, Thank you very much to all who responded to my inquiry, but did anyone has the experience with the Shandon Excelsior? thans again, Nena Dimaano,MT/HT(ASCP) Advanced Technology Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com From DonnaWillis <@t> texashealth.org Thu Aug 5 07:54:35 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Microwave Processing Message-ID: Brianna, We currently have 2 older Energy Beam units that we use for biopsy samples, 2 RHS1 Milestone units that are used for large (4mm)thick and biopsy samples and 5 routine tissue processors. It isn't that we couldn't do everything in the Milestone units, it is a staffing issue. One of the Milestone units is used in the Surgical Pathology area for fixation and the other one is used in Histology for fixation and processing. Everything that is dissected before 10:30 in the morning goes on the Milestone unit for processing. The sections usually fill the cassettes but biopsy samples are processed on the run also if they are not STATS. The run is ready for embedding at 4:30 in the afternoon. Works great. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of UniPath IHC Sent: Wednesday, August 04, 2004 4:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave Processing I don't mean to beat a dead horse, but I have some questions about microwave processing and what I found in the archives didn't exactly answer them. Does anyone use the microwave to process ALL of their tissues? If so do you have different runs for large vs. small bxs, prostates, GIs, etc.? Do you find the quality to be as good as or better than conventional processing, or just quicker? Thank you, Brianna Jackson UniPath, LLC Denver, CO 303-512-2220 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From Don.Birgerson <@t> leica-microsystems.com Thu Aug 5 07:56:43 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers Message-ID: Hi Deb, Coming from a company with a large interest in microscopes, the physics of film vs. glass would limit the use of advanced optical techniques. Interference contrast, and plain cross polars for crystal screening would be difficult if not impossible. The use of objectives above the 4X and 10X would be compromised. With the optical problems of film, Leica has waited till we could produce a glass coverslipper. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 WWmn916@aol.com Sent by: To: histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] Glass vs. Tape Coverslippers 08/04/2004 10:04 PM Hello again, I'm looking for opinions on the subject of glass coverslippers versus tape coverslipping. I have the opportunity to decide on a system. My only experience has been with tape coverslipping. I understand machines that glass coverslip are slower than tape systems. Is the refractive index better with glass coverslips under the microscope? Opinions pros/cons are appreciated. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From dardofer <@t> hotmail.com Thu Aug 5 09:01:25 2004 From: dardofer <@t> hotmail.com (Dardo Ferrara) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Staining for Macrophages Message-ID: mac-3 rat antimouse(BD pharmingen) at 1:500-1:200 works perfectly in IHC samples. >From: Jennifer Sipes >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Staining for Macrophages >Date: Wed, 4 Aug 2004 07:28:56 -0700 (PDT) > >I had previously asked about MAC-387 staining. Unfortunately, we can't use >the staining because we are staining mouse kidney and it doesn't look right >if you stain with a mouse anti-body. Does anyone know of a way to stain >for macrophages with out an antibody??? Or perhaps a polyclonal >antibody???? Thanks a bunch! > > >Jennifer K. Sipes, RALAT >Sr. Laboratory Technician >Johns Hopkins University >Ross 929 >720 Rutland Avenue >Baltimore, MD 21205 >phone: 410-614-0131 >cell: 443-413-0853 >e-mail: jengirl1014@yahoo.com > > > > > > > > > > > >--------------------------------- >Do you Yahoo!? >Yahoo! Mail - 50x more storage than other providers! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ ?Cu?nto vale tu auto? Tips para mantener tu carro. ?De todo en MSN Latino Autos! http://latino.msn.com/autos/ From GDawson <@t> Milw.Dynacare.com Thu Aug 5 09:03:25 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Private responses Message-ID: Baowei & All, If you don't want to deal with large amounts of grief, I would encourage you to respond privately. Due to threats of calls to my employer, lawsuits, profanity, constant flaming of every response I post and attaining my own little histonet stalker (you know who you are), I keep 99% of all my responses private. There are a number of people & vendors on this listserver that revel in tearing down the posts of others rather than creating helpful or insightful posts themselves. It is not a bad thing for you to post privately to avoid these problems. I actually used to post publicly up to the point that I was dealing with personal attacks more than I was gleaning useful information from the histonet. Engaging in those types of interactions was too time consuming, draining & enfuriating. Expressing one's own view is nice and, yes, it should be OK, but it isn't. I see no value in "shaming" someone into posting publicly. If you don't want to...don't. The funny thing is that I'll be getting flamed for this post which is whining about being flamed...isn't it ironic? Just My OPINION, Glen Dawson BS, HT & QIHC (ASCP) IHC Coordinator Milwaukee, WI From DDittus787 <@t> aol.com Thu Aug 5 09:10:27 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] electrical charge slides Message-ID: <0B2E10D9.3909FF86.0A1F969F@aol.com> To my colleagues: I have a question.Is anyone out there having occasional staining(orlack of staining on IHc slides?) I was told that the charge or the way the slides are charged can have (pardon the pun) a negative effect. Does anyone know /experience or have the scientific data for this? Dana From dardofer <@t> hotmail.com Thu Aug 5 09:12:29 2004 From: dardofer <@t> hotmail.com (Dardo Ferrara) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Secondary Antibodies dilutions Message-ID: I have had problem with biotinylated Abs .I have used 10ng/ml and it seems to be too much based on a lot of background in addition to some nonspecific binding even when using streptavidin conjugates... Dardo Ferrara Emory University >From: Gayle Callis >To: Laboratorio de Histolog?a >,Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Secondary Antibodies dilutions >Date: Tue, 03 Aug 2004 07:59:31 -0600 > >A rule of thumb is to use 2 - 10 ug/ml, do the calculation from >concentration of antibody found as mg/ml in the specification sheets. Some >people even use a lesser concentration - we have done dilution panels on >our secondaries at times to determine the optimal concentration needed in >ug/ml. We tend to average around 1:250 dilution on 0.5mg/ml >concentration. Specification sheets usually give the recommended dilution >for immunohistochemistry usage. > >At 05:41 PM 8/2/2004, you wrote: >>I wanted to know if some of you uses the following Antibodies , and to >>that dilution uses them in immunohistochemistry. >> >>Biotin-Goat anti-Rabbit IgG (Zymed 65-6140 >> >>Biotin-Goat anti-Mouse (Chemicon AP181B) >> >>Thanks >> >>Dr. Hugo H. Ortega (DMV, PhD) >> >>Departament of Cellular Biology >> >>Faculty of Veterinary Sciences >> >>Universidad Nacional del Litoral >> >>R.P. Kreder 2805 - Esperanza (3080) >> >>Santa Fe - ARGENTINA >> >>Tel. (54)3496-420639 >> >>Fax. (54)3496-426304 >> >> http://fcv.unl.edu.ar/histolog/ >> >> http://fcv.unl.edu.ar/bioterio/ >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ MSN Latino: el sitio MSN para los hispanos en EE.UU. http://latino.msn.com/ From juan.gutierrez <@t> christushealth.org Thu Aug 5 09:28:23 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] electrical charge slides Message-ID: Yes, I had this problem before. Turns out that the slides we were using, StatLab's Histobond, were supercharged and the stain was repelled from the sections. We use the Ventana Benchmarks and we were getting no staining in the center of the slide. The way we tested the slides was by taking a few slides from different suppliers and putting some buffer on them while they are laying flat. Take some hematoxylin or NFR and put a drop in the middle of the slide with the buffer on it. If the slides are overcharged you will see the stain migrate to the edges of the slides rather than mixing evenly like they are supposed to. Hope this helps. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor Christus Santa Rosa Hospital 333 N. Santa Rosa Ave. San Antonio, TX 78207 (210)704-2533 -----Original Message----- From: DDittus787@aol.com [mailto:DDittus787@aol.com] Sent: Thursday, August 05, 2004 9:10 AM To: histonet@pathology.swmed.edu Subject: [Histonet] electrical charge slides To my colleagues: I have a question.Is anyone out there having occasional staining(orlack of staining on IHc slides?) I was told that the charge or the way the slides are charged can have (pardon the pun) a negative effect. Does anyone know /experience or have the scientific data for this? Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bill501 <@t> mindspring.com Thu Aug 5 09:36:01 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers In-Reply-To: References: Message-ID: At 7:56 AM -0500 8/5/04, Don.Birgerson@leica-microsystems.com wrote: >Coming from a company with a large interest in microscopes What an understatement! ;-) I have never seen a taped slide. My 1st thoughts were are they usable for polarization studies, fluorescence, oil lenses etc. Thanks of answering. BB -- ______________ Bill Blank, MD Heartland Lab From subratab <@t> bdonline.com Thu Aug 5 09:57:16 2004 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Re: difference between the several Giemsa stain. Message-ID: <200408051502.i75F2bCa029937@mailout.proshikanet.com> Dear list users (and Baowei Peng) Thank you for your mail. Yes its not good to post private reply to histonet. However I expect histonet as an open forum for discussion. It helps a lot to learn so many important things. I request you all to send a copy of your reply to histonet while sending reply privately. I think most (if not all) list users will appreciate it. Thank you again Subrata Biswas University of Campinas, Brazil From gcallis <@t> montana.edu Thu Aug 5 09:57:05 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Staining for Macrophages In-Reply-To: References: <20040804142856.20519.qmail@web60608.mail.yahoo.com> Message-ID: <6.0.0.22.1.20040805084532.01b12ef0@gemini.msu.montana.edu> Sorry, but ED-1 (CD68) is a Mouse anti-Rat antibody for a subset of rat macrophages. It may cross react to mouse but you would be doing doing mouse on mouse IHC. The CD68 for mouse is rat anti-mouse, clone FA-11 for murine peritoneal macrophages. SEROTEC is a source of these antibodies, including the F4/80 (Rat anti-mouse). A sidenote: SEROTEC has a free macrophage differentiation poster available upon request. At 06:58 PM 8/4/2004, you wrote: You can use ED-1 from Serotec. It works on FFPE tissue with pH 6.0 citrate antigen retrieval and on frozen tissue without AR. You can also use F4/80 on frozen tissue. Both are rat monoclonals and you can use an anti-rat secondary Ab cross adsorbed against mouse IgG. You could also also use ricin 1 from Vector or E-Y Laboratories which binds mouse macrophages. Ricin 1 is not the dangerous ricin. I don't know details of how to use it (I did once, but I don't remember). There are books available or the manufacturers may know or maybe some other Histonet folks. I have used ED1 and F4/80 on mouse tissues and I think it's your best bet. Sharon I had previously asked about MAC-387 staining. Unfortunately, we can't use the staining because we are staining mouse kidney and it doesn't look right if you stain with a mouse anti-body. Does anyone know of a way to stain for macrophages with out an antibody??? Or perhaps a polyclonal antibody???? Thanks a bunch! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu [1]http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu [2]http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) References 1. http://lists.utsouthwestern.edu/mailman/listinfo/histonet 2. http://lists.utsouthwestern.edu/mailman/listinfo/histonet From timothy_m_coskran <@t> groton.pfizer.com Thu Aug 5 10:01:16 2004 From: timothy_m_coskran <@t> groton.pfizer.com (Coskran, Timothy M) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] cytokeratin 19 Message-ID: <75FBD3E83B69D711A9E600080261980C02BC9F08@groexmb04bak.pfizer.com> Hello, Has anyone had success using cytokeratin 19 on formalin fixed mouse and/or rat tissues? Thanks, Tim Coskran Pfizer LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From Barry.R.Rittman <@t> uth.tmc.edu Thu Aug 5 10:02:10 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Private responses Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0E2E02A@UTHEVS3.mail.uthouston.edu> I hate to see the word "flamed" as here in sunny Houston it has been in the upper 90s for about 2 weeks. While I appreciate people's perception about getting "flamed" when they have posted items, I have a real problem with posting most items privately. Histonet is an extremely valuable resource and there is a vast amount of expertise available. It is only logical for this knowledge and experience to be freely shared. I can't say that I have been "flamed" (or even mildly scorched) but sometimes I make mistakes and post incorrect information. It is a valuable learning experience for me and all interested parties to have any errors corrected. Sometimes it is matter of perception and how criticism is leveled (being brutally frank is not a good attitude). "Flaming" may be perceived by one individual to have occurred while another may regard a response as constructive criticism. If anyone here gets "flamed" or has their employer contacted by a company about a posting then please let us know. Companies do respond positively to public opinion. Any companies with such an attitude this should be held accountable and so should employers for not telling the company or their representative to take a hike. I do believe that if someone requests private responses that this be respected. Second, I believe that when responding one should be careful when criticizing a piece of equipment or a company. Negative criticism sticks and while this may be true for a particular individual it could be an isolated incident and due to many factors. Not everyone knows how to deal effectively with companies (and not every company knows how to deal responsibly with customers). I think that such listings should be private. The individual who receives these could then let us know about the final results in the form of a summary. If I am to get "flamed" for this or any other postings please include "flamed" at the start of the message so that I don't' just think that the response is constructive criticism. Barry From JNocito <@t> Pathreflab.com Thu Aug 5 10:10:50 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Private responses In-Reply-To: Message-ID: Glen, You just burst my bubble!! I thought I was the only one getting calls, lawsuits, sworn at, etc. Man, I thought I was special. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dawson, Glen Sent: Thursday, August 05, 2004 9:03 AM Cc: 'HistoNet Server' Subject: [Histonet] Private responses Baowei & All, If you don't want to deal with large amounts of grief, I would encourage you to respond privately. Due to threats of calls to my employer, lawsuits, profanity, constant flaming of every response I post and attaining my own little histonet stalker (you know who you are), I keep 99% of all my responses private. There are a number of people & vendors on this listserver that revel in tearing down the posts of others rather than creating helpful or insightful posts themselves. It is not a bad thing for you to post privately to avoid these problems. I actually used to post publicly up to the point that I was dealing with personal attacks more than I was gleaning useful information from the histonet. Engaging in those types of interactions was too time consuming, draining & enfuriating. Expressing one's own view is nice and, yes, it should be OK, but it isn't. I see no value in "shaming" someone into posting publicly. If you don't want to...don't. The funny thing is that I'll be getting flamed for this post which is whining about being flamed...isn't it ironic? Just My OPINION, Glen Dawson BS, HT & QIHC (ASCP) IHC Coordinator Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From browning <@t> HHSC.CA Thu Aug 5 10:19:35 2004 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers Message-ID: <3AADFB88753AD31189C100902786B91C0E27836A@hch_nt_exchange.hhsc.ca> We currently use the tape system at one of our sites since we have to ship the slides immediately. That is the ONLY benefit I have seen, the slides are clean, dry, and ready to ship within minutes. The tape does not give the same refractive index, sometimes gives a cracked or creased appearance, and falls off the slide after about 2-5 years, often taking the tissue with it. I would not recommend the tape system. -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Wednesday, August 04, 2004 11:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass vs. Tape Coverslippers Hello again, I'm looking for opinions on the subject of glass coverslippers versus tape coverslipping. I have the opportunity to decide on a system. My only experience has been with tape coverslipping. I understand machines that glass coverslip are slower than tape systems. Is the refractive index better with glass coverslips under the microscope? Opinions pros/cons are appreciated. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From laurie.colbert <@t> huntingtonhospital.com Thu Aug 5 10:23:25 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BFDB@EXCHANGE1.huntingtonhospital.com> We have a tape coverslipper, and we love it. It is fast and we have had very few problems with it. Our pathologists have no problem reading the slides, and as far as I know, there's never been a problem photographing a slide. We did demo the glass coverslippers when we were first looking for a new coverslipper, and there were too many problems with slides sticking together, air bubbles, and it was just all-around not as "user friendly." Laurie Colbert Huntington Hospital -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Wednesday, August 04, 2004 8:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass vs. Tape Coverslippers Hello again, I'm looking for opinions on the subject of glass coverslippers versus tape coverslipping. I have the opportunity to decide on a system. My only experience has been with tape coverslipping. I understand machines that glass coverslip are slower than tape systems. Is the refractive index better with glass coverslips under the microscope? Opinions pros/cons are appreciated. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Thu Aug 5 10:29:23 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] electrical charge slides In-Reply-To: <0B2E10D9.3909FF86.0A1F969F@aol.com> Message-ID: Dana, We have experienced situations where the slides had too much of a positive charge that it made the slides hydrophobic. When the reagents were placed on the slide, they just sit there and bead up. Now I expect to get "FLAMED" for this, but since Fisher purchased Erie Scientific (this is what was told to me), I'm beginning to wonder how this acquisition is going to play out not just with positive slides, but all slides. Sorry, back to the question. We've had problems with positive charged slides not holding the tissue on also. I think it must be a fine line when these slides are produced as to how much chemicals are mixed with the glass. Just my experience. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of DDittus787@aol.com Sent: Thursday, August 05, 2004 9:10 AM To: histonet@pathology.swmed.edu Subject: [Histonet] electrical charge slides To my colleagues: I have a question.Is anyone out there having occasional staining(orlack of staining on IHc slides?) I was told that the charge or the way the slides are charged can have (pardon the pun) a negative effect. Does anyone know /experience or have the scientific data for this? Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Aug 5 10:37:24 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Plus charge slides and negative effect In-Reply-To: <0B2E10D9.3909FF86.0A1F969F@aol.com> References: <0B2E10D9.3909FF86.0A1F969F@aol.com> Message-ID: <6.0.0.22.1.20040805092616.01b51530@gemini.msu.montana.edu> Dana, The plus charge, whether silane or poly l lysine can create some hydrophobic bonding with proteins and/or chromogen to the charge slide surface. This does shows up as background crud on slide away around section and that ugly colored "ring around the section". This is discussed by Boenisch in DAKO handbook, the freebie. Using a detergent e.g. Tween 20 or Triton X 100 eliminates this problem. We are very careful to have a good positive control to insure staining, as you do - if we have lack of staining, look to something other than plus charge slide being the culprit for cause. You posed an interesting question. I guess one could test this question on the negative effect and use a regular slide side by side with charged slide, but loss of section is always lurking (bummer). If you ever do it, let us know the results. so At 08:10 AM 8/5/2004, you wrote: >To my colleagues: > >I have a question.Is anyone out there having occasional staining(orlack of >staining on IHc slides?) I was told that the charge or the way the slides >are charged can have (pardon the pun) a negative effect. Does anyone know >/experience or have the scientific data for this? > Dana > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From tpmorken <@t> labvision.com Thu Aug 5 10:51:32 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2D94B23@usca0082k08.labvision.apogent.com> Deb my experience with both types is that tape is fine for daily work for the vast majority of slides - those that will be looked at once and filed away forever. Tape does have some waviness that affects critical microscopy - high power and photography to some extent. I've gotten excellent pictures with tape-coverslipped slides, but glass-coverslipped slides are much more consistent for photography. The newer glass coverslippers are more reliable and faster than the old ones so speed is not such an issue. Tim Morken -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Wednesday, August 04, 2004 8:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass vs. Tape Coverslippers Hello again, I'm looking for opinions on the subject of glass coverslippers versus tape coverslipping. I have the opportunity to decide on a system. My only experience has been with tape coverslipping. I understand machines that glass coverslip are slower than tape systems. Is the refractive index better with glass coverslips under the microscope? Opinions pros/cons are appreciated. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Thu Aug 5 10:53:24 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers Message-ID: Plastic is no substitute for glass. Just because one doesn't "see" a difference doesn't mean there isn't a difference. Whether the difference makes a real difference in outcomes is another story. ASTM specs for cover glasses apply to glass, not to tape. The higher the numerical aperture of an objective, and the better the objective quality (i.e., achromat, fluorite, apochromat -- plan and non-plan), the more likely that one will see imaging differences when tape is used. Of course, using glass doesn't ensure good quality. Practical stuff like mounting medium and cover glass thickness, clean lenses, and Kohler illumination also play a real role. Image quality can't be better than the weakest link. The specs are relative to the impact of the physical and optical properties of glass on light as it passes through the mounting medium and glass through the objective. For this reason, it's not nice to fool Mother Nature and use plastic. Gary Gill -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Thursday, August 05, 2004 10:23 AM To: WWmn916@aol.com; Histonet (E-mail) Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass vs. Tape Coverslippers We have a tape coverslipper, and we love it. It is fast and we have had very few problems with it. Our pathologists have no problem reading the slides, and as far as I know, there's never been a problem photographing a slide. We did demo the glass coverslippers when we were first looking for a new coverslipper, and there were too many problems with slides sticking together, air bubbles, and it was just all-around not as "user friendly." Laurie Colbert Huntington Hospital -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Wednesday, August 04, 2004 8:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass vs. Tape Coverslippers Hello again, I'm looking for opinions on the subject of glass coverslippers versus tape coverslipping. I have the opportunity to decide on a system. My only experience has been with tape coverslipping. I understand machines that glass coverslip are slower than tape systems. Is the refractive index better with glass coverslips under the microscope? Opinions pros/cons are appreciated. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From julien_lambreydesouza <@t> uqar.qc.ca Thu Aug 5 11:08:33 2004 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Tissue Processor References: <78F91CA4B8F3504E8FE5F13AAF122ED804A02CA6@nihexchange27.nih.gov> Message-ID: <005401c47b06$75621940$9310d784@uqar.qc.ca> We have the Shandon Citadel in our lab and love it. It works like a charm and I would recomend it to others. Julien Lambrey de Souza Biologie ?volutive, Universit? du Qu?bec ? Rimouski, (418) 723-1986 #1438 ----- Original Message ----- From: "Faucette, Lawrence (NIH/NIAID)" To: "'Ian'" ; "'Dimaano, Nena'" ; Sent: Thursday, August 05, 2004 6:38 AM Subject: RE: [Histonet] Tissue Processor > I have always been a strong fan of the Sakura tissue processors. But I have > to admit, the Shandon unit, with its on-board QA/QC features combined with > their system of rotating reagents have caught my eye and would have be be > seriously evaluated > > Lawrence J Faucette > > Contractor > > HT ASCP > > > http://www.niaid.nih.gov/dir/services/animalcare/VetPathology/VetPathology-i > ndex.html > > Infectious Disease Pathogenesis Section > > Comparative Medicine Branch > > Division of Intramural Research, NIAID, NIH > > Twinbrook III, Room 2W-01A, MSC 8135 > > 12735 Twinbrook Parkway > > Bethesda, MD 20892-8135 > > Telephone 301-451-1056 > > Fax 301-480-2343 > > SoBran, Inc. > > > > Disclaimer: The information in this e-mail and any of its attachments is > confidential and may contain sensitive information. It should not be used > by anyone who is not the original intended recipient. If you have received > this e-mail in error please inform the sender and delete it from your > mailbox or any other storage devices. The National Institute of Allergy and > Infectious Diseases (NIAID) shall not accept liability for any statement > made that are the sender's own and not expressly made on behalf of the NIAID > by one of its representatives. > > > -----Original Message----- > From: Ian [mailto:Ianbernard@netzero.com] > Sent: Wednesday, August 04, 2004 10:31 PM > To: 'Dimaano, Nena'; Histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Tissue Processor > > The new VIP's > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dimaano, > Nena > Sent: Wednesday, August 04, 2004 1:04 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Tissue Processor > > We are in the process of buying a new tissue processor. Can anyone recommend > a good and reliable processor good for R&D purposes? > > thanks, > Nena Dimaano > Advanced Technology > Stryker Orthopaedics > 325 Corporate Drive > Mahwah, NJ 07430 > tel: 201-831-5338 > fax: 201-831-6224 > email: Nena.Dimaano@stryker.com > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From gcallis <@t> montana.edu Thu Aug 5 10:57:57 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Private responses - well said, Barry! In-Reply-To: <566FB0B522443D43AF02D2ADBE35A6F0E2E02A@UTHEVS3.mail.uthous ton.edu> References: <566FB0B522443D43AF02D2ADBE35A6F0E2E02A@UTHEVS3.mail.uthouston.edu> Message-ID: <6.0.0.22.1.20040805094302.01b5ee70@gemini.msu.montana.edu> Thank you Barry - very well said. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman Bozeman MT 59717-3610 At 09:02 AM 8/5/2004, you wrote: >I hate to see the word "flamed" as here in sunny Houston it has been in >the upper 90s for about 2 weeks. > >While I appreciate people's perception about getting "flamed" when they >have posted items, I have a real problem with posting most items >privately. >Histonet is an extremely valuable resource and there is a vast amount of >expertise available. It is only logical for this knowledge and >experience to be freely shared. >I can't say that I have been "flamed" (or even mildly scorched) but >sometimes I make mistakes and post incorrect information. It is a >valuable learning experience for me and all interested parties to have >any errors corrected. Sometimes it is matter of perception and how >criticism is leveled (being brutally frank is not a good attitude). >"Flaming" may be perceived by one individual to have occurred while >another may regard a response as constructive criticism. >If anyone here gets "flamed" or has their employer contacted by a >company about a posting then please let us know. Companies do respond >positively to public opinion. Any companies with such an attitude this >should be held accountable and so should employers for not telling the >company or their representative to take a hike. >I do believe that if someone requests private responses that this be >respected. Second, I believe that when responding one should be careful >when criticizing a piece of equipment or a company. Negative criticism >sticks and while this may be true for a particular individual it could >be an isolated incident and due to many factors. Not everyone knows >how to deal effectively with companies (and not every company knows how >to deal responsibly with customers). I think that such listings should >be private. The individual who receives these could then let us know >about the final results in the form of a summary. >If I am to get "flamed" for this or any other postings please include >"flamed" at the start of the message so that I don't' just think that >the response is constructive criticism. >Barry > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JQB7 <@t> CDC.GOV Thu Aug 5 11:03:47 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Private responses Message-ID: Not at all. If you knew how often I get slammed for "wasting taxpayers dollars" you'd know you were no more special than the rest of us! :) Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, August 05, 2004 11:11 AM To: Dawson, Glen Cc: 'HistoNet Server' Subject: RE: [Histonet] Private responses Glen, You just burst my bubble!! I thought I was the only one getting calls, lawsuits, sworn at, etc. Man, I thought I was special. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dawson, Glen Sent: Thursday, August 05, 2004 9:03 AM Cc: 'HistoNet Server' Subject: [Histonet] Private responses Baowei & All, If you don't want to deal with large amounts of grief, I would encourage you to respond privately. Due to threats of calls to my employer, lawsuits, profanity, constant flaming of every response I post and attaining my own little histonet stalker (you know who you are), I keep 99% of all my responses private. There are a number of people & vendors on this listserver that revel in tearing down the posts of others rather than creating helpful or insightful posts themselves. It is not a bad thing for you to post privately to avoid these problems. I actually used to post publicly up to the point that I was dealing with personal attacks more than I was gleaning useful information from the histonet. Engaging in those types of interactions was too time consuming, draining & enfuriating. Expressing one's own view is nice and, yes, it should be OK, but it isn't. I see no value in "shaming" someone into posting publicly. If you don't want to...don't. The funny thing is that I'll be getting flamed for this post which is whining about being flamed...isn't it ironic? Just My OPINION, Glen Dawson BS, HT & QIHC (ASCP) IHC Coordinator Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Thu Aug 5 11:07:43 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2D94B24@usca0082k08.labvision.apogent.com> Gary, We did find that plastic was no good for cytology specimens - at least the lumpy ones - it wouldn't form a flat surface. For thin-prep it worked great. I think the quality is OK for routine histology. In our lab we did testing in the lab and when satisfied it was working well we switched over without telling the pathologists. About two weeks later the Chief Pathologist asked me when we would start using the plastic coverslips! Tim Morken -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Thursday, August 05, 2004 8:53 AM To: 'Laurie Colbert'; WWmn916@aol.com; Histonet (E-mail) Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass vs. Tape Coverslippers Plastic is no substitute for glass. Just because one doesn't "see" a difference doesn't mean there isn't a difference. Whether the difference makes a real difference in outcomes is another story. ASTM specs for cover glasses apply to glass, not to tape. The higher the numerical aperture of an objective, and the better the objective quality (i.e., achromat, fluorite, apochromat -- plan and non-plan), the more likely that one will see imaging differences when tape is used. Of course, using glass doesn't ensure good quality. Practical stuff like mounting medium and cover glass thickness, clean lenses, and Kohler illumination also play a real role. Image quality can't be better than the weakest link. The specs are relative to the impact of the physical and optical properties of glass on light as it passes through the mounting medium and glass through the objective. For this reason, it's not nice to fool Mother Nature and use plastic. Gary Gill -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Thursday, August 05, 2004 10:23 AM To: WWmn916@aol.com; Histonet (E-mail) Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass vs. Tape Coverslippers We have a tape coverslipper, and we love it. It is fast and we have had very few problems with it. Our pathologists have no problem reading the slides, and as far as I know, there's never been a problem photographing a slide. We did demo the glass coverslippers when we were first looking for a new coverslipper, and there were too many problems with slides sticking together, air bubbles, and it was just all-around not as "user friendly." Laurie Colbert Huntington Hospital -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Wednesday, August 04, 2004 8:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass vs. Tape Coverslippers Hello again, I'm looking for opinions on the subject of glass coverslippers versus tape coverslipping. I have the opportunity to decide on a system. My only experience has been with tape coverslipping. I understand machines that glass coverslip are slower than tape systems. Is the refractive index better with glass coverslips under the microscope? Opinions pros/cons are appreciated. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Thu Aug 5 11:12:48 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2D94B25@usca0082k08.labvision.apogent.com> Bill, Plastic is fine for polarazatin (at least the limited use of polarization in a routine pathology lab) and oil immersion is no problem. Can't be used for fluorescence, however, because the tape is adhered with xylene. The tape coverslipping machine is primarily for speed - 20 slides per minute - so it is intended for routine pathology and cranking out slides very quickly. Tim Morken -----Original Message----- From: Bill Blank [mailto:bill501@mindspring.com] Sent: Thursday, August 05, 2004 7:36 AM To: histonet@lists.utsouthwestern.edu Cc: Don.Birgerson@leica-microsystems.com Subject: Re: [Histonet] Glass vs. Tape Coverslippers At 7:56 AM -0500 8/5/04, Don.Birgerson@leica-microsystems.com wrote: >Coming from a company with a large interest in microscopes What an understatement! ;-) I have never seen a taped slide. My 1st thoughts were are they usable for polarization studies, fluorescence, oil lenses etc. Thanks of answering. BB -- ______________ Bill Blank, MD Heartland Lab _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Thu Aug 5 11:13:10 2004 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Private responses - well said, Barry! Message-ID: <080520041613.7189.41125C950005EF5000001C152200763704CECE030E9D0C9A03@comcast.net> I agree with both Gayle and Barry yet have one reservation. I was hit several times on this server for answering very general questions and told salespeople should not be on HistoNet. Although my answers had nothing to do with any product we sell and by the way I am not a sales person, just technical and product development. After the second incident that several others came on and agreed we were not welcome I stay off 99% of the time. It is difficult to walk the line here and not be on privately only. I do agree that if it is a sales pitch it should not be on the line as a reply that should be private answers only. Thanks for letting me say this and I hope no one is offended. Pam Marcum -------------- Original message -------------- > Thank you Barry - very well said. > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > Bozeman MT 59717-3610 > > At 09:02 AM 8/5/2004, you wrote: > > >I hate to see the word "flamed" as here in sunny Houston it has been in > >the upper 90s for about 2 weeks. > > > >While I appreciate people's perception about getting "flamed" when they > >have posted items, I have a real problem with posting most items > >privately. > >Histonet is an extremely valuable resource and there is a vast amount of > >expertise available. It is only logical for this knowledge and > >experience to be freely shared. > >I can't say that I have been "flamed" (or even mildly scorched) but > >sometimes I make mistakes and post incorrect information. It is a > >valuable learning experience for me and all interested parties to have > >any errors corrected. Sometimes it is matter of perception and how > >criticism is leveled (being brutally frank is not a good attitude). > >"Flaming" may be perceived by one individual to have occurred while > >another may regard a response as constructive criticism. > >If anyone here gets "flamed" or has their employer contacted by a > >company about a posting then please let us know. Companies do respond > >positively to public opinion. Any companies with such an attitude this > >should be held accountable and so should employers for not telling the > >company or their representative to take a hike. > >I do believe that if someone requests private responses that this be > >respected. Second, I believe that when responding one should be careful > >when criticizing a piece of equipment or a company. Negative criticism > >sticks and while this may be true for a particular individual it could > >be an isolated incident and due to many factors. Not everyone knows > >how to deal effectively with companies (and not every company knows how > >to deal responsibly with customers). I think that such listings should > >be private. The individual who receives these could then let us know > >about the final results in the form of a summary. > >If I am to get "flamed" for this or any other postings please include > >"flamed" at the start of the message so that I don't' just think that > >the response is constructive criticism. > >Barry > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cleger <@t> seattlecca.org Thu Aug 5 11:27:49 2004 From: cleger <@t> seattlecca.org (Leger, Carolyn A) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Gram stains Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B94841BAD7@wala01.seattlecca.org> I am looking for a Grams Iodine and a Basic Fuchsin stain that is commercially made that is comparable to ours that is made in-house. We already use the commercial Genetian Violet for our Gram stain. Can anyone please give me some opinions on their experience with either stain? Thanks, Carolyn Carolyn A. Leger Pathology Department 206/288-2232 206/288-1355 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please notify me by telephone at (206) 288-2232 or by electronic reply, and delete this message. Thank you This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From JWEEMS <@t> sjha.org Thu Aug 5 12:09:25 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Glass vs. Tape Coverslippers Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A626B@sjhaexc02.sjha.org> I was demoing coverslippers and didn't tell the pathologists. Thought I'd see if they could tell the difference. One of our pathologists made us remove the tape from her entire days workload and recoverslip them by hand. Oh well..... Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Morken, Tim - Labvision Sent: Thursday, August 05, 2004 12:08 PM To: 'Histonet (E-mail)' Subject: RE: [Histonet] Glass vs. Tape Coverslippers Gary, We did find that plastic was no good for cytology specimens - at least the lumpy ones - it wouldn't form a flat surface. For thin-prep it worked great. I think the quality is OK for routine histology. In our lab we did testing in the lab and when satisfied it was working well we switched over without telling the pathologists. About two weeks later the Chief Pathologist asked me when we would start using the plastic coverslips! Tim Morken -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Thursday, August 05, 2004 8:53 AM To: 'Laurie Colbert'; WWmn916@aol.com; Histonet (E-mail) Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass vs. Tape Coverslippers Plastic is no substitute for glass. Just because one doesn't "see" a difference doesn't mean there isn't a difference. Whether the difference makes a real difference in outcomes is another story. ASTM specs for cover glasses apply to glass, not to tape. The higher the numerical aperture of an objective, and the better the objective quality (i.e., achromat, fluorite, apochromat -- plan and non-plan), the more likely that one will see imaging differences when tape is used. Of course, using glass doesn't ensure good quality. Practical stuff like mounting medium and cover glass thickness, clean lenses, and Kohler illumination also play a real role. Image quality can't be better than the weakest link. The specs are relative to the impact of the physical and optical properties of glass on light as it passes through the mounting medium and glass through the objective. For this reason, it's not nice to fool Mother Nature and use plastic. Gary Gill -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Thursday, August 05, 2004 10:23 AM To: WWmn916@aol.com; Histonet (E-mail) Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass vs. Tape Coverslippers We have a tape coverslipper, and we love it. It is fast and we have had very few problems with it. Our pathologists have no problem reading the slides, and as far as I know, there's never been a problem photographing a slide. We did demo the glass coverslippers when we were first looking for a new coverslipper, and there were too many problems with slides sticking together, air bubbles, and it was just all-around not as "user friendly." Laurie Colbert Huntington Hospital -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Wednesday, August 04, 2004 8:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass vs. Tape Coverslippers Hello again, I'm looking for opinions on the subject of glass coverslippers versus tape coverslipping. I have the opportunity to decide on a system. My only experience has been with tape coverslipping. I understand machines that glass coverslip are slower than tape systems. Is the refractive index better with glass coverslips under the microscope? Opinions pros/cons are appreciated. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From scoop <@t> mail.nih.gov Thu Aug 5 12:24:18 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Staining for Macrophages In-Reply-To: <6.0.0.22.1.20040805084532.01b12ef0@gemini.msu.montana.edu> References: <20040804142856.20519.qmail@web60608.mail.yahoo.com> <6.0.0.22.1.20040805084532.01b12ef0@gemini.msu.montana.edu> Message-ID: Thanks, Gail, you're right. Does FA-11 work on FFPE tissue and do you need HIER? Serotec doesn't say. Thanks, Sharon >Sorry, but ED-1 (CD68) is a Mouse anti-Rat antibody for a subset of >rat macrophages. It may cross react to mouse but you would be doing >doing mouse on mouse IHC. The CD68 for mouse is rat anti-mouse, >clone FA-11 for murine peritoneal macrophages. SEROTEC is a source >of these antibodies, including the F4/80 (Rat anti-mouse). > >A sidenote: SEROTEC has a free macrophage differentiation poster >available upon request. > >At 06:58 PM 8/4/2004, you wrote: > >>You can use ED-1 from Serotec. It works on FFPE tissue with pH 6.0 >>citrate antigen retrieval and on frozen tissue without AR. You can >>also use F4/80 on frozen tissue. Both are rat monoclonals and you >>can use an anti-rat secondary Ab cross adsorbed against mouse IgG. >>You could also also use ricin 1 from Vector or E-Y Laboratories >>which binds mouse macrophages. Ricin 1 is not the dangerous ricin. >>I don't know details of how to use it (I did once, but I don't >>remember). There are books available or the manufacturers may know >>or maybe some other Histonet folks. I have used ED1 and F4/80 on >>mouse tissues and I think it's your best bet. >> >>Sharon >> >>>I had previously asked about MAC-387 staining. Unfortunately, we >>>can't use the staining because we are staining mouse kidney and it >>>doesn't look right if you stain with a mouse anti-body. Does >>>anyone know of a way to stain for macrophages with out an >>>antibody??? Or perhaps a polyclonal antibody???? Thanks a bunch! >>> >>> >>>Jennifer K. Sipes, RALAT >>>Sr. Laboratory Technician >>>Johns Hopkins University >>>Ross 929 >>>720 Rutland Avenue >>>Baltimore, MD 21205 >>>phone: 410-614-0131 >>>cell: 443-413-0853 >>>e-mail: jengirl1014@yahoo.com >>> >>> >>> >>> >>> >>> >>> >>> >>> >>> >>> >>>--------------------------------- >>>Do you Yahoo!? >>>Yahoo! Mail - 50x more storage than other providers! >>>_______________________________________________ >>>Histonet mailing list >>>Histonet@lists.utsouthwestern.edu >>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >> >> >>-- >>Sharon Cooperman >>NIH, NICHD, CBMB 301.435-7735 >>Building 18T, room 101 301.402-0078 fax >>Bethesda, MD 20892 >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From gcallis <@t> montana.edu Thu Aug 5 12:41:52 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Private responses - well said, Barry! In-Reply-To: <080520041613.7189.41125C950005EF5000001C152200763704CECE03 0E9D0C9A03@comcast.net> References: <080520041613.7189.41125C950005EF5000001C152200763704CECE030E9D0C9A03@comcast.net> Message-ID: <6.0.0.22.1.20040805112541.01afad70@gemini.msu.montana.edu> Pam and all, Too many people forget or ignore the fact that some individuals who are experts in the field of histotechnology are not employed in just clinical or research laboratories. If one is involved with a company doing technical research and development of a product destined for histoland, then they are should be very welcome to comment. Knowledge should not be smothered (flames!) I don't see you as a salesperson, but a valuable resource providing me with information I can choose to use or not. All part of the learning experience. Complaints = delete button. At 10:13 AM 8/5/2004, you wrote: I agree with both Gayle and Barry yet have one reservation. I was hit several times on this server for answering very general questions and told salespeople should not be on HistoNet. Although my answers had nothing to do with any product we sell and by the way I am not a sales person, just technical and product development. After the second incident that several others came on and agreed we were not welcome I stay off 99% of the time. It is difficult to walk the line here and not be on privately only. I do agree that if it is a sales pitch it should not be on the line as a reply that should be private answers only. Thanks for letting me say this and I hope no one is offended. Pam Marcum -------------- Original message -------------- > Thank you Barry - very well said. > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > Bozeman MT 59717-3610 > > At 09:02 AM 8/5/2004, you wrote: > > >I hate to see the word "flamed" as here in sunny Houston it has been in > >the upper 90s for about 2 weeks. > > > >While I appreciate people's perception about getting "flamed" when they > >have posted items, I have a real problem with posting most items > >privately. > >Histonet is an extremely valuable resource and there is a vast amount of > >expertise available. It is only logical for this knowledge and > >experience to be f! reely shared. > >I can't say that I have been "flamed" (or even mildly scorched) but > >sometimes I make mistakes and post incorrect information. It is a > >valuable learning experience for me and all interested parties to have > >any errors corrected. Sometimes it is matter of perception and how > >criticism is leveled (being brutally frank is not a good attitude). > >"Flaming" may be perceived by one individual to have occurred while > >another may regard a response as constructive criticism. > >If anyone here gets "flamed" or has their employer contacted by a > >company about a posting then please let us know. Companies do respond > >positively to public opinion. Any companies with such an attitude this > >should be held accountable and so should employers for not telling the > >company or their representative to take a hike. > >I do believe t! hat if someone requests private responses that this be >! ; >re spected. Second, I believe that when responding one should be careful > >when criticizing a piece of equipment or a company. Negative criticism > >sticks and while this may be true for a particular individual it could > >be an isolated incident and due to many factors. Not everyone knows > >how to deal effectively with companies (and not every company knows how > >to deal responsibly with customers). I think that such listings should > >be private. The individual who receives these could then let us know > >about the final results in the form of a summary. > >If I am to get "flamed" for this or any other postings please include > >"flamed" at the start of the message so that I don't' just think that > >the response is constructive criticism. > >Barry > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >[1]http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > [2]http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) References 1. http://lists.utsouthwestern.edu/mailman/listinfo/histonet 2. http://lists.utsouthwestern.edu/mailman/listinfo/histonet From susan.wells <@t> bms.com Thu Aug 5 13:20:13 2004 From: susan.wells <@t> bms.com (Susan Q Wells) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Private responses - well said, Barry! In-Reply-To: <6.0.0.22.1.20040805112541.01afad70@gemini.msu.montana.edu> References: <080520041613.7189.41125C950005EF5000001C152200763704CECE030E9D0C9A03@comcast.net> <6.0.0.22.1.20040805112541.01afad70@gemini.msu.montana.edu> Message-ID: <41127A5D.4080406@bms.com> I completely agree with Gayle! This list is invaluable to most of us working in the field. Gayle Callis wrote: > Pam and all, > Too many people forget or ignore the fact that some individuals who > are experts in the field of histotechnology are not employed in just > clinical or research laboratories. If one is involved with a company > doing technical research and development of a product destined for > histoland, then they are should be very welcome to comment. > Knowledge should not be smothered (flames!) I don't see you as a > salesperson, but a valuable resource providing me with information I > can choose to use or not. All part of the learning experience. > Complaints = delete button. > > > At 10:13 AM 8/5/2004, you wrote: > > I agree with both Gayle and Barry yet have one reservation. I was > hit several times on this server for answering very general > questions and told salespeople should not be on HistoNet. Although > my answers had nothing to do with any product we sell and by the > way I am not a sales person, just technical and product > development. After the second incident that several others came on > and agreed we were not welcome I stay off 99% of the time. It is > difficult to walk the line here and not be on privately only. I do > agree that if it is a sales pitch it should not be on the line as a > reply that should be private answers only. > Thanks for letting me say this and I hope no one is offended. > Pam Marcum > > -------------- Original message -------------- > > Thank you Barry - very well said. > > > > Gayle Callis > > MT,HT,HTL(ASCP) > > Research Histopathology Supervisor > > Veterinary Molecular Biology > > Montana State University - Bozeman > > Bozeman MT 59717-3610 > > > > At 09:02 AM 8/5/2004, you wrote: > > > > >I hate to see the word "flamed" as here in sunny Houston it > has been in > > >the upper 90s for about 2 weeks. > > > > > >While I appreciate people's perception about getting > "flamed" when they > > >have posted items, I have a real problem with posting most > items > > >privately. > > >Histonet is an extremely valuable resource and there is a > vast amount of > > >expertise available. It is only logical for this knowledge > and > > >experience to be f! reely shared. > > >I can't say that I have been "flamed" (or even mildly > scorched) but > > >sometimes I make mistakes and post incorrect information. It > is a > > >valuable learning experience for me and all interested > parties to have > > >any errors corrected. Sometimes it is matter of perception > and how > > >criticism is leveled (being brutally frank is not a good > attitude). > > >"Flaming" may be perceived by one individual to have > occurred while > > >another may regard a response as constructive criticism. > > >If anyone here gets "flamed" or has their employer contacted > by a > > >company about a posting then please let us know. Companies > do respond > > >positively to public opinion. Any companies with such an > attitude this > > >should be held accountable and so should employers for not > telling the > > >company or their representative to take a hike. > > >I do believe t! hat if someone requests private responses > that this be > >! > ; >re spected. Second, I believe that when responding one > should be careful > > >when criticizing a piece of equipment or a company. Negative > criticism > > >sticks and while this may be true for a particular > individual it could > > >be an isolated incident and due to many factors. Not > everyone knows > > >how to deal effectively with companies (and not every > company knows how > > >to deal responsibly with customers). I think that such > listings should > > >be private. The individual who receives these could then let > us know > > >about the final results in the form of a summary. > > >If I am to get "flamed" for this or any other postings > please include > > >"flamed" at the start of the message so that I don't' just > think that > > >the response is constructive criticism. > > >Barry > > > > > > > > >_______________________________________________ > > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > > >[1]http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > [2]http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > > MT,HT,HTL(ASCP) > > Research Histopathology Supervisor > > Veterinary Molecular Biology > > Montana State University - Bozeman > > PO Box 173610 > > Bozeman MT 59717-3610 > > 406 994-6367 (lab with voice mail) > > 406 994-4303 (FAX) > >References > > 1. http://lists.utsouthwestern.edu/mailman/listinfo/histonet > 2. http://lists.utsouthwestern.edu/mailman/listinfo/histonet >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From jstaruk <@t> masshistology.com Thu Aug 5 13:33:59 2004 From: jstaruk <@t> masshistology.com (Mass Histology Service) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Private responses In-Reply-To: <6.0.0.22.1.20040805112541.01afad70@gemini.msu.montana.edu> Message-ID: Especially the archives! Quite often a topic is of no interest to me at the present time yet a week later, I have to stain for that certain CD antibody that I remember was discussed in detail the prior week. I use the archives several times a week and if all responses were private, there would be no archives! Jim ____________________ James Staruk, HT(ASCP) Mass Histology Service www.masshistology.com From browning <@t> HHSC.CA Thu Aug 5 14:05:35 2004 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] (no subject) Message-ID: <3AADFB88753AD31189C100902786B91C0E27836D@hch_nt_exchange.hhsc.ca> Here is a puzzle. In order to streamline the IHC workload, I have had the techs organize the slides in vertical racks that can be used for dewaxing, heat retrieval, and quenching (meth/px), instead of the old way of dewaxing in one set of racks, switching to other racks for retrieval, and then into coplin jars for quenching. Sounds great. We ended up with an unusual vertical staining pattern with central staining of the tissues, but both edges that corresponded with the edges of the slide racks had no staining. The slides were heated at 60 deg. overnight. Slides that did not require heat retrieval were not affected, therefore it was not a dewaxing problem. All slides lay flat for application of antibodies and detection system. I know of another site that vertically handles their slides, and they do not get this artefact, the only difference between them and us is quenching before retrieval vs after. Any suggestions? We went back to the old way, and the artefact is gone, nothing changed except the position (vertical vs horizontal) of the slides. Looking forward to scientific explanations, inquiring minds want to know. Debra Browning, ART Technical Specialist, Immunohistochemistry Anatomic Pathology Hamilton Health Sciences phone: (905) 527-4322 ext 46131 e-mail: browning@hhsc.ca fax: (905) 524-2681 This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From browning <@t> HHSC.CA Thu Aug 5 14:07:12 2004 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Recall: Message-ID: <3AADFB88753AD31189C100902786B91C0E27836E@hch_nt_exchange.hhsc.ca> Browning Deb would like to recall the message, "". This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From browning <@t> HHSC.CA Thu Aug 5 14:07:41 2004 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] retrieval puzzle, forgot this line on last message Message-ID: <3AADFB88753AD31189C100902786B91C0E27836F@hch_nt_exchange.hhsc.ca> Here is a puzzle. In order to streamline the IHC workload, I have had the techs organize the slides in vertical racks that can be used for dewaxing, heat retrieval, and quenching (meth/px), instead of the old way of dewaxing in one set of racks, switching to other racks for retrieval, and then into coplin jars for quenching. Sounds great. We ended up with an unusual vertical staining pattern with central staining of the tissues, but both edges that corresponded with the edges of the slide racks had no staining. The slides were heated at 60 deg. overnight. Slides that did not require heat retrieval were not affected, therefore it was not a dewaxing problem. All slides lay flat for application of antibodies and detection system. I know of another site that vertically handles their slides, and they do not get this artefact, the only difference between them and us is quenching before retrieval vs after. Any suggestions? We went back to the old way, and the artefact is gone, nothing changed except the position (vertical vs horizontal) of the slides. Looking forward to scientific explanations, inquiring minds want to know. Debra Browning, ART Technical Specialist, Immunohistochemistry Anatomic Pathology Hamilton Health Sciences phone: (905) 527-4322 ext 46131 e-mail: browning@hhsc.ca fax: (905) 524-2681 This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From jwatson <@t> gnf.org Thu Aug 5 14:31:21 2004 From: jwatson <@t> gnf.org (James Watson) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Jeff Lowen Message-ID: <7BA50F61CB491B4692B8BCFBB656B5F903428986@EXCHCLUSTER01.lj.gnf.org> Jeff Lowen, Please contact me. Thank you. James Watson HT, ASCP Facilities Manager of Histology GNF, C015 858-332-4647 jwatson@gnf.org From lab <@t> mcpathology.com Thu Aug 5 13:50:20 2004 From: lab <@t> mcpathology.com (Lab) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] Benchmark XT Message-ID: <001101c47b1d$0f0f5e70$70cfa8c0@LAB> Our lab has had 2 ventana nexes systems and we are looking to upgrade to 2 benchmark xt units.We have had one in our lab to demo for approx. 3 months.During this time we have had 2 thermal pad issues and several file corruptions. Has anyone with a benchmark xt had similar problems and if so how often? Any opions about the instrument would be helpful. Thanks, Amy Boan,HT From kmjacobs <@t> vcu.edu Thu Aug 5 15:32:43 2004 From: kmjacobs <@t> vcu.edu (Kimberle M. Jacobs) Date: Fri Sep 16 15:23:50 2005 Subject: [Histonet] BRDU free floating vibratome sections Message-ID: <5.1.0.14.2.20040805162931.01c9f2a8@mail1.vcu.edu> Hi I have searched the archives and found a lot of discussion of BRDU, but no answer to the question if there is a protocol out there for formaldehyde-fixed, vibratome-cut, free floating sections. Actually we could mount them on slides as well, I just don't want to do the whole paraffin embedding thing. Also is it really necessary to have thin sections if you are going to use pepsin and acid, etc. ?? We could probably cut as thin as 20 um, but would prefer to use 40 or 60 um. We will be doing this in pregnant ferrets (E36, gestation is ~42 days) and examining the brains at ages P0-P15. Thanks very much for any help!! Kimberle M. Jacobs, Ph.D. Assistant Professor Department of Anatomy and Neurobiology Virginia Commonwealth University P.O. Box 980709 Richmond, VA 23298-0709 (804) 827-2135 FAX: (804) 828-9477 kmjacobs@vcu.edu From kmjacobs <@t> vcu.edu Thu Aug 5 15:36:57 2004 From: kmjacobs <@t> vcu.edu (Kimberle M. Jacobs) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] ID-2 Message-ID: <5.1.0.14.2.20040805163247.01c9d160@mail1.vcu.edu> Has anyone used the antibody to ID-2? We purchased it from Santa Cruz Biotech. They had some recommendations for western blot, but none for immunohistochemistry. Our goal is simply to label the superficial layers of ferret visual cortex. But on our first try using 1:50, 1:100, and 1:200 primary dilutions - it didn't work at all. We were wondering if we need to do some antigen revealing step. Or perhaps it will not be found this caudal? Anyone know of any antibodies that label a specific layer in visual cortex? I've heard that the otx1 antibody is not working well right now. We are going to get RORbeta to attempt to stain layer IV, but again - no specific protocol for it. Thanks very much for any help! Kimberle M. Jacobs, Ph.D. Assistant Professor Department of Anatomy and Neurobiology Virginia Commonwealth University P.O. Box 980709 Richmond, VA 23298-0709 (804) 827-2135 FAX: (804) 828-9477 kmjacobs@vcu.edu From JNocito <@t> Pathreflab.com Thu Aug 5 15:44:15 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Benchmark XT In-Reply-To: <001101c47b1d$0f0f5e70$70cfa8c0@LAB> Message-ID: Amy, I have a new carousal waiting to be installed as we speak. I have 2 XT's, the oldest one since February of this year. I've had 8 different heating pads go bad on three different occasions since February. Most of the time, we place our control on the same slide as the patient, so if the control worked, there is no question that everything worked okay. If the control is negative, then we repeat the slide on the newer machine. In all fairness to Vetnana, they have responded well to our phone calls and emails. I think this is an issue that they are trying to work on in house. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Lab Sent: Thursday, August 05, 2004 1:50 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Benchmark XT Our lab has had 2 ventana nexes systems and we are looking to upgrade to 2 benchmark xt units.We have had one in our lab to demo for approx. 3 months.During this time we have had 2 thermal pad issues and several file corruptions. Has anyone with a benchmark xt had similar problems and if so how often? Any opions about the instrument would be helpful. Thanks, Amy Boan,HT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjones <@t> cvm.tamu.edu Thu Aug 5 16:57:22 2004 From: sjones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] BRDU free floating vibratome sections Message-ID: Hi Kimberle, I've done quite a bit of vibratome sectioning and free floating staining. It's quite challenging! Two ways I can think of that might help you. One would be to use the Shandon coverplate system. The sections would be floated onto slides and then the coverplate carefully placed on to the slide. It then fits into a holder and it gravity feeds the reagents over the slide. It uses a very small amount of reagent. The other way is to handle the free floating sections in netwells or you can make your own using open cassettes and small petri dishes. The most important steps would be making sure the rinsing steps were sufficient. The netwell/petri dish method would use a much greater amount of reagent. Hope this helps. Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Kimberle M. Jacobs" 08/05/04 3:32 PM >>> Hi I have searched the archives and found a lot of discussion of BRDU, but no answer to the question if there is a protocol out there for formaldehyde-fixed, vibratome-cut, free floating sections. Actually we could mount them on slides as well, I just don't want to do the whole paraffin embedding thing. Also is it really necessary to have thin sections if you are going to use pepsin and acid, etc. ?? We could probably cut as thin as 20 um, but would prefer to use 40 or 60 um. We will be doing this in pregnant ferrets (E36, gestation is ~42 days) and examining the brains at ages P0-P15. Thanks very much for any help!! Kimberle M. Jacobs, Ph.D. Assistant Professor Department of Anatomy and Neurobiology Virginia Commonwealth University P.O. Box 980709 Richmond, VA 23298-0709 (804) 827-2135 FAX: (804) 828-9477 kmjacobs@vcu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Thu Aug 5 18:39:09 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] H. pylori control tissue Message-ID: Is anyone looking for H. pylori control tissue? We just had a partial resection of stomach and it is loaded with bugs. If there is an overwhelming response, I may just send control blocks to the NSH tissue bank for distribution. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Hartford Hospital Hartford, CT 06102 (860) 545-1596 From AnthonyH <@t> chw.edu.au Thu Aug 5 21:31:42 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Private responses Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E230@simba.kids> Glen, I respect your right to free choice and accept your views. Keep well, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Dawson, Glen [mailto:GDawson@Milw.Dynacare.com] Sent: Friday, 6 August 2004 12:03 AM Cc: 'HistoNet Server' Subject: [Histonet] Private responses Baowei & All, If you don't want to deal with large amounts of grief, I would encourage you to respond privately. Due to threats of calls to my employer, lawsuits, profanity, constant flaming of every response I post and attaining my own little histonet stalker (you know who you are), I keep 99% of all my responses private. There are a number of people & vendors on this listserver that revel in tearing down the posts of others rather than creating helpful or insightful posts themselves. It is not a bad thing for you to post privately to avoid these problems. I actually used to post publicly up to the point that I was dealing with personal attacks more than I was gleaning useful information from the histonet. Engaging in those types of interactions was too time consuming, draining & enfuriating. Expressing one's own view is nice and, yes, it should be OK, but it isn't. I see no value in "shaming" someone into posting publicly. If you don't want to...don't. The funny thing is that I'll be getting flamed for this post which is whining about being flamed...isn't it ironic? Just My OPINION, Glen Dawson BS, HT & QIHC (ASCP) IHC Coordinator Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From bills <@t> icpmr.wsahs.nsw.gov.au Thu Aug 5 21:54:41 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] electrical charge slides In-Reply-To: <0B2E10D9.3909FF86.0A1F969F@wsahs.nsw.gov.au> Message-ID: <000201c47b60$b8b82810$e1ce080a@wsahs.nsw.gov.au> Dana, There was heaps written about this on Histonet previously, look in the archives. We found that the problem was associated with the Plus slides when used in conjunction with an EDTA buffer as the material used to coat the slides is a calcium salt and the EDTA was affecting the coating by chelating the calcium!!! All the best Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of DDittus787@aol.com Sent: Friday, 06 August 2004 12:10 AM To: histonet@pathology.swmed.edu Subject: [Histonet] electrical charge slides To my colleagues: I have a question.Is anyone out there having occasional staining(orlack of staining on IHc slides?) I was told that the charge or the way the slides are charged can have (pardon the pun) a negative effect. Does anyone know /experience or have the scientific data for this? Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From p_bourne_14526 <@t> yahoo.com Thu Aug 5 21:55:56 2004 From: p_bourne_14526 <@t> yahoo.com (Patricia Bourne) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Rank and Rank L Message-ID: <20040806025556.77653.qmail@web53706.mail.yahoo.com> Hello everyone....Has anyone worked successfully with Rank and Rank L? I've been through three (3) antibody sets and still not great results.....Help is really needed on this one. Thanks in advance for the help and information. --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From Bdeer24 <@t> aol.com Fri Aug 6 02:19:37 2004 From: Bdeer24 <@t> aol.com (Bdeer24@aol.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Problem with Masson's Trichrome Message-ID: <10A0ACD8.74118519.0015B53C@aol.com> Hi Everyone! I was recommended to try the Masson's Trichrome stainings on the slides of uterine tissue that I have been collecting (Previously I had only performed Hematoxylin & Eosin stains). This is a new method to our lab and thus far, staining has not gone well. Although my sections contain a significant amount of muscle, I have only been able to see the aniline blue stain. On a couple of the slides, there was a slight dark purple coloration on the epithelial lining. I have noticed upon comparing the protocol we received to others found on the internet that ours lacks the step in which the slides are placed in Bouin's fluid. There was also no specification in our protocol for the type of hematoxylin, so we had been using the same hematoxylin (Harris's modified with acetic acid) that we use in our H & E stainings. I was wondering if any of these might have led to the stainings not going as planned. The tissue sections also looked slightly brittle, perhaps due to a harsher stain. I am wondering if this has anything to do with my fixation process in which I use a 4% paraformaldehyde/PBS mixture, ethanol and xylene steps, as well as embedding the tissues in paraffin. Is Bouin's a crucial step, would I have better staining with a different tissue fixation, or should I be looking in a different direction? Any suggestions would be very much appreciated! Thanks! Amber Greenbank Arizona State University From lpwenk <@t> sbcglobal.net Fri Aug 6 03:44:36 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] H. pylori control tissue References: Message-ID: <007d01c47b91$9b6e7bc0$7434d445@domainnotset.invalid> I believe how the NSH Control bank works is that you let NSH control block coordinator know what you are willing to share. You then hang onto the blocks, and if anyone contacts NSH asking for a specific control, they receive a list of the people willing to share. That person then calls someone on the list, asks for the control, and the person with the control sends it out. Below is the NSH control bank person, if I remember correctly. Quality Control Ethel Macrea Ventana Medical Systems 1910 E. Innovation Park Drive Tucson, AZ 85739 800-227-2155 ext. 3856 emacrea@ventanamed.com Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI ----- Original Message ----- From: "Richard Cartun" To: Sent: Thursday, August 05, 2004 7:39 PM Subject: [Histonet] H. pylori control tissue > Is anyone looking for H. pylori control tissue? We just had a partial > resection of stomach and it is loaded with bugs. If there is an > overwhelming response, I may just send control blocks to the NSH tissue > bank for distribution. > > Richard > > Richard W. Cartun, Ph.D. > Director, Immunopathology & Histology > Hartford Hospital > Hartford, CT 06102 > (860) 545-1596 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Myri37 <@t> aol.com Fri Aug 6 04:34:20 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Fixative for FACS Message-ID: <69986EA4.0E45DDBE.0005167B@aol.com> hello Do you know which fixative can i use for intracellular immunostaining for FACS ? We use saponine 0.1 % for permeabilisation. Thank you for your help Myriam Natural implant From Tbarnhart <@t> primecare.org Fri Aug 6 06:11:22 2004 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Oncocytoma vs. Chromophobe Message-ID: <1779904B5E82D511914C00D0B793339205BFD830@exchangent> I was wondering if there is(are) any immunohistochemical antibodies or special stains besides Hale's Colloidal Iron for the differentiation of Oncocytoma and Chromophobe in kidney? I am not able to make the dialysised (sp?) iron needed for the Hale's but would like to offer my pathologist an alternate. Any suggestions? Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. From rschoon <@t> email.unc.edu Fri Aug 6 07:25:17 2004 From: rschoon <@t> email.unc.edu (rschoon) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Problem with Masson's Trichrome In-Reply-To: <10A0ACD8.74118519.0015B53C@aol.com> References: <10A0ACD8.74118519.0015B53C@aol.com> Message-ID: <411378AD.8020806@email.unc.edu> Amber, Don't know where you got your protocol but I would suggest ANY of the standerd Histology books. As for an answer to your problems. 1- mordent in either Boin's fluid or saturated picric acid for 60 min. at 60oC (there are other time, temp.'s and concentrations if one looks them up). The method simply will not work without the modent. 2- use Weigert's Iron Hematoxylin as the nuclear stain. Robert Schoonhoven Bdeer24@aol.com wrote: >Hi Everyone! > >I was recommended to try the Masson's Trichrome stainings on the slides of uterine tissue that I have been collecting (Previously I had only performed Hematoxylin & Eosin stains). This is a new method to our lab and thus far, staining has not gone well. Although my sections contain a significant amount of muscle, I have only been able to see the aniline blue stain. On a couple of the slides, there was a slight dark purple coloration on the epithelial lining. > >I have noticed upon comparing the protocol we received to others found on the internet that ours lacks the step in which the slides are placed in Bouin's fluid. There was also no specification in our protocol for the type of hematoxylin, so we had been using the same hematoxylin (Harris's modified with acetic acid) that we use in our H & E stainings. I was wondering if any of these might have led to the stainings not going as planned. The tissue sections also looked slightly brittle, perhaps due to a harsher stain. I am wondering if this has anything to do with my fixation process in which I use a 4% paraformaldehyde/PBS mixture, ethanol and xylene steps, as well as embedding the tissues in paraffin. > >Is Bouin's a crucial step, would I have better staining with a different tissue fixation, or should I be looking in a different direction? Any suggestions would be very much appreciated! >Thanks! > >Amber Greenbank > >Arizona State University > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From hasselblatt <@t> uni-muenster.de Fri Aug 6 08:08:47 2004 From: hasselblatt <@t> uni-muenster.de (Martin Hasselblatt) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Cytochrome oxidase (COX) staining: non-carcinogenic alternative to DAB? Message-ID: <411382DF.4030602@uni-muenster.de> Dear Histonet members, staining of snap frozen muscle tissue for cytochrome oxidase (COX) using diaminobenzidine (DAB) works nicely in our laboratory. We would be happy to know, however, if anybody is aware of an non-carcinogenic alternative method to visualize cytochrome oxidase activity. Thank you very much for your help Martin Hasselblatt Martin Hasselblatt Institute of Neuropathology University Hospital M?nster, Germany http://www.klinikum.uni-muenster.de/institute/npatho/mitarbeiter/hasselblatt/index.html From hborgeri <@t> wfubmc.edu Fri Aug 6 08:15:39 2004 From: hborgeri <@t> wfubmc.edu (Hermina Borgerink) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Masson Trichrome Message-ID: <9AEEF1FB6254224AA355ED285F8491650999C853@EXCHVS2.medctr.ad.wfubmc.edu> Amber, Below is a modified version of the original Masson's Trichrome which I have successfully used for over 40 years. Instead of using Woodstain-Scarlet, which I don't think is available anymore, substitute Crocein Scarlet which Sigma sells. Masson's Trichrome (Modified) Post-fix in Bouin's fixative overnight if primary fixation was not done using Bouin's 1. Bring sections to water and wash for 5 - 10 minutes. 2. Stain nuclei for 15 minutes in Weigert's iron hematoxylin 3. Wash for 5 - 10 minutes in running water and rinse in 1% acetic acid. 4. Place slides in Woodstain Scarlet-Acid Fuchsin mixture for 5 minutes. 5. Rinse briefly in 1% acetic acid and differentiate in 5% phosphomolybdic acid. Differentiate to the point where collagen and ground substances are pale pink to colorless, usually about 30 seconds to 2 minutes. 6. Rinse in 1% acetic acid. 7. Place for 15 - 20 seconds in the Aniline Blue solution (Aniline Blue - Woodstain Scarlet - Acid Fuchsin mixture). 8. Place for a quick rinse in 1% acetic acid. 9. Dehydrate in four changes of alcohol. 10. Clear in xylene and mount in permount. Preparation of Staining Solutions: Weigert's Hematoxylin: Solution A: 29% ferric chloride - 4 ml Distilled water - 95 ml Conc. HCl - 1 ml Solution B: Hematoxylin - 1 gm 95% ethanol - 100 ml For use, mix equal parts of solutions A and B. Use fresh each time Woodstain Scarlet-Acid Fuchsin: prepare a 1% solution of Woodstain Scarlet NS and a 1% solution of Acid Fuchsin (C.I. no 692). Mix 8 parts of Woodstain Scarlet solution with 2 parts of Acid Fuchsin solution and add 1ml acetic acid to 99ml of the mixed dye solution. Aniline blue: To 85ml of 1% acetic acid add 5ml of a saturated solution of Aniline Blue (C.I. no 707) and 10ml of the above Woodstain Scarlet-Acid Fuchsin mixture. Hermina M. Borgerink, BA, HTL(ASCP)QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax (336) 716-1515 e-mail hborgeri@wfubmc.edu From Barbara_Lentz <@t> dahlchase.com Fri Aug 6 08:38:20 2004 From: Barbara_Lentz <@t> dahlchase.com (Barbara Lentz) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Benchmark XT Message-ID: We have had 2 Benchmarks and 1 XT for a year now. We have had Thermal Pad problems with one of the Benchmarks only. No file corruptions have occurred. The main problem we've had with the XT involved a sensor to the slide drawer. Barb >>> "Lab" 08/05/04 02:50PM >>> Our lab has had 2 ventana nexes systems and we are looking to upgrade to 2 benchmark xt units.We have had one in our lab to demo for approx. 3 months.During this time we have had 2 thermal pad issues and several file corruptions. Has anyone with a benchmark xt had similar problems and if so how often? Any opions about the instrument would be helpful. Thanks, Amy Boan,HT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMacDougall <@t> CuraGen.com Fri Aug 6 09:14:32 2004 From: JMacDougall <@t> CuraGen.com (MacDougall, John) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Dephosphorylation in FFPE sections Message-ID: Dear Histonet We are using an anti-receptor tyrosine kinase phosphospecific antibody with good success in FFPE IHC experiments. We wanted to confirm our staining by removing the phosphate and hopefully seeing the staining disappear. Has anyone performed this type of reaction in FFPE sections? If so, please share protocols. Thanks for any help. John R. MacDougall, Ph.D. Project Leader Preclinical Development - Toxicology/Pathology CuraGen Corporation 322 East Main St. Branford, CT, 06405 Voice: 203-871-4320 Fax: 203-315-2735 From cwscouten <@t> myneurolab.com Fri Aug 6 09:17:40 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] BRDU free floating vibratome sections Message-ID: The Histodipper from the Vibratome Company is designed for free floating sections cut on the Vibratome(TM). It also conserves reagents. See the following link: http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=464101&catdesc=Histology+Equipment&CatThreeID=635&CatOneID=4&subcatdesc=Tissue+Staining&idsubcategory=192 Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sarah Jones Sent: Thursday, August 05, 2004 4:57 PM To: histonet@lists.utsouthwestern.edu; kmjacobs@vcu.edu Subject: Re: [Histonet] BRDU free floating vibratome sections Hi Kimberle, I've done quite a bit of vibratome sectioning and free floating staining. It's quite challenging! Two ways I can think of that might help you. One would be to use the Shandon coverplate system. The sections would be floated onto slides and then the coverplate carefully placed on to the slide. It then fits into a holder and it gravity feeds the reagents over the slide. It uses a very small amount of reagent. The other way is to handle the free floating sections in netwells or you can make your own using open cassettes and small petri dishes. The most important steps would be making sure the rinsing steps were sufficient. The netwell/petri dish method would use a much greater amount of reagent. Hope this helps. Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Kimberle M. Jacobs" 08/05/04 3:32 PM >>> Hi I have searched the archives and found a lot of discussion of BRDU, but no answer to the question if there is a protocol out there for formaldehyde-fixed, vibratome-cut, free floating sections. Actually we could mount them on slides as well, I just don't want to do the whole paraffin embedding thing. Also is it really necessary to have thin sections if you are going to use pepsin and acid, etc. ?? We could probably cut as thin as 20 um, but would prefer to use 40 or 60 um. We will be doing this in pregnant ferrets (E36, gestation is ~42 days) and examining the brains at ages P0-P15. Thanks very much for any help!! Kimberle M. Jacobs, Ph.D. Assistant Professor Department of Anatomy and Neurobiology Virginia Commonwealth University P.O. Box 980709 Richmond, VA 23298-0709 (804) 827-2135 FAX: (804) 828-9477 kmjacobs@vcu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Myri37 <@t> aol.com Fri Aug 6 09:26:03 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] rapid procedure for methylmetacrylate Message-ID: <3D27A9DD.4779BE3A.0005167B@aol.com> Hi everyone i am using a methylmetacrylate embedding procedure on thin and soft resconstructed tissue on titane biomaterial, this protocole i found is adapted for bone specimen with long time for dehydratation (1 day for each alcohol) and impregnation (2 days for each bath). Could you give me please a successfull rapid procedure for embedding this type of tissue in MMA ? Any advices on reducing times are welcome. thanking you in advance. Myriam baali From asmith <@t> mail.barry.edu Fri Aug 6 09:28:30 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Woodstain scarlet in trichrome stains Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3BB7@exchsrv01.barrynet.barry.edu> To the best of my knowledge, Woodstain Scarlet, Crocein Scarlet N, and Brilliant Crocein MOO are the same dye (Acid Red 73, C.I. 27290). Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From JQB7 <@t> CDC.GOV Fri Aug 6 08:23:25 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Problem with Masson's Trichrome Message-ID: The Gomori's One-Step trichrome is faster than the Masson and should suit your needs beautifully. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of rschoon Sent: Friday, August 06, 2004 8:25 AM To: Bdeer24@aol.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Problem with Masson's Trichrome Amber, Don't know where you got your protocol but I would suggest ANY of the standerd Histology books. As for an answer to your problems. 1- mordent in either Boin's fluid or saturated picric acid for 60 min. at 60oC (there are other time, temp.'s and concentrations if one looks them up). The method simply will not work without the modent. 2- use Weigert's Iron Hematoxylin as the nuclear stain. Robert Schoonhoven Bdeer24@aol.com wrote: >Hi Everyone! > >I was recommended to try the Masson's Trichrome stainings on the slides >of uterine tissue that I have been collecting (Previously I had only performed Hematoxylin & Eosin stains). This is a new method to our lab and thus far, staining has not gone well. Although my sections contain a significant amount of muscle, I have only been able to see the aniline blue stain. On a couple of the slides, there was a slight dark purple coloration on the epithelial lining. > >I have noticed upon comparing the protocol we received to others found >on the internet that ours lacks the step in which the slides are placed in Bouin's fluid. There was also no specification in our protocol for the type of hematoxylin, so we had been using the same hematoxylin (Harris's modified with acetic acid) that we use in our H & E stainings. I was wondering if any of these might have led to the stainings not going as planned. The tissue sections also looked slightly brittle, perhaps due to a harsher stain. I am wondering if this has anything to do with my fixation process in which I use a 4% paraformaldehyde/PBS mixture, ethanol and xylene steps, as well as embedding the tissues in paraffin. > >Is Bouin's a crucial step, would I have better staining with a >different tissue fixation, or should I be looking in a different >direction? Any suggestions would be very much appreciated! Thanks! > >Amber Greenbank > >Arizona State University > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From John.Auld <@t> whnt.nhs.uk Fri Aug 6 10:31:58 2004 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Re:MSB stain Message-ID: Amber A long time ago in a galaxy far away, OK when I was a trainee and in Bonnie Scotland, we used Lendrum's method. It worked every time on formalin fixed tissue with no Bouin's or Picric acid pretreatment. It was as follows Dewax Stain nuclei with Celestine blue haematoxylin differentiate as required rinse in 95% alcohol stain in Martius Yellow 2 Mins ( 0.5 gms Martius Yellow, 2 gms Phoshotungstic acid, 100 ml 95% ethanol) Rinse in water stain in Brilliant Crystal Scarlet 10 mins ( 1gm Brilliant Crystal Scarlet 6R, 100mls 2.5% aqueous acetic acid) Rinse in water treat with 1% aqueous Phosphotungstic acid 5 mins rinse in water Stain in Soluble Blue 10 mins (0.5gms Soluble Blue, 100 mls 1% aqueous acetic acid) Rinse in water Blot dehydrate in 100% alcohol Clear in xylene and coverslip Worked every time and gave brilliant staining Good luck John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral UK External Tel 0151 604 7025 From gcallis <@t> montana.edu Fri Aug 6 10:30:00 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Problem with Masson's Trichrome In-Reply-To: <10A0ACD8.74118519.0015B53C@aol.com> References: <10A0ACD8.74118519.0015B53C@aol.com> Message-ID: <6.0.0.22.1.20040806090459.01aefae0@gemini.msu.montana.edu> One thing, I do not think you have a correct Massons Trichrome staining procedure as Bouins is absolutely crucial to success of Massons Trichrome - Weigerts Iron hematoxylin is the hematoxylin of choice (black nuclei). In fact, most histotechnology books say Bouins is fixative of choice for Mass Tri, or if your tissues are fixed with PFA you are using, you MUST postfix with this fixative prior to staining at either 56C for 1 hour or let sections sit in this overnight at RT (this is nice as you do not have heated Bouins fumes). You can continue to use your fixative, but must do the Bouins step. You can buy Bouins ready to use from Sigma and other supplies - advisable as Bouins contains picric acid, saves you time and having to store a hazardous substance. The Weigerts iron hematoxylin is best when made up fresh each time. It continues to oxidize, and if kept a long time as a working solution, loses strength, resulting in weak nuclei staining. You should access a histotechnology textbook and read the theory of Masson Trichrome stain (your university library). The reason for Bouins and acidified staining solutions (aniline blue) is the acid pH is necessary to increase the selectivity for the collagen fibers. It is hard to explain, maybe John Kiernan or someone else will expound on this. Hrapchak and Sheehan Theory and Practice of Histotechnology has a good discussion on mechanism of Mass Trichrome staining. The stain is straight forward and easy to do, uterine tissue stained with Mass Tri is beautiful - a good example of how the stain works - blue collagen and mucin, red muscle fibers, cytokeratin and cytoplasm, and black nuclei. If I remember correctly, many, many years ago, Mass tri stained uterus was required for ASCP HTL exam. Your brittle tissue may be do to over exposure to alcohols, clearants, or heat of paraffin and not because of stain protocol. If the tissue is not totally fixed prior to processing, alcohols will complete fixation and dry the tissue excessively. Check fixation time and your processing schedule - you did not indicate which species your were working with? At 01:19 AM 8/6/2004, you wrote: >Hi Everyone! > >I was recommended to try the Masson's Trichrome stainings on the slides of >uterine tissue that I have been collecting (Previously I had only >performed Hematoxylin & Eosin stains). This is a new method to our lab and >thus far, staining has not gone well. Although my sections contain a >significant amount of muscle, I have only been able to see the aniline >blue stain. On a couple of the slides, there was a slight dark purple >coloration on the epithelial lining. > >I have noticed upon comparing the protocol we received to others found on >the internet that ours lacks the step in which the slides are placed in >Bouin's fluid. There was also no specification in our protocol for the >type of hematoxylin, so we had been using the same hematoxylin (Harris's >modified with acetic acid) that we use in our H & E stainings. I was >wondering if any of these might have led to the stainings not going as >planned. The tissue sections also looked slightly brittle, perhaps due to >a harsher stain. I am wondering if this has anything to do with my >fixation process in which I use a 4% paraformaldehyde/PBS mixture, ethanol >and xylene steps, as well as embedding the tissues in paraffin. > >Is Bouin's a crucial step, would I have better staining with a different >tissue fixation, or should I be looking in a different direction? Any >suggestions would be very much appreciated! >Thanks! > >Amber Greenbank > >Arizona State University > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From ynwang <@t> u.washington.edu Fri Aug 6 10:37:10 2004 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Activated macrophages in porcine tissue (again!) In-Reply-To: <6.0.0.22.1.20040804105633.01b2e008@gemini.msu.montana.edu> References: <6.0.0.22.1.20040804105633.01b2e008@gemini.msu.montana.edu> Message-ID: All, I am still looking for an antibody for activated macrophages that can be used for immunohistochemistry in porcine tissue. I have contacted SEROTEC as well as several other companies but without success and was wondering if anyone had any ideas. Just a brief background: We want to identify resident and activated macrophages in porcine skin samples (frozen sections). We already have a general macrophage label but are having trouble finding specific ones. Any other ideas on how we could distingush between the two would also be greatly appreciated. As a side note. Can anyone explain if ED1 or ED2 denotes activated macrophages as the literature I have come across is very conflicted! Thank you very much for your help Yak-Nam Wang Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 On Wed, 4 Aug 2004, Gayle Callis wrote: > Contact SEROTEC, they have cross reaction charts to all kinds of species, > including porcine. If anything, they will do a search for you. > > At 10:33 AM 8/4/2004, you wrote: > >Does anyone know of an antibody that can be used to label (IHC) activated > >macrophages in porcine tissue? > > > >Thank you > > > >Yak-Nam Wang > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > From gcallis <@t> montana.edu Fri Aug 6 10:44:17 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] rapid procedure for methylmetacrylate In-Reply-To: <3D27A9DD.4779BE3A.0005167B@aol.com> References: <3D27A9DD.4779BE3A.0005167B@aol.com> Message-ID: <6.0.0.22.1.20040806093356.01b2d278@gemini.msu.montana.edu> If the tissue is soft, just cut back on the times in your solutions. You can process your tissues up through 1 change of xylene and 1 change Clearite 3 on your tissue processor - nice an easy, DO NOT GO THROUGH THE parafffin stations. If the tissues are small, 1 hour per station is acceptable. 70%, 80%, 95% x 2, 100% x 2, Xylene X 2 OR Xylene x 1, Clearite 3 X 1 - these are number of changes. This eliminates tedious hand processing. Start infiltration with MMA mixtures (2 mixtures) 4 hours each with last change overnight in a final mixture to insure proper infiltration, then embed, let sit overnight at RT, then polymerize in a 37C waterbath (not an oven!) I will be happy to send via private attachment, a protocol by Diane Sterchi for MMA mixtures, etc. She did short schedules for MMA with great success. This still is not a truly rapid protocol, but probably faster than what you use now. Her MMA mixtures infiltrated nicely, were not overly thick and polymerized like a dream. At 08:26 AM 8/6/2004, you wrote: Hi everyone i am using a methylmetacrylate embedding procedure on thin and soft resconstructed tissue on titane biomaterial, this protocole i found is adapted for bone specimen with long time for dehydratation (1 day for each alcohol) and impregnation (2 days for each bath). Could you give me please a successfull rapid procedure for embedding this type of tissue in MMA ? Any advices on reducing times are welcome. thanking you in advance. Myriam baali _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu [1]http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) References 1. http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Aug 6 11:12:06 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] electrical charge slides In-Reply-To: <000201c47b60$b8b82810$e1ce080a@wsahs.nsw.gov.au> References: <0B2E10D9.3909FF86.0A1F969F@wsahs.nsw.gov.au> <000201c47b60$b8b82810$e1ce080a@wsahs.nsw.gov.au> Message-ID: <6.0.0.22.1.20040806100903.01b322a0@gemini.msu.montana.edu> Bill, Would you explain what is used for coating that contains calcium? Does Silane contain calcium? Poly L lysine? If there another coating we are not aware of, the one containing calcium? I have never heard this theory before and am fascinated by your comments. - At 08:54 PM 8/5/2004, you wrote: >Dana, > >There was heaps written about this on Histonet previously, look in the >archives. >We found that the problem was associated with the Plus slides when used in >conjunction with an EDTA buffer as the material used to coat the slides is a >calcium salt and the EDTA was affecting the coating by chelating the >calcium!!! > >All the best >Bill Sinai >Laboratory Manager >Tissue Pathology, ICPMR >Westmead NSW 2145 >Australia >Ph 02 9845 7774 > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >DDittus787@aol.com >Sent: Friday, 06 August 2004 12:10 AM >To: histonet@pathology.swmed.edu >Subject: [Histonet] electrical charge slides > > >To my colleagues: > >I have a question.Is anyone out there having occasional staining(orlack of >staining on IHc slides?) I was told that the charge or the way the slides >are charged can have (pardon the pun) a negative effect. Does anyone know >/experience or have the scientific data for this? > Dana > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >__________________________________________________________________ > >This electronic message and any attachments may be confidential. If you >are not the intended recipient of this message would you please delete the >message and any attachments and advise the sender. Western Sydney >Area Health Services (WSAHS) uses virus scanning software but excludes >any liability for viruses contained in any email or attachment. > >This email may contain privileged and confidential information intended >only for the use of the addressees named above. If you are not the >intended recipient of this email, you are hereby notified that any use, >dissemination, distribution, or reproduction of this email is prohibited. If >you have received this email in error, please notify WSAHS >immediately. > >Any views expressed in this email are those of the individual sender >except where the sender expressly and with authority states them >to be the views of WSAHS. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From dcrippen <@t> buckinstitute.org Fri Aug 6 11:14:21 2004 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] hand processing rat brain-problem Message-ID: <4AA34A707932424EBE2D973764D226A901D92EBA@inverness.buckcenter.org> Dear All, We are having a puzzling issue in paraffin processing rat brains from one particular investigator. We do all our paraffin processing by hand because we don't have the volume yet to require an automatic processor. Our standard protocol which has worked for every other tissue (rat brain, mouse brain, human brain, mouse embryo, bovine adrenal gland etc...) over the years is the following: 1.) Following fixation or perfusion (performed by the investigator or tissue bank--not us) with PFA or Bouin's: 30min-2hour wash in PBS 2.) 2x22 min 50% EtOH 3.) 2x22 min 70% EtOH 4.) 2x22 min 80% EtOH 5.) 2x22 min 90% EtOH 6.) 2x30 min 95% EtOH 7.) 2x60 min 100% EtOH 8.) 3x15min xylene 9.) 2x60 min in melted Paraplast Plus under vacuum (-50kPa = -15mmHg) at 600C 10.) 1x2h in melted Paraplast Plus under vacuum (-50kPa = -15mmHg) at 600C 11.) Embed When we followed this protocol with these particular rat brains, the center of the tissue (including ventricles, striatum, hippocampus....) completely fell apart while sectioning 7um coronal sections. The outer portion of the tissue (cortex etc.) seems very much intact. We thought perhaps we had incomplete processing. The next time we received rat brains from this investigator we dissected for hippocampus (removing occipital lobe and cerebellum regions) in hopes this would increase the rate of infusion of all processing reagents. Then, we also left the tissue in paraffin under vacuum at 600C for 8hours with 3 changes, instead of 4 hours with 3 changes. BUT the same thing happens when we section. Can it be a perfusion issue??? My apologies for the long winded message, but we are really perplexed!! Many thanks in advance!! Danielle Crippen Morphology Core Manager Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen@buckinstitute.org From dcrippen <@t> buckinstitute.org Fri Aug 6 11:26:27 2004 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] hand processing rat brain-problem Message-ID: <4AA34A707932424EBE2D973764D226A901D7577C@inverness.buckcenter.org> Sorry--the protocol should read 60 degrees C. My degree symbol turned into a zero when I sent the message!! Thanks, Danielle Crippen Morphology Core Manager Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen@buckinstitute.org -----Original Message----- From: Danielle Crippen Sent: Friday, August 06, 2004 9:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] hand processing rat brain-problem Dear All, We are having a puzzling issue in paraffin processing rat brains from one particular investigator. We do all our paraffin processing by hand because we don't have the volume yet to require an automatic processor. Our standard protocol which has worked for every other tissue (rat brain, mouse brain, human brain, mouse embryo, bovine adrenal gland etc...) over the years is the following: 1.) Following fixation or perfusion (performed by the investigator or tissue bank--not us) with PFA or Bouin's: 30min-2hour wash in PBS 2.) 2x22 min 50% EtOH 3.) 2x22 min 70% EtOH 4.) 2x22 min 80% EtOH 5.) 2x22 min 90% EtOH 6.) 2x30 min 95% EtOH 7.) 2x60 min 100% EtOH 8.) 3x15min xylene 9.) 2x60 min in melted Paraplast Plus under vacuum (-50kPa = -15mmHg) at 600C 10.) 1x2h in melted Paraplast Plus under vacuum (-50kPa = -15mmHg) at 600C 11.) Embed When we followed this protocol with these particular rat brains, the center of the tissue (including ventricles, striatum, hippocampus....) completely fell apart while sectioning 7um coronal sections. The outer portion of the tissue (cortex etc.) seems very much intact. We thought perhaps we had incomplete processing. The next time we received rat brains from this investigator we dissected for hippocampus (removing occipital lobe and cerebellum regions) in hopes this would increase the rate of infusion of all processing reagents. Then, we also left the tissue in paraffin under vacuum at 600C for 8hours with 3 changes, instead of 4 hours with 3 changes. BUT the same thing happens when we section. Can it be a perfusion issue??? My apologies for the long winded message, but we are really perplexed!! Many thanks in advance!! Danielle Crippen Morphology Core Manager Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen@buckinstitute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JShaw <@t> rockdale.org Fri Aug 6 11:45:35 2004 From: JShaw <@t> rockdale.org (John Shaw) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] (no subject) Message-ID: Can anyone tell me, is there a regulation pertaining to the "Minimum" amount of time placenta specimens must be held after sign-out? Thanks John L. Shaw, HT(ASCP) Histology Supv. Rockdale Medical Center Conyers, Ga. 30012 From cwscouten <@t> myneurolab.com Fri Aug 6 11:56:38 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] hand processing rat brain-problem Message-ID: It sounds like this tissue was immersion fixed, and the core was not reached by formaldehyde before considerable autolysis had set in. Or the perfusion was inadeqate to get formaldehyde everywhere. It definitely sounds like a perfusion/fixation issue. Do you know if the tissue was perfused, how long, what prewash, etc? Consider the following link for a commercial product for reliable thorough perfusion. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Danielle Crippen Sent: Friday, August 06, 2004 11:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] hand processing rat brain-problem Dear All, We are having a puzzling issue in paraffin processing rat brains from one particular investigator. We do all our paraffin processing by hand because we don't have the volume yet to require an automatic processor. Our standard protocol which has worked for every other tissue (rat brain, mouse brain, human brain, mouse embryo, bovine adrenal gland etc...) over the years is the following: 1.) Following fixation or perfusion (performed by the investigator or tissue bank--not us) with PFA or Bouin's: 30min-2hour wash in PBS 2.) 2x22 min 50% EtOH 3.) 2x22 min 70% EtOH 4.) 2x22 min 80% EtOH 5.) 2x22 min 90% EtOH 6.) 2x30 min 95% EtOH 7.) 2x60 min 100% EtOH 8.) 3x15min xylene 9.) 2x60 min in melted Paraplast Plus under vacuum (-50kPa = -15mmHg) at 600C 10.) 1x2h in melted Paraplast Plus under vacuum (-50kPa = -15mmHg) at 600C 11.) Embed When we followed this protocol with these particular rat brains, the center of the tissue (including ventricles, striatum, hippocampus....) completely fell apart while sectioning 7um coronal sections. The outer portion of the tissue (cortex etc.) seems very much intact. We thought perhaps we had incomplete processing. The next time we received rat brains from this investigator we dissected for hippocampus (removing occipital lobe and cerebellum regions) in hopes this would increase the rate of infusion of all processing reagents. Then, we also left the tissue in paraffin under vacuum at 600C for 8hours with 3 changes, instead of 4 hours with 3 changes. BUT the same thing happens when we section. Can it be a perfusion issue??? My apologies for the long winded message, but we are really perplexed!! Many thanks in advance!! Danielle Crippen Morphology Core Manager Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen@buckinstitute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lfaucette <@t> niaid.nih.gov Fri Aug 6 12:00:56 2004 From: lfaucette <@t> niaid.nih.gov (Faucette, Lawrence (NIH/NIAID)) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] (no subject) Message-ID: <78F91CA4B8F3504E8FE5F13AAF122ED804A02CAD@nihexchange27.nih.gov> RETENTION OF LABORATORY RECORDS AND MATERIALS The College of American Pathologists makes the following recommendations for the minimum requirements for the retention of laboratory records and materials. They meet or exceed the regulatory requirements specified in the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88). The College of American Pathologists urges laboratories to retain records and/or materials for a longer period of time than specified when such would be appropriate for patient care, education or quality improvement needs. Some state regulations as well as other federal mandates may require retention of records and/or materials for a longer time period than that specified in the CLIA-88 regulations; therefore any applicable state or federal laws should be reviewed carefully when individual laboratories develop their record retention policies. MATERIAL/RECORD PERIOD OF RETENTION General Laboratory Accession log records 2 years Maintenance/instrument maintenance 2 years Quality control records 2 years Surgical Pathology (including bone marrows) Wet tissue 2 weeks after final report Paraffin blocks 10 years Slides 10 years Reports 10 years This appendix is from the CAP guidelines and was dated 2001. Lawrence J Faucette Contractor HT ASCP http://www.niaid.nih.gov/dir/services/animalcare/VetPathology/VetPathology-i ndex.html Infectious Disease Pathogenesis Section Comparative Medicine Branch Division of Intramural Research, NIAID, NIH Twinbrook III, Room 2W-01A, MSC 8135 12735 Twinbrook Parkway Bethesda, MD 20892-8135 Telephone 301-451-1056 Fax 301-480-2343 SoBran, Inc. Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. The National Institute of Allergy and Infectious Diseases (NIAID) shall not accept liability for any statement made that are the sender's own and not expressly made on behalf of the NIAID by one of its representatives. -----Original Message----- From: John Shaw [mailto:JShaw@rockdale.org] Sent: Friday, August 06, 2004 12:46 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] (no subject) Can anyone tell me, is there a regulation pertaining to the "Minimum" amount of time placenta specimens must be held after sign-out? Thanks John L. Shaw, HT(ASCP) Histology Supv. Rockdale Medical Center Conyers, Ga. 30012 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From subratab <@t> bdonline.com Fri Aug 6 12:23:48 2004 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Cytochrome oxidase (COX) staining Message-ID: <200408061729.i76HTOCa017998@mailout.proshikanet.com> Dear Martin Your mail has drawn my attention. I am also interested to know if there is any non-DAB method for COX staining. I expect that someone from histoland may reply. However, we were unable to get a good COX staining in frozen kidney tissue (rat), may be due to excess endo peroxidase in kidney. Could you please e-mail me your protocol? (PS: I have seen your nice website, although in Deutsch.) Best regards Subrata Biswas University of Campinas, Brazil. http://www.histosearch.com/homepages/DrSubrataKumarBiswas.html Martin wrote: Dear Histonet members, staining of snap frozen muscle tissue for cytochrome oxidase (COX) using diaminobenzidine (DAB) works nicely in our laboratory. We would be happy to know, however, if anybody is aware of an non-carcinogenic alternative method to visualize cytochrome oxidase activity. Thank you very much for your help Martin Hasselblatt Martin Hasselblatt Institute of Neuropathology University Hospital M?nster, Germany From gcallis <@t> montana.edu Fri Aug 6 12:16:14 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] hand processing rat brain-problem In-Reply-To: <4AA34A707932424EBE2D973764D226A901D7577C@inverness.buckcen ter.org> References: <4AA34A707932424EBE2D973764D226A901D7577C@inverness.buckcenter.org> Message-ID: <6.0.0.22.1.20040806111448.01b63c90@gemini.msu.montana.edu> Danielle, I have replied with a private message and attached protocol for hamster brain, not much bigger than rat - this has been discussed recently on Histonet, so is basically a duplicate of previous messaging. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From LuckG <@t> empirehealth.org Fri Aug 6 12:17:06 2004 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Lymph Node Revealing Solution Message-ID: Laurie, We use "Tri-Fix" which I have made for us by American Mastertech, Inc. It is a trichloroacetic acid based reagent that works well to "light up" lymphoid tissue in fatty specimens. Catalog item is "FXTIGAL". Good luck, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Monday, August 02, 2004 11:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Lymph Node Revealing Solution Someone told one of my pathologists about a lymph node revealing solution that is made by Sigma than can be disposed of down the drain. I called Sigma, and no one there knows what I'm talking about. Has anyone out there heard of this product? Laurie Colbert Huntington Memorial Hospital Pasadena, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Fri Aug 6 12:50:48 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] (no subject) In-Reply-To: Message-ID: John The March 2003 Checklist, question ANP.12500 states wet tissue, at a minimum, must be stored two weeks after the case is completed. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of John Shaw Sent: Friday, August 06, 2004 11:46 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] (no subject) Can anyone tell me, is there a regulation pertaining to the "Minimum" amount of time placenta specimens must be held after sign-out? Thanks John L. Shaw, HT(ASCP) Histology Supv. Rockdale Medical Center Conyers, Ga. 30012 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From flemons <@t> bhset.org Fri Aug 6 13:07:39 2004 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Study guides for exam Message-ID: I tried looking at the ASCP website, but had trouble locating study guides to purchase for the HT exam. Does anyone know where I can locate a source for this? When I got my cert. there was a study guide with practice questions available that was helpful I know NSH has self-assessments, but I was looking for something specifically from ASCP. Thanks in advance for your help! Fran Walker From JQB7 <@t> CDC.GOV Fri Aug 6 13:16:49 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Study guides for exam Message-ID: http://www.ascp.org/511live/timssnet/search/collectionsearchaction.cfm Try this. There are also online practice tests at www.ascp.org/bor -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fran Lemons Sent: Friday, August 06, 2004 2:08 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Study guides for exam I tried looking at the ASCP website, but had trouble locating study guides to purchase for the HT exam. Does anyone know where I can locate a source for this? When I got my cert. there was a study guide with practice questions available that was helpful I know NSH has self-assessments, but I was looking for something specifically from ASCP. Thanks in advance for your help! Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JSCHUMA1 <@t> FAIRVIEW.ORG Fri Aug 6 13:20:47 2004 From: JSCHUMA1 <@t> FAIRVIEW.ORG (JENNIFER SCHUMACHER) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Standard abbreviations Message-ID: Does anybody know if there is a list somewhere of standard medical abbreviations? Such as "RUL=Right Upper Lobe". Thanks. Jennifer The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including “protected health information.” If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> From JQB7 <@t> CDC.GOV Fri Aug 6 13:21:14 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Study guides for exam Message-ID: For some reason the link doesn't seem to want to open. I found the information by going to www.ascp.org and in the Search box I typed "study guide". -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine Sent: Friday, August 06, 2004 2:17 PM To: Fran Lemons; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Study guides for exam http://www.ascp.org/511live/timssnet/search/collectionsearchaction.cfm Try this. There are also online practice tests at www.ascp.org/bor -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fran Lemons Sent: Friday, August 06, 2004 2:08 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Study guides for exam I tried looking at the ASCP website, but had trouble locating study guides to purchase for the HT exam. Does anyone know where I can locate a source for this? When I got my cert. there was a study guide with practice questions available that was helpful I know NSH has self-assessments, but I was looking for something specifically from ASCP. Thanks in advance for your help! Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Fri Aug 6 13:28:15 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Standard abbreviations Message-ID: Just 2 of 62,500 Google hits for "medical abbreviations": http://www.pharma-lexicon.com/medicalabbreviations.php http://www.jdmd.com/glossary/medabbr.pdf Gary Gill -----Original Message----- From: JENNIFER SCHUMACHER [mailto:JSCHUMA1@FAIRVIEW.ORG] Sent: Friday, August 06, 2004 1:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Standard abbreviations Does anybody know if there is a list somewhere of standard medical abbreviations? Such as "RUL=Right Upper Lobe". Thanks. Jennifer The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including "protected health information." If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Gloria.Niehans <@t> med.va.gov Fri Aug 6 13:29:53 2004 From: Gloria.Niehans <@t> med.va.gov (Gloria.Niehans@med.va.gov) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Oncocytoma vs. Chromophobe Message-ID: <2C7B953912C2D511B6470000F8033DA50146D4A7@VHAMINEXC1> We use an immunohistochemical antibody to cytokeratin 7. Chromophobe carcinomas are positive for CK7, while oncocytomas are negative (or minimally positive). [Leroy et al, "Utility of cytokeratin 7 for distinguishing chromophobe renal cell carcinoma from renal oncocytoma." Eur Urol 2000; 37: 484-487, also in Dabbs immunohistochemistry text, pp 477-481.] We've found it a lot easier to do and interpret than colloidal iron. We get our antibody from Biocare (clone K72.7), and run it on a Ventana immunostainer with protease predigestion, but I think a lot of immunohistochemistry companies carry cytokeratin 7 antibodies, so you could check your catalogs and see what's out there. Gloria Niehans Minneapolis VAMC -----Original Message----- From: Barnhart, Tammy [mailto:Tbarnhart@primecare.org] Sent: Friday, August 06, 2004 6:11 AM To: Histonet (E-mail) Subject: [Histonet] Oncocytoma vs. Chromophobe I was wondering if there is(are) any immunohistochemical antibodies or special stains besides Hale's Colloidal Iron for the differentiation of Oncocytoma and Chromophobe in kidney? I am not able to make the dialysised (sp?) iron needed for the Hale's but would like to offer my pathologist an alternate. Any suggestions? Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JQB7 <@t> CDC.GOV Fri Aug 6 13:28:52 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Standard abbreviations Message-ID: Try this: http://www.media4u.com/bbp/medabb_a.html -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JENNIFER SCHUMACHER Sent: Friday, August 06, 2004 2:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Standard abbreviations Does anybody know if there is a list somewhere of standard medical abbreviations? Such as "RUL=Right Upper Lobe". Thanks. Jennifer The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including "protected health information." If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Fri Aug 6 13:37:27 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Histo dipper from Vibratome Company Message-ID: Charles, Can you bring one of these to the exhibition table at the NSH meeting in Toronto? I just can't quite get a feel for how this works by looking at a picture on the website. Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From carl.hobbs <@t> kcl.ac.uk Fri Aug 6 14:44:03 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] re electrical charge slides Message-ID: <002d01c47bed$ba985eb0$63412cd9@home> I've asked this before but with no replies: The Superfrost Plus slides that many of us buy, instead of APES - coating our own - they state that they are electrically charged......do they achieve this by performing APES- coating? Or is it a proprietary secret? Thanks. --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.726 / Virus Database: 481 - Release Date: 22/07/2004 From cwscouten <@t> myneurolab.com Fri Aug 6 14:45:49 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Histo dipper from Vibratome Company Message-ID: I will not be there personally this year, conflicting meetings, but I will ask that a Histodipper be taken and be on display. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Friday, August 06, 2004 1:37 PM To: Histonet Subject: [Histonet] Histo dipper from Vibratome Company Charles, Can you bring one of these to the exhibition table at the NSH meeting in Toronto? I just can't quite get a feel for how this works by looking at a picture on the website. Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Fri Aug 6 14:59:16 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] re hand processing rat brain-problem Message-ID: <003601c47bef$dada5b40$63412cd9@home> I think it's probably the wax infiltration stage at 600C that's causing the problem ;-)) Seriously...don't know. I process rat and mouse brain( perfused and immersion-fixed) but with much longer times in alcohol and xylene ( my earlier post). No probs so far. It may be inadequate perfusion- fixation...I've had tissue disintegration with this before. May be someone's cocked up the processing reagents...........( checking the block surface after a week will confirm any inadequacy in processing ) Re processing by hand: I don't have high volume either but certainly justify having an automated 'dip'n'dunk' processor......put it on and walk away. I still deal with people who refuse to use a processing machine.....why? Fine if it's costs ie you can't afford it. Otherwise.......reminds me again of the old argument regarding the inadequacies of synthetic mounting media when compared to Canada Balsam for example.......who uses it now? ( think of the trees.........) --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.726 / Virus Database: 481 - Release Date: 22/07/2004 From Jackie.O'Connor <@t> abbott.com Fri Aug 6 15:17:56 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Standard abbreviations Message-ID: My favorite one is from the Emergency Room - LOLFDGB. i.e. Little old lady fall down go boom. "Bartlett, Jeanine" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/06/2004 01:28 PM To: "JENNIFER SCHUMACHER" , cc: Subject: RE: [Histonet] Standard abbreviations Try this: http://www.media4u.com/bbp/medabb_a.html -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JENNIFER SCHUMACHER Sent: Friday, August 06, 2004 2:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Standard abbreviations Does anybody know if there is a list somewhere of standard medical abbreviations? Such as "RUL=Right Upper Lobe". Thanks. Jennifer The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including "protected health information." If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cdan1204 <@t> yahoo.com.cn Sat Aug 7 03:24:44 2004 From: cdan1204 <@t> yahoo.com.cn (=?gb2312?q?d=20c?=) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] one question Message-ID: <20040807082444.34504.qmail@web15607.mail.cnb.yahoo.com> My name is danc, our laboratory stores all of the paraffin-embeded specimens at room temperature.Since we learned that eighteen months is considered a reasonable estimate for storage of tissue for optimal quality.Our director wants to improve our storage techniques in order to extend the storage time.So now I am responsible for the techniques problem.I would greatly appreciate any instructions for paraffin-embeded specimens storage that you folks could give me.Thank you very much. danc Jinling Hospital Nanjing China --------------------------------- Do You Yahoo!? 150ÍňÇúMP3ˇčżńËŃŁŹ´řÄú´łČëŇôŔÖľîĚĂ ĂŔĹŽĂ÷ĐÇÓŚÓĐžĄÓĐŁŹËŃąéĂŔÍźĄ˘ŃŢÍźşÍżáÍź 1GžÍĘÇ1000ŐףŹŃĹť˘ľçÓĘ×ÔÖúŔŠČÝŁĄ From cdan1204 <@t> yahoo.com.cn Sat Aug 7 08:37:23 2004 From: cdan1204 <@t> yahoo.com.cn (=?gb2312?q?d=20c?=) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] one question Message-ID: <20040807133723.71233.qmail@web15604.mail.cnb.yahoo.com> Note: forwarded message attached. --------------------------------- Do You Yahoo!? 150ÍňÇúMP3ˇčżńËŃŁŹ´řÄú´łČëŇôŔÖľîĚĂ ĂŔĹŽĂ÷ĐÇÓŚÓĐžĄÓĐŁŹËŃąéĂŔÍźĄ˘ŃŢÍźşÍżáÍź 1GžÍĘÇ1000ŐףŹŃĹť˘ľçÓĘ×ÔÖúŔŠČÝŁĄ From tahseen <@t> brain.net.pk Sat Aug 7 13:19:48 2004 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] proposal for post graduate diploma program in histology. Message-ID: <002d01c47cab$264abaa0$972bfea9@m7c0y4> Dear Histoneter, My medical director has given me a task to prepare a proposal for post graduate diploma program in histology. Would any one like to share with me in preparation of a comprehensive proposal for post graduate diploma program. Thanks in advance Muhammad Tahseen Histology Supervisor SKMCH & RC. > From tahseen <@t> brain.net.pk Sat Aug 7 13:32:36 2004 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Migrating to Canada - Looking for Job Opportunity Message-ID: <005101c47cac$e9fb5bc0$972bfea9@m7c0y4> Dear Histoneter, Mr. Sohail Asif, a very close friend of mine has got Canadian Immigration and moving there later this month. Basically he is a Medical Technologist of clinical side (specially Flowcytometry, Hematology and Clinical Chemistry), who worked in our hospital for about ten years. I would like to request all of you that if you or your organization can consider him for the job of Medical Technologist. He is a multiskilled technologist and can work anywhere in pathology. His qualification and experience is almost in the final phase of registration with CSMLS, he'll get this in a month or so. Please find below his brief CV, if you find any opportunity, please feel free to contact him for detailed CV/information. Thanks in advance. Muhammad Tahseen I. Personal Information Name : Sohail Asif Date of Birth : 05 July 1973 E-MAIL : sasiff@hotmail.com II. Working Experience A. July 1995 to August 2004 Sr. Medical Technologist (Hematology/ Clinical Chemistry) Shaukat Khanum Memorial Cancer Hospital & Research Center, Lahore - Pakistan. B. December 1993 to June 1995 Jr. Medical Technologist Lahore Lab, Lahore - Pakistan. III. SKILLS A. Clinical Chemistry B. Hematology C. Blood Banking D. Serology Hands on experience on following Instruments: Chemistry Vitros 550, 750, 950 Haematology Bayer Technicon H*3 RTC Kodak Ektachem Cell Dyne 2000 and 800 (DT 60 II, DTSC II, & DTE II.) XE-2100 (Sysmex). Roche/Hitachi 912 MLA Electra 900C & 750 Bayer Technicon OPERA RA2010 Sysmex K1000, SE9500 DPC Immulite Miles Hematek Slide Stainer Abbott TDx and Imx Sysmex CA500 for Coagulation CIBA Corning 248 Spectrophotometer POINTE 180 CIBA Corning 410C Flame Photometer SEAC Screen Master & CH100 NOVA Plus Blood Gas Analyzer MERCK Micro Lab SPOTCHEM SE - 1510 BM 4010 Cytometry FACS Callibur (Four color) - BD Transfusion Services Baxter, CS-3000 Blood cell separator IV. ACADEMIC QUALIFICATIONS> A. B.B.A, 1999-2001. B. Medical Laboratory Technology (M.L.T), 1990-1992. C. Diploma in computer application, 1994-1995. D. Short course on Hospital Management, October 2002. V. Training Overseas Key operator training for Vitros 950 by Ms. Christina Winneroth of Ortho-Clinical Diagnostics, Strasbourg, France. In House A. Operators Trouble Shooting training for H3 Hematology. B. Operators Trouble Shooting training for MLA 900c/750C Coagulation Timer C. Operators Trouble Shooting training for Opera 2000 Chemistry System. D. Key operator training for Vitros 550, 750 (Ektachem Kodak). E. Key operator training on FACS-Caliber (Becton Dickinson). F. Internal Quality Auditing for ISO 9001:2000 Quality Management System. G. Theoretical and Practical Training on Sysmex Coagulation Analyzer Model CA-550. VII. Awards A. A certificate of dedicated service for the Hospital by SKMCH & RC. B. Best performance award for five years of service from SKMCH & RC. C. Best effort award for achievement of ISO 9002 Certification. D. Long Service Award October 2002. E. Quality Management System Certification of Pathology Department VIII. MEMBERSHIPS A. An Affiliate Member of IBMS. (UK), Membership no: 330248 B. A. P. M. L. T. (Pakistan), Membership no: 652 C. Subscribed Member of CSMLS (Canada), Membership no: 1007463/S From ccdub <@t> earthlink.net Sun Aug 8 07:49:33 2004 From: ccdub <@t> earthlink.net (Cindy DuBois) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Placenta disposal Message-ID: We have a CAP standard which states all processed tissues blocks are to be kept 5 years. Gross only specimens are to be kept 2 weeks. We keep the wet tissue 3 weeks (just to make sure). However, our lab manager was kind enough to offer an "archival" system to the hospitals we service as an added incentive to keep their business. This means we now receive, process and block every single placenta and keep the blocks 20 years !! This has created a huge storage problem for us. We only cut the blocks on the cases where the placenta is abnormal, or there was a birthing or baby problem. All others have a "tag" in them and we store them w/o cutting. I can only remember on 2 occasions(we have been doing this for 3 years) where we had to go back and process the archived blocks. Cindy DuBois, HT ASCP Delta Pathology Assoc. Stockton CA From philip.bergin <@t> microbio.gu.se Sun Aug 8 11:11:19 2004 From: philip.bergin <@t> microbio.gu.se (Philip Bergin) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Private responses - well said, Barry! In-Reply-To: <41127A5D.4080406@bms.com> Message-ID: My 2 cents worth too! I have just finished a research PhD and have found the Histonet to be of great help (consequently I seem to hardly ever have anything helpful to say myself!). A lot of the stuff that was helpful hadn't even thought of- it was just in the answers to anothers questions. So I find the responses really helpful..... As for salespeople- they seem to be great as well. The only time I remember them being annoying was a few years back when someone kept on advertising new products all the time. I still find answers from salespeople incredibly helpful- even if they do talk about their own product! Histonet has been great- I think I have learn't more about histology/cytometry/antibodies/beer/ and the weather in different countries then I have from all my time at university (and its been a long time!). I have also learn't a great respect for your sense of community (shown by the great response to the Steve Slapp tragedy). I do believe that if people stop responding and being able to give their own opinion then a lot of people won't have the chance to get from Histonet what I have and that is a tragedy. If you disagree with someone opinion, but character attacks and flaming ruin it for everyone- delete is always there! Also, I personally would like to know what vendors are so ashamed and unable to sell their equipment when people give their honest opinions of it. If the equipment is so bad that you have to complain to a persons boss when they describe their experience with it, then it must be really bad! Personally I like peoples opinions, if they are wrong, then could you please explain to the rest of us why they are wrong- much more useful! If you've made it this far then thanks to everyone who spends their time helping out the rest of us that really don't know that much! Cheer, Phil ----------------------------------------------------------------- Philip Bergin PhD G?teborg University Department of Medical Microbiology and Immunology Box 435, SE-405 30 G?teborg, Sweden Phone: +46-31-7736213 Fax: +46-31-7736205 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Susan Q Wells Sent: den 5 augusti 2004 20:20 To: Gayle Callis Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Private responses - well said, Barry! I completely agree with Gayle! This list is invaluable to most of us working in the field. Gayle Callis wrote: > Pam and all, > Too many people forget or ignore the fact that some individuals who > are experts in the field of histotechnology are not employed in just > clinical or research laboratories. If one is involved with a company > doing technical research and development of a product destined for > histoland, then they are should be very welcome to comment. > Knowledge should not be smothered (flames!) I don't see you as a > salesperson, but a valuable resource providing me with information I > can choose to use or not. All part of the learning experience. > Complaints = delete button. > > > At 10:13 AM 8/5/2004, you wrote: > > I agree with both Gayle and Barry yet have one reservation. I was > hit several times on this server for answering very general > questions and told salespeople should not be on HistoNet. Although > my answers had nothing to do with any product we sell and by the > way I am not a sales person, just technical and product > development. After the second incident that several others came on > and agreed we were not welcome I stay off 99% of the time. It is > difficult to walk the line here and not be on privately only. I do > agree that if it is a sales pitch it should not be on the line as a > reply that should be private answers only. > Thanks for letting me say this and I hope no one is offended. > Pam Marcum > > -------------- Original message -------------- > > Thank you Barry - very well said. > > > > Gayle Callis > > MT,HT,HTL(ASCP) > > Research Histopathology Supervisor > > Veterinary Molecular Biology > > Montana State University - Bozeman > > Bozeman MT 59717-3610 > > > > At 09:02 AM 8/5/2004, you wrote: > > > > >I hate to see the word "flamed" as here in sunny Houston it > has been in > > >the upper 90s for about 2 weeks. > > > > > >While I appreciate people's perception about getting > "flamed" when they > > >have posted items, I have a real problem with posting most > items > > >privately. > > >Histonet is an extremely valuable resource and there is a > vast amount of > > >expertise available. It is only logical for this knowledge > and > > >experience to be f! reely shared. > > >I can't say that I have been "flamed" (or even mildly > scorched) but > > >sometimes I make mistakes and post incorrect information. It > is a > > >valuable learning experience for me and all interested > parties to have > > >any errors corrected. Sometimes it is matter of perception > and how > > >criticism is leveled (being brutally frank is not a good > attitude). > > >"Flaming" may be perceived by one individual to have > occurred while > > >another may regard a response as constructive criticism. > > >If anyone here gets "flamed" or has their employer contacted > by a > > >company about a posting then please let us know. Companies > do respond > > >positively to public opinion. Any companies with such an > attitude this > > >should be held accountable and so should employers for not > telling the > > >company or their representative to take a hike. > > >I do believe t! hat if someone requests private responses > that this be > >! > ; >re spected. Second, I believe that when responding one > should be careful > > >when criticizing a piece of equipment or a company. Negative > criticism > > >sticks and while this may be true for a particular > individual it could > > >be an isolated incident and due to many factors. Not > everyone knows > > >how to deal effectively with companies (and not every > company knows how > > >to deal responsibly with customers). I think that such > listings should > > >be private. The individual who receives these could then let > us know > > >about the final results in the form of a summary. > > >If I am to get "flamed" for this or any other postings > please include > > >"flamed" at the start of the message so that I don't' just > think that > > >the response is constructive criticism. > > >Barry > > > > > > > > >_______________________________________________ > > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > > >[1]http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > [2]http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > > MT,HT,HTL(ASCP) > > Research Histopathology Supervisor > > Veterinary Molecular Biology > > Montana State University - Bozeman > > PO Box 173610 > > Bozeman MT 59717-3610 > > 406 994-6367 (lab with voice mail) > > 406 994-4303 (FAX) > >References > > 1. http://lists.utsouthwestern.edu/mailman/listinfo/histonet > 2. http://lists.utsouthwestern.edu/mailman/listinfo/histonet >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From slimwillie <@t> cox.net Sun Aug 8 15:19:22 2004 From: slimwillie <@t> cox.net (Jerry Wilson) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Placenta disposal References: Message-ID: <003001c47d84$fe3bbf20$f2340e44@no.cox.net> I thought the CAP had changed the time frame back to 10 years for block retention. It seems they had changed from 10 to 5 a couple of years ago and then more recently changed back to 10 years. Can anyone clarify this? The wet tissue should be kept for 2 weeks after the final report- we keep ours one month just to be safe Jerry Wilson ----- Original Message ----- From: "Cindy DuBois" To: Cc: "Histonet" Sent: Sunday, August 08, 2004 7:49 AM Subject: [Histonet] Placenta disposal > We have a CAP standard which states all processed tissues blocks are to be > kept 5 years. Gross only specimens are to be kept 2 weeks. > > We keep the wet tissue 3 weeks (just to make sure). However, our lab > manager was kind enough to offer an "archival" system to the hospitals we > service as an added incentive to keep their business. This means we now > receive, process and block every single placenta and keep the blocks 20 > years !! This has created a huge storage problem for us. We only cut the > blocks on the cases where the placenta is abnormal, or there was a birthing > or baby problem. All others have a "tag" in them and we store them w/o > cutting. I can only remember on 2 occasions(we have been doing this for 3 > years) where we had to go back and process the archived blocks. > > > Cindy DuBois, HT ASCP > Delta Pathology Assoc. > Stockton CA > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bills <@t> icpmr.wsahs.nsw.gov.au Sun Aug 8 16:39:12 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] electrical charge slides In-Reply-To: <6.0.0.22.1.20040806100903.01b322a0@wsahs.nsw.gov.au> Message-ID: <000401c47d90$25d2ef80$e1ce080a@wsahs.nsw.gov.au> Gayle, I have no idea what the substance they use. I was told by Menzel in Germany that the process is a secret. When I enquired about the "New" Superfrost Plus Ultra they informed me it was a different process than the Superfrost Plus. I wish I knew more. The Superfrost Plus Ultra have to date eliminated the staining problems with the Ventana Benchmark. Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Saturday, 07 August 2004 2:12 AM To: Bill Sinai; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] electrical charge slides Bill, Would you explain what is used for coating that contains calcium? Does Silane contain calcium? Poly L lysine? If there another coating we are not aware of, the one containing calcium? I have never heard this theory before and am fascinated by your comments. - At 08:54 PM 8/5/2004, you wrote: >Dana, > >There was heaps written about this on Histonet previously, look in the >archives. >We found that the problem was associated with the Plus slides when used in >conjunction with an EDTA buffer as the material used to coat the slides is a >calcium salt and the EDTA was affecting the coating by chelating the >calcium!!! > >All the best >Bill Sinai >Laboratory Manager >Tissue Pathology, ICPMR >Westmead NSW 2145 >Australia >Ph 02 9845 7774 > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >DDittus787@aol.com >Sent: Friday, 06 August 2004 12:10 AM >To: histonet@pathology.swmed.edu >Subject: [Histonet] electrical charge slides > > >To my colleagues: > >I have a question.Is anyone out there having occasional staining(orlack of >staining on IHc slides?) I was told that the charge or the way the slides >are charged can have (pardon the pun) a negative effect. Does anyone know >/experience or have the scientific data for this? > Dana > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >__________________________________________________________________ > >This electronic message and any attachments may be confidential. If you >are not the intended recipient of this message would you please delete the >message and any attachments and advise the sender. Western Sydney >Area Health Services (WSAHS) uses virus scanning software but excludes >any liability for viruses contained in any email or attachment. > >This email may contain privileged and confidential information intended >only for the use of the addressees named above. If you are not the >intended recipient of this email, you are hereby notified that any use, >dissemination, distribution, or reproduction of this email is prohibited. If >you have received this email in error, please notify WSAHS >immediately. > >Any views expressed in this email are those of the individual sender >except where the sender expressly and with authority states them >to be the views of WSAHS. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From jnocito <@t> satx.rr.com Sun Aug 8 18:32:02 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Placenta disposal References: <003001c47d84$fe3bbf20$f2340e44@no.cox.net> Message-ID: <001a01c47d9f$e8fbe1b0$5d6bce44@yourxhtr8hvc4p> Jerry, Yes, CAP did change block retention back to 10 years. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Jerry Wilson" To: "Cindy DuBois" ; Cc: "Histonet" Sent: Sunday, August 08, 2004 3:19 PM Subject: Re: [Histonet] Placenta disposal > I thought the CAP had changed the time frame back to 10 years for block > retention. It seems they had changed from 10 to 5 a couple of years ago and > then more recently changed back to 10 years. Can anyone clarify this? The > wet tissue should be kept for 2 weeks after the final report- we keep ours > one month just to be safe > > Jerry Wilson > ----- Original Message ----- > From: "Cindy DuBois" > To: > Cc: "Histonet" > Sent: Sunday, August 08, 2004 7:49 AM > Subject: [Histonet] Placenta disposal > > > > We have a CAP standard which states all processed tissues blocks are to be > > kept 5 years. Gross only specimens are to be kept 2 weeks. > > > > We keep the wet tissue 3 weeks (just to make sure). However, our lab > > manager was kind enough to offer an "archival" system to the hospitals we > > service as an added incentive to keep their business. This means we now > > receive, process and block every single placenta and keep the blocks 20 > > years !! This has created a huge storage problem for us. We only cut the > > blocks on the cases where the placenta is abnormal, or there was a > birthing > > or baby problem. All others have a "tag" in them and we store them w/o > > cutting. I can only remember on 2 occasions(we have been doing this for 3 > > years) where we had to go back and process the archived blocks. > > > > > > Cindy DuBois, HT ASCP > > Delta Pathology Assoc. > > Stockton CA > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From JCarpenter764 <@t> aol.com Sun Aug 8 18:39:22 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Has anyone received HT practical for the fall exam yet? Message-ID: <67.2ff9afb5.2e4813aa@aol.com> Hey everyone...it's me the famous worry wart. I still have not recieved my HT practical by mail yet am I the only one? Thanks Jennell From lpwenk <@t> sbcglobal.net Mon Aug 9 03:40:29 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Has anyone received HT practical for the fall exam yet? References: <67.2ff9afb5.2e4813aa@aol.com> Message-ID: <001b01c47dec$87d73b80$ae2dd445@domainnotset.invalid> I don't know if they have been sent out or not (someone else who has signed up for this cycle should answer this), but the list of tissues/stains required for the practical exams can be found on the ASCP web page. http://www.ascp.org/bor/certification/index.asp Click on HT or HTL Practical exam, on the upper right side. Both are due in by October 1, 2004. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Sunday, August 08, 2004 7:39 PM Subject: [Histonet] Has anyone received HT practical for the fall exam yet? > Hey everyone...it's me the famous worry wart. I still have not recieved my > HT practical by mail yet am I the only one? Thanks Jennell > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Mon Aug 9 04:05:05 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] WAGE/VACANCY SURVEY Message-ID: <002101c47def$f6f62a00$ae2dd445@domainnotset.invalid> ASCP has their preliminary latest survey on line http://www.ascp.org/bor/center/center_research.asp Click on 2003. (2002 has the complete, 2 part report, if you are still interested) Usually, ASCP does the survey every other year (on the even years 2000, 2002) and reports the findings in the following year (2001, 2003). However, ASCP felt that the market was changing so fast that they did another survey on the off year, last year (2003) and are reporting is out this year (2004). In general, vacancies are less than the previous year, and wages increased a small percent (2-5% for most). When I did a review, histotechs (HT) are still one of the most needed, when compared to the rest of the labs. PBT = 6.1% (of labs reporting shortages) HT = 6.0% MLT = 5.9% MT = 4.3% CT = 4.3% HTL = 3.6% For Supervisors, histology is leading the vacancy rate: HT/HTL - Supervisor = 4.9% (of labs reporting shortages) MT - Supervisor = 3.3% PBT - Supervisor = 2.7% MT - Manager = 1.9% CT - Supervisor = 1.6% MLT - Supervisor = 1.2% One change appears that ASCP is sampling the CMS (Centers for Medicare and Medicaid) database, rather than using ASCP's list of supervisors who are ASCP registered and still ASCP members. That gave the survey access to more labs. Hope some of you can use this information. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From JCarpenter764 <@t> aol.com Mon Aug 9 05:44:35 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Fall practical? clarifying first question Message-ID: <1B887239.6781B566.40C6E687@aol.com> I'm sorry i think i might have been misunderstood. I have applied to take the practical in June for the July 1st deadline. They say to wait 30 days before recieving your practical by mail, and i still haven't recieved it yet. I did pull it off of the internet though so i was able to collect my tissue and stuff. I just want to know if anyone has received the exam by mail b/c it is due October 1st. From JCarpenter764 <@t> aol.com Mon Aug 9 05:53:41 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Question about practical...or suggestions Message-ID: <562A2C76.41ECCF5F.40C6E687@aol.com> Does anyone know how many sections to a slide we are suppose to have. ?My supervisor says one but i just want to check. Of course she would tell me this after cutting several slides with two pieces of lung on each. ?Anyways someone PLEASE HELP!!!!! thanks the worry wart From j_gorenstein <@t> yahoo.com Mon Aug 9 06:58:41 2004 From: j_gorenstein <@t> yahoo.com (Juile Gorenstein) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Lac Z staining Message-ID: <20040809115841.73062.qmail@web42008.mail.yahoo.com> Hello everyone, Thank you to those who has responded to my embryo processing e-mail! I was wondering if anyone has a protocol for lac Z staining on paraffin sections. I have been staining frozen sections, but have been having a little bit of trouble keeping the tissue attached to the slides (I use Superfrost Plus slides: I allow the tissue to dry for about an hr after I attach the section to the slide, store them at -80oC and then dry them again before staining -- I stain by hand, it's not an instrument issue). Please let me know if a protocol for paraffin sections exists. Thank you in advance, Julie Gorenstein Novartis, Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! From jengirl1014 <@t> yahoo.com Mon Aug 9 07:15:33 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Processing Kidneys Message-ID: <20040809121533.42729.qmail@web60602.mail.yahoo.com> Any recommendations for a protocol for processing kidneys out of the capsule? I don't have a tissue processor, so it's done the old fashioned way. Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - 100MB free storage! From Myri37 <@t> aol.com Mon Aug 9 07:12:03 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] (no subject) Message-ID: <17C24F79.3FEC69A6.0005167B@aol.com> Dear histonetters i have a question about dehydration of tissues before embedding in methylmetacrylate. i think that most of people dehydrate in ethanol 100% some others in pure 2-propanol, do you know the difference beetwen them, and between a dehydration with 2-propanol, and with 1-propanol ? Thank you very much myriam baali Natural implant From Myri37 <@t> aol.com Mon Aug 9 09:03:30 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Dehydration for MMA Message-ID: <1ECDCC13.0C446001.0005167B@aol.com> Dear histonetters i have a question about dehydration of tissues before embedding in methylmetacrylate. i think that most of people dehydrate in ethanol 100% some others in pure 2-propanol, do you know the difference beetwen them, and between a dehydration with 2-propanol, and with 1-propanol ? Thank you very much myriam baali Natural implant From meyenhmf <@t> umdnj.edu Mon Aug 9 04:40:55 2004 From: meyenhmf <@t> umdnj.edu (meyenhmf@umdnj.edu) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Processing Kidneys In-Reply-To: <20040809121533.42729.qmail@web60602.mail.yahoo.com> References: <20040809121533.42729.qmail@web60602.mail.yahoo.com> Message-ID: <1092062455.41178cf7a1417@webmail.umdnj.edu> Open a gauze pad, make a loose bag by holding the 4 corners, insert kidney and small paper with # (use pencil)tie with string and process. Done this way before tissue capsules were available. Quoting Jennifer Sipes : > Any recommendations for a protocol for processing kidneys out of the > capsule? I don't have a tissue processor, so it's done the old fashioned > way. > > > Jennifer K. Sipes, RALAT > Sr. Laboratory Technician > Johns Hopkins University > Ross 929 > 720 Rutland Avenue > Baltimore, MD 21205 > phone: 410-614-0131 > cell: 443-413-0853 > e-mail: jengirl1014@yahoo.com > > > > > > > > > > > > --------------------------------- > Do you Yahoo!? > New and Improved Yahoo! Mail - 100MB free storage! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- From BWinters <@t> NCH.ORG Mon Aug 9 09:44:03 2004 From: BWinters <@t> NCH.ORG (Winters, Bert) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] CONTROL FOR GIARDIA Message-ID: <270614B321ACB44D8C1D91F4F921FDC362777A@NCH01EX02.nch.org> We have been doing trichrome stains for giardia on tissue blocks. We have been using smear controls that we obtain from micrbiology. Are pathologist wonder if this is an appropriate control because it is not a tissue control. If anyone out there can tell me if using this type of control is a problem. Thank you for any information you can give. From pam <@t> ategra.com Mon Aug 9 10:24:05 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Job Opportunities in Histology Latest Update 08/09/04 Message-ID: Hi Histonetters, Here is the latest update on opportunities with some of my best clients throughout the US who are seeking Histology Supervisors, Histo Technologists and Histo Technicians. These are positions as direct employees of our client. There are fulltime 40 hour per week positions.. As a direct employee of one of our clients you will be provided with full benefits including Health Insurance, Vacation, Sick Pay, Relocation money and a lucrative sign-on bonus. I have part time and temporary positions as well. I have supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are some of my HOTTEST Histology Supervisor positions: 1. Alabama - Histology Supervisor 2. Colorado - Histology Manager/PA (I also need a cytology manager/supervisor for this client if you know anyone) 3. Virginia - Histology Supervisor 4. California - Immunohistochemistry Supervisor Here are some of my HOTTEST Histo Tech and Histology Supervisor positions: 1. Georgia - Histo Tech 2. Texas - Histo Tech temp to perm 3. Maine - Histo Tech 4. Maine - IHC Tech 5. N. California - Histo Tech 6. NYC - Lead Histo Tech - evenings 2p-11p 7. NYC - IHC Tech - nights 10p-7a 8. Virginia - Histo Tech 9. Michigan - Histo Tech 10. Nebraska - Histo Tech 11. Florida - Histo Tech 12. Rhode Island - Histo Tech 13. Massachusetts - Histo Tech If you are interested in these jobs, please CALL ME ASAP at 800 466 9919 x234. To speed things up, please also send me a copy of your resume, (if you haven't already done so). If you are interested in jobs outside the above-mentioned areas, please send me your resume as well. I have clients throughout the US. I will keep your resume confidential and will not release it to anyone without your permission (This is Ategra policy as well as my own). My services are at no charge to you. Of course, you may be happy in your present job, but it never hurts to to keep an eye open. Also, if you have friends/peers who might be interested as well, if you could pass my query & name on to them I'd be very grateful. However, if you are interested in any of the jobs above, please call me. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Pam Barker Senior Lab Recruiter Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd. Winter Park, FL 32792 VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com To Learn More About Ategra: http://www.ategra.com -------------------------------------------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. -------------------------------------------------------------------------------------------------- From Rcartun <@t> harthosp.org Mon Aug 9 10:53:26 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] H. pylori control tissue #2 Message-ID: I am amazed by the number of people who have contacted me about getting a control block of H. pylori-infected stomach. Why is there such a demand for this tissue? Are your pathologists not helping you identify control tissue from internal positive cases? In any event, the number of requests exceeds what I can possibly handle. I will send a block (or two) to anyone who provides me with their name, complete mailing address, telephone number, and a Federal Express or UPS account number (or other prepayment form for the shipping/mailing). I will also contact the person in charge of the NSH tissue bank to see if they can accommodate the other requests. Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Hartford Hospital Hartford, CT 06102 (860) 545-1596 From Bauer.Karen <@t> mayo.edu Mon Aug 9 11:35:48 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Color-Coding Biopsies Message-ID: <8C6E05FA69571948B461F1327CBB893E0629F0F7@lmmail2.ad.lmmhs.org> Hi Joyce, We routinely use green cassettes for all of our tissues, but when we get prostate and breast cores, we use yellow cassettes. This tells the embedder and the cutter that they are dealing with prostate or breast and can handle them accordingly. Of course, all breast and prostate cases are always staggered between other tissues. If for some reason two cases are accessioned consecutively (like at the end of the day and there are no other tissues to put between them), the cut slides are placed on two separate slide trays and the Pathologist is notified. We also use a blue dye on our prostate biopsies and sometimes on the breast cores if needed. The dye we use is Mrs. Stewart's Bluing Solution. It's purchased at any grocery store, in the laundry isle. It never gets thick, gritty, or clumpy and stays on through the entire processing and staining cycles. I believe the cost is around $2.00. We love it and have used it for years and years (20+ or so). Every 3 months, I do a search of all the prostate and breast cases to make sure they were staggered between other cases. My director wants me to do this to make sure everyone is following procedure and we keep these on file as part of our annual QI/QA. Good luck to you, Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Wednesday, August 04, 2004 9:45 AM To: Histonet Subject: [Histonet] Color-Coding Biopsies Hello Group, Is anyone color coding prostate and/or breast biopsies to help with identification of these little critters? The color would be dictated into the gross description and would aid in being certain multiple biopsies of like tissue did not get mixed up. We're thinking of doing this, and I would like to get your ideas. Thanks in advance. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From Angelicola <@t> uchc.edu Mon Aug 9 13:27:34 2004 From: Angelicola <@t> uchc.edu (Angelicola,Lisa) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] F4/80 antibody Message-ID: I'm working on a research project that involves demonstrating macrophages in formalin fixed paraffin embedded mouse skin wounds. The researcher has purchased the F4/80 rat-anti-mouse antibody from abcam. Has anyone out there in histoland had any experience working on fixed, processed tissue with this antibody? The spec sheet states that it will work with pretreatment of the tissue (protein digestion, which I would like to avoid if possible since the tissue is susceptible to falling off). Also where do you purchase your anti-rat secondary from? I would appreciate any advice anyone might have as far as pretreatment, dilutions, ect. Thanks in advance, Lisa Angelicola Immunohistochemistry Lab UCONN Health Center Farmington, Ct. From lfaucette <@t> niaid.nih.gov Mon Aug 9 13:46:16 2004 From: lfaucette <@t> niaid.nih.gov (Faucette, Lawrence (NIH/NIAID)) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] F4/80 antibody Message-ID: <78F91CA4B8F3504E8FE5F13AAF122ED804A02CB3@nihexchange27.nih.gov> I just recently finished a research project staining for macrophages in mouse lungs. I used an F4/80 Rat anti-mouse from Serotec I used positive charged slides and enzyme pretreatment and did not have any trouble with tissue washing off, but lungs can be more forgiving than skin. I used the Pro-K from Dako for pretreatment, Primary was diluted 1:50 fro 60min, and the Vector Elite Rat setup. Lawrence J Faucette Contractor HT ASCP http://www.niaid.nih.gov/dir/services/animalcare/VetPathology/VetPathology-i ndex.html Infectious Disease Pathogenesis Section Comparative Medicine Branch Division of Intramural Research, NIAID, NIH Twinbrook III, Room 2W-01A, MSC 8135 12735 Twinbrook Parkway Bethesda, MD 20892-8135 Telephone 301-451-1056 Fax 301-480-2343 SoBran, Inc. Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. The National Institute of Allergy and Infectious Diseases (NIAID) shall not accept liability for any statement made that are the sender's own and not expressly made on behalf of the NIAID by one of its representatives. -----Original Message----- From: Angelicola,Lisa [mailto:Angelicola@uchc.edu] Sent: Monday, August 09, 2004 2:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] F4/80 antibody I'm working on a research project that involves demonstrating macrophages in formalin fixed paraffin embedded mouse skin wounds. The researcher has purchased the F4/80 rat-anti-mouse antibody from abcam. Has anyone out there in histoland had any experience working on fixed, processed tissue with this antibody? The spec sheet states that it will work with pretreatment of the tissue (protein digestion, which I would like to avoid if possible since the tissue is susceptible to falling off). Also where do you purchase your anti-rat secondary from? I would appreciate any advice anyone might have as far as pretreatment, dilutions, ect. Thanks in advance, Lisa Angelicola Immunohistochemistry Lab UCONN Health Center Farmington, Ct. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mark.bliek <@t> bolton-works.com Mon Aug 9 14:38:49 2004 From: mark.bliek <@t> bolton-works.com (Mark Bliek) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Leica / Reichert Polycut M S or E wanted Message-ID: <01C47E26.F81AF5F0.mark.bliek@bolton-works.com> Looking for used (read: low-cost .....) Leica / Reichert Polycut M S or E microtome for research project, working or non-working condition. Email (mark.bliek@bolton-works.com) or call (860) 646-3948. Mark Bliek Bolton Works LLC Vernon, CT From jtsonger <@t> vt.edu Mon Aug 9 14:40:54 2004 From: jtsonger <@t> vt.edu (Jill Songer) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Problem with VIP 5 Message-ID: <6.0.0.22.0.20040809153650.01b06628@pop.vt.edu> We are having a problem with our VIP 5 having a strong formalin odor at the end of the run when we open the retort to remove the tissues. There is also a large amount of fluid-formalin?? on the lid and in the chamber. Has anyone else experienced this? I have called Sakura and they are sending a field service tech but I was curious if anyone has had this problem. Whenever I call a vendor it seems they always say, well, we haven't had THIS problem before. Thanks. Jill Songer HT (ASCP) Histopathology Lab Virginia-Maryland Regional College of Veterinary Medicine Teaching Hospital Virginia Tech Blacksburg, VA 24061-0443 www.vetmed.vt.edu From gcallis <@t> montana.edu Mon Aug 9 14:56:51 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] F4/80 antibody In-Reply-To: References: Message-ID: <6.0.0.22.1.20040809134502.01b20eb0@gemini.msu.montana.edu> Histonet archives (www.histosearch.org) has informtation on F4/80 (you should do a dilution panel on tissue expressing mature macrophages to determine optimal working concentration adn start at 10 ug/ml.) From one Histonet message, Barb Wright did Pretreatment: 0.4% Pepsin in 0.1N HCl (5 minutes at 37?C). As for secondary, we use TAGO/Biosource goat antirat F(ab')2 frag of IgG, biotinylated adsorbed to mouse or Jackson Donkey antiRat F(ab')2 frag of IgG-biotinylated adsorbed to mouse (and other species) 0.5 mg/ml dilutrd 1:250. If you do not have success with the ABCAM antibody, buy SEROTEC - it works very well. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From cfavara <@t> niaid.nih.gov Mon Aug 9 15:36:54 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] F4/80 antibody Message-ID: I have used this antibody successfully in FFPE mouse tissue using pretreatment with citrate buffer pH 6.0 120c for 20 minutes. Can't remember the dil I used I suggest a dilution curve, you can use liver as F4/80 will stain Kupher cells nicely. I use vector biotinylated anti-rat, mouse absorbed, any reliable supplier should work well followed by streptavidin and AEC as a chromogen. Relatively pain free! Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Angelicola,Lisa [mailto:Angelicola@uchc.edu] Sent: Monday, August 09, 2004 12:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] F4/80 antibody I'm working on a research project that involves demonstrating macrophages in formalin fixed paraffin embedded mouse skin wounds. The researcher has purchased the F4/80 rat-anti-mouse antibody from abcam. Has anyone out there in histoland had any experience working on fixed, processed tissue with this antibody? The spec sheet states that it will work with pretreatment of the tissue (protein digestion, which I would like to avoid if possible since the tissue is susceptible to falling off). Also where do you purchase your anti-rat secondary from? I would appreciate any advice anyone might have as far as pretreatment, dilutions, ect. Thanks in advance, Lisa Angelicola Immunohistochemistry Lab UCONN Health Center Farmington, Ct. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Aug 9 15:39:56 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] VIR members at Toronto for committee table Message-ID: <6.0.0.22.1.20040809142838.01aec510@gemini.msu.montana.edu> To VIR'/NSH members attending Toronto NSH convention/symposium, the VIR committee needs volunteers to "man" the table during some coffee and lunch breaks from Saturday to Tuesday. If you can help out it would be greatly appreciated, please contact the VIR chairperson, Diane Sterchi at sterchi_diane_l@lilly.com or me (Gayle Callis) so I can forward your volunteered services to Diane. The Veterinary, Industry and Research Committee meeting will be Monday, Sept 20th from 5 to 6 pm. We invite both nonmembers and members to bring your ideas and participate in some of the new things being discussed, planned and updated for the VIR committee. Thank you, Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Jackie.O'Connor <@t> abbott.com Mon Aug 9 15:38:50 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Problem with VIP 5 Message-ID: Yes - I remember it well - - maybe not so well - but sort of - - - Had the same problem about 12 years ago with a VIP - there was a ceramic piece (valve changer?) in machine that spins to allow different solutions into the retort. This piece wore down and didn't seat accurately - allowing other solutions to backwash into the lines. I don't know if they've changed this design over the years - but your story struck a familar chord . . . . Jackie O'Connor Abbott Labs Discovery Chemotherapeutics Jill Songer Sent by: histonet-bounces@lists.utsouthwestern.edu 08/09/2004 02:40 PM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Problem with VIP 5 We are having a problem with our VIP 5 having a strong formalin odor at the end of the run when we open the retort to remove the tissues. There is also a large amount of fluid-formalin?? on the lid and in the chamber. Has anyone else experienced this? I have called Sakura and they are sending a field service tech but I was curious if anyone has had this problem. Whenever I call a vendor it seems they always say, well, we haven't had THIS problem before. Thanks. Jill Songer HT (ASCP) Histopathology Lab Virginia-Maryland Regional College of Veterinary Medicine Teaching Hospital Virginia Tech Blacksburg, VA 24061-0443 www.vetmed.vt.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kennedya <@t> email.cs.nsw.gov.au Mon Aug 9 16:01:21 2004 From: kennedya <@t> email.cs.nsw.gov.au (Andrew Kennedy) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Medical Abbreviations Message-ID: <200408100658109.SM01312@crgcsls813> There was an earlier thread asking about common medical abbreviations. An editorial in Human Pathology talks about some "Do not use" abbreviations and "Dangerous Pathology Abbreviations". It's worth a read if you can get hold of it. Reference: Booker, D.L. Berman, J.J. Dangerous Abbreviations. Human Pathology Vol 35 No.5. May 2004. pp529-531 Andrew Kennedy Senior Science Officer Anatomical Pathology Concord Repatriation General Hospital Hospital Road Concord NSW 2139 ph: +612 9767 6115 Fax +612 9767 8427 "corpora non agunt nisi fixata" From kennedya <@t> email.cs.nsw.gov.au Mon Aug 9 16:19:03 2004 From: kennedya <@t> email.cs.nsw.gov.au (Andrew Kennedy) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] TB in Lung Specimens Message-ID: <200408100715107.SM01312@crgcsls813> Hi Histonetters, A recent article in Human Pathology suggests that Mycobacteria in formalin fixed tissue can remain viable and therefore there is a risk of contracting TB from these specimens. It suggests that formalin fixed tissue from suspected TB cases should be handled with gloves, gown and mask. I wonder if we should be using these precautions for every lung specimen at every step in the histological process! If the organisms are still viable, trimmings from blocks should probably be bagged and disposed of in infectious waste. It could quite possibly end up being the same as with CJD brain specimens. What do you all think about this? Reference: Gerston, K.F. Blumberg, L. Tshabalala, V.A. Murray, J. Viability of Mycobacteria in Formalin Fixed Lungs. Human Pathology, Vol 35, No 5. May 2004. pp 571-575 Andrew Kennedy Senior Science Officer Anatomical Pathology Concord Repatriation General Hospital Hospital Road Concord NSW 2139 ph: +612 9767 6115 Fax +612 9767 8427 "corpora non agunt nisi fixata" From JCarpenter764 <@t> aol.com Mon Aug 9 16:32:00 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Thanks to everyone's response to all my silly questions... Message-ID: <9f.4b26c70c.2e494750@aol.com> To All on the Histonet Mailing List: Thank you to everyone that has given me great responses to ALL of my questions...I don't know where I would be without all of you...Love Ya All Jennell From Patty.Lott <@t> ORTHO.UAB.EDU Mon Aug 9 16:46:54 2004 From: Patty.Lott <@t> ORTHO.UAB.EDU (Patty Lott) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] unsubscribe Message-ID: <85F6C7A1330E794DB8540AFD001CC77E0532891A@rosco.ortho.uab.edu> Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 From gcallis <@t> montana.edu Mon Aug 9 16:59:53 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Thanks to everyone's response to all my silly questions... In-Reply-To: <9f.4b26c70c.2e494750@aol.com> References: <9f.4b26c70c.2e494750@aol.com> Message-ID: <6.0.0.22.1.20040809155831.01b22f40@gemini.msu.montana.edu> Jennell, There are no silly nor dumb questions! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From stema_cba <@t> yahoo.it Mon Aug 9 17:03:33 2004 From: stema_cba <@t> yahoo.it (=?iso-8859-1?q?Stefano=20Mantero?=) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Lac Z staining In-Reply-To: <20040809115841.73062.qmail@web42008.mail.yahoo.com> Message-ID: <20040809220333.55480.qmail@web60405.mail.yahoo.com> Hello Julie, Unfotunately the beta Galactosidase are inactivated from heat. If you want to use the paraffin you have two possibilities: 1 reveal the beta galactosidase with an antibodies (Chemicon works well) 2 use a very low melting point paraffin (42-44 C), in this case you preserve the activity of the enzyme but the section is not simple to obtain, it is necessary a cryostat setted at +4 C Good luck Stefano Stefano Mantero Dulbecco Telethon Institute Via fratelli Cervi, 93 Segrate (MI) Italy --- Juile Gorenstein ha scritto: > Hello everyone, > Thank you to those who has responded to my embryo > processing e-mail! > I was wondering if anyone has a protocol for lac Z > staining on paraffin sections. I have been staining > frozen sections, but have been having a little bit > of trouble keeping the tissue attached to the slides > (I use Superfrost Plus slides: I allow the tissue > to dry for about an hr after I attach the section to > the slide, store them at -80oC and then dry them > again before staining -- I stain by hand, it's not > an instrument issue). Please let me know if a > protocol for paraffin sections exists. > > Thank you in advance, > Julie Gorenstein > Novartis, Cambridge, MA > > > --------------------------------- > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ____________________________________________________________ Yahoo! Companion - Scarica gratis la toolbar di Ricerca di Yahoo! http://companion.yahoo.it From jnocito <@t> satx.rr.com Mon Aug 9 17:26:59 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Problem with VIP 5 References: <6.0.0.22.0.20040809153650.01b06628@pop.vt.edu> Message-ID: <00d101c47e5f$fcb13000$5d6bce44@yourxhtr8hvc4p> Jill, how long have you had the VIP 5? I've had my machine almost 3 years and haven't had that problem. If you don't close the lid properly, when it pumps in fluid, the lid pops off, but we haven't had any smells. Please let us know what the service rep says. I always like new stories. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Jill Songer" To: Sent: Monday, August 09, 2004 2:40 PM Subject: [Histonet] Problem with VIP 5 > We are having a problem with our VIP 5 having a strong formalin odor at the > end of the run when we open the retort to remove the tissues. There is also > a large amount of fluid-formalin?? on the lid and in the chamber. > > Has anyone else experienced this? I have called Sakura and they are sending > a field service tech but I was curious if anyone has had this problem. > Whenever I call a vendor it seems they always say, well, we haven't had > THIS problem before. > > Thanks. > > Jill Songer HT (ASCP) > Histopathology Lab > Virginia-Maryland Regional College of Veterinary Medicine > Teaching Hospital > Virginia Tech > Blacksburg, VA 24061-0443 > www.vetmed.vt.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From neuroant <@t> hotmail.com Mon Aug 9 17:35:35 2004 From: neuroant <@t> hotmail.com (Ant S.) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] ASCP exam...anyone there? Message-ID: I have not yet recieved even a notice that I am qualified to take the exam, much less the practical exam information. Has anyone else at least gotten a letter saying --thank you for applying, here's more information?-- Antoinette Swensson Univeristy of Washington/Harborview Medical Center Neuropathology 325 9th Ave MS 359-791 Seattle, WA 98104 (206) 731-3910 _________________________________________________________________ [1]Check out Election 2004 for up-to-date election news, plus voter tools and more! References 1. http://g.msn.com/8HMBENUS/2746??PS=47575 From Gervaip <@t> aol.com Mon Aug 9 17:50:02 2004 From: Gervaip <@t> aol.com (Gervaip@aol.com) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] Problem with VIP 5 Message-ID: The only time I smelled formalin when I opened the retort.... the rotor head was cracked. Is there formalin in the paraffin tanks? Pearl From laurie.colbert <@t> huntingtonhospital.com Mon Aug 9 17:53:18 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] 2N HCL Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BFE7@EXCHANGE1.huntingtonhospital.com> Can anyone tell me how to make a 2N solution of HCL from concentrated HCL? Laurie Colbert From tstella1 <@t> tampabay.rr.com Mon Aug 9 18:47:03 2004 From: tstella1 <@t> tampabay.rr.com (Terri Stella-Vega) Date: Fri Sep 16 15:23:51 2005 Subject: [Histonet] ASCP exam...anyone there? References: Message-ID: <000b01c47e6b$2d77e890$667ba8c0@Terri> I am currently working with several candidates. If you have not received a call/paper by now you should call ASCP. One of our candidates had not received papers yet; had placed a phone call to them to discover she was in the system, check cashed, but no notice. She mentioned they had explained there are hundreds that have applied. ----- Original Message ----- From: "Ant S." To: Sent: Monday, August 09, 2004 6:35 PM Subject: [Histonet] ASCP exam...anyone there? > > I have not yet recieved even a notice that I am qualified to take the > exam, much less the practical exam information. Has anyone else at > least gotten a letter saying --thank you for applying, here's more > information?-- > > > Antoinette Swensson > Univeristy of Washington/Harborview Medical Center > Neuropathology > 325 9th Ave MS 359-791 > Seattle, WA 98104 > (206) 731-3910 > _________________________________________________________________ > > [1]Check out Election 2004 for up-to-date election news, plus voter > tools and more! > > References > > 1. http://g.msn.com/8HMBENUS/2746??PS=47575 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rschoon <@t> email.unc.edu Mon Aug 9 20:03:16 2004 From: rschoon <@t> email.unc.edu (Robert Schoonhoven) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] 2N HCL In-Reply-To: <0BE6ADFAE4E7E04496BF21ABD346628001C5BFE7@EXCHANGE1.huntingtonhospital.com> References: <0BE6ADFAE4E7E04496BF21ABD346628001C5BFE7@EXCHANGE1.huntingtonhospital.com> Message-ID: <41181ED4.9000404@email.unc.edu> Concentrated HCl is about 12N. Should be able to make your dilution from that. Laurie Colbert wrote: > Can anyone tell me how to make a 2N solution of HCL from concentrated HCL? > > Laurie Colbert > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ianbernard <@t> netzero.com Mon Aug 9 21:02:23 2004 From: Ianbernard <@t> netzero.com (Ian) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Problem with VIP 5 In-Reply-To: <00d101c47e5f$fcb13000$5d6bce44@yourxhtr8hvc4p> Message-ID: Hey, you all don't scare me WE just purchase a new VIP 5 and have not have used it yet! Hey Joe, how are you doing? SSgt Bernard 59th Clinical Research Squadron "The USAF's largest Clinical Research facility" Lackland AFB, Texas -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Monday, August 09, 2004 3:27 PM To: Jill Songer; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Problem with VIP 5 Jill, how long have you had the VIP 5? I've had my machine almost 3 years and haven't had that problem. If you don't close the lid properly, when it pumps in fluid, the lid pops off, but we haven't had any smells. Please let us know what the service rep says. I always like new stories. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Jill Songer" To: Sent: Monday, August 09, 2004 2:40 PM Subject: [Histonet] Problem with VIP 5 > We are having a problem with our VIP 5 having a strong formalin odor at the > end of the run when we open the retort to remove the tissues. There is also > a large amount of fluid-formalin?? on the lid and in the chamber. > > Has anyone else experienced this? I have called Sakura and they are sending > a field service tech but I was curious if anyone has had this problem. > Whenever I call a vendor it seems they always say, well, we haven't had > THIS problem before. > > Thanks. > > Jill Songer HT (ASCP) > Histopathology Lab > Virginia-Maryland Regional College of Veterinary Medicine > Teaching Hospital > Virginia Tech > Blacksburg, VA 24061-0443 > www.vetmed.vt.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From aldo.anile <@t> HDScientific.com.au Mon Aug 9 21:21:27 2004 From: aldo.anile <@t> HDScientific.com.au (Aldo Anile) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] RE: Glass vs Tape coverslippers Message-ID: There is little question that if all things being equal, glass would be the preferred option for coverslipping. Some glass coverslippers today have addressed the main advantages that tape has had over glass, speed and wet slides. The Medite RCM7000 (Meisei Promounter) has the ability to coverslip at a rate of 450 slides/hr. It also has a slide drying fan that produces touch dry slides, eliminating the messy wet slide issue that has historically been associated with glass coverslipping. Anyone that has gone back to slides years after being coverslipped with tape would be familiar with the deteriorating nature of tape coverslipping over time. Aldo Anile Applications Consultant HD SCIENTIFIC SUPPLIES PTY LTD (Aust) E-mail: aldo.anile@hdscientific.com.au Web: www.hdscientific.com.au -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Friday, 6 August 2004 23:29 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 9, Issue 8 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: Glass vs. Tape Coverslippers (Weems, Joyce) 2. Re: Staining for Macrophages (Sharon Cooperman) 3. Re: Private responses - well said, Barry! (Gayle Callis) 4. Re: Private responses - well said, Barry! (Susan Q Wells) 5. RE: Private responses (Mass Histology Service) 6. (no subject) (Browning Deb) 7. Recall: (Browning Deb) 8. retrieval puzzle, forgot this line on last message (Browning Deb) 9. Jeff Lowen (James Watson) 10. Benchmark XT (Lab) 11. BRDU free floating vibratome sections (Kimberle M. Jacobs) 12. ID-2 (Kimberle M. Jacobs) 13. RE: Benchmark XT (Joe Nocito) 14. Re: BRDU free floating vibratome sections (Sarah Jones) 15. H. pylori control tissue (Richard Cartun) 16. RE: Private responses (Tony Henwood) 17. RE: electrical charge slides (Bill Sinai) 18. Rank and Rank L (Patricia Bourne) 19. Problem with Masson's Trichrome (Bdeer24@aol.com) 20. Re: H. pylori control tissue (lpwenk@sbcglobal.net) 21. Fixative for FACS (Myri37@aol.com) 22. Oncocytoma vs. Chromophobe (Barnhart, Tammy) 23. Re: Problem with Masson's Trichrome (rschoon) 24. Cytochrome oxidase (COX) staining: non-carcinogenic alternative to DAB? (Martin Hasselblatt) 25. Masson Trichrome (Hermina Borgerink) ---------------------------------------------------------------------- Message: 1 Date: Thu, 5 Aug 2004 13:09:25 -0400 From: "Weems, Joyce" Subject: RE: [Histonet] Glass vs. Tape Coverslippers To: "Morken, Tim - Labvision" , "Histonet \(E-mail\)" Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A626B@sjhaexc02.sjha.org> Content-Type: text/plain; charset="iso-8859-1" I was demoing coverslippers and didn't tell the pathologists. Thought I'd see if they could tell the difference. One of our pathologists made us remove the tape from her entire days workload and recoverslip them by hand. Oh well..... Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Morken, Tim - Labvision Sent: Thursday, August 05, 2004 12:08 PM To: 'Histonet (E-mail)' Subject: RE: [Histonet] Glass vs. Tape Coverslippers Gary, We did find that plastic was no good for cytology specimens - at least the lumpy ones - it wouldn't form a flat surface. For thin-prep it worked great. I think the quality is OK for routine histology. In our lab we did testing in the lab and when satisfied it was working well we switched over without telling the pathologists. About two weeks later the Chief Pathologist asked me when we would start using the plastic coverslips! Tim Morken -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Thursday, August 05, 2004 8:53 AM To: 'Laurie Colbert'; WWmn916@aol.com; Histonet (E-mail) Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass vs. Tape Coverslippers Plastic is no substitute for glass. Just because one doesn't "see" a difference doesn't mean there isn't a difference. Whether the difference makes a real difference in outcomes is another story. ASTM specs for cover glasses apply to glass, not to tape. The higher the numerical aperture of an objective, and the better the objective quality (i.e., achromat, fluorite, apochromat -- plan and non-plan), the more likely that one will see imaging differences when tape is used. Of course, using glass doesn't ensure good quality. Practical stuff like mounting medium and cover glass thickness, clean lenses, and Kohler illumination also play a real role. Image quality can't be better than the weakest link. The specs are relative to the impact of the physical and optical properties of glass on light as it passes through the mounting medium and glass through the objective. For this reason, it's not nice to fool Mother Nature and use plastic. Gary Gill -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Thursday, August 05, 2004 10:23 AM To: WWmn916@aol.com; Histonet (E-mail) Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass vs. Tape Coverslippers We have a tape coverslipper, and we love it. It is fast and we have had very few problems with it. Our pathologists have no problem reading the slides, and as far as I know, there's never been a problem photographing a slide. We did demo the glass coverslippers when we were first looking for a new coverslipper, and there were too many problems with slides sticking together, air bubbles, and it was just all-around not as "user friendly." Laurie Colbert Huntington Hospital -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Wednesday, August 04, 2004 8:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass vs. Tape Coverslippers Hello again, I'm looking for opinions on the subject of glass coverslippers versus tape coverslipping. I have the opportunity to decide on a system. My only experience has been with tape coverslipping. I understand machines that glass coverslip are slower than tape systems. Is the refractive index better with glass coverslips under the microscope? Opinions pros/cons are appreciated. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. ------------------------------ Message: 2 Date: Thu, 5 Aug 2004 13:24:18 -0400 From: Sharon Cooperman Subject: Re: [Histonet] Staining for Macrophages To: Gayle Callis Cc: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Thanks, Gail, you're right. Does FA-11 work on FFPE tissue and do you need HIER? Serotec doesn't say. Thanks, Sharon >Sorry, but ED-1 (CD68) is a Mouse anti-Rat antibody for a subset of >rat macrophages. It may cross react to mouse but you would be doing >doing mouse on mouse IHC. The CD68 for mouse is rat anti-mouse, >clone FA-11 for murine peritoneal macrophages. SEROTEC is a source >of these antibodies, including the F4/80 (Rat anti-mouse). > >A sidenote: SEROTEC has a free macrophage differentiation poster >available upon request. > >At 06:58 PM 8/4/2004, you wrote: > >>You can use ED-1 from Serotec. It works on FFPE tissue with pH 6.0 >>citrate antigen retrieval and on frozen tissue without AR. You can >>also use F4/80 on frozen tissue. Both are rat monoclonals and you >>can use an anti-rat secondary Ab cross adsorbed against mouse IgG. >>You could also also use ricin 1 from Vector or E-Y Laboratories >>which binds mouse macrophages. Ricin 1 is not the dangerous ricin. >>I don't know details of how to use it (I did once, but I don't >>remember). There are books available or the manufacturers may know >>or maybe some other Histonet folks. I have used ED1 and F4/80 on >>mouse tissues and I think it's your best bet. >> >>Sharon >> >>>I had previously asked about MAC-387 staining. Unfortunately, we >>>can't use the staining because we are staining mouse kidney and it >>>doesn't look right if you stain with a mouse anti-body. Does >>>anyone know of a way to stain for macrophages with out an >>>antibody??? Or perhaps a polyclonal antibody???? Thanks a bunch! >>> >>> >>>Jennifer K. Sipes, RALAT >>>Sr. Laboratory Technician >>>Johns Hopkins University >>>Ross 929 >>>720 Rutland Avenue >>>Baltimore, MD 21205 >>>phone: 410-614-0131 >>>cell: 443-413-0853 >>>e-mail: jengirl1014@yahoo.com >>> >>> >>> >>> >>> >>> >>> >>> >>> >>> >>> >>>--------------------------------- >>>Do you Yahoo!? >>>Yahoo! Mail - 50x more storage than other providers! >>>_______________________________________________ >>>Histonet mailing list >>>Histonet@lists.utsouthwestern.edu >>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >> >> >>-- >>Sharon Cooperman >>NIH, NICHD, CBMB 301.435-7735 >>Building 18T, room 101 301.402-0078 fax >>Bethesda, MD 20892 >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 ------------------------------ Message: 3 Date: Thu, 05 Aug 2004 11:41:52 -0600 From: Gayle Callis Subject: Re: [Histonet] Private responses - well said, Barry! To: mucram11@comcast.net, Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20040805112541.01afad70@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" Pam and all, Too many people forget or ignore the fact that some individuals who are experts in the field of histotechnology are not employed in just clinical or research laboratories. If one is involved with a company doing technical research and development of a product destined for histoland, then they are should be very welcome to comment. Knowledge should not be smothered (flames!) I don't see you as a salesperson, but a valuable resource providing me with information I can choose to use or not. All part of the learning experience. Complaints = delete button. At 10:13 AM 8/5/2004, you wrote: I agree with both Gayle and Barry yet have one reservation. I was hit several times on this server for answering very general questions and told salespeople should not be on HistoNet. Although my answers had nothing to do with any product we sell and by the way I am not a sales person, just technical and product development. After the second incident that several others came on and agreed we were not welcome I stay off 99% of the time. It is difficult to walk the line here and not be on privately only. I do agree that if it is a sales pitch it should not be on the line as a reply that should be private answers only. Thanks for letting me say this and I hope no one is offended. Pam Marcum -------------- Original message -------------- > Thank you Barry - very well said. > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > Bozeman MT 59717-3610 > > At 09:02 AM 8/5/2004, you wrote: > > >I hate to see the word "flamed" as here in sunny Houston it has been in > >the upper 90s for about 2 weeks. > > > >While I appreciate people's perception about getting "flamed" when they > >have posted items, I have a real problem with posting most items > >privately. > >Histonet is an extremely valuable resource and there is a vast amount of > >expertise available. It is only logical for this knowledge and > >experience to be f! reely shared. > >I can't say that I have been "flamed" (or even mildly scorched) but > >sometimes I make mistakes and post incorrect information. It is a > >valuable learning experience for me and all interested parties to have > >any errors corrected. Sometimes it is matter of perception and how > >criticism is leveled (being brutally frank is not a good attitude). > >"Flaming" may be perceived by one individual to have occurred while > >another may regard a response as constructive criticism. > >If anyone here gets "flamed" or has their employer contacted by a > >company about a posting then please let us know. Companies do respond > >positively to public opinion. Any companies with such an attitude this > >should be held accountable and so should employers for not telling the > >company or their representative to take a hike. > >I do believe t! hat if someone requests private responses that this be >! ; >re spected. Second, I believe that when responding one should be careful > >when criticizing a piece of equipment or a company. Negative criticism > >sticks and while this may be true for a particular individual it could > >be an isolated incident and due to many factors. Not everyone knows > >how to deal effectively with companies (and not every company knows how > >to deal responsibly with customers). I think that such listings should > >be private. The individual who receives these could then let us know > >about the final results in the form of a summary. > >If I am to get "flamed" for this or any other postings please include > >"flamed" at the start of the message so that I don't' just think that > >the response is constructive criticism. > >Barry > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >[1]http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > [2]http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) References 1. http://lists.utsouthwestern.edu/mailman/listinfo/histonet 2. http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Thu, 05 Aug 2004 14:20:13 -0400 From: Susan Q Wells Subject: Re: [Histonet] Private responses - well said, Barry! To: Gayle Callis Cc: Histonet@lists.utsouthwestern.edu Message-ID: <41127A5D.4080406@bms.com> Content-Type: text/plain; format=flowed; charset=us-ascii I completely agree with Gayle! This list is invaluable to most of us working in the field. Gayle Callis wrote: > Pam and all, > Too many people forget or ignore the fact that some individuals who > are experts in the field of histotechnology are not employed in just > clinical or research laboratories. If one is involved with a company > doing technical research and development of a product destined for > histoland, then they are should be very welcome to comment. > Knowledge should not be smothered (flames!) I don't see you as a > salesperson, but a valuable resource providing me with information I > can choose to use or not. All part of the learning experience. > Complaints = delete button. > > > At 10:13 AM 8/5/2004, you wrote: > > I agree with both Gayle and Barry yet have one reservation. I was > hit several times on this server for answering very general > questions and told salespeople should not be on HistoNet. Although > my answers had nothing to do with any product we sell and by the > way I am not a sales person, just technical and product > development. After the second incident that several others came on > and agreed we were not welcome I stay off 99% of the time. It is > difficult to walk the line here and not be on privately only. I do > agree that if it is a sales pitch it should not be on the line as a > reply that should be private answers only. > Thanks for letting me say this and I hope no one is offended. > Pam Marcum > > -------------- Original message -------------- > > Thank you Barry - very well said. > > > > Gayle Callis > > MT,HT,HTL(ASCP) > > Research Histopathology Supervisor > > Veterinary Molecular Biology > > Montana State University - Bozeman > > Bozeman MT 59717-3610 > > > > At 09:02 AM 8/5/2004, you wrote: > > > > >I hate to see the word "flamed" as here in sunny Houston it > has been in > > >the upper 90s for about 2 weeks. > > > > > >While I appreciate people's perception about getting > "flamed" when they > > >have posted items, I have a real problem with posting most > items > > >privately. > > >Histonet is an extremely valuable resource and there is a > vast amount of > > >expertise available. It is only logical for this knowledge > and > > >experience to be f! reely shared. > > >I can't say that I have been "flamed" (or even mildly > scorched) but > > >sometimes I make mistakes and post incorrect information. It > is a > > >valuable learning experience for me and all interested > parties to have > > >any errors corrected. Sometimes it is matter of perception > and how > > >criticism is leveled (being brutally frank is not a good > attitude). > > >"Flaming" may be perceived by one individual to have > occurred while > > >another may regard a response as constructive criticism. > > >If anyone here gets "flamed" or has their employer contacted > by a > > >company about a posting then please let us know. Companies > do respond > > >positively to public opinion. Any companies with such an > attitude this > > >should be held accountable and so should employers for not > telling the > > >company or their representative to take a hike. > > >I do believe t! hat if someone requests private responses > that this be > >! > ; >re spected. Second, I believe that when responding one > should be careful > > >when criticizing a piece of equipment or a company. Negative > criticism > > >sticks and while this may be true for a particular > individual it could > > >be an isolated incident and due to many factors. Not > everyone knows > > >how to deal effectively with companies (and not every > company knows how > > >to deal responsibly with customers). I think that such > listings should > > >be private. The individual who receives these could then let > us know > > >about the final results in the form of a summary. > > >If I am to get "flamed" for this or any other postings > please include > > >"flamed" at the start of the message so that I don't' just > think that > > >the response is constructive criticism. > > >Barry > > > > > > > > >_______________________________________________ > > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > > >[1]http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > [2]http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > > MT,HT,HTL(ASCP) > > Research Histopathology Supervisor > > Veterinary Molecular Biology > > Montana State University - Bozeman > > PO Box 173610 > > Bozeman MT 59717-3610 > > 406 994-6367 (lab with voice mail) > > 406 994-4303 (FAX) > >References > > 1. http://lists.utsouthwestern.edu/mailman/listinfo/histonet > 2. http://lists.utsouthwestern.edu/mailman/listinfo/histonet >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 5 Date: Thu, 5 Aug 2004 14:33:59 -0400 From: "Mass Histology Service" Subject: RE: [Histonet] Private responses To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Especially the archives! Quite often a topic is of no interest to me at the present time yet a week later, I have to stain for that certain CD antibody that I remember was discussed in detail the prior week. I use the archives several times a week and if all responses were private, there would be no archives! Jim ____________________ James Staruk, HT(ASCP) Mass Histology Service www.masshistology.com ------------------------------ Message: 6 Date: Thu, 5 Aug 2004 15:05:35 -0400 From: Browning Deb Subject: [Histonet] (no subject) To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <3AADFB88753AD31189C100902786B91C0E27836D@hch_nt_exchange.hhsc.ca> Content-Type: text/plain; charset="iso-8859-1" Here is a puzzle. In order to streamline the IHC workload, I have had the techs organize the slides in vertical racks that can be used for dewaxing, heat retrieval, and quenching (meth/px), instead of the old way of dewaxing in one set of racks, switching to other racks for retrieval, and then into coplin jars for quenching. Sounds great. We ended up with an unusual vertical staining pattern with central staining of the tissues, but both edges that corresponded with the edges of the slide racks had no staining. The slides were heated at 60 deg. overnight. Slides that did not require heat retrieval were not affected, therefore it was not a dewaxing problem. All slides lay flat for application of antibodies and detection system. I know of another site that vertically handles their slides, and they do not get this artefact, the only difference between them and us is quenching before retrieval vs after. Any suggestions? We went back to the old way, and the artefact is gone, nothing changed except the position (vertical vs horizontal) of the slides. Looking forward to scientific explanations, inquiring minds want to know. Debra Browning, ART Technical Specialist, Immunohistochemistry Anatomic Pathology Hamilton Health Sciences phone: (905) 527-4322 ext 46131 e-mail: browning@hhsc.ca fax: (905) 524-2681 This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. ------------------------------ Message: 7 Date: Thu, 5 Aug 2004 15:07:12 -0400 From: Browning Deb Subject: [Histonet] Recall: To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <3AADFB88753AD31189C100902786B91C0E27836E@hch_nt_exchange.hhsc.ca> Content-Type: text/plain Browning Deb would like to recall the message, "". This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. ------------------------------ Message: 8 Date: Thu, 5 Aug 2004 15:07:41 -0400 From: Browning Deb Subject: [Histonet] retrieval puzzle, forgot this line on last message To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <3AADFB88753AD31189C100902786B91C0E27836F@hch_nt_exchange.hhsc.ca> Content-Type: text/plain; charset="iso-8859-1" Here is a puzzle. In order to streamline the IHC workload, I have had the techs organize the slides in vertical racks that can be used for dewaxing, heat retrieval, and quenching (meth/px), instead of the old way of dewaxing in one set of racks, switching to other racks for retrieval, and then into coplin jars for quenching. Sounds great. We ended up with an unusual vertical staining pattern with central staining of the tissues, but both edges that corresponded with the edges of the slide racks had no staining. The slides were heated at 60 deg. overnight. Slides that did not require heat retrieval were not affected, therefore it was not a dewaxing problem. All slides lay flat for application of antibodies and detection system. I know of another site that vertically handles their slides, and they do not get this artefact, the only difference between them and us is quenching before retrieval vs after. Any suggestions? We went back to the old way, and the artefact is gone, nothing changed except the position (vertical vs horizontal) of the slides. Looking forward to scientific explanations, inquiring minds want to know. Debra Browning, ART Technical Specialist, Immunohistochemistry Anatomic Pathology Hamilton Health Sciences phone: (905) 527-4322 ext 46131 e-mail: browning@hhsc.ca fax: (905) 524-2681 This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. ------------------------------ Message: 9 Date: Thu, 5 Aug 2004 12:31:21 -0700 From: "James Watson" Subject: [Histonet] Jeff Lowen To: Message-ID: <7BA50F61CB491B4692B8BCFBB656B5F903428986@EXCHCLUSTER01.lj.gnf.org> Content-Type: text/plain; charset="us-ascii" Jeff Lowen, Please contact me. Thank you. James Watson HT, ASCP Facilities Manager of Histology GNF, C015 858-332-4647 jwatson@gnf.org ------------------------------ Message: 10 Date: Thu, 5 Aug 2004 14:50:20 -0400 From: "Lab" Subject: [Histonet] Benchmark XT To: Message-ID: <001101c47b1d$0f0f5e70$70cfa8c0@LAB> Content-Type: text/plain; charset="iso-8859-1" Our lab has had 2 ventana nexes systems and we are looking to upgrade to 2 benchmark xt units.We have had one in our lab to demo for approx. 3 months.During this time we have had 2 thermal pad issues and several file corruptions. Has anyone with a benchmark xt had similar problems and if so how often? Any opions about the instrument would be helpful. Thanks, Amy Boan,HT ------------------------------ Message: 11 Date: Thu, 05 Aug 2004 16:32:43 -0400 From: "Kimberle M. Jacobs" Subject: [Histonet] BRDU free floating vibratome sections To: histonet@lists.utsouthwestern.edu Message-ID: <5.1.0.14.2.20040805162931.01c9f2a8@mail1.vcu.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Hi I have searched the archives and found a lot of discussion of BRDU, but no answer to the question if there is a protocol out there for formaldehyde-fixed, vibratome-cut, free floating sections. Actually we could mount them on slides as well, I just don't want to do the whole paraffin embedding thing. Also is it really necessary to have thin sections if you are going to use pepsin and acid, etc. ?? We could probably cut as thin as 20 um, but would prefer to use 40 or 60 um. We will be doing this in pregnant ferrets (E36, gestation is ~42 days) and examining the brains at ages P0-P15. Thanks very much for any help!! Kimberle M. Jacobs, Ph.D. Assistant Professor Department of Anatomy and Neurobiology Virginia Commonwealth University P.O. Box 980709 Richmond, VA 23298-0709 (804) 827-2135 FAX: (804) 828-9477 kmjacobs@vcu.edu ------------------------------ Message: 12 Date: Thu, 05 Aug 2004 16:36:57 -0400 From: "Kimberle M. Jacobs" Subject: [Histonet] ID-2 To: histonet@lists.utsouthwestern.edu Message-ID: <5.1.0.14.2.20040805163247.01c9d160@mail1.vcu.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Has anyone used the antibody to ID-2? We purchased it from Santa Cruz Biotech. They had some recommendations for western blot, but none for immunohistochemistry. Our goal is simply to label the superficial layers of ferret visual cortex. But on our first try using 1:50, 1:100, and 1:200 primary dilutions - it didn't work at all. We were wondering if we need to do some antigen revealing step. Or perhaps it will not be found this caudal? Anyone know of any antibodies that label a specific layer in visual cortex? I've heard that the otx1 antibody is not working well right now. We are going to get RORbeta to attempt to stain layer IV, but again - no specific protocol for it. Thanks very much for any help! Kimberle M. Jacobs, Ph.D. Assistant Professor Department of Anatomy and Neurobiology Virginia Commonwealth University P.O. Box 980709 Richmond, VA 23298-0709 (804) 827-2135 FAX: (804) 828-9477 kmjacobs@vcu.edu ------------------------------ Message: 13 Date: Thu, 5 Aug 2004 15:44:15 -0500 From: "Joe Nocito" Subject: RE: [Histonet] Benchmark XT To: "Lab" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Amy, I have a new carousal waiting to be installed as we speak. I have 2 XT's, the oldest one since February of this year. I've had 8 different heating pads go bad on three different occasions since February. Most of the time, we place our control on the same slide as the patient, so if the control worked, there is no question that everything worked okay. If the control is negative, then we repeat the slide on the newer machine. In all fairness to Vetnana, they have responded well to our phone calls and emails. I think this is an issue that they are trying to work on in house. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Lab Sent: Thursday, August 05, 2004 1:50 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Benchmark XT Our lab has had 2 ventana nexes systems and we are looking to upgrade to 2 benchmark xt units.We have had one in our lab to demo for approx. 3 months.During this time we have had 2 thermal pad issues and several file corruptions. Has anyone with a benchmark xt had similar problems and if so how often? Any opions about the instrument would be helpful. Thanks, Amy Boan,HT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Thu, 05 Aug 2004 16:57:22 -0500 From: "Sarah Jones" Subject: Re: [Histonet] BRDU free floating vibratome sections To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Kimberle, I've done quite a bit of vibratome sectioning and free floating staining. It's quite challenging! Two ways I can think of that might help you. One would be to use the Shandon coverplate system. The sections would be floated onto slides and then the coverplate carefully placed on to the slide. It then fits into a holder and it gravity feeds the reagents over the slide. It uses a very small amount of reagent. The other way is to handle the free floating sections in netwells or you can make your own using open cassettes and small petri dishes. The most important steps would be making sure the rinsing steps were sufficient. The netwell/petri dish method would use a much greater amount of reagent. Hope this helps. Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Kimberle M. Jacobs" 08/05/04 3:32 PM >>> Hi I have searched the archives and found a lot of discussion of BRDU, but no answer to the question if there is a protocol out there for formaldehyde-fixed, vibratome-cut, free floating sections. Actually we could mount them on slides as well, I just don't want to do the whole paraffin embedding thing. Also is it really necessary to have thin sections if you are going to use pepsin and acid, etc. ?? We could probably cut as thin as 20 um, but would prefer to use 40 or 60 um. We will be doing this in pregnant ferrets (E36, gestation is ~42 days) and examining the brains at ages P0-P15. Thanks very much for any help!! Kimberle M. Jacobs, Ph.D. Assistant Professor Department of Anatomy and Neurobiology Virginia Commonwealth University P.O. Box 980709 Richmond, VA 23298-0709 (804) 827-2135 FAX: (804) 828-9477 kmjacobs@vcu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Thu, 05 Aug 2004 19:39:09 -0400 From: "Richard Cartun" Subject: [Histonet] H. pylori control tissue To: Message-ID: Content-Type: text/plain; charset=US-ASCII Is anyone looking for H. pylori control tissue? We just had a partial resection of stomach and it is loaded with bugs. If there is an overwhelming response, I may just send control blocks to the NSH tissue bank for distribution. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Hartford Hospital Hartford, CT 06102 (860) 545-1596 ------------------------------ Message: 16 Date: Fri, 6 Aug 2004 12:31:42 +1000 From: Tony Henwood Subject: RE: [Histonet] Private responses To: "'Dawson, Glen'" Cc: 'HistoNet Server' Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E230@simba.kids> Content-Type: text/plain Glen, I respect your right to free choice and accept your views. Keep well, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Dawson, Glen [mailto:GDawson@Milw.Dynacare.com] Sent: Friday, 6 August 2004 12:03 AM Cc: 'HistoNet Server' Subject: [Histonet] Private responses Baowei & All, If you don't want to deal with large amounts of grief, I would encourage you to respond privately. Due to threats of calls to my employer, lawsuits, profanity, constant flaming of every response I post and attaining my own little histonet stalker (you know who you are), I keep 99% of all my responses private. There are a number of people & vendors on this listserver that revel in tearing down the posts of others rather than creating helpful or insightful posts themselves. It is not a bad thing for you to post privately to avoid these problems. I actually used to post publicly up to the point that I was dealing with personal attacks more than I was gleaning useful information from the histonet. Engaging in those types of interactions was too time consuming, draining & enfuriating. Expressing one's own view is nice and, yes, it should be OK, but it isn't. I see no value in "shaming" someone into posting publicly. If you don't want to...don't. The funny thing is that I'll be getting flamed for this post which is whining about being flamed...isn't it ironic? Just My OPINION, Glen Dawson BS, HT & QIHC (ASCP) IHC Coordinator Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** ------------------------------ Message: 17 Date: Fri, 6 Aug 2004 12:54:41 +1000 From: "Bill Sinai" Subject: RE: [Histonet] electrical charge slides To: , "histonet \(E-mail\)" Message-ID: <000201c47b60$b8b82810$e1ce080a@wsahs.nsw.gov.au> Content-Type: text/plain; charset="iso-8859-1" Dana, There was heaps written about this on Histonet previously, look in the archives. We found that the problem was associated with the Plus slides when used in conjunction with an EDTA buffer as the material used to coat the slides is a calcium salt and the EDTA was affecting the coating by chelating the calcium!!! All the best Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of DDittus787@aol.com Sent: Friday, 06 August 2004 12:10 AM To: histonet@pathology.swmed.edu Subject: [Histonet] electrical charge slides To my colleagues: I have a question.Is anyone out there having occasional staining(orlack of staining on IHc slides?) I was told that the charge or the way the slides are charged can have (pardon the pun) a negative effect. Does anyone know /experience or have the scientific data for this? Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. ------------------------------ Message: 18 Date: Thu, 5 Aug 2004 19:55:56 -0700 (PDT) From: Patricia Bourne Subject: [Histonet] Rank and Rank L To: HistoNet Server Message-ID: <20040806025556.77653.qmail@web53706.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hello everyone....Has anyone worked successfully with Rank and Rank L? I've been through three (3) antibody sets and still not great results.....Help is really needed on this one. Thanks in advance for the help and information. --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. ------------------------------ Message: 19 Date: Fri, 06 Aug 2004 03:19:37 -0400 From: Bdeer24@aol.com Subject: [Histonet] Problem with Masson's Trichrome To: histonet@lists.utsouthwestern.edu Message-ID: <10A0ACD8.74118519.0015B53C@aol.com> Content-Type: text/plain; charset=iso-8859-1 Hi Everyone! I was recommended to try the Masson's Trichrome stainings on the slides of uterine tissue that I have been collecting (Previously I had only performed Hematoxylin & Eosin stains). This is a new method to our lab and thus far, staining has not gone well. Although my sections contain a significant amount of muscle, I have only been able to see the aniline blue stain. On a couple of the slides, there was a slight dark purple coloration on the epithelial lining. I have noticed upon comparing the protocol we received to others found on the internet that ours lacks the step in which the slides are placed in Bouin's fluid. There was also no specification in our protocol for the type of hematoxylin, so we had been using the same hematoxylin (Harris's modified with acetic acid) that we use in our H & E stainings. I was wondering if any of these might have led to the stainings not going as planned. The tissue sections also looked slightly brittle, perhaps due to a harsher stain. I am wondering if this has anything to do with my fixation process in which I use a 4% paraformaldehyde/PBS mixture, ethanol and xylene steps, as well as embedding the tissues in paraffin. Is Bouin's a crucial step, would I have better staining with a different tissue fixation, or should I be looking in a different direction? Any suggestions would be very much appreciated! Thanks! Amber Greenbank Arizona State University ------------------------------ Message: 20 Date: Fri, 6 Aug 2004 04:44:36 -0400 From: Subject: Re: [Histonet] H. pylori control tissue To: "Richard Cartun" , Message-ID: <007d01c47b91$9b6e7bc0$7434d445@domainnotset.invalid> Content-Type: text/plain; charset="iso-8859-1" I believe how the NSH Control bank works is that you let NSH control block coordinator know what you are willing to share. You then hang onto the blocks, and if anyone contacts NSH asking for a specific control, they receive a list of the people willing to share. That person then calls someone on the list, asks for the control, and the person with the control sends it out. Below is the NSH control bank person, if I remember correctly. Quality Control Ethel Macrea Ventana Medical Systems 1910 E. Innovation Park Drive Tucson, AZ 85739 800-227-2155 ext. 3856 emacrea@ventanamed.com Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI ----- Original Message ----- From: "Richard Cartun" To: Sent: Thursday, August 05, 2004 7:39 PM Subject: [Histonet] H. pylori control tissue > Is anyone looking for H. pylori control tissue? We just had a partial > resection of stomach and it is loaded with bugs. If there is an > overwhelming response, I may just send control blocks to the NSH tissue > bank for distribution. > > Richard > > Richard W. Cartun, Ph.D. > Director, Immunopathology & Histology > Hartford Hospital > Hartford, CT 06102 > (860) 545-1596 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Fri, 06 Aug 2004 05:34:20 -0400 From: Myri37@aol.com Subject: [Histonet] Fixative for FACS To: histonet@pathology.swmed.edu Message-ID: <69986EA4.0E45DDBE.0005167B@aol.com> Content-Type: text/plain; charset=iso-8859-1 hello Do you know which fixative can i use for intracellular immunostaining for FACS ? We use saponine 0.1 % for permeabilisation. Thank you for your help Myriam Natural implant ------------------------------ Message: 22 Date: Fri, 6 Aug 2004 06:11:22 -0500 From: "Barnhart, Tammy" Subject: [Histonet] Oncocytoma vs. Chromophobe To: "Histonet (E-mail)" Message-ID: <1779904B5E82D511914C00D0B793339205BFD830@exchangent> Content-Type: text/plain; charset="iso-8859-1" I was wondering if there is(are) any immunohistochemical antibodies or special stains besides Hale's Colloidal Iron for the differentiation of Oncocytoma and Chromophobe in kidney? I am not able to make the dialysised (sp?) iron needed for the Hale's but would like to offer my pathologist an alternate. Any suggestions? Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. ------------------------------ Message: 23 Date: Fri, 06 Aug 2004 08:25:17 -0400 From: rschoon Subject: Re: [Histonet] Problem with Masson's Trichrome To: Bdeer24@aol.com Cc: histonet@lists.utsouthwestern.edu Message-ID: <411378AD.8020806@email.unc.edu> Content-Type: text/plain; charset=us-ascii; format=flowed Amber, Don't know where you got your protocol but I would suggest ANY of the standerd Histology books. As for an answer to your problems. 1- mordent in either Boin's fluid or saturated picric acid for 60 min. at 60oC (there are other time, temp.'s and concentrations if one looks them up). The method simply will not work without the modent. 2- use Weigert's Iron Hematoxylin as the nuclear stain. Robert Schoonhoven Bdeer24@aol.com wrote: >Hi Everyone! > >I was recommended to try the Masson's Trichrome stainings on the slides of uterine tissue that I have been collecting (Previously I had only performed Hematoxylin & Eosin stains). This is a new method to our lab and thus far, staining has not gone well. Although my sections contain a significant amount of muscle, I have only been able to see the aniline blue stain. On a couple of the slides, there was a slight dark purple coloration on the epithelial lining. > >I have noticed upon comparing the protocol we received to others found on the internet that ours lacks the step in which the slides are placed in Bouin's fluid. There was also no specification in our protocol for the type of hematoxylin, so we had been using the same hematoxylin (Harris's modified with acetic acid) that we use in our H & E stainings. I was wondering if any of these might have led to the stainings not going as planned. The tissue sections also looked slightly brittle, perhaps due to a harsher stain. I am wondering if this has anything to do with my fixation process in which I use a 4% paraformaldehyde/PBS mixture, ethanol and xylene steps, as well as embedding the tissues in paraffin. > >Is Bouin's a crucial step, would I have better staining with a different tissue fixation, or should I be looking in a different direction? Any suggestions would be very much appreciated! >Thanks! > >Amber Greenbank > >Arizona State University > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 24 Date: Fri, 06 Aug 2004 15:08:47 +0200 From: Martin Hasselblatt Subject: [Histonet] Cytochrome oxidase (COX) staining: non-carcinogenic alternative to DAB? To: histonet@lists.utsouthwestern.edu Message-ID: <411382DF.4030602@uni-muenster.de> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Dear Histonet members, staining of snap frozen muscle tissue for cytochrome oxidase (COX) using diaminobenzidine (DAB) works nicely in our laboratory. We would be happy to know, however, if anybody is aware of an non-carcinogenic alternative method to visualize cytochrome oxidase activity. Thank you very much for your help Martin Hasselblatt Martin Hasselblatt Institute of Neuropathology University Hospital M?nster, Germany http://www.klinikum.uni-muenster.de/institute/npatho/mitarbeiter/hasselblatt/index.html ------------------------------ Message: 25 Date: Fri, 6 Aug 2004 09:15:39 -0400 From: "Hermina Borgerink" Subject: [Histonet] Masson Trichrome To: , Message-ID: <9AEEF1FB6254224AA355ED285F8491650999C853@EXCHVS2.medctr.ad.wfubmc.edu> Content-Type: text/plain; charset="US-ASCII" Amber, Below is a modified version of the original Masson's Trichrome which I have successfully used for over 40 years. Instead of using Woodstain-Scarlet, which I don't think is available anymore, substitute Crocein Scarlet which Sigma sells. Masson's Trichrome (Modified) Post-fix in Bouin's fixative overnight if primary fixation was not done using Bouin's 1. Bring sections to water and wash for 5 - 10 minutes. 2. Stain nuclei for 15 minutes in Weigert's iron hematoxylin 3. Wash for 5 - 10 minutes in running water and rinse in 1% acetic acid. 4. Place slides in Woodstain Scarlet-Acid Fuchsin mixture for 5 minutes. 5. Rinse briefly in 1% acetic acid and differentiate in 5% phosphomolybdic acid. Differentiate to the point where collagen and ground substances are pale pink to colorless, usually about 30 seconds to 2 minutes. 6. Rinse in 1% acetic acid. 7. Place for 15 - 20 seconds in the Aniline Blue solution (Aniline Blue - Woodstain Scarlet - Acid Fuchsin mixture). 8. Place for a quick rinse in 1% acetic acid. 9. Dehydrate in four changes of alcohol. 10. Clear in xylene and mount in permount. Preparation of Staining Solutions: Weigert's Hematoxylin: Solution A: 29% ferric chloride - 4 ml Distilled water - 95 ml Conc. HCl - 1 ml Solution B: Hematoxylin - 1 gm 95% ethanol - 100 ml For use, mix equal parts of solutions A and B. Use fresh each time Woodstain Scarlet-Acid Fuchsin: prepare a 1% solution of Woodstain Scarlet NS and a 1% solution of Acid Fuchsin (C.I. no 692). Mix 8 parts of Woodstain Scarlet solution with 2 parts of Acid Fuchsin solution and add 1ml acetic acid to 99ml of the mixed dye solution. Aniline blue: To 85ml of 1% acetic acid add 5ml of a saturated solution of Aniline Blue (C.I. no 707) and 10ml of the above Woodstain Scarlet-Acid Fuchsin mixture. Hermina M. Borgerink, BA, HTL(ASCP)QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax (336) 716-1515 e-mail hborgeri@wfubmc.edu ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 9, Issue 8 ************************************** From JWEEMS <@t> sjha.org Mon Aug 9 21:45:53 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Problem with VIP 5 Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0B8D7E@sjhaexc02.sjha.org> Don't be scared. We had this problem once. The formalin was where the xylene should have been. It'll do it every time. (Check that first!) Good luck, Joyce :>) ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ian Sent: Mon 8/9/2004 10:02 PM To: 'Joe Nocito'; 'Jill Songer'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Problem with VIP 5 Hey, you all don't scare me WE just purchase a new VIP 5 and have not have used it yet! Hey Joe, how are you doing? SSgt Bernard 59th Clinical Research Squadron "The USAF's largest Clinical Research facility" Lackland AFB, Texas -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Monday, August 09, 2004 3:27 PM To: Jill Songer; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Problem with VIP 5 Jill, how long have you had the VIP 5? I've had my machine almost 3 years and haven't had that problem. If you don't close the lid properly, when it pumps in fluid, the lid pops off, but we haven't had any smells. Please let us know what the service rep says. I always like new stories. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Jill Songer" To: Sent: Monday, August 09, 2004 2:40 PM Subject: [Histonet] Problem with VIP 5 > We are having a problem with our VIP 5 having a strong formalin odor at the > end of the run when we open the retort to remove the tissues. There is also > a large amount of fluid-formalin?? on the lid and in the chamber. > > Has anyone else experienced this? I have called Sakura and they are sending > a field service tech but I was curious if anyone has had this problem. > Whenever I call a vendor it seems they always say, well, we haven't had > THIS problem before. > > Thanks. > > Jill Songer HT (ASCP) > Histopathology Lab > Virginia-Maryland Regional College of Veterinary Medicine > Teaching Hospital > Virginia Tech > Blacksburg, VA 24061-0443 > www.vetmed.vt.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From arme <@t> optonline.net Tue Aug 10 00:19:26 2004 From: arme <@t> optonline.net (Mark Sofferman) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] for sale - tissue processor Shandon Pathcentre In-Reply-To: <0I2700B7QKWBMB@mta12.srv.hcvlny.cv.net> Message-ID: One tissue processor is available and can be delivered refurbished and warrantied. This is a current unit high volume applications. Unit will be about half of new cost. Serious inquiries only please. ARME Tel. 201-833-1550 From arvind <@t> nbrc.res.in Tue Aug 10 02:41:08 2004 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] job querry Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A6B3@mail.nbrc.res.in> re3r43rf From JColCLEFA <@t> aol.com Tue Aug 10 03:24:49 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] IHC artefact puzzle Message-ID: <75.30829084.2e49e051@aol.com> If you are using heat retrieval with slides in a vertical rack and you remove the rack from the hot retrieval solution , the solution will evaporate from the edges of the section first, drying the tissue edges and ruining the reactivity on the tissue edges. If you leave the racks in the hot solution vessel after the time is up for retrieval and cool your retrieval solution, racks and all, by sitting them in a cool water bath without removing them from the solution until cool (less than 40C) the evaporation will be slowed, the edges will not dry out, and antigenicity should be preserved. JPJC HT,QIHC From AFeatherstone <@t> KaleidaHealth.Org Tue Aug 10 05:43:41 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] 2N HCL Message-ID: 1N HCL contains 85ml concentrated solution per liter. The AFIP shows how to calculate normal solutions in Laboratory Calculations. Annette Featherstone HT/MLT -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Monday, August 09, 2004 18:53 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] 2N HCL Can anyone tell me how to make a 2N solution of HCL from concentrated HCL? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From Sarah.Blachford <@t> ngh.nhs.uk Tue Aug 10 06:55:17 2004 From: Sarah.Blachford <@t> ngh.nhs.uk (Blachford, Sarah - Histopath Main) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] blistering skin condotions Message-ID: <854FA06D8BE2D611ABA30030052D1DF6FA58F1@NGH_EXCHANGE1> Dear All, I am in the process of setting up a reliable technique(s) to demonstrate blistering skin conditions. The gold standard is to use flourescence. What method do people use, and do you use conjugated FITC antibodies or separate antibodies and fluorochromes. Is any body using tTgase as part of their panel along with the normal IG's, C3 Thanks Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. From Dorothy.L.Webb <@t> HealthPartners.Com Tue Aug 10 06:52:41 2004 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] gram iodine Message-ID: To the person who asked about commercially prepared Gram's iodine, we use the Gram stain kit from Sigma and you can also purchase the iodine separately from them. If you do not have access to the catalogue, I can give you the order numbers. ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From Tony.Crick <@t> rli.mbht.nhs.uk Tue Aug 10 07:04:32 2004 From: Tony.Crick <@t> rli.mbht.nhs.uk (Crick Tony) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] blistering skin condotions Message-ID: Hi Sarah We use a direct method, with FITC conjugated IgG, IgA, IgM etc. It is a simples yet quick and effective method with results available within 1hr. Tony Crick Morecambe Bay Hospitals -----Original Message----- From: Blachford, Sarah - Histopath Main [mailto:Sarah.Blachford@ngh.nhs.uk] Sent: 10 August 2004 12:55 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] blistering skin condotions Dear All, I am in the process of setting up a reliable technique(s) to demonstrate blistering skin conditions. The gold standard is to use flourescence. What method do people use, and do you use conjugated FITC antibodies or separate antibodies and fluorochromes. Is any body using tTgase as part of their panel along with the normal IG's, C3 Thanks Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From JNocito <@t> Pathreflab.com Tue Aug 10 07:22:42 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Thanks to everyone's response to all my sillyquestions... In-Reply-To: <9f.4b26c70c.2e494750@aol.com> Message-ID: Jennell, The only silly question is one that is not asked. We all have different levels of experiences, different problems we've encountered, etc. The time to get out of this job is when you can't learn anything any more. I've been in histology over 25 years and I'm still learning. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of JCarpenter764@aol.com Sent: Monday, August 09, 2004 4:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Thanks to everyone's response to all my sillyquestions... To All on the Histonet Mailing List: Thank you to everyone that has given me great responses to ALL of my questions...I don't know where I would be without all of you...Love Ya All Jennell _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cmrgalle <@t> usc.es Tue Aug 10 07:34:23 2004 From: cmrgalle <@t> usc.es (cmrgalle@usc.es) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] HCl 2N Message-ID: <1092141263.4118c0cf7c6e8@correoweb.usc.es> 16.7 cc of commercial HCl in 100 cc distilled water (McManus book of histological techniques) From frouwke <@t> sci.kun.nl Tue Aug 10 07:42:20 2004 From: frouwke <@t> sci.kun.nl (Frouwke Kuijpers) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Some questions about cryosectioning Message-ID: <004201c47ed7$7bf775a0$4a8aae83@sci.kun.nl> Hallo Everyone I have some questions about cryosectioning. We use the following protocol for brain tissue of xenopus (a kind of frog) a.. Bouinn fixation by perfusion, one night postfixation b.. Wash in alc 70%, washing away picricacid c.. sucrose d.. section and mount on glass. e.. immunostaining Just a normal routine, I think. I am used to do everything in one week, I begin with the perfusion and at the end of the week I have my sections stained. Now the question: because I have to do many tissue's I cannot handle it in one week. The immunostaining has to be done for all tissue's at the same time etc.. a.. What is the best step to leave the tissue in for a while? Bouinn, alc 70%, sucrose, and still have good immunostainings and good sectioning. What is for each different step the maximum time, does it mather anyway? b.. Already mounted on the slides , how long can you leave them (RT or 4C) on the slide and still have good immunoreactivity. Thanks Frouwke F.J.Kuijpers-Kwant Dept. Cellular Animal Physiology University of Nijmegen Toernooiveld 1 6525 ED Nijmegen the Netherlands frouwke@sci.kun.nl From cdan1204 <@t> yahoo.com.cn Tue Aug 10 08:09:39 2004 From: cdan1204 <@t> yahoo.com.cn (=?gb2312?q?d=20c?=) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] storage issues of serum and tissue Message-ID: <20040810130939.9280.qmail@web15606.mail.cnb.yahoo.com> Dear histoneter, Since stored samples may be very fragile and degrade over time,reliable storage conditions of serum and tissue are important.Could you provide any guideline about it? Thanks in advance. danc Jinling Hospital Nanjing , China --------------------------------- Do You Yahoo!? 150ÍňÇúMP3ˇčżńËŃŁŹ´řÄú´łČëŇôŔÖľîĚĂ ĂŔĹŽĂ÷ĐÇÓŚÓĐžĄÓĐŁŹËŃąéĂŔÍźĄ˘ŃŢÍźşÍżáÍź 1GžÍĘÇ1000ŐףŹŃĹť˘ľçÓĘ×ÔÖúŔŠČÝŁĄ From muololi <@t> umdnj.edu Tue Aug 10 08:39:32 2004 From: muololi <@t> umdnj.edu (Lilia Muolo) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] ASCP exam...anyone there? Message-ID: Hi, in our lab only one person rec'd their paperwork on Saturday. So hopefully the rest of the paperwork is on the way. Lil Cancer Institute of New Jersey >>> "Ant S." 8/9/2004 6:35:35 PM >>> I have not yet recieved even a notice that I am qualified to take the exam, much less the practical exam information. Has anyone else at least gotten a letter saying --thank you for applying, here's more information?-- Antoinette Swensson Univeristy of Washington/Harborview Medical Center Neuropathology 325 9th Ave MS 359-791 Seattle, WA 98104 (206) 731-3910 _________________________________________________________________ [1]Check out Election 2004 for up-to-date election news, plus voter tools and more! References 1. http://g.msn.com/8HMBENUS/2746??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Samuel.Jones2 <@t> med.va.gov Tue Aug 10 08:52:36 2004 From: Samuel.Jones2 <@t> med.va.gov (Jones, Samuel) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Dallas Job (2 Positions) Message-ID: <18EE321ED15F364D89A880C2509F2088D984A5@VHANTXEXC1> <<...OLE_Obj...>> Department of Veterans Affairs VACANCY ANNOUNCEMENT STATION 549 ADDRESS (City, State and ZIP Code) VA North Texas Health Care System 4500 South Lancaster Road Dallas, Texas 75216 AREA OF PROMOTION CONSIDERATION NATIONWIDE ANNOUNCEMENT NUMBER 04-B14-221 POSITION TITLE HISTOPATHOLOGY TECHNICIAN SERIES AND GRADE Pay Plan/Series: GS-646 Grade: 8 NO. OF POSITIONS 1 PROMOTION POTENTIAL TO N/A OPENING DATE 8/9/04 LOCATION VANTHCS Pathology & Laboratory Medicine Dallas, TX SALARY RANGE $37,600- $48,881 CLOSING DATE 8/27/04 FOR INFORMATION CONTACT Donna Robinson HR Specialist 214-857-1681 HOURS OF WORK 6:00am to 2:30pm Monday - Friday NOTE: This announcement is a solicitation for applications from current VA employees for competitive promotion consideration. It does not, however, restrict the right to consider or select applicants from any other recruitment source such as reassignment, appointment, demotion, transfer, reinstatement or special appointing authorities such as those for disabled veterans, veterans recruitment appointment (VRA) eligibles, severely handicapped individuals, etc. NOTE: Current permanent Veterans Canteen Service employees may apply for consideration under this vacancy announcement. NOTE: If you are selected for this position, you may be required to undergo a physical examination if the physical requirements are different from the position you now occupy. NOTE: Performance Based Interviewing (PBI) may be used. For information on the basics of PBI visit the PBI website at www.va.gov/pbi . EQUAL EMPLOYMENT OPPORTUNITY: All applicants will receive consideration regardless of race, color, age, religion, sex, national origin, political affiliation, sexual orientation, marital status, status as a parent, and non-disqualifying physical handicap. DRUG TESTING: All applicants tentatively selected for VA employment (in a testing designated position) are subject to urinalysis to screen for illegal drug use prior to appointment. Applicants who refuse to be tested will be denied employment with the VA. RESPONSIBILITIES: The position is in the Histology Unit of the Anatomic Pathology Section, Pathology and Laboratory Medicine Service. Incumbent's duties consist of receiving specimens, and accessioning specimen and patient data into histology logbook. Processing tissues, using automatic tissue processing equipment; orientation, positioning, and embedding of tissue samples using paraffin or other less commonly employed embedding media; sectioning of blocks of paraffin-embedded tissues; staining of tissue sections with routine stains; immunohistochemical staining. Maintaining accuracy of tissue/patient identification throughout the special handling procedures. Using independent judgement and close interaction with the supervisor, pathologists and residents to ensure that the final product is of appropriate quality. Incumbent properly stores and safely handles all chemicals. Using safely and appropriately a variety of instruments including, but not restricted to, analytical balances, pH meters, automatic tissue processors, microtomes, automatic stainer, cryostats, automatic coverslipper, photographic equipment, conventional light microscopes, fluorescence microscopes, immuno stainer, cytospins, and various computer-assisted work stations. QUALIFICATION REQUIREMENTS: Must be serving in a permanent Title 5 appointment. In addition to meeting the Basic Requirements outlined in the Office of Personnel Management (OPM) Qualification Standards Operating Manual, applicants must meet the following experience requirements: EXPERIENCE: Applicants must demonstrate 1 year of specialized experience which is directly related to the position to be filled and which has equipped the candidate with the particular knowledge, skills and abilities to successfully perform the duties of the position. To be credible, specialized experience must have been at least equivalent to the next lower level. SPECIALIZED EXPERIENCE: Experience which has equipped the applicant with the particular knowledge, skills, and abilities to perform successfully the duties of the position, and that is in Anatomical pathology technician work in: A. Cutting and staining very thin sections of human tissue specimens for microscopic examination. B. Testing and examining body fluids, etc., for abnormalities in cell structure. To be creditable, this experience must be comparable to the next lowest level positions in Federal service. OR EDUCATION: One and one-half (1 1/2) years of graduate education, which is directly related to the work of the position meets the requirements for a GS-8. TIME-IN-GRADE RESTRICTIONS: Applicants must have completed at least 52 weeks of service at the GS-6 level. RATING FACTORS FOR HISTOPATHOLOGY TECHNICIAN, GS-646-8 For each of the following knowledge, skills, and/or abilities, please indicate your major accomplishments as they relate to the above position. Since your rating will be based primarily on your responses, please be specific for actual duties performed or education received. Each item will be evaluated separately, so please do not provide one narrative summary covering all items. 1. Ability to operate, calibrate, perform preventive maintenance, and troubleshoot a wide variety of histology laboratory instruments. 2. Knowledge of tissue processing techniques. 3. Skill in orienting and positioning minute and delicate tissue specimens. 4. Ability to operate and maintain a micro tome and cryostat. 5. Ability to work independently and to complete tasks within time constraints. NOTE: Questions concerning this announcement may be referred to Donna Robinson, Human Resources Specialist, at 214-857-1681. Applicants for the positions with direct patient care must meet language proficiency as required by Public Law 95-201. HOW TO APPLY: Applicants must submit VAF 5-4078, Application for Promotion or Reassignment by 4:30 pm on 8/27/04, and VAF 5-4676a, Employee Supplemental Qualifications Statement, and VAF 5-4667b, Supervisory Appraisal of Employee for Promotion by 4:30 pm on 9/3/04, to Human Resources Management Service, Room A-133. Forms can be obtained in the Human Resources Management Service or on VANTHCS home page under local job announcements. UPDATING OF OPF: All employees are reminded of their responsibility to insure that all pertinent experience, education, training, and outside activities are included in their Official Personnel Folders (OPF). If an amendment is necessary for this particular position, use SF-172, Amendment to Application for Federal Employment, and submit no later than the closing date of this announcement. DISTRIBUTION: "B Samuel E. Jones, MS, HT (ASCP) HTL, QIHC Supervisor Anatomic Pathology VA North Texas Healthcare System 4500 South Lancaster Road 113 Dallas, Texas 75216 Phone: 214-857-0659 Fax: 214-302-1457 E-mail: samuel.jones2@med.va.gov From jfountain <@t> enviroguard.com Tue Aug 10 09:29:34 2004 From: jfountain <@t> enviroguard.com (Jody Fountain) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] RE: Histonet Digest, Vol 9, Issue 13 Message-ID: Questions on new product we are looking at. HISTO-CLEAR, Xylene substitute. 1. Who has used it and how effective is it? Seems to good to be true. 2. Dangers & reactions with using this product? MSDA seems too vague. 3. Monitoring of vapors for co-workers, what are you using? Appreciate your feedback, J Fountain, Sensors Analytical Lab jfountain@enviroguard.com --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.729 / Virus Database: 484 - Release Date: 7/27/2004 From escott8 <@t> houston.rr.com Tue Aug 10 10:03:24 2004 From: escott8 <@t> houston.rr.com (Edward Scott) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Pass Through Refrigerator Message-ID: <000601c47eeb$2f2959f0$042a1942@thescotts> I have a pass through refrigerator that is between the OR and our Gross room. The OR people put the specimens in there during the day and at night. It is double sided where you can reach through it from either side. It is 14 years old and is showing signs of wear. I have contacted every vendor that we do business with and then some. No one seems to know where we can find one. The company that the hospital purchased it from is no longer in business, and our course this took place before I became the supervisor. We ordered one from Jewitt, and it was the wrong one , even though they had the dimensions that we requested. It wound up going to blood bank. If any one has any suggestions of a staring point please let me know. Thanks in advance. Allison Scott HT(ASCP) Histology Supervisor LBJ Hospital Houston, Texas 77026 From lance.erickson <@t> ihc.com Tue Aug 10 10:10:19 2004 From: lance.erickson <@t> ihc.com (Lance Erickson) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Problem with VIP 5 Message-ID: Jill, We have had this same problem. The fix we have come up with is that too much fluid is condensing on the lid and making it's way through the whole processing cycle. The retort lid is not insulated at all and solutions will condense on the lid inside the retort. Our solution was to place a towel on top of the processor to insulate the lid. This seemed to solve the problem. We are custom making a cover to fit the lid that should work better than the towel. Try it and see if it works. Let me know if it was the same problem. Lance Erickson Anatomic Pathology Supervisor Primary Children's Medical Center Salt Lake City, UT >>> "Joe Nocito" 08/09/04 04:26PM >>> Jill, how long have you had the VIP 5? I've had my machine almost 3 years and haven't had that problem. If you don't close the lid properly, when it pumps in fluid, the lid pops off, but we haven't had any smells. Please let us know what the service rep says. I always like new stories. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Jill Songer" To: Sent: Monday, August 09, 2004 2:40 PM Subject: [Histonet] Problem with VIP 5 > We are having a problem with our VIP 5 having a strong formalin odor at the > end of the run when we open the retort to remove the tissues. There is also > a large amount of fluid-formalin?? on the lid and in the chamber. > > Has anyone else experienced this? I have called Sakura and they are sending > a field service tech but I was curious if anyone has had this problem. > Whenever I call a vendor it seems they always say, well, we haven't had > THIS problem before. > > Thanks. > > Jill Songer HT (ASCP) > Histopathology Lab > Virginia-Maryland Regional College of Veterinary Medicine > Teaching Hospital > Virginia Tech > Blacksburg, VA 24061-0443 > www.vetmed.vt.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Tue Aug 10 10:29:46 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Ventana Discovery units for sale? Message-ID: A while back (could be 6 months ago or so?) somebody posted that they were selling their Ventana Discovery units. I believe they had 5 of the staining modules available. Are these still available, and if so would you please contact me via email? Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From abright <@t> brightinstruments.com Tue Aug 10 10:45:45 2004 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Pass Through Refrigerator Message-ID: Dear Allison, I cannot see that you would have a problem to replace your Pass Through Refrigerator, any Cold room or refrigerated display cabinet manufacturer should be able to make this for you to the specifications you require without a problem. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Edward Scott [mailto:escott8@houston.rr.com] Sent: 10 August 2004 16:03 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pass Through Refrigerator I have a pass through refrigerator that is between the OR and our Gross room. The OR people put the specimens in there during the day and at night. It is double sided where you can reach through it from either side. It is 14 years old and is showing signs of wear. I have contacted every vendor that we do business with and then some. No one seems to know where we can find one. The company that the hospital purchased it from is no longer in business, and our course this took place before I became the supervisor. We ordered one from Jewitt, and it was the wrong one , even though they had the dimensions that we requested. It wound up going to blood bank. If any one has any suggestions of a staring point please let me know. Thanks in advance. Allison Scott HT(ASCP) Histology Supervisor LBJ Hospital Houston, Texas 77026 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Aug 10 11:02:17 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Some questions about cryosectioning In-Reply-To: <004201c47ed7$7bf775a0$4a8aae83@sci.kun.nl> References: <004201c47ed7$7bf775a0$4a8aae83@sci.kun.nl> Message-ID: <6.0.0.22.1.20040810095017.01b585b0@gemini.msu.montana.edu> I would not want to overfix in Bouins, do not change that step - to keep your fixation optimal. Getting rid of Bouins in 70% alcohol puts your tissue in an 'antifreeze' state with the alcohol (any alcohol in tissue can intefer with proper snap freezing) so the sucrose cryoprotection step should probably be done with at least one or two changes to insure the NO alcohol is in the tissue at snap freezing time, basically you are rehydrating with sucrose in aqueous solution. If you dissolve the sucrose in PBS, and keep tissues in refrigerator, you may get away with longer storage - however, snap frozen tissues in the block are probably the most stable way to store your tissues. Just store your frozen blocks in a -80C freezer, inside a tube or ziplock plastic food bag until cryosectioning time. Sections should probably be air dried and stored in a -80C freezer with dessicant (16 mesh silica gel in a bag) and brought to RT in an enclosed box for 30 min prior to immunostaining. Protect your antigen as much as possible. It will depend on the stability of antigen whether you can store them at RT before IHC, this is also true of other storage temperatures. You may have to determine that or someone may know the answer already. We NEVER store frozen sections at 4C, ideally -80C, or -27C for a shorter time. One thing we do avoid is any water condensation that may form on a section going from cold to warm temperature, also never store the FS inside a cryostat. OR better yet, do the immunostaining as soon as possible after sectioning. At 06:42 AM 8/10/2004, you wrote: >Hallo Everyone > >I have some questions about cryosectioning. >We use the following protocol for brain tissue of xenopus (a kind of frog) > > a.. Bouinn fixation by perfusion, one night postfixation > b.. Wash in alc 70%, washing away picricacid > c.. sucrose > d.. section and mount on glass. > e.. immunostaining >Just a normal routine, I think. I am used to do everything in one week, I >begin with the perfusion and at the end of the week I have my sections >stained. >Now the question: because I have to do many tissue's I cannot handle it >in one week. The immunostaining has to be done for all tissue's at the >same time etc.. > a.. What is the best step to leave the tissue in for a while? > Bouinn, alc 70%, sucrose, and still have good immunostainings and good > sectioning. > What is for each different step the maximum time, does it mather anyway? > b.. Already mounted on the slides , how long can you leave them (RT or > 4C) on the slide and still have good immunoreactivity. >Thanks Frouwke > >F.J.Kuijpers-Kwant >Dept. Cellular Animal Physiology >University of Nijmegen >Toernooiveld 1 >6525 ED Nijmegen >the Netherlands >frouwke@sci.kun.nl >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From marytedo <@t> hotmail.com Tue Aug 10 11:04:37 2004 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] To George Cole Message-ID: If george Cole or someone knows him, please . I want to give him thanks! Because I have received the parcel he sent me two months ago. About muscle and nerve biopsies rutine. I apreciate it very much. HT. Maria Teresa Dominguez Pathology Service, Río Grande's Regional Hospital Tierra del Fuego, Argentina. _________________________________________________________________ STOP MORE SPAM with [1]the new MSN 8 and get 2 months FREE* References 1. http://g.msn.com/8HMAEN/2728??PS=47575 From diane <@t> discoverysolution.com Tue Aug 10 12:10:18 2004 From: diane <@t> discoverysolution.com (Discovery Solutions) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Immunohistochemical Research Position Opening in NY In-Reply-To: Message-ID: <001801c47efc$ee3ab620$6401a8c0@DISCOVERYWORK> Hi, I don't mean to interrupt but I have a histology position open in NY if anyone is interested. Here are the particulars: This opportunity based in New York, NY is with a well established biopharmaceutical company focused on developing chimerized (made from mouse and human antibodies) cancer therapies. With successful products on the market and more in the pipeline they offer growth potential and great benefits. We are currently seeking a research scientist with expertise in immunohistochemical techniques to run a program testing the ability of antibodies to be used as immunohistochemical staining reagents in tissues from a variety of species. In addition this individual will perform general staining technique development for a variety of cancer related proteins. You would be responsible for screening antibodies for binding to tissues from multiple species, developing and utilizing immunohistochemical staining techniques for various cancer related proteins, harvesting mouse tissue samples and other basic histology research duties. The minimum requirements for this position are a B.S. degree plus at least 5 years experience in an active histology lab. We are looking for someone proficient at frozen and paraffin sectioning and immunohistochemistry with experience in all aspects of processing patient tissue samples for immunohistochemistry. Plus experience harvesting and processing rodent tissues for immunohistochemistry is required. Also a history of published immunohistochemical work is a plus. Give me a call if you or anyone you know would be interested in learning more about this. Thanks, Diane K. Ranney Discovery Solutions Specializing in Drug Discovery Searches 410-838-4557 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, August 10, 2004 1:09 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 9, Issue 14 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." From funderwood <@t> mcohio.org Tue Aug 10 12:57:37 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:23:52 2005 Subject: [BULK] - RE: [Histonet] Problem with VIP 5 Message-ID: I am looking into a new processor this next year. I thought that the older VIPs were good units. However, reading about the problems people are having with the VIP 5, I'm having doubts. I'm curious if those of you having problems were given a mulligan, would you still chose a VIP or something else? Fred >>> "Ian" 08/09/04 10:02PM >>> Hey, you all don't scare me WE just purchase a new VIP 5 and have not have used it yet! Hey Joe, how are you doing? SSgt Bernard 59th Clinical Research Squadron "The USAF's largest Clinical Research facility" Lackland AFB, Texas -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Monday, August 09, 2004 3:27 PM To: Jill Songer; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Problem with VIP 5 Jill, how long have you had the VIP 5? I've had my machine almost 3 years and haven't had that problem. If you don't close the lid properly, when it pumps in fluid, the lid pops off, but we haven't had any smells. Please let us know what the service rep says. I always like new stories. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Jill Songer" To: Sent: Monday, August 09, 2004 2:40 PM Subject: [Histonet] Problem with VIP 5 > We are having a problem with our VIP 5 having a strong formalin odor at the > end of the run when we open the retort to remove the tissues. There is also > a large amount of fluid-formalin?? on the lid and in the chamber. > > Has anyone else experienced this? I have called Sakura and they are sending > a field service tech but I was curious if anyone has had this problem. > Whenever I call a vendor it seems they always say, well, we haven't had > THIS problem before. > > Thanks. > > Jill Songer HT (ASCP) > Histopathology Lab > Virginia-Maryland Regional College of Veterinary Medicine > Teaching Hospital > Virginia Tech > Blacksburg, VA 24061-0443 > www.vetmed.vt.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JPCOLEMA <@t> sentara.com Tue Aug 10 13:06:12 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Skin blisters Message-ID: We use diluted concentrates of FITC conjugated antibodies, direct procedure, on michel's fixed skins. IgG, IgA, IgM, C3c, C4c, fibrinogen. From michael_keysock <@t> merck.com Tue Aug 10 13:37:37 2004 From: michael_keysock <@t> merck.com (Keysock, Michael A) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] (no subject) Message-ID: I am sending out an inquiry about Sakura Finetek Coverslipping Mounting Medium. Our lab has chosen the Sakura Finetek Glas Coverslipper, and our current mounting medium, Fisher brand Permount, is not compatible with it because it is Toluene based. We were looking into using the Sakura brand Mounting Medium, but do not have any background on it. I am wondering if anyone has any experience with this mounting medium. Does the mounting medium give you air bubbles; if you archive slides, does it last of an extended amount of time; any other information someone may have on it. Thank you. Michael A. Keysock Staff Biologist Merck Research Labs (215) 652-9397 michael_keysock@merck.com h ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From Jackie.O'Connor <@t> abbott.com Tue Aug 10 13:50:17 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:52 2005 Subject: [BULK] - RE: [Histonet] Problem with VIP 5 Message-ID: I don't know - - what's a mulligan? Is it something like a henway? Jackie O' "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/10/2004 12:57 PM To: cc: Subject: [BULK] - RE: [Histonet] Problem with VIP 5 I am looking into a new processor this next year. I thought that the older VIPs were good units. However, reading about the problems people are having with the VIP 5, I'm having doubts. I'm curious if those of you having problems were given a mulligan, would you still chose a VIP or something else? Fred >>> "Ian" 08/09/04 10:02PM >>> Hey, you all don't scare me WE just purchase a new VIP 5 and have not have used it yet! Hey Joe, how are you doing? SSgt Bernard 59th Clinical Research Squadron "The USAF's largest Clinical Research facility" Lackland AFB, Texas -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Monday, August 09, 2004 3:27 PM To: Jill Songer; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Problem with VIP 5 Jill, how long have you had the VIP 5? I've had my machine almost 3 years and haven't had that problem. If you don't close the lid properly, when it pumps in fluid, the lid pops off, but we haven't had any smells. Please let us know what the service rep says. I always like new stories. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Jill Songer" To: Sent: Monday, August 09, 2004 2:40 PM Subject: [Histonet] Problem with VIP 5 > We are having a problem with our VIP 5 having a strong formalin odor at the > end of the run when we open the retort to remove the tissues. There is also > a large amount of fluid-formalin?? on the lid and in the chamber. > > Has anyone else experienced this? I have called Sakura and they are sending > a field service tech but I was curious if anyone has had this problem. > Whenever I call a vendor it seems they always say, well, we haven't had > THIS problem before. > > Thanks. > > Jill Songer HT (ASCP) > Histopathology Lab > Virginia-Maryland Regional College of Veterinary Medicine > Teaching Hospital > Virginia Tech > Blacksburg, VA 24061-0443 > www.vetmed.vt.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lfaucette <@t> niaid.nih.gov Tue Aug 10 13:59:22 2004 From: lfaucette <@t> niaid.nih.gov (Faucette, Lawrence (NIH/NIAID)) Date: Fri Sep 16 15:23:52 2005 Subject: [BULK] - RE: [Histonet] Problem with VIP 5 Message-ID: <78F91CA4B8F3504E8FE5F13AAF122ED804A02CC0@nihexchange27.nih.gov> ROFL, Leave it to the USAF to speak in golf terms :) ( just kidding ) Mulligan would be a golf shot that you pretend never really happened :) Lawrence J Faucette Contractor HT ASCP http://www.niaid.nih.gov/dir/services/animalcare/VetPathology/VetPathology-i ndex.html Infectious Disease Pathogenesis Section Comparative Medicine Branch Division of Intramural Research, NIAID, NIH Twinbrook III, Room 2W-01A, MSC 8135 12735 Twinbrook Parkway Bethesda, MD 20892-8135 Telephone 301-451-1056 Fax 301-480-2343 SoBran, Inc. Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. The National Institute of Allergy and Infectious Diseases (NIAID) shall not accept liability for any statement made that are the sender's own and not expressly made on behalf of the NIAID by one of its representatives. -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Tuesday, August 10, 2004 2:50 PM To: Fred Underwood Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [BULK] - RE: [Histonet] Problem with VIP 5 I don't know - - what's a mulligan? Is it something like a henway? Jackie O' "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/10/2004 12:57 PM To: cc: Subject: [BULK] - RE: [Histonet] Problem with VIP 5 I am looking into a new processor this next year. I thought that the older VIPs were good units. However, reading about the problems people are having with the VIP 5, I'm having doubts. I'm curious if those of you having problems were given a mulligan, would you still chose a VIP or something else? Fred >>> "Ian" 08/09/04 10:02PM >>> Hey, you all don't scare me WE just purchase a new VIP 5 and have not have used it yet! Hey Joe, how are you doing? SSgt Bernard 59th Clinical Research Squadron "The USAF's largest Clinical Research facility" Lackland AFB, Texas -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Monday, August 09, 2004 3:27 PM To: Jill Songer; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Problem with VIP 5 Jill, how long have you had the VIP 5? I've had my machine almost 3 years and haven't had that problem. If you don't close the lid properly, when it pumps in fluid, the lid pops off, but we haven't had any smells. Please let us know what the service rep says. I always like new stories. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Jill Songer" To: Sent: Monday, August 09, 2004 2:40 PM Subject: [Histonet] Problem with VIP 5 > We are having a problem with our VIP 5 having a strong formalin odor at the > end of the run when we open the retort to remove the tissues. There is also > a large amount of fluid-formalin?? on the lid and in the chamber. > > Has anyone else experienced this? I have called Sakura and they are sending > a field service tech but I was curious if anyone has had this problem. > Whenever I call a vendor it seems they always say, well, we haven't had > THIS problem before. > > Thanks. > > Jill Songer HT (ASCP) > Histopathology Lab > Virginia-Maryland Regional College of Veterinary Medicine > Teaching Hospital > Virginia Tech > Blacksburg, VA 24061-0443 > www.vetmed.vt.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jtsonger <@t> vt.edu Tue Aug 10 14:08:07 2004 From: jtsonger <@t> vt.edu (Jill Songer) Date: Fri Sep 16 15:23:52 2005 Subject: [BULK] - RE: [Histonet] Problem with VIP 5 Message-ID: <6.0.0.22.0.20040810150646.01b94c00@pop.vt.edu> I don't know what a mulligan is-but, yes, if I had a do-over, I would still choose the VIP. At 02:50 PM 8/10/2004, Jackie.O'Connor@abbott.com wrote: >I don't know - - what's a mulligan? Is it something like a henway? >Jackie O' > > > > >"Fred Underwood" >Sent by: histonet-bounces@lists.utsouthwestern.edu >08/10/2004 12:57 PM > > > To: > cc: > Subject: [BULK] - RE: [Histonet] Problem with VIP 5 > > >I am looking into a new processor this next year. I thought that the >older VIPs were good units. However, reading about the problems people >are having with the VIP 5, I'm having doubts. I'm curious if those of >you having problems were given a mulligan, would you still chose a VIP >or something else? > >Fred > > >>> "Ian" 08/09/04 10:02PM >>> >Hey, you all don't scare me WE just purchase a new VIP 5 and have not >have >used it yet! > >Hey Joe, how are you doing? > >SSgt Bernard >59th Clinical Research Squadron >"The USAF's largest Clinical Research facility" >Lackland AFB, Texas > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe >Nocito >Sent: Monday, August 09, 2004 3:27 PM >To: Jill Songer; histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Problem with VIP 5 > >Jill, >how long have you had the VIP 5? I've had my machine almost 3 years >and >haven't had that problem. If you don't close the lid properly, when it >pumps >in fluid, the lid pops off, but we haven't had any smells. > Please let us know what the service rep says. I always like new >stories. > >Joe Nocito BS, HT(ASCP)QIHC >Histology Manager >Pathology Reference Lab >San Antonio, TX >----- Original Message ----- >From: "Jill Songer" >To: >Sent: Monday, August 09, 2004 2:40 PM >Subject: [Histonet] Problem with VIP 5 > > > > We are having a problem with our VIP 5 having a strong formalin odor >at >the > > end of the run when we open the retort to remove the tissues. There >is >also > > a large amount of fluid-formalin?? on the lid and in the chamber. > > > > Has anyone else experienced this? I have called Sakura and they are >sending > > a field service tech but I was curious if anyone has had this >problem. > > Whenever I call a vendor it seems they always say, well, we haven't >had > > THIS problem before. > > > > Thanks. > > > > Jill Songer HT (ASCP) > > Histopathology Lab > > Virginia-Maryland Regional College of Veterinary Medicine > > Teaching Hospital > > Virginia Tech > > Blacksburg, VA 24061-0443 > > www.vetmed.vt.edu > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Jill Songer HT (ASCP) Histopathology Lab Virginia-Maryland Regional College of Veterinary Medicine Teaching Hospital Virginia Tech Blacksburg, VA 24061-0443 www.vetmed.vt.edu From pmarcum <@t> polysciences.com Tue Aug 10 14:04:38 2004 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:23:52 2005 Subject: [BULK] - RE: [Histonet] Problem with VIP 5 In-Reply-To: Message-ID: <001301c47f0c$e283cc80$4f00a8c0@PMARCUM2K> Hi Jackie, Depends on what a henway is. It is another term for lemon or poorly preforming new car or other mechanical equipment. Pam Marcum > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of > Jackie.O'Connor@abbott.com > Sent: Tuesday, August 10, 2004 2:50 PM > To: Fred Underwood > Cc: histonet@lists.utsouthwestern.edu; > histonet-bounces@lists.utsouthwestern.edu > Subject: Re: [BULK] - RE: [Histonet] Problem with VIP 5 > > > I don't know - - what's a mulligan? Is it something like a henway? > Jackie O' > > > > > "Fred Underwood" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 08/10/2004 12:57 PM > > > To: > cc: > Subject: [BULK] - RE: [Histonet] Problem with VIP 5 > > > I am looking into a new processor this next year. I thought that the > older VIPs were good units. However, reading about the problems people > are having with the VIP 5, I'm having doubts. I'm curious if those of > you having problems were given a mulligan, would you still chose a VIP > or something else? > > Fred > > >>> "Ian" 08/09/04 10:02PM >>> > Hey, you all don't scare me WE just purchase a new VIP 5 and have not > have > used it yet! > > Hey Joe, how are you doing? > > SSgt Bernard > 59th Clinical Research Squadron > "The USAF's largest Clinical Research facility" > Lackland AFB, Texas > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe > Nocito > Sent: Monday, August 09, 2004 3:27 PM > To: Jill Songer; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Problem with VIP 5 > > Jill, > how long have you had the VIP 5? I've had my machine almost 3 years > and > haven't had that problem. If you don't close the lid properly, when it > pumps > in fluid, the lid pops off, but we haven't had any smells. > Please let us know what the service rep says. I always like new > stories. > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > ----- Original Message ----- > From: "Jill Songer" > To: > Sent: Monday, August 09, 2004 2:40 PM > Subject: [Histonet] Problem with VIP 5 > > > > We are having a problem with our VIP 5 having a strong formalin odor > at > the > > end of the run when we open the retort to remove the tissues. There > is > also > > a large amount of fluid-formalin?? on the lid and in the chamber. > > > > Has anyone else experienced this? I have called Sakura and they are > sending > > a field service tech but I was curious if anyone has had this > problem. > > Whenever I call a vendor it seems they always say, well, we haven't > had > > THIS problem before. > > > > Thanks. > > > > Jill Songer HT (ASCP) > > Histopathology Lab > > Virginia-Maryland Regional College of Veterinary Medicine > > Teaching Hospital > > Virginia Tech > > Blacksburg, VA 24061-0443 > > www.vetmed.vt.edu > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From georgecole <@t> ev1.net Tue Aug 10 14:28:54 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] fixing immuno and antibody tissues Message-ID: <000001c47f10$49a9de10$064dbad0@hppav> Dear Histotechs; Placate an old retiree and get a quizzical question out of my head: In 1974, I was assigned muscle and nerve biopsy work and immunofluorescence work on kidneys. Fixation ruined just about everything in all the procedures involved with those studies, so fixatives were OUT in all of my work. After moving to another hospital with my pathologist, I continued the muscle and nerve work but not the immunos. Over the years, news sort of trickled down that you histotechs were doing antibody work on fixed tissues. I guessed you had found some way to repair the tissues after being fixed. The Histonet has many messages sent back and forth between histotechs doing antibody work, and they always specified what fixative they used. A nd that always bothered me. It seemed like a ring-around-the-rosie to fix, then, Unfix tissues for antibody work. To up and fix the tissues, then turn around and repair SOME of the damage done to the tissues-because I thought the tissues would not come away unscathed from being bombarded with a fixative. And if it was necessary to return the fixed tissues to an unfixed-like state, why not just leave them unfixed in the first place? Does fixation do something good to the tissues, making them better for antibody studies than fresh frozen tissues? Muscle tissues are totally ruined for histochemistry, and there is no way to repair the harm done to them by fixation. It seemed like something out The Peterkin Papers: One story from that wonderful book involved a cup of tea that had too much sugar in it.The lady who wanted to drink it went up and down the lane gettjng advice as to what to do, involved every herb, spice and remedy which just kept making the tea worse. But the little old lady at the end of the lane suggested that she make a fresh cup of tea. I wonder if the little old lady at the end of the lane had been consulted in this vase, she might have laughed merrily and suggested you just not fix your tissues in the first place. Is there is some improvement in the tissues brought about by fixation? As fixation totally ruined all my work, I find it hard to believe that the tissues are going to come out unscathed from fixation. Anyway, this has been a mystery working in the back of my head for years .Can you put this matter to rest for this 76 year old? georgecole@ev1.net From akbitting <@t> geisinger.edu Tue Aug 10 14:33:57 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:23:52 2005 Subject: [BULK] - RE: [Histonet] Problem with VIP 5 Message-ID: Although I am hearing that the VIP5 has some problems. I would not steer anyone away from the older VIP models. They are true blue work-horses! >>> "Fred Underwood" 08/10/04 01:57PM >>> I am looking into a new processor this next year. I thought that the older VIPs were good units. However, reading about the problems people are having with the VIP 5, I'm having doubts. I'm curious if those of you having problems were given a mulligan, would you still chose a VIP or something else? Fred >>> "Ian" 08/09/04 10:02PM >>> Hey, you all don't scare me WE just purchase a new VIP 5 and have not have used it yet! Hey Joe, how are you doing? SSgt Bernard 59th Clinical Research Squadron "The USAF's largest Clinical Research facility" Lackland AFB, Texas -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Monday, August 09, 2004 3:27 PM To: Jill Songer; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Problem with VIP 5 Jill, how long have you had the VIP 5? I've had my machine almost 3 years and haven't had that problem. If you don't close the lid properly, when it pumps in fluid, the lid pops off, but we haven't had any smells. Please let us know what the service rep says. I always like new stories. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Jill Songer" To: Sent: Monday, August 09, 2004 2:40 PM Subject: [Histonet] Problem with VIP 5 > We are having a problem with our VIP 5 having a strong formalin odor at the > end of the run when we open the retort to remove the tissues. There is also > a large amount of fluid-formalin?? on the lid and in the chamber. > > Has anyone else experienced this? I have called Sakura and they are sending > a field service tech but I was curious if anyone has had this problem. > Whenever I call a vendor it seems they always say, well, we haven't had > THIS problem before. > > Thanks. > > Jill Songer HT (ASCP) > Histopathology Lab > Virginia-Maryland Regional College of Veterinary Medicine > Teaching Hospital > Virginia Tech > Blacksburg, VA 24061-0443 > www.vetmed.vt.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From lagehan <@t> hotmail.com Tue Aug 10 14:38:13 2004 From: lagehan <@t> hotmail.com (LORALEE GEHAN) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Position in Northern VA Message-ID: I have a position opening in Northern Virginia. It is a small private lab that does mostly skin samples and a little GYN cytology. We are looking for someone who has any experience in microtomy. The lab does all of the grossing of the tissue and also performs special stains and immunohistochemical stains. Experience in this is not necessary. We will teach. The hours are Monday thourgh Friday from 7a to 3p. Possibly could be a part time position, if that is what you are looking for. If you are interested please send me an e-mail with your resume. Or if you perfer I'll give you a fax number. Loralee Gehan, HTL Histology Supervisor _________________________________________________________________ [1]Dont just search. Find. Check out the new MSN Search! References 1. http://g.msn.com/8HMBENUS/2740??PS=47575 From Angelicola <@t> uchc.edu Tue Aug 10 15:08:55 2004 From: Angelicola <@t> uchc.edu (Angelicola,Lisa) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] F4/80 antibody Message-ID: Thanks to everyone who responded to my question concerning F4/80. It will make it much easier to work it out with all of your advice. Don't you just love the histonet!!! Lisa Angelicola From kgrobert <@t> rci.rutgers.edu Tue Aug 10 15:24:55 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Problem with VIP 5 In-Reply-To: References: Message-ID: <41192F17.9070807@rci.rutgers.edu> Our VIP 5 has the same condensation problem, but as we do not use formalin in the processor-we fix on the bench, then put them in the processor-we have not had any problems so far. But, out of curiosity, what kind of towel? I'm assuming the terrycloth kind, but how big? Thanks, Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Rutgers University Lance Erickson wrote: >Jill, >We have had this same problem. The fix we have come up with is that too >much fluid is condensing on the lid and making it's way through the >whole processing cycle. The retort lid is not insulated at all and >solutions will condense on the lid inside the retort. Our solution was >to place a towel on top of the processor to insulate the lid. This >seemed to solve the problem. We are custom making a cover to fit the lid >that should work better than the towel. Try it and see if it works. Let >me know if it was the same problem. > >Lance Erickson >Anatomic Pathology Supervisor >Primary Children's Medical Center >Salt Lake City, UT > > > >>>>"Joe Nocito" 08/09/04 04:26PM >>> >>>> >>>> >Jill, >how long have you had the VIP 5? I've had my machine almost 3 years >and >haven't had that problem. If you don't close the lid properly, when it >pumps >in fluid, the lid pops off, but we haven't had any smells. > Please let us know what the service rep says. I always like new >stories. > >Joe Nocito BS, HT(ASCP)QIHC >Histology Manager >Pathology Reference Lab >San Antonio, TX >----- Original Message ----- >From: "Jill Songer" >To: >Sent: Monday, August 09, 2004 2:40 PM >Subject: [Histonet] Problem with VIP 5 > > > > >>We are having a problem with our VIP 5 having a strong formalin odor >> >> >at >the > > >>end of the run when we open the retort to remove the tissues. There >> >> >is >also > > >>a large amount of fluid-formalin?? on the lid and in the chamber. >> >>Has anyone else experienced this? I have called Sakura and they are >> >> >sending > > >>a field service tech but I was curious if anyone has had this >> >> >problem. > > >>Whenever I call a vendor it seems they always say, well, we haven't >> >> >had > > >>THIS problem before. >> >>Thanks. >> >>Jill Songer HT (ASCP) >>Histopathology Lab >>Virginia-Maryland Regional College of Veterinary Medicine >>Teaching Hospital >>Virginia Tech >>Blacksburg, VA 24061-0443 >>www.vetmed.vt.edu >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From pathrm35 <@t> adelphia.net Tue Aug 10 17:03:09 2004 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Tampa,FL positions Message-ID: <20040810220309.EGCC2583.mta9.adelphia.net@mail.adelphia.net> Fellow techs, Does anyone know of any histo positions available in the Tampa, Fl area? Thanks, Ron Martin From ploykasek <@t> phenopath.com Tue Aug 10 17:54:24 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:52 2005 Subject: FW: [Histonet] fixing immuno and antibody tissues In-Reply-To: Message-ID: ------ Forwarded Message From: Patti Loykasek Date: Tue, 10 Aug 2004 15:06:23 -0700 To: George Cole Subject: Re: [Histonet] fixing immuno and antibody tissues What a wonderful viewpoint on this conundrum. I don't pretend to have all the answers, but here's what I think I know! The pathologists prefer to make diagnoses on formalin fixed tissues - it's the "gold standard" for morphology. The pathologists are familiar with the artifacts caused by routine formalin & processing. In many instances these days they are making diagnoses on needle biopsies, and there is simply not enough tissue to freeze & submit for routine processing. It is in this milieu that recovering antigens became important. First with enzymes, and more recently with heat pretreatment employing various buffers. Definitely have to be careful that you are getting actual staining & not some type of artifact caused by the heat pretreatment. Sometimes, I think we are too quick to pretreat. It?s always good to try to get an antibody signal with no pretreatment. Many of the new & more esoteric antibodies require pretreatment. When possible, it?s great to work up an antibody on both fresh & formalin fixed tissue. Well, now my brain is beat. Hope this explains at least part of the reasoning behind pretreatment. Patti Loykasek PhenoPath Laboratories Seattle, WA > Dear Histotechs; > Placate an old retiree and get a quizzical question out of my head: In > 1974, I was assigned muscle and nerve biopsy work and immunofluorescence > work on kidneys. Fixation ruined just about everything in all the > procedures involved with those studies, so fixatives were OUT in all of > my work. After moving to another hospital with my pathologist, I > continued the muscle and nerve work but not the immunos. Over the years, > news sort of trickled down that you histotechs were doing antibody work > on fixed tissues. I guessed you had found some way to repair the > tissues after being fixed. The Histonet has many messages sent back and > forth between histotechs doing antibody work, and they always specified > what fixative they used. A nd that always bothered me. It seemed like > a ring-around-the-rosie to fix, then, Unfix tissues for antibody work. > To up and fix the tissues, then turn around and repair SOME of the > damage done to the tissues-because I thought the tissues would not come > away unscathed from being bombarded with a fixative. And if it was > necessary to return the fixed tissues to an unfixed-like state, why not > just leave them unfixed in the first place? Does fixation do something > good to the tissues, making them better for antibody studies than fresh > frozen tissues? Muscle tissues are totally ruined for histochemistry, > and there is no way to repair the harm done to them by fixation. It > seemed like something out The Peterkin Papers: One story from that > wonderful book involved a cup of tea that had too much sugar in it.The > lady who wanted to drink it went up and down the lane gettjng advice as > to what to do, involved every herb, spice and remedy which just kept > making the tea worse. But the little old lady at the end of the lane > suggested that she make a fresh cup of tea. I wonder if the little old > lady at the end of the lane had been consulted in this vase, she might > have laughed merrily and suggested you just not fix your tissues in the > first place. Is there is some improvement in the tissues brought about > by fixation? As fixation totally ruined all my work, I find it hard to > believe that the tissues are going to come out unscathed from fixation. > Anyway, this has been a mystery working in the back of my head for years > .Can you put this matter to rest for this 76 year old? > georgecole@ev1.net > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------ End of Forwarded Message From PWebster <@t> hei.org Tue Aug 10 19:06:43 2004 From: PWebster <@t> hei.org (Webster, Paul) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] fixing immuno and antibody tissues Message-ID: <4E8C1F1E4E8FA748B5487C50C91695F0862F85@heimail.hei.org> Hi George, I think your questions will provoke more questions than answers. The problem is two-fold - good morphology with unlimited accessibility. We want to preserve morphology so that we can immobilize antigens and localize them to specific, identifiable regions in the tissue, but we also want to give antibodies the maximum access to antigens. Sometimes these two aims cannot be met and often the end result will depend on the physical properties of the antigen. For example, if an antigen is embedded deep inside a membrane, then gaining access will require strong extraction treatments. Conversely, a soluble antigen will require stronger fixation to immobilize it in the tissue. Unfixed tissues are usually immobilized by freezing. The frozen tissue is then sectioned, stuck to a slide and fixed, either by chemical fixation (aldehyde or acohol) or by drying. Sometimes it is a combination of these treatments. The reason why different treatments produce different results is because they offer varying degrees of accessibility. It is interesting that your experience rules out fixatives because they "totally ruined your work". I guess that fixation treatments you performed were interfered with your antibody labeling in some way. However, in electron microscopy, chemical fixation is almost always a prerequisite for immunolabeling. Many antigens survive embedding in acrylic or epoxy resins. Similarly, there are many enzymes that remain active after aldehyde treatment and form the basis of the very successful field of EM cytochemistry. I suspect that antibody production technology has greatly improved over the last 40 years and may be one reason why immunolabeling has become more versatile and successful, even on fixed tissues. It is interesting that you perceive fixation as doing damage and then the subsequent treatments, which often involve boiling sections in citrate buffer in a pressure cooker (!), as repairing the damage done by the fixative. I would suspect that a good fixation (e.g. in phosphate-buffered formaldehyde) will immobilize most proteins in a tissue and form cross-links between the proteins that may interfere with antibody access to antigens. Only the subsequent hydrolysis that damages the tissue section and which is often called antigen retrieval, is able to restore the access to antigens. Using unfixed tissues may offer easy access to antigens but then you run the risk of antigen migration if there is no immobilization treatment. This may not be a problem in histology where tissue sections are examined at relatively low magnification, but it is an important problem in electron microscopy, where antigens moving distances of less than 1?m can produce meaningless results. This brings me to a question of my own. If anyone is stillrreading, maybe they could offer and explanation of why histologists use shorter processing times for smaller tissue blocks. I have been reading the recent thread about smaller blocks being brittle because they were being processed for too long a time. Does anyone know why the tissue becomes brittle? I am going to stick out my neck and say it is probably not because of over-fixation in formaldehyde because I routinely cut thin sections (100nm) of tissues that have been left in formaldehyde for weeks. The tissues are infiltrated with sucrose, frozen and sectioned at -120 degrees and there is no difference between them and tissues fixed for only a few hours. Could the problem with brittle blocks be a result of exposure to alcohol or xylene? Thanks for your time. Regards, Paul Webster. Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 phone (213) 273 8026 fax (213) 413 6739 email: pwebster@hei.org Disclaimer: I am an electron microscopist who has been hanging out here because it currently seems more fun than the MSA listserver. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of George Cole > Sent: Tuesday, August 10, 2004 12:28 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] fixing immuno and antibody tissues > > Dear Histotechs; > Placate an old retiree and get a quizzical question out of my head: In > 1974, I was assigned muscle and nerve biopsy work and immunofluorescence > work on kidneys. Fixation ruined just about everything in all the > procedures involved with those studies, so fixatives were OUT in all of > my work. After moving to another hospital with my pathologist, I > continued the muscle and nerve work but not the immunos. Over the years, > news sort of trickled down that you histotechs were doing antibody work > on fixed tissues. I guessed you had found some way to repair the > tissues after being fixed. The Histonet has many messages sent back and > forth between histotechs doing antibody work, and they always specified > what fixative they used. A nd that always bothered me. It seemed like > a ring-around-the-rosie to fix, then, Unfix tissues for antibody work. > To up and fix the tissues, then turn around and repair SOME of the > damage done to the tissues-because I thought the tissues would not come > away unscathed from being bombarded with a fixative. And if it was > necessary to return the fixed tissues to an unfixed-like state, why not > just leave them unfixed in the first place? Does fixation do something > good to the tissues, making them better for antibody studies than fresh > frozen tissues? Muscle tissues are totally ruined for histochemistry, > and there is no way to repair the harm done to them by fixation. It > seemed like something out The Peterkin Papers: One story from that > wonderful book involved a cup of tea that had too much sugar in it.The > lady who wanted to drink it went up and down the lane gettjng advice as > to what to do, involved every herb, spice and remedy which just kept > making the tea worse. But the little old lady at the end of the lane > suggested that she make a fresh cup of tea. I wonder if the little old > lady at the end of the lane had been consulted in this vase, she might > have laughed merrily and suggested you just not fix your tissues in the > first place. Is there is some improvement in the tissues brought about > by fixation? As fixation totally ruined all my work, I find it hard to > believe that the tissues are going to come out unscathed from fixation. > Anyway, this has been a mystery working in the back of my head for years > .Can you put this matter to rest for this 76 year old? > georgecole@ev1.net > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From failm <@t> musc.edu Wed Aug 11 04:08:46 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] fixing immuno and antibody tissues Message-ID: Extended periods of time in alcohol and/or xylene will cause brittleness in small biopsies Rena Fail >>> "Webster, Paul" 08/10/04 20:06 PM >>> Hi George, I think your questions will provoke more questions than answers. The problem is two-fold - good morphology with unlimited accessibility. We want to preserve morphology so that we can immobilize antigens and localize them to specific, identifiable regions in the tissue, but we also want to give antibodies the maximum access to antigens. Sometimes these two aims cannot be met and often the end result will depend on the physical properties of the antigen. For example, if an antigen is embedded deep inside a membrane, then gaining access will require strong extraction treatments. Conversely, a soluble antigen will require stronger fixation to immobilize it in the tissue. Unfixed tissues are usually immobilized by freezing. The frozen tissue is then sectioned, stuck to a slide and fixed, either by chemical fixation (aldehyde or acohol) or by drying. Sometimes it is a combination of these treatments. The reason why different treatments produce different results is because they offer varying degrees of accessibility. It is interesting that your experience rules out fixatives because they "totally ruined your work". I guess that fixation treatments you performed were interfered with your antibody labeling in some way. However, in electron microscopy, chemical fixation is almost always a prerequisite for immunolabeling. Many antigens survive embedding in acrylic or epoxy resins. Similarly, there are many enzymes that remain active after aldehyde treatment and form the basis of the very successful field of EM cytochemistry. I suspect that antibody production technology has greatly improved over the last 40 years and may be one reason why immunolabeling has become more versatile and successful, even on fixed tissues. It is interesting that you perceive fixation as doing damage and then the subsequent treatments, which often involve boiling sections in citrate buffer in a pressure cooker (!), as repairing the damage done by the fixative. I would suspect that a good fixation (e.g. in phosphate-buffered formaldehyde) will immobilize most proteins in a tissue and form cross-links between the proteins that may interfere with antibody access to antigens. Only the subsequent hydrolysis that damages the tissue section and which is often called antigen retrieval, is able to restore the access to antigens. Using unfixed tissues may offer easy access to antigens but then you run the risk of antigen migration if there is no immobilization treatment. This may not be a problem in histology where tissue sections are examined at relatively low magnification, but it is an important problem in electron microscopy, where antigens moving distances of less than 1?m can produce meaningless results. This brings me to a question of my own. If anyone is stillrreading, maybe they could offer and explanation of why histologists use shorter processing times for smaller tissue blocks. I have been reading the recent thread about smaller blocks being brittle because they were being processed for too long a time. Does anyone know why the tissue becomes brittle? I am going to stick out my neck and say it is probably not because of over-fixation in formaldehyde because I routinely cut thin sections (100nm) of tissues that have been left in formaldehyde for weeks. The tissues are infiltrated with sucrose, frozen and sectioned at -120 degrees and there is no difference between them and tissues fixed for only a few hours. Could the problem with brittle blocks be a result of exposure to alcohol or xylene? Thanks for your time. Regards, Paul Webster. Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 phone (213) 273 8026 fax (213) 413 6739 email: pwebster@hei.org Disclaimer: I am an electron microscopist who has been hanging out here because it currently seems more fun than the MSA listserver. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of George Cole > Sent: Tuesday, August 10, 2004 12:28 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] fixing immuno and antibody tissues > > Dear Histotechs; > Placate an old retiree and get a quizzical question out of my head: In > 1974, I was assigned muscle and nerve biopsy work and immunofluorescence > work on kidneys. Fixation ruined just about everything in all the > procedures involved with those studies, so fixatives were OUT in all of > my work. After moving to another hospital with my pathologist, I > continued the muscle and nerve work but not the immunos. Over the years, > news sort of trickled down that you histotechs were doing antibody work > on fixed tissues. I guessed you had found some way to repair the > tissues after being fixed. The Histonet has many messages sent back and > forth between histotechs doing antibody work, and they always specified > what fixative they used. A nd that always bothered me. It seemed like > a ring-around-the-rosie to fix, then, Unfix tissues for antibody work. > To up and fix the tissues, then turn around and repair SOME of the > damage done to the tissues-because I thought the tissues would not come > away unscathed from being bombarded with a fixative. And if it was > necessary to return the fixed tissues to an unfixed-like state, why not > just leave them unfixed in the first place? Does fixation do something > good to the tissues, making them better for antibody studies than fresh > frozen tissues? Muscle tissues are totally ruined for histochemistry, > and there is no way to repair the harm done to them by fixation. It > seemed like something out The Peterkin Papers: One story from that > wonderful book involved a cup of tea that had too much sugar in it.The > lady who wanted to drink it went up and down the lane gettjng advice as > to what to do, involved every herb, spice and remedy which just kept > making the tea worse. But the little old lady at the end of the lane > suggested that she make a fresh cup of tea. I wonder if the little old > lady at the end of the lane had been consulted in this vase, she might > have laughed merrily and suggested you just not fix your tissues in the > first place. Is there is some improvement in the tissues brought about > by fixation? As fixation totally ruined all my work, I find it hard to > believe that the tissues are going to come out unscathed from fixation. > Anyway, this has been a mystery working in the back of my head for years > .Can you put this matter to rest for this 76 year old? > georgecole@ev1.net > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lfaucette <@t> niaid.nih.gov Wed Aug 11 06:05:00 2004 From: lfaucette <@t> niaid.nih.gov (Faucette, Lawrence (NIH/NIAID)) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] fixing immuno and antibody tissues Message-ID: <78F91CA4B8F3504E8FE5F13AAF122ED804A02CC1@nihexchange27.nih.gov> Good Morning, It has been my experience that the excess drying of tissue while processing is from both the alcohol and xylene. Additionally leaving them at temp in melted paraffin will cause extra hardening. On the original topic of fixation vs unfixed tissue for enzyme/antibody testing. Both in research and diagnostic pathology (although more often in diagnostic work) may enzyme/antibody workups are derived after the fact. That is to say once the original work is done and further studies are deemed valuable. At that point the choice to fix or not to fix has often already been made. Lawrence J Faucette Contractor HT ASCP http://www.niaid.nih.gov/dir/services/animalcare/VetPathology/VetPathology-i ndex.html Infectious Disease Pathogenesis Section Comparative Medicine Branch Division of Intramural Research, NIAID, NIH Twinbrook III, Room 2W-01A, MSC 8135 12735 Twinbrook Parkway Bethesda, MD 20892-8135 Telephone 301-451-1056 Fax 301-480-2343 SoBran, Inc. Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. The National Institute of Allergy and Infectious Diseases (NIAID) shall not accept liability for any statement made that are the sender's own and not expressly made on behalf of the NIAID by one of its representatives. -----Original Message----- From: Mildred Fail [mailto:failm@musc.edu] Sent: Wednesday, August 11, 2004 5:09 AM To: georgecole@ev1.net; PWebster@hei.org; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] fixing immuno and antibody tissues Extended periods of time in alcohol and/or xylene will cause brittleness in small biopsies Rena Fail >>> "Webster, Paul" 08/10/04 20:06 PM >>> Hi George, I think your questions will provoke more questions than answers. The problem is two-fold - good morphology with unlimited accessibility. We want to preserve morphology so that we can immobilize antigens and localize them to specific, identifiable regions in the tissue, but we also want to give antibodies the maximum access to antigens. Sometimes these two aims cannot be met and often the end result will depend on the physical properties of the antigen. For example, if an antigen is embedded deep inside a membrane, then gaining access will require strong extraction treatments. Conversely, a soluble antigen will require stronger fixation to immobilize it in the tissue. Unfixed tissues are usually immobilized by freezing. The frozen tissue is then sectioned, stuck to a slide and fixed, either by chemical fixation (aldehyde or acohol) or by drying. Sometimes it is a combination of these treatments. The reason why different treatments produce different results is because they offer varying degrees of accessibility. It is interesting that your experience rules out fixatives because they "totally ruined your work". I guess that fixation treatments you performed were interfered with your antibody labeling in some way. However, in electron microscopy, chemical fixation is almost always a prerequisite for immunolabeling. Many antigens survive embedding in acrylic or epoxy resins. Similarly, there are many enzymes that remain active after aldehyde treatment and form the basis of the very successful field of EM cytochemistry. I suspect that antibody production technology has greatly improved over the last 40 years and may be one reason why immunolabeling has become more versatile and successful, even on fixed tissues. It is interesting that you perceive fixation as doing damage and then the subsequent treatments, which often involve boiling sections in citrate buffer in a pressure cooker (!), as repairing the damage done by the fixative. I would suspect that a good fixation (e.g. in phosphate-buffered formaldehyde) will immobilize most proteins in a tissue and form cross-links between the proteins that may interfere with antibody access to antigens. Only the subsequent hydrolysis that damages the tissue section and which is often called antigen retrieval, is able to restore the access to antigens. Using unfixed tissues may offer easy access to antigens but then you run the risk of antigen migration if there is no immobilization treatment. This may not be a problem in histology where tissue sections are examined at relatively low magnification, but it is an important problem in electron microscopy, where antigens moving distances of less than 1?m can produce meaningless results. This brings me to a question of my own. If anyone is stillrreading, maybe they could offer and explanation of why histologists use shorter processing times for smaller tissue blocks. I have been reading the recent thread about smaller blocks being brittle because they were being processed for too long a time. Does anyone know why the tissue becomes brittle? I am going to stick out my neck and say it is probably not because of over-fixation in formaldehyde because I routinely cut thin sections (100nm) of tissues that have been left in formaldehyde for weeks. The tissues are infiltrated with sucrose, frozen and sectioned at -120 degrees and there is no difference between them and tissues fixed for only a few hours. Could the problem with brittle blocks be a result of exposure to alcohol or xylene? Thanks for your time. Regards, Paul Webster. Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 phone (213) 273 8026 fax (213) 413 6739 email: pwebster@hei.org Disclaimer: I am an electron microscopist who has been hanging out here because it currently seems more fun than the MSA listserver. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of George Cole > Sent: Tuesday, August 10, 2004 12:28 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] fixing immuno and antibody tissues > > Dear Histotechs; > Placate an old retiree and get a quizzical question out of my head: In > 1974, I was assigned muscle and nerve biopsy work and immunofluorescence > work on kidneys. Fixation ruined just about everything in all the > procedures involved with those studies, so fixatives were OUT in all of > my work. After moving to another hospital with my pathologist, I > continued the muscle and nerve work but not the immunos. Over the years, > news sort of trickled down that you histotechs were doing antibody work > on fixed tissues. I guessed you had found some way to repair the > tissues after being fixed. The Histonet has many messages sent back and > forth between histotechs doing antibody work, and they always specified > what fixative they used. A nd that always bothered me. It seemed like > a ring-around-the-rosie to fix, then, Unfix tissues for antibody work. > To up and fix the tissues, then turn around and repair SOME of the > damage done to the tissues-because I thought the tissues would not come > away unscathed from being bombarded with a fixative. And if it was > necessary to return the fixed tissues to an unfixed-like state, why not > just leave them unfixed in the first place? Does fixation do something > good to the tissues, making them better for antibody studies than fresh > frozen tissues? Muscle tissues are totally ruined for histochemistry, > and there is no way to repair the harm done to them by fixation. It > seemed like something out The Peterkin Papers: One story from that > wonderful book involved a cup of tea that had too much sugar in it.The > lady who wanted to drink it went up and down the lane gettjng advice as > to what to do, involved every herb, spice and remedy which just kept > making the tea worse. But the little old lady at the end of the lane > suggested that she make a fresh cup of tea. I wonder if the little old > lady at the end of the lane had been consulted in this vase, she might > have laughed merrily and suggested you just not fix your tissues in the > first place. Is there is some improvement in the tissues brought about > by fixation? As fixation totally ruined all my work, I find it hard to > believe that the tissues are going to come out unscathed from fixation. > Anyway, this has been a mystery working in the back of my head for years > .Can you put this matter to rest for this 76 year old? > georgecole@ev1.net > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dencrowl <@t> MIT.EDU Wed Aug 11 07:47:01 2004 From: dencrowl <@t> MIT.EDU (Denise Crowley) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] employment at MIT in Cambridge MA Message-ID: We are looking to add a histotech to our Histology Core Facility in the Center for Cancer Research at Massachusetts Institute of Technology. Our staff consists of 2 techs and a facility manager working for 20 different laboratories all interested in researching the genetic basis for cancer. Our work consists of processing, sectioning, and H&E staining of animal tissues (mostly mouse, but some zebrafish as well). We have the latest in automated equipment including a ThermoElectron Excelsior processor, ThermoElectron Finesse microtomes, ThermoElectron Cryotome, ThermoElectron Gemini Stainer, and ThermoElectron Consul Coverslipper We are looking for a full-time tech with Histology experience. While certification is not required, we need someone who is committed to staying in Histology and who can serve as a resource for grad students and postdoctoral fellows in helping to design their experiments. Hours are flexible and benefits are great. If you are interested in a challenging environment where your expertise is truly appreciated, send me a resume and we will talk. Denise Crowley, Facility Manager Histology Massachusetts Institute of Technology Center for Cancer Research 40 Ames St. E17-230 Cambridge MA 02139 dencrowl@mit.edu From GDawson <@t> Milw.Dynacare.com Wed Aug 11 07:54:52 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] RE: Sakura Mounting Medium Message-ID: Michael, My lab has been using the Sakura mounting medium for the Finetek coverslipper for some time now. We haven't had problems with bubbles, durability, cloudiness, etc... The quality is excellent and I'd say it's only detraction is the price. Permount is significantly cheaper than the Sakura mounting medium but, as you say, it's not compatable with the coverslipper. The Sakura is by far the best coverslipper we have so it is worth the added expense. Glen Dawson BS, HT & QIHC IHC Coordinator Milwaukee, WI From TJJ <@t> Stowers-Institute.org Wed Aug 11 08:56:05 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] RE: fixing immuno and antibody tissues Message-ID: Paul Webster gave a brilliant answer to George Cole's question about fixation and its role in immunohistochemistry. Patti Loykasek also gave an excellent response, especially considering the IHC done on routine surgical pathology specimens. I totally agree with Patti in doing everything you can to make sure you are not obtaining false positives resulting from digestion or unmasking techniques. Controls are critical at this point! Regardless of what is recommended on the antibody data sheet, I will always work up a new antibody using no pretreatment, an enzyme digestion (usually trypsin), and a HIER method using citrate buffer pH 6.0 in the electric pressure cooker or waterbath. If none of those conditions work, I'll try with different pretreatment regimens, i.e. pepsin, protease, and/or high pH HIER. Additionally, most of the antibodies that are tested for use in IHC have been tested using cryosections. These tissues are generally not fixed, but snap frozen, sectioned, allowed to air dry, and then fixed in acetone (or a mixture of acetone/alcohol) before immunostaining. Some labs will use formalin-fixed, sucrose cryopreserved cryosections with good results, but if formalin fixation renders the antigen inaccessible or not well preserved, then this is no improvement from using formalin fixed, paraffin embedded (FFPE) samples. Luckily, for most clinical applications, many of the antibodies are well characterized and optimized for use in FFPE samples. Research applications tend to be a little more challenging on this front. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From adelsyscarol <@t> yahoo.com Wed Aug 11 09:39:27 2004 From: adelsyscarol <@t> yahoo.com (Carol Wilson) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Looking for a paraffin pot from an old autotechnicon or similar Message-ID: <20040811143927.76864.qmail@web50606.mail.yahoo.com> Hello all, Does anyone out there have an old paraffin pot in working condition from an old style processor that they are willing to sell? It needs to be the kind that can plug into a normal wall socket away from the processor. Please respond to me directly. Thank you. Carol Wilson Adelsys, Inc. --------------------------------- Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. From funderwood <@t> mcohio.org Wed Aug 11 09:58:12 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:23:52 2005 Subject: [BULK] - RE: [Histonet] Problem with VIP 5 Message-ID: Thanks for your responses. If I had a Mulligan, I wouldn't take up golf. Fred >>> "Faucette, Lawrence (NIH/NIAID)" 08/10/04 02:59PM >>> ROFL, Leave it to the USAF to speak in golf terms :) ( just kidding ) Mulligan would be a golf shot that you pretend never really happened :) Lawrence J Faucette Contractor HT ASCP http://www.niaid.nih.gov/dir/services/animalcare/VetPathology/VetPathology-i ndex.html Infectious Disease Pathogenesis Section Comparative Medicine Branch Division of Intramural Research, NIAID, NIH Twinbrook III, Room 2W-01A, MSC 8135 12735 Twinbrook Parkway Bethesda, MD 20892-8135 Telephone 301-451-1056 Fax 301-480-2343 SoBran, Inc. Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. The National Institute of Allergy and Infectious Diseases (NIAID) shall not accept liability for any statement made that are the sender's own and not expressly made on behalf of the NIAID by one of its representatives. -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Tuesday, August 10, 2004 2:50 PM To: Fred Underwood Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [BULK] - RE: [Histonet] Problem with VIP 5 I don't know - - what's a mulligan? Is it something like a henway? Jackie O' "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/10/2004 12:57 PM To: cc: Subject: [BULK] - RE: [Histonet] Problem with VIP 5 I am looking into a new processor this next year. I thought that the older VIPs were good units. However, reading about the problems people are having with the VIP 5, I'm having doubts. I'm curious if those of you having problems were given a mulligan, would you still chose a VIP or something else? Fred >>> "Ian" 08/09/04 10:02PM >>> Hey, you all don't scare me WE just purchase a new VIP 5 and have not have used it yet! Hey Joe, how are you doing? SSgt Bernard 59th Clinical Research Squadron "The USAF's largest Clinical Research facility" Lackland AFB, Texas -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Monday, August 09, 2004 3:27 PM To: Jill Songer; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Problem with VIP 5 Jill, how long have you had the VIP 5? I've had my machine almost 3 years and haven't had that problem. If you don't close the lid properly, when it pumps in fluid, the lid pops off, but we haven't had any smells. Please let us know what the service rep says. I always like new stories. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Jill Songer" To: Sent: Monday, August 09, 2004 2:40 PM Subject: [Histonet] Problem with VIP 5 > We are having a problem with our VIP 5 having a strong formalin odor at the > end of the run when we open the retort to remove the tissues. There is also > a large amount of fluid-formalin?? on the lid and in the chamber. > > Has anyone else experienced this? I have called Sakura and they are sending > a field service tech but I was curious if anyone has had this problem. > Whenever I call a vendor it seems they always say, well, we haven't had > THIS problem before. > > Thanks. > > Jill Songer HT (ASCP) > Histopathology Lab > Virginia-Maryland Regional College of Veterinary Medicine > Teaching Hospital > Virginia Tech > Blacksburg, VA 24061-0443 > www.vetmed.vt.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KTennako <@t> odu.edu Wed Aug 11 10:13:39 2004 From: KTennako <@t> odu.edu (Kushan U Tennakoon) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] wax embedded section mounting on slides Message-ID: Dear All, I'm trying to mount wax embedded plant material on glass slides to differentiate tissue tyes using theToludine blue stain. Can you please advice me what is the best subbing medium that I should use to mount the wax ribbons. Has any one used just "egg albumine" as the subbing medium? Many thanks, Sincerely, Kushan Tennakoon ktennako@odu.edu From georgecole <@t> ev1.net Wed Aug 11 10:52:16 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] fixation/no fixation Message-ID: <000001c47fbb$30d2cdb0$074dbad0@hppav> Histotechs; Thank you for your responses to my question. They reflect a much broader range of tissues and entities studied than my muscle/nerve/kidney studies. My questions arose from my experience over 27 years with biopsy tissues of muscles, nerves and kidneys. The question simply was, given, that once fixation was considered impossible for use with immunofluorescent work and, I thought, all antibody techniques. Now the histonet carries constant references to fixation in these studies. The questions were 1) what do you do to make fixed tissues useable for these studies and, 2) what is the comparable results between fixed and non fixed tissues in the results? I am impressed with the broad responses reflecting much expertise in work done. But questions 1 and 2 have been whiffed aside by an acceptance of the fixed tissue work, and this retiree is still in the dark about what has altered a once Absolute Rejection of fixation, and allowed histotechs of today an acceptance of fixation in certain areas of histochemistry. Like the food ladeller and Oliver Twist: When Oliver asks, "Please, sir. May I have some more?", the man serving the children shouts; "More! The boy wants more! Surely the boy shall hang!!" That I should question fixatives seems worse than Oliver asking for more oatmeal! The question about fixatives seems foreign to the current generalized mind set with strong habits of use of them. I ask these questions after a working lifetime in which fixatives caused absolute failure in all but a few processes in muscle work and it was taken as an additional absolute, that fixatives ruined imnunofluorescnce in kidney work. Have these absolutes evaporated? georgecole@ev1.net From tpmorken <@t> labvision.com Wed Aug 11 11:09:15 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:52 2005 Subject: Brittle tissues ...was...RE: [Histonet] fixing immuno and antibod y tissues Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2D94B5F@usca0082k08.labvision.apogent.com> Geroge asks (yes I read it through!) <> On the order of 4% of water in tissue is "bound" (MAGMA. 1996 Mar;4(1):55-9. Improved estimation of tissue hydration and bound water fraction in rat liver tissue). Removing bound water is apparantly the reason tissue becomes hard and brittle. Alcohol is the main culprit. You might want to read this article about the effect of removing bound water in tissue (see the part on dehydration). This is something Dick Dapson from Anatech has been emphasising at NSH meetings for many years. http://www.americanhistology.com/literature/library/minimizs.htm This is reproduced from J Histotechnol 16:71, 1993 (note that the reference given on the webpage is incorrect - it reads 1992; it is actually from the march 1993 issue). Tim Morken Lab Vision - Neomarkers www.labvision.com -----Original Message----- From: Webster, Paul [mailto:PWebster@hei.org] Sent: Tuesday, August 10, 2004 5:07 PM To: histonet@lists.utsouthwestern.edu; George Cole Subject: RE: [Histonet] fixing immuno and antibody tissues Hi George, I think your questions will provoke more questions than answers. The problem is two-fold - good morphology with unlimited accessibility. We want to preserve morphology so that we can immobilize antigens and localize them to specific, identifiable regions in the tissue, but we also want to give antibodies the maximum access to antigens. Sometimes these two aims cannot be met and often the end result will depend on the physical properties of the antigen. For example, if an antigen is embedded deep inside a membrane, then gaining access will require strong extraction treatments. Conversely, a soluble antigen will require stronger fixation to immobilize it in the tissue. Unfixed tissues are usually immobilized by freezing. The frozen tissue is then sectioned, stuck to a slide and fixed, either by chemical fixation (aldehyde or acohol) or by drying. Sometimes it is a combination of these treatments. The reason why different treatments produce different results is because they offer varying degrees of accessibility. It is interesting that your experience rules out fixatives because they "totally ruined your work". I guess that fixation treatments you performed were interfered with your antibody labeling in some way. However, in electron microscopy, chemical fixation is almost always a prerequisite for immunolabeling. Many antigens survive embedding in acrylic or epoxy resins. Similarly, there are many enzymes that remain active after aldehyde treatment and form the basis of the very successful field of EM cytochemistry. I suspect that antibody production technology has greatly improved over the last 40 years and may be one reason why immunolabeling has become more versatile and successful, even on fixed tissues. It is interesting that you perceive fixation as doing damage and then the subsequent treatments, which often involve boiling sections in citrate buffer in a pressure cooker (!), as repairing the damage done by the fixative. I would suspect that a good fixation (e.g. in phosphate-buffered formaldehyde) will immobilize most proteins in a tissue and form cross-links between the proteins that may interfere with antibody access to antigens. Only the subsequent hydrolysis that damages the tissue section and which is often called antigen retrieval, is able to restore the access to antigens. Using unfixed tissues may offer easy access to antigens but then you run the risk of antigen migration if there is no immobilization treatment. This may not be a problem in histology where tissue sections are examined at relatively low magnification, but it is an important problem in electron microscopy, where antigens moving distances of less than 1?m can produce meaningless results. This brings me to a question of my own. If anyone is stillrreading, maybe they could offer and explanation of why histologists use shorter processing times for smaller tissue blocks. I have been reading the recent thread about smaller blocks being brittle because they were being processed for too long a time. Does anyone know why the tissue becomes brittle? I am going to stick out my neck and say it is probably not because of over-fixation in formaldehyde because I routinely cut thin sections (100nm) of tissues that have been left in formaldehyde for weeks. The tissues are infiltrated with sucrose, frozen and sectioned at -120 degrees and there is no difference between them and tissues fixed for only a few hours. Could the problem with brittle blocks be a result of exposure to alcohol or xylene? Thanks for your time. Regards, Paul Webster. Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 phone (213) 273 8026 fax (213) 413 6739 email: pwebster@hei.org Disclaimer: I am an electron microscopist who has been hanging out here because it currently seems more fun than the MSA listserver. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of George Cole > Sent: Tuesday, August 10, 2004 12:28 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] fixing immuno and antibody tissues > > Dear Histotechs; > Placate an old retiree and get a quizzical question out of my head: > In 1974, I was assigned muscle and nerve biopsy work and > immunofluorescence work on kidneys. Fixation ruined just about > everything in all the procedures involved with those studies, so > fixatives were OUT in all of my work. After moving to another hospital > with my pathologist, I continued the muscle and nerve work but not the > immunos. Over the years, news sort of trickled down that you > histotechs were doing antibody work on fixed tissues. I guessed you > had found some way to repair the tissues after being fixed. The > Histonet has many messages sent back and forth between histotechs > doing antibody work, and they always specified what fixative they > used. A nd that always bothered me. It seemed like a > ring-around-the-rosie to fix, then, Unfix tissues for antibody work. > To up and fix the tissues, then turn around and repair SOME of the > damage done to the tissues-because I thought the tissues would not > come away unscathed from being bombarded with a fixative. And if it > was necessary to return the fixed tissues to an unfixed-like state, > why not just leave them unfixed in the first place? Does fixation do > something good to the tissues, making them better for antibody studies > than fresh frozen tissues? Muscle tissues are totally ruined for > histochemistry, and there is no way to repair the harm done to them by > fixation. It seemed like something out The Peterkin Papers: One story > from that wonderful book involved a cup of tea that had too much sugar > in it.The lady who wanted to drink it went up and down the lane > gettjng advice as to what to do, involved every herb, spice and remedy > which just kept making the tea worse. But the little old lady at the > end of the lane suggested that she make a fresh cup of tea. I wonder > if the little old lady at the end of the lane had been consulted in > this vase, she might have laughed merrily and suggested you just not > fix your tissues in the first place. Is there is some improvement in > the tissues brought about by fixation? As fixation totally ruined all > my work, I find it hard to believe that the tissues are going to come > out unscathed from fixation. Anyway, this has been a mystery working in the back of my head for years .Can you put this matter to rest for this 76 year old? > georgecole@ev1.net > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Marjorie.Lehman <@t> unilever.com Wed Aug 11 11:19:58 2004 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] OT: Telephone Consulting Message-ID: Hi, Has anyone ever done this? If so, would you be kind enough to tell me what your arrangements were/are. I have been approached about being "available to help people" and I can't see myself sitting around waiting for the phone to ring without getting some "reward". Thanks, Marge From gcallis <@t> montana.edu Wed Aug 11 11:22:39 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] fixing immuno and antibody tissues In-Reply-To: <4E8C1F1E4E8FA748B5487C50C91695F0862F85@heimail.hei.org> References: <4E8C1F1E4E8FA748B5487C50C91695F0862F85@heimail.hei.org> Message-ID: <6.0.0.22.1.20040811100541.01b18918@gemini.msu.montana.edu> Paul, In response to your last question about smaller pieces of tissue: Jerry Fredenburgh (Richard Allan) gave an explanation (chemistry is one of his hobbies!) that bound water on protein can be removed along with free water found in tissue spaces. The object is to retain bound water, and only remove the free water. If you overexpose to alcohol during dehydration or add heat to processing during dehydration steps, bound water is removed, tissue becomes more brittle, dry, difficult to section. I think you answered your own question with your last sentence, although clearing agent, e.g. xylene is there to remove alcohol in preparation for infiltration by paraffin and/or some plastics (methylmethacrylate). Xylene is considered hardening to tissues also, more so than some of the xylene substitutes, eg Propar and Clearite 3 - single aliphatic hydrocarbons. Heat during paraffin infiltration also dries tissue and contributes to brittleness (many of us have had paraffin bath temperature failures, and the tissue is "cooked" into rockhard nuggets when the paraffin is too hot. Just some thoughts - also, in our work with murine CD marker immunohistochemistry, any aldehyde fixative creates chaos, and immnuonstaining is never achieved even with minimal fixation time, or after any kind of enzyme digestion and retrieval method - CD4 and CD8 are notorious (how many times have I said this- ad nauseum!?) . We then rely on fresh tissue frozen sections fixed after air drying and with acetone alcohol mixture OR acetone fixation) precipitating these antigens rather than aldehyde crosslinkage. Maybe some day there will be CD4 and CD8clones that work with FFPE mouse tissue ----- waiting -------------- At 06:06 PM 8/10/2004, you wrote: >If anyone is stillrreading, maybe they could offer and explanation of why >histologists use shorter processing times for smaller tissue blocks. I >have been reading the recent thread about smaller blocks being brittle >because they were being processed for too long a time. Does anyone know >why the tissue becomes brittle? I am going to stick out my neck and say it >is probably not because of over-fixation in formaldehyde because I >routinely cut thin sections (100nm) of tissues that have been left in >formaldehyde for weeks. The tissues are infiltrated with sucrose, frozen >and sectioned at -120 degrees and there is no difference between them and >tissues fixed for only a few hours. Could the problem with brittle blocks >be a result of exposure to alcohol or xylene? > >Thanks for your time. > >Regards, > >Paul Webster. > > > > >Paul Webster, Ph.D. >House Ear Institute >2100 West Third Street >Los Angeles >CA 90057 >phone (213) 273 8026 >fax (213) 413 6739 >email: pwebster@hei.org > > >Disclaimer: I am an electron microscopist who has been hanging out here >because it currently seems more fun than the MSA listserver. > > > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > George Cole > > Sent: Tuesday, August 10, 2004 12:28 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] fixing immuno and antibody tissues > > > > Dear Histotechs; > > Placate an old retiree and get a quizzical question out of my head: In > > 1974, I was assigned muscle and nerve biopsy work and immunofluorescence > > work on kidneys. Fixation ruined just about everything in all the > > procedures involved with those studies, so fixatives were OUT in all of > > my work. After moving to another hospital with my pathologist, I > > continued the muscle and nerve work but not the immunos. Over the years, > > news sort of trickled down that you histotechs were doing antibody work > > on fixed tissues. I guessed you had found some way to repair the > > tissues after being fixed. The Histonet has many messages sent back and > > forth between histotechs doing antibody work, and they always specified > > what fixative they used. A nd that always bothered me. It seemed like > > a ring-around-the-rosie to fix, then, Unfix tissues for antibody work. > > To up and fix the tissues, then turn around and repair SOME of the > > damage done to the tissues-because I thought the tissues would not come > > away unscathed from being bombarded with a fixative. And if it was > > necessary to return the fixed tissues to an unfixed-like state, why not > > just leave them unfixed in the first place? Does fixation do something > > good to the tissues, making them better for antibody studies than fresh > > frozen tissues? Muscle tissues are totally ruined for histochemistry, > > and there is no way to repair the harm done to them by fixation. It > > seemed like something out The Peterkin Papers: One story from that > > wonderful book involved a cup of tea that had too much sugar in it.The > > lady who wanted to drink it went up and down the lane gettjng advice as > > to what to do, involved every herb, spice and remedy which just kept > > making the tea worse. But the little old lady at the end of the lane > > suggested that she make a fresh cup of tea. I wonder if the little old > > lady at the end of the lane had been consulted in this vase, she might > > have laughed merrily and suggested you just not fix your tissues in the > > first place. Is there is some improvement in the tissues brought about > > by fixation? As fixation totally ruined all my work, I find it hard to > > believe that the tissues are going to come out unscathed from fixation. > > Anyway, this has been a mystery working in the back of my head for years > > .Can you put this matter to rest for this 76 year old? > > georgecole@ev1.net > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From tpmorken <@t> labvision.com Wed Aug 11 11:30:29 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] fixation/no fixation Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2D94B60@usca0082k08.labvision.apogent.com> George asks (again) << The questions were 1) what do you do to make fixed tissues useable for these studies There's two parts to this, one involving the tissue, one the antibody First, fixed tissues, specifically formalin-fixed tissues, are being "treated" with various enzyems (trypsin, proteinase K, etc) or heated buffers at various pH's and temperatures in order to "somehow" "unmask" the epitopes. Very vague, really, but there is some research being done and various explanations, from breaking formalin-induced bonds to restoring electrical charges. I was just at a histochemistry meeting in San Diego where all these explanations went head-to-head. It was very interesting and I think the "restoring charges" explanation is edging ahead. Second, we are screening antibodies to find the ones that work best on formalin-fixed tissue although they may still require some "pretreatment." Indeed, many of these antibodies don't work at all on frozen tissue. and, 2) what is the comparable results between fixed and non fixed tissues in the results? >> Since formalin-fixed tissue shows decent morphology, it is preferred over frozens if the target can be detected. Many times we really do want to see the tissue structure where an antibody is found, not just the general location of an antibody. Since freezing can destroy cell and tissue boundaries, many protiens (and their target epitopes) are translocated during the thawing that occurs after the freezing. So location is not necessarily exact in frozen tissue. In my experience, tissue that is properly fixed gives great results - strong signal, exact location and excellent morphology. And many of the targets once detected by histochemistry can now be detected by antibodies, thus making frozens even less necessary. But the primary moving force for fixed tissue is the clinical pathology need - if a test can be done on fixed tissue it makes life easier for everyone, plus you may have a (nearly) permanent paraffin block to use in the future for other tests. Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: George Cole [mailto:georgecole@ev1.net] Sent: Wednesday, August 11, 2004 8:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fixation/no fixation Histotechs; Thank you for your responses to my question. They reflect a much broader range of tissues and entities studied than my muscle/nerve/kidney studies. My questions arose from my experience over 27 years with biopsy tissues of muscles, nerves and kidneys. The question simply was, given, that once fixation was considered impossible for use with immunofluorescent work and, I thought, all antibody techniques. Now the histonet carries constant references to fixation in these studies. The questions were 1) what do you do to make fixed tissues useable for these studies and, 2) what is the comparable results between fixed and non fixed tissues in the results? I am impressed with the broad responses reflecting much expertise in work done. But questions 1 and 2 have been whiffed aside by an acceptance of the fixed tissue work, and this retiree is still in the dark about what has altered a once Absolute Rejection of fixation, and allowed histotechs of today an acceptance of fixation in certain areas of histochemistry. Like the food ladeller and Oliver Twist: When Oliver asks, "Please, sir. May I have some more?", the man serving the children shouts; "More! The boy wants more! Surely the boy shall hang!!" That I should question fixatives seems worse than Oliver asking for more oatmeal! The question about fixatives seems foreign to the current generalized mind set with strong habits of use of them. I ask these questions after a working lifetime in which fixatives caused absolute failure in all but a few processes in muscle work and it was taken as an additional absolute, that fixatives ruined imnunofluorescnce in kidney work. Have these absolutes evaporated? georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vbaker60 <@t> yahoo.com Wed Aug 11 12:04:26 2004 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] p16 INK 4a antibody on FFPE rat tissue Message-ID: <20040811170426.84953.qmail@web50103.mail.yahoo.com> Hi Is anyone doing the above antibody on rat tissue? If so, could you please tell me who you are getting it from? Thanks. Vikki Baker Institute for Cancer Prevention Valhalla, NY 10595 __________________________________ Do you Yahoo!? New and Improved Yahoo! Mail - 100MB free storage! http://promotions.yahoo.com/new_mail From gcallis <@t> montana.edu Wed Aug 11 12:53:59 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Re: OT: Telephone Consulting In-Reply-To: References: Message-ID: <6.0.0.22.1.20040811112435.01b278b0@gemini.msu.montana.edu> By OT, do you mean "overtime, out side of job consulting? Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From ihc <@t> unipathllc.com Wed Aug 11 13:09:07 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] HTL Practical Message-ID: <000001c47fce$4c787610$4500a8c0@unipath02> Hello all, Does anyone know if it is permitted to ask questions on the Histonet regarding troubleshooting stains for our practicals? I don't want to ask if I'm not supposed to. Thanks, Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO From RCazares <@t> schosp.org Wed Aug 11 15:47:42 2004 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Thymidine Synthetase on prostate biopsies Message-ID: <229A3566B9F0D311826E00D0B7441D7907E0AFF1@swedish_nt1.schosp.org> Hello Histonetters, Is anyone doing Thymidine Synthetase (IHC) on prostate biopsies? If it is part of a panel, what other antibodies are run? And finally, is Thymidylate Synthase different than Thymidine Synthetase? Any and all help is appreciated ! Thank you, Ruth Cazares Swedish Covenant Hospital Chicago, Illinois *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ************************* From jmitchell <@t> neurology.wisc.edu Wed Aug 11 16:34:57 2004 From: jmitchell <@t> neurology.wisc.edu (Mitchell (Jean)) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Caveolin 3 Message-ID: Anyone staining for caveolin 3 in muscle & have a good source for this antibody? Any & all input would be greatly appreciated! Thanks much! Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI From TJasper <@t> smdc.org Wed Aug 11 17:18:53 2004 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Position Opportunity Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E44F6D@harrier> Opportunity available in Duluth, MN. SMDC (St. Mary's/Duluth Clinic) seeks to fill a position in the Histology Department. This is a full-time day shift position at a progressive tertiary care facility. SMDC is located near the shore of scenic Lake Superior. Duluth is a clean and green community within minutes of wilderness in both northern Minnesota and Wisconsin. Greater urban amenities are available in Minneapolis/St. Paul 150 miles south on I-35. Duluth offers much in the way of culture, education and entertainment in the greater western Lake Superior area. SMDC serves northern Minnesota, northern Wisconsin and the western Upper Peninsula of Michigan. Consider joining our respected and respectful pathology service, here at our hidden gem. For further information and application information log on to www.smdc.org . Phone 218/786-4564 Fax 218/786-4018. Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org From JColCLEFA <@t> aol.com Wed Aug 11 18:38:09 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Fixed IF's, small brittle bx's Message-ID: George, the absolutes, much like absolute alcohol would if left open, have certainly evaporated. Borrowing from my total formalin fixation technique IHC lab, I have successfully and routinely stained skin biopsies as well as kidney biopsies with FITC conjugated antibodies after enzyme pretreatment with Protease 0.1%, 30 minutes@ 37C. The kidneys were fixed in Zamboni's fixative, the skin in Formalin, both paraffin processed. The only "artefact" was a reduction in non-specific background staining. ( a plus in my book!) In the kidney arena, we now prefer fixed tissue to fresh frozen. J Coleman HT(ASCP)QIHC we have found that overdehydration of small biopsies is the culprit. From info <@t> ihcworld.com Wed Aug 11 20:18:40 2004 From: info <@t> ihcworld.com (IHC World) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] p16 INK 4a antibody on FFPE rat tissue In-Reply-To: <20040811170426.84953.qmail@web50103.mail.yahoo.com> Message-ID: Hi Vikki, There was a nice long discussion about p16INK4a antibody in IHC World Forum. Here is the link: http://www.ihcworld.com/forum/viewtopic.php?t=322&highlight=p16+ink Hope this helps. Richard IHC World http://www.ihcworld.com -----Original Message----- From: Victoria Baker [mailto:vbaker60@yahoo.com] Sent: Wednesday, August 11, 2004 12:04 PM To: HistoNet Server Subject: [Histonet] p16 INK 4a antibody on FFPE rat tissue Hi Is anyone doing the above antibody on rat tissue? If so, could you please tell me who you are getting it from? Thanks. Vikki Baker Institute for Cancer Prevention Valhalla, NY 10595 __________________________________ Do you Yahoo!? New and Improved Yahoo! Mail - 100MB free storage! http://promotions.yahoo.com/new_mail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jason.PALMER <@t> svhm.org.au Wed Aug 11 23:04:36 2004 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] immunostaining with Technovit 8100 / glycol methacrylate Message-ID: Hi Histonetters. I'm trying to immunostain some sections of rat tissue embedded in Technovit 8100 (a glycol methacrylate) with an ED1 antibody (Serotec rat anti mouse), and have had no success to this point. Have tried various antibody concentrations and antigen retrievals including pronase, trypsin (which we use successfully for immunostaining with the same Ab on paraffin-embedded tissues) and high temperature citric acid buffer retrieval. Our samples are paraformaldehyde fixed overnight, and we use a labelled streptavidin-biotin system. The technical information that comes with the product suggests it is suitable for immunohistochemistry (and this is why we purchased it, as well as its water miscibility and lack of requirement for heat or solvents during processing / embedding) but I been unable to find any reference to its use in this way in the scientific literature, or on the net in general, so far. Can anybody give me any suggestions at all as what I might try, or how I might be able to find useful information on immunos with glycol methacrylate in general or Technovit 8100 in particular? I did see recently an archival Histonet posting on "etching" of glycol methacrylate sections using a cured (ie aged) NaOH solution - does anybody have an opinion on this? Many thanks, Jason Palmer Bernard O'Brien Institute of Microsurgery 42 Fitzroy St, Fitzroy Victoria 3065 Australia tel +61 3 9288 4018 fax +61 3 9416 0926 email: palmerj@svhm.org.au -------------- next part -------------- Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. From failm <@t> musc.edu Thu Aug 12 06:15:10 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Immunofluorescence/slide storage Message-ID: I have several questions related to storage of slides stained for immunofluorescence. I do know they should be viewed as soon after staining as possible, but it is not always possible for our pathologist to do so. How long can they be stored without loss of reactivity? Do you store them in the fridge? After storage for several days, do you see more background, less background? After storage does your slide appear dirty when viewing microscopically? Thank you Rena Fail From MAUGER <@t> email.chop.edu Thu Aug 12 08:43:29 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Caveolin 3 Message-ID: Jean, I use caveolin 3 from BD Transduction Labs, #610420. A dilution of 1:1000 for 1 hr inc. works well. I use an anti mouse fitc secondary for muscles at 1:100 for 1 hr. Jo >>> "Mitchell (Jean)" 08/11/04 05:34PM >>> Anyone staining for caveolin 3 in muscle & have a good source for this antibody? Any & all input would be greatly appreciated! Thanks much! Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dmikita <@t> wmcnet.org Thu Aug 12 09:00:07 2004 From: dmikita <@t> wmcnet.org (Daryl Mikita) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Problems with VIP 5 Message-ID: Hello, We have had our VIP 5 for about a year now, and have not had any problems with it (functional) at all. The only problem we have found is that the counter for the amount of times each solution is used doesn't keep track of the last paraffin correctly. We only use the first two baths of paraffin and the # 12 paraffin count is usually not correct. The machine processes correctly and the tissue is fine. If we would have to buy a new machine again, we would buy the VIP 5 again, with no second thoughts. TTYL, Daryl Mikita From cmrgalle <@t> usc.es Thu Aug 12 09:03:22 2004 From: cmrgalle <@t> usc.es (Rosalia Gallego Gomez) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] immunofluorescenceslide storage Message-ID: <411B78A9.D6CE573D@usc.es> We always store immunofluorescence stained slides at -20?C in dark box and we have found nearly no loss of fluorescence in weeks. Rosal?a Gallego From gcallis <@t> montana.edu Thu Aug 12 10:13:55 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] Repy on immunostaining with Technovit 8100 / glycol methacrylate In-Reply-To: References: Message-ID: <6.0.0.22.1.20040812083750.01b10de0@gemini.msu.montana.edu> GMA is not a very ideal embedding media for immunohistochemistry, you would be far better off with paraffin using this antibody. GMA cannot be removed once polymerized nor sodium ethoxide (generally reserved for electron microscopy resins) etched with much success, you would be better off embedding in methylmethacrylate and remove the plastic entirely. This has been done with great success by Neil Hand (he has publications) using warm xylene, but he used stringent pressure cooker retrieval and worked with human tissue. The problem with GMA is it prevents the immunoglobulins from reaching antigenic sites, as the GMA is hydrophobic plus it can't be removed once polymerized. It will soften in the presence of water. Also, as GMA polymerizes it becomes very hot due to exothermic reaction unless you control this temperature by letting your blocks polymerize on ice in a refrigerator?? GMA with IHC problems have been discussed at length on Histonet many times, go to Histonet archives and search at www.histosearch.org. Personally, I don't think the product "suggestion" is correct, as it only "suggests" but does not say it WILL work. That is probably the reason you don't find it in the literature! Many people have experienced failure of IHC on GMA embedded tissues. I think some have had success with immunofluroescence using immunoglobulins and not a lot of antibodies either. It was a tedious stringent protocol described in a symposium talk. I think you will find in Histonet commentary, that most people attempting IHC/GMA suggest going to another embedding media. Also, CD68 ED1 is a Mouse antiRat, rather than a rat antiMouse (clone FA-11). ED1 and other rat macrophage markers, ED2, ED3, have been discussed on Histonet as FFPE tissues including retrieval for IHC. The staining was very straightforward as was retrieval described and solvents used plus heat of paraffin processing did not damage antigen. ED1, when used in on rat tissue, works well on paraffin sections, although we prefer frozen sections to avoid all aldehyde fixation and no retrieval. When we do frozen sections, the ED1 is diluted out 1:3000 or so, it will not be so dilute for paraffin sections and we detected with secondary to mouse IgG1 isotype (Southern Biotechnology) SA-HRP, and AEC chromogen. Staining pattern is spectacular in a rat spleen, normal positive control. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 At 10:04 PM 8/11/2004, you wrote: >Hi Histonetters. > >I'm trying to immunostain some sections of rat tissue embedded in >Technovit 8100 (a glycol methacrylate) with an ED1 antibody (Serotec rat >anti mouse), and have had no success to this point. Have tried various >antibody concentrations and antigen retrievals including pronase, trypsin >(which we use successfully for immunostaining with the same Ab on >paraffin-embedded tissues) and high temperature citric acid buffer >retrieval. Our samples are paraformaldehyde fixed overnight, and we use a >labelled streptavidin-biotin system. The technical information that comes >with the product suggests it is suitable for immunohistochemistry (and >this is why we purchased it, as well as its water miscibility and lack of >requirement for heat or solvents during processing / embedding) but I been >unable to find any reference to its use in this way in the scientific >literature, or on the net in general, so far. Can anybody give me any >suggestions at all as what I might try, or how I might be able to find >useful information on immunos with glycol methacrylate in general or >Technovit 8100 in particular? I did see recently an archival Histonet >posting on "etching" of glycol methacrylate sections using a cured (ie >aged) NaOH solution - does anybody have an opinion on this? > >Many thanks, > >Jason Palmer > >Bernard O'Brien Institute of Microsurgery >42 Fitzroy St, Fitzroy Victoria 3065 >Australia >tel +61 3 9288 4018 >fax +61 3 9416 0926 >email: palmerj@svhm.org.au > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From hej01 <@t> health.state.ny.us Thu Aug 12 10:38:05 2004 From: hej01 <@t> health.state.ny.us (Helen E Johnson) Date: Fri Sep 16 15:23:52 2005 Subject: [Histonet] special stain Message-ID: I'm looking for protocols for FluoroJade stain and Cupric Silver stain. From JWEEMS <@t> sjha.org Thu Aug 12 10:39:51 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Artisan Stainer Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A62DB@sjhaexc02.sjha.org> Hi Gang, Anyone using this instrument? I'd like to hear from those who are. Thanks, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From DDittus787 <@t> aol.com Thu Aug 12 10:45:15 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Artisan Stainer Message-ID: <393C6805.3C06A542.0A1F969F@aol.com> joyce: we were the first site on the east coast to get the artisan, we have 2 instruments, they produce gorgeous stains, our volume increased and the machines handle well, eay to use, good quality,have saved our butts when people out on vacation and sick. can't say enough good about them. dana From gcallis <@t> montana.edu Thu Aug 12 10:56:19 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Long discussion on Immunofluorescence/slide storage In-Reply-To: References: Message-ID: <6.0.0.22.1.20040812091449.01b135b8@gemini.msu.montana.edu> Rena, We do a great deal of immunofluorescent (IFA) and GFP work for both fluorescent and confocal laser scanning microscopes. At 05:15 AM 8/12/2004, you wrote: > I have several questions related to storage of slides stained for > immunofluorescence. I do know they should be viewed as soon after > staining as possible, but it is not always possible for our pathologist > to do so. > How long can they be stored without loss of reactivity? **Preferably, look at them the same day and depends on what fluorophore you are using. Some fluorophores will fade faster than others, FITC photobleaches faster than Alexa 488. Rhodamines come in several derivatives, rhodamine (TRITC) photobleaches faster than Rhodamine RX or RRX from Jackson Immunoresearch. We now use Alexa fluorophores as much as possible, due to less photobleaching plus extremely bright - or use a Strepavidin Alexa conjugate with biotinylated primary or secondary. ****Hopefully IFA staining is done under cover of a dark towel or foiled staining chamber and NEVER on an autostainer where you cannot protect fluorophore from light. Don't start photobleaching before viewing or storage! > Do you store them in the fridge? **You can. We have stored them in slide cardboard folder, in dark and on occasion in a freezer. The latter is not popular since slides must come room temperature for viewing. We often look at them the next day, try not to prolong this time too much - expensive in terms of time and material, plus painful to lose fluorescent signal. The biggest help is use antifade mounting media that reduces loss of fluorescence aka photobleaching. Molecular Probes, Prolong Gold ready to use is ideal for this purpose. It must be stored in -20C freezer with dessicant, thaw (in my pocket!), use, then return to freezer after use, but the benefits outweigh handling and fluorescence loss. You must seal edges of coverslip with diluted permanent mounting media (dilute with toluene or xylene to consistency of thin top coat nail polish, but do NOT use nail polish itself.) This media is pricey, but worth its weight in "gold". It sets up hard and since it is protective, sections can be viewed again. Sections still need protection from all light - never on an open slide tray. Go to www.probes.com/pl/prolong and look at the chart on fluorophores with %loss using other media and Prolong Gold. It also works with GFP nicely. **Vector has VectaShield Hard Set that is supposed to work also. It is nice to have an aqueous mounting media for fluorescent work that sets up hard!! > After storage for several days, do you see more background, less > background? No more background than before, there are different reasons for background, do you mean autofluorescence? or background from staining? If the fluorophore photobleaches (fades) yes the autoflourescence background would be more apparent. Preventing background due to crossreaction of immunoglobulins to tissues, or aldehyde fixation is possible. Proper blocking, dilutions, etc are part of this. > After storage does your slide appear dirty when viewing microscopically? **No, if you have a dirty looking slide (I do not know what you mean exactly, particles that sit on a section and fluoresce? If the latter happens (glowing garbage!) then spin a diluted fluorophore conjugated antibody just before application and take aliquot off top and apply that to section. Antibodies conjugated to fluorophores, even sitting around in refrigerator OR when thawed, have protein aggregates (sp?) with fluorohore attached, and this bright junk will sit around on top of section - microcentrifuging briefly eliminates this problem. If you do what we do, mount a coverslip on a slide taken from buffer, the buffer salts sometimes leave a tidge cloudy film on back of slide. After coverslipping, wipe back with kimwipe damp with water - takes off buffer salts, all is clear. Last suggestion, go to Molecular Probes and get their Handbook - a wonderful, educational, free book on handling and storage of fluorophores, and multitidbits on fluorescence related products. I believe you can download it too. Oh my, what a long discussion, been there - done it too many times Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From lkbauer <@t> unmc.edu Thu Aug 12 10:56:56 2004 From: lkbauer <@t> unmc.edu (lkbauer@unmc.edu) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Murine ISH Message-ID: Does anyone have a good protocol for mouse embryo ISH. We are open to working with either paraffin or frozen sections. I promise not to flame anyone as I am a novice and just appriciate all the shared knowledge from those of you who have been around the block. Linda(Lin) Bauer 985455 Nebraska Medical Center Omaha, NE 68198-5455 phone: (402) 559-2863 fax: (402) 559-4001 email: lkbauer@unmc.edu From browning <@t> HHSC.CA Thu Aug 12 11:10:23 2004 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Caveolin 3 Message-ID: <3AADFB88753AD31189C100902786B91C0E27839D@hch_nt_exchange.hhsc.ca> Caveolin 3 from Transduction, 1/100 for 1 hour, LSAB detection. -----Original Message----- From: Mitchell (Jean) [mailto:jmitchell@neurology.wisc.edu] Sent: Wednesday, August 11, 2004 5:35 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Caveolin 3 Anyone staining for caveolin 3 in muscle & have a good source for this antibody? Any & all input would be greatly appreciated! Thanks much! Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From Barbara_Lentz <@t> dahlchase.com Thu Aug 12 11:15:44 2004 From: Barbara_Lentz <@t> dahlchase.com (Barbara Lentz) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Artisan Stainer Message-ID: We have been using the Artisan for about 5 years now. Anything in particular you'd like to know? Barb >>> "Weems, Joyce" 08/12/04 11:39AM >>> Hi Gang, Anyone using this instrument? I'd like to hear from those who are. Thanks, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From WWmn916 <@t> aol.com Thu Aug 12 11:16:06 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Re: Artisan stainer Message-ID: <14.30a6c089.2e4cf1c6@aol.com> Hello everyone, I'm interested in more feedback from the Artisan stainer too. Mostly, because in my very limited experience with it, it was disappointing. We seemed to have a problem with precipitation on everything we ran. Then other times, the stains came out beautifully. I made sure we did valve rinses after every run, but still couldn't problem solve to any reliability with the stains. I wanted so much for this machine to work, but decided in the end it wasn't worth putting so much time into problem solving precipitated slides (especially on busy, stressful days!) I feel there is a "missing link" in my experience, because more often than not, I see histonet posting about how much others in histoland love the Artisan. Deb King, HT(ASCP) Sacramento, CA From failm <@t> musc.edu Thu Aug 12 11:15:50 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Re: Long discussion on Immunofluorescence/slide storage Message-ID: Gayle, We cover the autostainer mindful the slides should not be exposed to light. We coverslip from water because of the salts in the buffers. The refrigerator light is burnt out and I refuse to replace it. (because of IF slides) It isn't trash on the slides exactly, it a light very light cloudiness. I'll check out molecular probes Thank you Rena >>> Gayle Callis 08/12/04 11:56AM >>> Rena, We do a great deal of immunofluorescent (IFA) and GFP work for both fluorescent and confocal laser scanning microscopes. At 05:15 AM 8/12/2004, you wrote: > I have several questions related to storage of slides stained for > immunofluorescence. I do know they should be viewed as soon after > staining as possible, but it is not always possible for our pathologist > to do so. > How long can they be stored without loss of reactivity? **Preferably, look at them the same day and depends on what fluorophore you are using. Some fluorophores will fade faster than others, FITC photobleaches faster than Alexa 488. Rhodamines come in several derivatives, rhodamine (TRITC) photobleaches faster than Rhodamine RX or RRX from Jackson Immunoresearch. We now use Alexa fluorophores as much as possible, due to less photobleaching plus extremely bright - or use a Strepavidin Alexa conjugate with biotinylated primary or secondary. ****Hopefully IFA staining is done under cover of a dark towel or foiled staining chamber and NEVER on an autostainer where you cannot protect fluorophore from light. Don't start photobleaching before viewing or storage! > Do you store them in the fridge? **You can. We have stored them in slide cardboard folder, in dark and on occasion in a freezer. The latter is not popular since slides must come room temperature for viewing. We often look at them the next day, try not to prolong this time too much - expensive in terms of time and material, plus painful to lose fluorescent signal. The biggest help is use antifade mounting media that reduces loss of fluorescence aka photobleaching. Molecular Probes, Prolong Gold ready to use is ideal for this purpose. It must be stored in -20C freezer with dessicant, thaw (in my pocket!), use, then return to freezer after use, but the benefits outweigh handling and fluorescence loss. You must seal edges of coverslip with diluted permanent mounting media (dilute with toluene or xylene to consistency of thin top coat nail polish, but do NOT use nail polish itself.) This media is pricey, but worth its weight in "gold". It sets up hard and since it is protective, sections can be viewed again. Sections still need protection from all light - never on an open slide tray. Go to www.probes.com/pl/prolong and look at the chart on fluorophores with %loss using other media and Prolong Gold. It also works with GFP nicely. **Vector has VectaShield Hard Set that is supposed to work also. It is nice to have an aqueous mounting media for fluorescent work that sets up hard!! > After storage for several days, do you see more background, less > background? No more background than before, there are different reasons for background, do you mean autofluorescence? or background from staining? If the fluorophore photobleaches (fades) yes the autoflourescence background would be more apparent. Preventing background due to crossreaction of immunoglobulins to tissues, or aldehyde fixation is possible. Proper blocking, dilutions, etc are part of this. > After storage does your slide appear dirty when viewing microscopically? **No, if you have a dirty looking slide (I do not know what you mean exactly, particles that sit on a section and fluoresce? If the latter happens (glowing garbage!) then spin a diluted fluorophore conjugated antibody just before application and take aliquot off top and apply that to section. Antibodies conjugated to fluorophores, even sitting around in refrigerator OR when thawed, have protein aggregates (sp?) with fluorohore attached, and this bright junk will sit around on top of section - microcentrifuging briefly eliminates this problem. If you do what we do, mount a coverslip on a slide taken from buffer, the buffer salts sometimes leave a tidge cloudy film on back of slide. After coverslipping, wipe back with kimwipe damp with water - takes off buffer salts, all is clear. Last suggestion, go to Molecular Probes and get their Handbook - a wonderful, educational, free book on handling and storage of fluorophores, and multitidbits on fluorescence related products. I believe you can download it too. Oh my, what a long discussion, been there - done it too many times Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From dcrippen <@t> buckinstitute.org Thu Aug 12 11:18:33 2004 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Long discussion on Immunofluorescence/slide storage Message-ID: <4AA34A707932424EBE2D973764D226A901D757B4@inverness.buckcenter.org> Hi Gayle, In terms of sealing the coverslips with permanent mounting media--do you mean something like permount? Also, can you explain why you advise NOT using nail polish itself?? Many thanks!! Danielle Crippen Morphology Core Manager Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen@buckinstitute.org -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Thursday, August 12, 2004 8:56 AM To: Mildred Fail; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Long discussion on Immunofluorescence/slide storage Rena, We do a great deal of immunofluorescent (IFA) and GFP work for both fluorescent and confocal laser scanning microscopes. At 05:15 AM 8/12/2004, you wrote: > I have several questions related to storage of slides stained for > immunofluorescence. I do know they should be viewed as soon after > staining as possible, but it is not always possible for our pathologist > to do so. > How long can they be stored without loss of reactivity? **Preferably, look at them the same day and depends on what fluorophore you are using. Some fluorophores will fade faster than others, FITC photobleaches faster than Alexa 488. Rhodamines come in several derivatives, rhodamine (TRITC) photobleaches faster than Rhodamine RX or RRX from Jackson Immunoresearch. We now use Alexa fluorophores as much as possible, due to less photobleaching plus extremely bright - or use a Strepavidin Alexa conjugate with biotinylated primary or secondary. ****Hopefully IFA staining is done under cover of a dark towel or foiled staining chamber and NEVER on an autostainer where you cannot protect fluorophore from light. Don't start photobleaching before viewing or storage! > Do you store them in the fridge? **You can. We have stored them in slide cardboard folder, in dark and on occasion in a freezer. The latter is not popular since slides must come room temperature for viewing. We often look at them the next day, try not to prolong this time too much - expensive in terms of time and material, plus painful to lose fluorescent signal. The biggest help is use antifade mounting media that reduces loss of fluorescence aka photobleaching. Molecular Probes, Prolong Gold ready to use is ideal for this purpose. It must be stored in -20C freezer with dessicant, thaw (in my pocket!), use, then return to freezer after use, but the benefits outweigh handling and fluorescence loss. You must seal edges of coverslip with diluted permanent mounting media (dilute with toluene or xylene to consistency of thin top coat nail polish, but do NOT use nail polish itself.) This media is pricey, but worth its weight in "gold". It sets up hard and since it is protective, sections can be viewed again. Sections still need protection from all light - never on an open slide tray. Go to www.probes.com/pl/prolong and look at the chart on fluorophores with %loss using other media and Prolong Gold. It also works with GFP nicely. **Vector has VectaShield Hard Set that is supposed to work also. It is nice to have an aqueous mounting media for fluorescent work that sets up hard!! > After storage for several days, do you see more background, less > background? No more background than before, there are different reasons for background, do you mean autofluorescence? or background from staining? If the fluorophore photobleaches (fades) yes the autoflourescence background would be more apparent. Preventing background due to crossreaction of immunoglobulins to tissues, or aldehyde fixation is possible. Proper blocking, dilutions, etc are part of this. > After storage does your slide appear dirty when viewing microscopically? **No, if you have a dirty looking slide (I do not know what you mean exactly, particles that sit on a section and fluoresce? If the latter happens (glowing garbage!) then spin a diluted fluorophore conjugated antibody just before application and take aliquot off top and apply that to section. Antibodies conjugated to fluorophores, even sitting around in refrigerator OR when thawed, have protein aggregates (sp?) with fluorohore attached, and this bright junk will sit around on top of section - microcentrifuging briefly eliminates this problem. If you do what we do, mount a coverslip on a slide taken from buffer, the buffer salts sometimes leave a tidge cloudy film on back of slide. After coverslipping, wipe back with kimwipe damp with water - takes off buffer salts, all is clear. Last suggestion, go to Molecular Probes and get their Handbook - a wonderful, educational, free book on handling and storage of fluorophores, and multitidbits on fluorescence related products. I believe you can download it too. Oh my, what a long discussion, been there - done it too many times Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DDittus787 <@t> aol.com Thu Aug 12 11:20:34 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Re: Artisan stainer Message-ID: <56B01023.3F81BAD1.0A1F969F@aol.com> deb: It is rare to have precipatate on the slides, i have found that etoh quality and good reagent grade water make the difference.our distilled water(that we made wash buffer with) had bacteria in it ,so our stains had a problem, not a machine issue. However since we fixed water source, things are beautiful. Dana From Barbara_Lentz <@t> dahlchase.com Thu Aug 12 11:42:31 2004 From: Barbara_Lentz <@t> dahlchase.com (Barbara Lentz) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Re: Artisan stainer Message-ID: The only "precipitation" we've had on our slides seems to come from the silver stains. I've tweaked the procedures to decrease time in the silver reagent and have been rather successful in eliminating the precip. Other times, when the slides appeared "dirty" on all the slides from a particular run, we found there was a blockage in the line from the bulk fluids (this was noticed when the inventory showed a great decrease in the fluid level, however, the actual fluid level was significantly higher. We instituted a few maintenance measures: twice a month, we run 95% ETOH through all the lines and once a month, emptying the bulk fluids and bleaching them (including the lids). This has also cut down on the fungal contamination we also experienced. We also experienced a few bad lots of reagents--they leaked excessively (left puddles). Dako was very good about replacing the kits. We have been looking for another stain system to replace the Artisan, since, after Dako took over, they do not reimburse for expired reagent kits. >>> 08/12/04 12:16PM >>> Hello everyone, I'm interested in more feedback from the Artisan stainer too. Mostly, because in my very limited experience with it, it was disappointing. We seemed to have a problem with precipitation on everything we ran. Then other times, the stains came out beautifully. I made sure we did valve rinses after every run, but still couldn't problem solve to any reliability with the stains. I wanted so much for this machine to work, but decided in the end it wasn't worth putting so much time into problem solving precipitated slides (especially on busy, stressful days!) I feel there is a "missing link" in my experience, because more often than not, I see histonet posting about how much others in histoland love the Artisan. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Thu Aug 12 11:51:48 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Artisan Stainer Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A62E0@sjhaexc02.sjha.org> I'm trying to justify for budget purposes. Did you justify increase in reagent cost? thanks, J -----Original Message----- From: Barbara Lentz [mailto:Barbara_Lentz@dahlchase.com] Sent: Thursday, August 12, 2004 12:16 PM To: histonet@lists.utsouthwestern.edu; Weems, Joyce Subject: Re: [Histonet] Artisan Stainer We have been using the Artisan for about 5 years now. Anything in particular you'd like to know? Barb >>> "Weems, Joyce" 08/12/04 11:39AM >>> Hi Gang, Anyone using this instrument? I'd like to hear from those who are. Thanks, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From gcallis <@t> montana.edu Thu Aug 12 11:52:01 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Nail polish seal vs diluted permanent mounting media) Why's, another freebie In-Reply-To: <4AA34A707932424EBE2D973764D226A901D757B4@inverness.buckcen ter.org> References: <4AA34A707932424EBE2D973764D226A901D757B4@inverness.buckcenter.org> Message-ID: <6.0.0.22.1.20040812102730.01b455a8@gemini.msu.montana.edu> Danielle, Yes, Permount or similar permanent media. Just use the same solvent e.g. toluene or xylene already contained in media. Nail polish contains isopropyl alcohol which leaches into aqueous mounting media under coverslip and/or PBS (we have used this). Nail polish avoidance is reported in literature (Science, GFP publication) Toluene and xylene are NOT water miscible, don't leach into aqueous medias under coverslip. Alcohol causes GFP protein and maybe other fluorophores(?) to not fluoresce. Another mounting media reported in literature that works (not sure how much photobleaching prevention it is capable of) is Mowiol. Molecular Probes has a comparison chart of several aqueous mounting medias showing which are superior for preserving fluorescence - very enlightening. Molecular Probes suggests paraffin sealing, NOT fun, ultra-messy, and if you use inverted fluorescent microscope with confocal set up or other wise, objectives get slimed with paraffin! MP said to avoid nail polish with Prolong Gold, but is may be because people use this for GFP work. Clontech (GFP gurus) suggests using rubber cement (ooey gooey messy too!), and suggested solvent diluted permanent mounting media. Another hint on free literature. Clontech website, go to literature and download Living Colours Manual -this freebie has tons of info on GFP, mounting media, fixation, and autofluorescence. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman >In terms of sealing the coverslips with permanent mounting media--do you >mean something like permount? Also, can you explain why you advise NOT >using nail polish itself?? > >Many thanks!! > >Danielle Crippen >Morphology Core Manager >Buck Institute for Age Research >8001 Redwood Blvd. >Novato, CA 94945 >415-209-2046 >dcrippen@buckinstitute.org > > >-----Original Message----- >From: Gayle Callis [mailto:gcallis@montana.edu] >Sent: Thursday, August 12, 2004 8:56 AM >To: Mildred Fail; Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Long discussion on Immunofluorescence/slide storage > > >Rena, > >We do a great deal of immunofluorescent (IFA) and GFP work for both >fluorescent and confocal laser scanning microscopes. > >At 05:15 AM 8/12/2004, you wrote: > > I have several questions related to storage of slides stained for > > immunofluorescence. I do know they should be viewed as soon after > > staining as possible, but it is not always possible for our pathologist > > to do so. > > How long can they be stored without loss of reactivity? > >**Preferably, look at them the same day and depends on what fluorophore you >are using. Some fluorophores will fade faster than others, FITC >photobleaches faster than Alexa 488. Rhodamines come in several >derivatives, rhodamine (TRITC) photobleaches faster than Rhodamine RX or >RRX from Jackson Immunoresearch. We now use Alexa fluorophores as much as >possible, due to less photobleaching plus extremely bright - or use a >Strepavidin Alexa conjugate with biotinylated primary or secondary. > >****Hopefully IFA staining is done under cover of a dark towel or foiled >staining chamber and NEVER on an autostainer where you cannot protect >fluorophore from light. Don't start photobleaching before viewing or storage! > > > Do you store them in the fridge? > > **You can. We have stored them in slide cardboard folder, in dark and on >occasion in a freezer. The latter is not popular since slides must come >room temperature for viewing. We often look at them the next day, try not >to prolong this time too much - expensive in terms of time and material, >plus painful to lose fluorescent signal. > >The biggest help is use antifade mounting media that reduces loss of >fluorescence aka photobleaching. Molecular Probes, Prolong Gold ready to >use is ideal for this purpose. It must be stored in -20C freezer with >dessicant, thaw (in my pocket!), use, then return to freezer after use, >but the benefits outweigh handling and fluorescence loss. You must seal >edges of coverslip with diluted permanent mounting media (dilute with >toluene or xylene to consistency of thin top coat nail polish, but do NOT >use nail polish itself.) This media is pricey, but worth its weight in >"gold". It sets up hard and since it is protective, sections can be viewed >again. Sections still need protection from all light - never on an open >slide tray. Go to www.probes.com/pl/prolong and look at the chart on >fluorophores with %loss using other media and Prolong Gold. It also works >with GFP nicely. > >**Vector has VectaShield Hard Set that is supposed to work also. It is >nice to have an aqueous mounting media for fluorescent work that sets up >hard!! > > > After storage for several days, do you see more background, less > > background? > >No more background than before, there are different reasons for background, >do you mean autofluorescence? or background from staining? If the >fluorophore photobleaches (fades) yes the autoflourescence background would >be more apparent. Preventing background due to crossreaction of >immunoglobulins to tissues, or aldehyde fixation is possible. Proper >blocking, dilutions, etc are part of this. > > > After storage does your slide appear dirty when viewing microscopically? >**No, if you have a dirty looking slide (I do not know what you mean >exactly, particles that sit on a section and fluoresce? If the latter >happens (glowing garbage!) then spin a diluted fluorophore conjugated >antibody just before application and take aliquot off top and apply that to >section. Antibodies conjugated to fluorophores, even sitting around in >refrigerator OR when thawed, have protein aggregates (sp?) with fluorohore >attached, and this bright junk will sit around on top of section - >microcentrifuging briefly eliminates this problem. > >If you do what we do, mount a coverslip on a slide taken from buffer, the >buffer salts sometimes leave a tidge cloudy film on back of slide. After >coverslipping, wipe back with kimwipe damp with water - takes off buffer >salts, all is clear. > >Last suggestion, go to Molecular Probes and get their Handbook - a >wonderful, educational, free book on handling and storage of fluorophores, >and multitidbits on fluorescence related products. I believe you can >download it too. > >Oh my, what a long discussion, been there - done it too many times > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barbara_Lentz <@t> dahlchase.com Thu Aug 12 12:27:36 2004 From: Barbara_Lentz <@t> dahlchase.com (Barbara Lentz) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Artisan Stainer Message-ID: At the time we were looking at automated stainers, we tried the Biogenex, the Artisan and the Ventana. We were not happy with the silver stains of the Biogenex and Ventana, so cost did not factor in too heavily. At the time, when the Artisan belonged to Cytologix, when reagent kits expired, the company would reimburse for unused tests. Also at that time, each kit was good for 50 tests. This was cost effective. Now that Dako merged with Cytologix, they no longer reimburse and they have increased the tests per kit to 100. We are now doing some tests manually, since we do not do enough of these to justify buying kits that expire with 50 to 95 tests left in the kit. We have recently revisited Ventana (do not like that it takes so many reagents for some stains and that it is not very efficient, time-wise) and will be looking at the Biogenex again. Barb >>> "Weems, Joyce" 08/12/04 12:51PM >>> I'm trying to justify for budget purposes. Did you justify increase in reagent cost? thanks, J -----Original Message----- From: Barbara Lentz [mailto:Barbara_Lentz@dahlchase.com] Sent: Thursday, August 12, 2004 12:16 PM To: histonet@lists.utsouthwestern.edu; Weems, Joyce Subject: Re: [Histonet] Artisan Stainer We have been using the Artisan for about 5 years now. Anything in particular you'd like to know? Barb >>> "Weems, Joyce" 08/12/04 11:39AM >>> Hi Gang, Anyone using this instrument? I'd like to hear from those who are. Thanks, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From kgrobert <@t> rci.rutgers.edu Thu Aug 12 12:53:16 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Long discussion on Immunofluorescence/slide storage In-Reply-To: <6.0.0.22.1.20040812091449.01b135b8@gemini.msu.montana.edu> References: <6.0.0.22.1.20040812091449.01b135b8@gemini.msu.montana.edu> Message-ID: <411BAE8C.1050800@rci.rutgers.edu> Here's a question: has anybody tried those Quantum Dot antibody or streptavidin conjugates? Supposedly this stuff DOESN'T fade, ever. I'm looking into it, but have not tried them yet. www.qdots.com, in case anybody is interested. Kathleen Roberts Principal Lab Technician Rutgers University Gayle Callis wrote: > Rena, > > We do a great deal of immunofluorescent (IFA) and GFP work for both > fluorescent and confocal laser scanning microscopes. > > At 05:15 AM 8/12/2004, you wrote: > >> I have several questions related to storage of slides stained for >> immunofluorescence. I do know they should be viewed as soon after >> staining as possible, but it is not always possible for our >> pathologist to do so. >> How long can they be stored without loss of reactivity? > > > **Preferably, look at them the same day and depends on what > fluorophore you are using. Some fluorophores will fade faster than > others, FITC photobleaches faster than Alexa 488. Rhodamines come in > several derivatives, rhodamine (TRITC) photobleaches faster than > Rhodamine RX or RRX from Jackson Immunoresearch. We now use Alexa > fluorophores as much as possible, due to less photobleaching plus > extremely bright - or use a Strepavidin Alexa conjugate with > biotinylated primary or secondary. > > ****Hopefully IFA staining is done under cover of a dark towel or > foiled staining chamber and NEVER on an autostainer where you cannot > protect fluorophore from light. Don't start photobleaching before > viewing or storage! > >> Do you store them in the fridge? > > > **You can. We have stored them in slide cardboard folder, in dark > and on occasion in a freezer. The latter is not popular since slides > must come room temperature for viewing. We often look at them the > next day, try not to prolong this time too much - expensive in terms > of time and material, plus painful to lose fluorescent signal. > > The biggest help is use antifade mounting media that reduces loss of > fluorescence aka photobleaching. Molecular Probes, Prolong Gold > ready to use is ideal for this purpose. It must be stored in -20C > freezer with dessicant, thaw (in my pocket!), use, then return to > freezer after use, but the benefits outweigh handling and fluorescence > loss. You must seal edges of coverslip with diluted permanent mounting > media (dilute with toluene or xylene to consistency of thin top coat > nail polish, but do NOT use nail polish itself.) This media is > pricey, but worth its weight in "gold". It sets up hard and since it > is protective, sections can be viewed again. Sections still need > protection from all light - never on an open slide tray. Go to > www.probes.com/pl/prolong and look at the chart on fluorophores with > %loss using other media and Prolong Gold. It also works with GFP nicely. > > **Vector has VectaShield Hard Set that is supposed to work also. It > is nice to have an aqueous mounting media for fluorescent work that > sets up hard!! > >> After storage for several days, do you see more background, less >> background? > > > No more background than before, there are different reasons for > background, do you mean autofluorescence? or background from > staining? If the fluorophore photobleaches (fades) yes the > autoflourescence background would be more apparent. Preventing > background due to crossreaction of immunoglobulins to tissues, or > aldehyde fixation is possible. Proper blocking, dilutions, etc are > part of this. > >> After storage does your slide appear dirty when viewing >> microscopically? > > **No, if you have a dirty looking slide (I do not know what you mean > exactly, particles that sit on a section and fluoresce? If the latter > happens (glowing garbage!) then spin a diluted fluorophore conjugated > antibody just before application and take aliquot off top and apply > that to section. Antibodies conjugated to fluorophores, even sitting > around in refrigerator OR when thawed, have protein aggregates (sp?) > with fluorohore attached, and this bright junk will sit around on top > of section - microcentrifuging briefly eliminates this problem. > > If you do what we do, mount a coverslip on a slide taken from buffer, > the buffer salts sometimes leave a tidge cloudy film on back of > slide. After coverslipping, wipe back with kimwipe damp with water - > takes off buffer salts, all is clear. > > Last suggestion, go to Molecular Probes and get their Handbook - a > wonderful, educational, free book on handling and storage of > fluorophores, and multitidbits on fluorescence related products. I > believe you can download it too. > > Oh my, what a long discussion, been there - done it too many times > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ihc <@t> unipathllc.com Thu Aug 12 12:56:08 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] HTL Practical-Mallory's PTAH Message-ID: <001601c48095$a64d4220$4500a8c0@unipath02> Okay, so here's my question. Is there a secret to getting a decent section of longitudinal muscle? I fixed my fresh muscle in Zenker's fluid for about 18 hrs., washed in many changes of water for about 6 hrs., and processed starting in 70% alcohol (skipping formalin). I embedded the pieces at a 45 degree angle. The sections I'm getting are terrible, all shredded and torn. I've tried soaking on a wet ice cube and soaking in ammonia water, but neither seems to help. Any suggestions would be greatly appreciated! Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO From gcallis <@t> montana.edu Thu Aug 12 12:58:57 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] QDots aka nanocrystals In-Reply-To: <411BAE8C.1050800@rci.rutgers.edu> References: <6.0.0.22.1.20040812091449.01b135b8@gemini.msu.montana.edu> <411BAE8C.1050800@rci.rutgers.edu> Message-ID: <6.0.0.22.1.20040812115000.01b2bfd8@gemini.msu.montana.edu> Kathleen, Would love to work with them, bet they are pricey although I am twisting arms hard! The initial work with breast cancer markers looked interesting, and I have heard buffer and pH requirements were picky (this is in past tense since I viewed staining protocol on website). BE sure to use Strepavidin/biotin blocking, from VECTOR. The joy of these nanocrystals is that they do not photobleach and are extremely bright. If you do work with them let us in on results, and good luck! There is a confocal listserver out of Buffalo, easy subscribing, etc. People who do confocal work are beginning to use these more and more, there will be questions/answers like Histonet on this listserver - a good place to see what is going on Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Rcartun <@t> harthosp.org Thu Aug 12 13:08:42 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Contact info for DIFCO Labs Message-ID: I realize this is outside the realm of histology, but does anyone know how to contact DIFCO Laboratories (a supplier of Microbiology products). I'm sure they have been taken-over or bought-out by someone else. Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From ihc <@t> unipathllc.com Thu Aug 12 13:13:00 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Mallory's PTAH Message-ID: <000001c48098$00735800$4500a8c0@unipath02> I actually already tried to use formalin-fixed muscle. You're exactly right, it sectioned great, but I could never get any decent staining. I tried post-fixing for 24 hrs. in Zenker's vs. post-fixing for 3 Hrs in Zenker's at 56 degrees. I tried all three protocols in Sheehan and the protocol in Carson. I'm at a loss for what to do next if I can't make the Zenker fixed muscle work. At least I've learned a lot about the stain. :-) Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO From ihc <@t> unipathllc.com Thu Aug 12 13:16:32 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Mallory's PTAH Message-ID: <000501c48098$7f214e50$4500a8c0@unipath02> I forgot to add- I'm cutting human muscle. I don't think its being overly dehydrated during processing, our processor runs are pretty standard. Would the dehydration steps be shorter after Zenker fixation? Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO From thewoodruffinstitute <@t> hotmail.com Thu Aug 12 13:18:40 2004 From: thewoodruffinstitute <@t> hotmail.com (Rebecca Lambert) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Mohs Histotech Message-ID: I am a Mohs surgeon in Naples, FL looking for a part time histotech with Mohs experience. If any one is interested or knows of a good resource to find a Mohs tech in this area, please let me know. 239-596-9337. Rebecca Lambert, MD _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From gcallis <@t> montana.edu Thu Aug 12 13:28:36 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Contact info for DIFCO Labs In-Reply-To: References: Message-ID: <6.0.0.22.1.20040812122250.01b18170@gemini.msu.montana.edu> Richard, They were bought by Becton Dickinson, and a search turned up a quick link to Difco & BBL Manual - BD Diagnostic Systems at BD website. Good luck. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Annette_hall <@t> pa-ucl.com Thu Aug 12 13:28:33 2004 From: Annette_hall <@t> pa-ucl.com (Annette Hall) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Contact info for DIFCO Labs Message-ID: <9FC023A4AB52BB4D87DC6456081A822C070061@mercury.pa-ucl.com> Try BD: www.difco.com Difco & BBL Manual - BD Diagnostic Systems The first edition of the Difco(tm) & BBL(tm) Manual marks the beginning of a new tradition. The new manual replaces the Difco Manual (11th edition) and the Manual of BBL Products and Laboratory Procedures (6th edition). The new Difco & BBL Manual is a comprehensive guide to our entire line of Difco and BBL brand media including dehydrated culture media (DCM) and prepared plated, tubed and bottled media. BD Diagnostic Systems 7 Loveton Circle Sparks, MD USA 21152 800.638.8663 (toll free in USA & Canada) 410.316.4000 Annette J Hall, MT Micro/Cyto/Histo Supervisor United Clinical Labs -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Thursday, August 12, 2004 1:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Contact info for DIFCO Labs I realize this is outside the realm of histology, but does anyone know how to contact DIFCO Laboratories (a supplier of Microbiology products). I'm sure they have been taken-over or bought-out by someone else. Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Thu Aug 12 14:25:25 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] DIFCO - Thank you! Message-ID: I now have the information I need. Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From JNocito <@t> Pathreflab.com Thu Aug 12 15:34:54 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this, then again, maybe not Message-ID: Afternoon Netter's, Remember about a week or two the topic was getting "flamed"? Well, here's a new one. Sometime last year, I made several postings about a certain company and their product that I used and it did not work for me. I wrote them a letter and returned all the merchandise back to them saying I was not happy with their product. I waited several months and never got a response, so I named the company on the Histonet. Since then, I have been throwing away their flyers, catalogs and any other correspondence I receive. Obviously, they are doing fine without my business and I am doing fine without their products. Today, I receive a letter from the company with a copy of my Histonet postings from over a year ago, telling me that this has been in customer service for about a year (hmmmmmmmm, I'm trying to find out where the customer service is in all of this. (One year, okay, I'll get back to that), that their customer service has tried several times to contact me but to no avail. (Hmmmmm, one way would have been to respond to my email address, but I'll come back to this also). So after all this time, they want to do business with me. (So close to the NSH meeting, hmmmmmmmmm, another point to ponder). My big beef is that these companies read this forum and try to use strong arm tactics on people who post negative comments about them or their products. I have many friends that are sales reps in this field (I'm talking like come over my house and eat type of friends) and I love these people. I must reiterate that this forum was set up to exchange ideas, theories and experiences, both positive and negative. If this was such a "hot item", point #1, why did it take a whole year to get a response when they had my company and address on file? Is this how customer service works? I don't think so!! Point #2 If you are a vendor and don't like a particular posting, why don't you reply back to that person via email. Point #3 don't wait until it's almost time for the NSH annual S/C to fix a problem, it's probably too late by then. Point #4 If you haven't had a purchase order or have not corresponded with a customer for over a year, chances are that customer does not want to do business with you. Let lying dogs lie and don't open old scars. All you did was get me going on a tangent. I know, there goes Nocito again, but I refuse to be treated like this from any one. Like they say in the movies, "Don't call me, I'll call you". Okay, I've said my piece now let the FLAMING BEGIN!!!!!!!!! As always, the opinions stated by this author in no way reflects the opinions of his employer, subordinates, friends or relatives. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX From garygill <@t> dcla.com Thu Aug 12 15:51:47 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this, t hen again, maybe not Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070459@HALIBUT.dcla.com> "eat type of friends" -- as in "dinner with" or "with dinner"? Sorry, couldn't resist. Gary Gill -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Thursday, August 12, 2004 3:35 PM To: Histonet Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] You may want to hit the delete button with this, then again, maybe not Afternoon Netter's, Remember about a week or two the topic was getting "flamed"? Well, here's a new one. Sometime last year, I made several postings about a certain company and their product that I used and it did not work for me. I wrote them a letter and returned all the merchandise back to them saying I was not happy with their product. I waited several months and never got a response, so I named the company on the Histonet. Since then, I have been throwing away their flyers, catalogs and any other correspondence I receive. Obviously, they are doing fine without my business and I am doing fine without their products. Today, I receive a letter from the company with a copy of my Histonet postings from over a year ago, telling me that this has been in customer service for about a year (hmmmmmmmm, I'm trying to find out where the customer service is in all of this. (One year, okay, I'll get back to that), that their customer service has tried several times to contact me but to no avail. (Hmmmmm, one way would have been to respond to my email address, but I'll come back to this also). So after all this time, they want to do business with me. (So close to the NSH meeting, hmmmmmmmmm, another point to ponder). My big beef is that these companies read this forum and try to use strong arm tactics on people who post negative comments about them or their products. I have many friends that are sales reps in this field (I'm talking like come over my house and eat type of friends) and I love these people. I must reiterate that this forum was set up to exchange ideas, theories and experiences, both positive and negative. If this was such a "hot item", point #1, why did it take a whole year to get a response when they had my company and address on file? Is this how customer service works? I don't think so!! Point #2 If you are a vendor and don't like a particular posting, why don't you reply back to that person via email. Point #3 don't wait until it's almost time for the NSH annual S/C to fix a problem, it's probably too late by then. Point #4 If you haven't had a purchase order or have not corresponded with a customer for over a year, chances are that customer does not want to do business with you. Let lying dogs lie and don't open old scars. All you did was get me going on a tangent. I know, there goes Nocito again, but I refuse to be treated like this from any one. Like they say in the movies, "Don't call me, I'll call you". Okay, I've said my piece now let the FLAMING BEGIN!!!!!!!!! As always, the opinions stated by this author in no way reflects the opinions of his employer, subordinates, friends or relatives. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Thu Aug 12 15:51:47 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this, t hen again, maybe not Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070459@HALIBUT.dcla.com> "eat type of friends" -- as in "dinner with" or "with dinner"? Sorry, couldn't resist. Gary Gill -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Thursday, August 12, 2004 3:35 PM To: Histonet Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] You may want to hit the delete button with this, then again, maybe not Afternoon Netter's, Remember about a week or two the topic was getting "flamed"? Well, here's a new one. Sometime last year, I made several postings about a certain company and their product that I used and it did not work for me. I wrote them a letter and returned all the merchandise back to them saying I was not happy with their product. I waited several months and never got a response, so I named the company on the Histonet. Since then, I have been throwing away their flyers, catalogs and any other correspondence I receive. Obviously, they are doing fine without my business and I am doing fine without their products. Today, I receive a letter from the company with a copy of my Histonet postings from over a year ago, telling me that this has been in customer service for about a year (hmmmmmmmm, I'm trying to find out where the customer service is in all of this. (One year, okay, I'll get back to that), that their customer service has tried several times to contact me but to no avail. (Hmmmmm, one way would have been to respond to my email address, but I'll come back to this also). So after all this time, they want to do business with me. (So close to the NSH meeting, hmmmmmmmmm, another point to ponder). My big beef is that these companies read this forum and try to use strong arm tactics on people who post negative comments about them or their products. I have many friends that are sales reps in this field (I'm talking like come over my house and eat type of friends) and I love these people. I must reiterate that this forum was set up to exchange ideas, theories and experiences, both positive and negative. If this was such a "hot item", point #1, why did it take a whole year to get a response when they had my company and address on file? Is this how customer service works? I don't think so!! Point #2 If you are a vendor and don't like a particular posting, why don't you reply back to that person via email. Point #3 don't wait until it's almost time for the NSH annual S/C to fix a problem, it's probably too late by then. Point #4 If you haven't had a purchase order or have not corresponded with a customer for over a year, chances are that customer does not want to do business with you. Let lying dogs lie and don't open old scars. All you did was get me going on a tangent. I know, there goes Nocito again, but I refuse to be treated like this from any one. Like they say in the movies, "Don't call me, I'll call you". Okay, I've said my piece now let the FLAMING BEGIN!!!!!!!!! As always, the opinions stated by this author in no way reflects the opinions of his employer, subordinates, friends or relatives. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pathrm35 <@t> adelphia.net Thu Aug 12 16:01:20 2004 From: pathrm35 <@t> adelphia.net (Ron Martin) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] proper disposal of specimen transpot biohazard bags Message-ID: <002001c480af$850dc200$5dedeb44@Pathrm35> Fellow techs, What is the proper disposal means for the specimen transport biohazard bags? The bags we use say "stat" and "biohazard" on them. I always thought that they were to be discarded in a larger biohazard bag with the other contaminated material. My manager says that they are to be discarded in the regular trash unless formalin or other fluid leaks in them. What is everyone else doing? Thanks, Ron Martin From DDittus787 <@t> aol.com Thu Aug 12 16:03:16 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this, then again, maybe not Message-ID: <1CEDD130.613A9B5A.0A1F969F@aol.com> gary- worst thing you can do is tease a guy who is mad. joe- go to it man, if you have an opinion you should express it, nobody should tell you, you can't. If a company has a problem hearing a customers truth(you might think they would learn what we want from it and fix the problems,then they could say so on Histonet and all would benefit.) I don't think any company is problem free and they seem to watch our conversations for product needs, and to make money from us, so if you want the cash then deal with it all!!!!!! my perspective only. detection kit 400.00 technical problems 1,000000's able to express ones opinion on Histonet---priceless From DDittus787 <@t> aol.com Thu Aug 12 16:03:16 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this, then again, maybe not Message-ID: <1CEDD130.613A9B5A.0A1F969F@aol.com> gary- worst thing you can do is tease a guy who is mad. joe- go to it man, if you have an opinion you should express it, nobody should tell you, you can't. If a company has a problem hearing a customers truth(you might think they would learn what we want from it and fix the problems,then they could say so on Histonet and all would benefit.) I don't think any company is problem free and they seem to watch our conversations for product needs, and to make money from us, so if you want the cash then deal with it all!!!!!! my perspective only. detection kit 400.00 technical problems 1,000000's able to express ones opinion on Histonet---priceless From tpmorken <@t> labvision.com Thu Aug 12 16:17:12 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] proper disposal of specimen transpot biohazard bag s Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2D94B7D@usca0082k08.labvision.apogent.com> Well... When working at a nameless lab many years ago we had a call from a local landfill that freaked when they found an empty biohazard bag in a load of our "regular" trash, so I don't think they like that kind of thing. Tim Morken -----Original Message----- From: Ron Martin [mailto:pathrm35@adelphia.net] Sent: Thursday, August 12, 2004 2:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] proper disposal of specimen transpot biohazard bags Fellow techs, What is the proper disposal means for the specimen transport biohazard bags? The bags we use say "stat" and "biohazard" on them. I always thought that they were to be discarded in a larger biohazard bag with the other contaminated material. My manager says that they are to be discarded in the regular trash unless formalin or other fluid leaks in them. What is everyone else doing? Thanks, Ron Martin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Thu Aug 12 16:17:46 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Fluorescence and photobleaching discussion Message-ID: Gayle writes: **Vector has VectaShield Hard Set that is supposed to work also. It is nice to have an aqueous mounting media for fluorescent work that sets up hard!! I have used this product and I don't like it. Initially I did because it is thin in consistency and spreads well. However, with time, thick specimens (12 microns) developed air pockets in some of them. Having learned this, I used more on the next batch of slides, and ended up with the same problem. Also, they state that you can easily remount if necessary by soaking in water to remove the coverslip. It took 3 days before the coverslip was loose enough to remove, and remounting wasn't really satisfactory for the coverslips that had cultured cells on it. Some of the cells adhered to the glass slide. Others in our facility use this stuff and love it. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From garygill <@t> dcla.com Thu Aug 12 16:06:12 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this, t hen again, maybe not Message-ID: <0B11620AEE5EFB4FA7A7558339972A8307045D@HALIBUT.dcla.com> Under the circumstances, I see humor as a defuser, not a detonator. Gary Gill -----Original Message----- From: DDittus787@aol.com [mailto:DDittus787@aol.com] Sent: Thursday, August 12, 2004 4:03 PM To: garygill@dcla.com; JNocito@Pathreflab.com; histonet@pathology.swmed.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] You may want to hit the delete button with this, then again, maybe not gary- worst thing you can do is tease a guy who is mad. joe- go to it man, if you have an opinion you should express it, nobody should tell you, you can't. If a company has a problem hearing a customers truth(you might think they would learn what we want from it and fix the problems,then they could say so on Histonet and all would benefit.) I don't think any company is problem free and they seem to watch our conversations for product needs, and to make money from us, so if you want the cash then deal with it all!!!!!! my perspective only. detection kit 400.00 technical problems 1,000000's able to express ones opinion on Histonet---priceless From garygill <@t> dcla.com Thu Aug 12 16:21:50 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] proper disposal of specimen transpot biohazard bag s Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070461@HALIBUT.dcla.com> "No one ever went broke underestimating the intelligence of the American people." -- H.L. Mencken Gary Gill -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Thursday, August 12, 2004 4:17 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] proper disposal of specimen transpot biohazard bag s Well... When working at a nameless lab many years ago we had a call from a local landfill that freaked when they found an empty biohazard bag in a load of our "regular" trash, so I don't think they like that kind of thing. Tim Morken -----Original Message----- From: Ron Martin [mailto:pathrm35@adelphia.net] Sent: Thursday, August 12, 2004 2:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] proper disposal of specimen transpot biohazard bags Fellow techs, What is the proper disposal means for the specimen transport biohazard bags? The bags we use say "stat" and "biohazard" on them. I always thought that they were to be discarded in a larger biohazard bag with the other contaminated material. My manager says that they are to be discarded in the regular trash unless formalin or other fluid leaks in them. What is everyone else doing? Thanks, Ron Martin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Thu Aug 12 16:47:33 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this, then again, maybe not References: <1CEDD130.613A9B5A.0A1F969F@aol.com> Message-ID: <028b01c480b5$f9c771d0$5d6bce44@yourxhtr8hvc4p> see, that's just it, I laughed at Gary's response, couldn't help it. Believe it or not, I do have a sense of humor, alibit a little off the wall, but to know me is to love me. Joe ----- Original Message ----- From: To: "Gary Gill" ; "'Joe Nocito'" ; "Histonet" Cc: Sent: Thursday, August 12, 2004 4:03 PM Subject: RE: [Histonet] You may want to hit the delete button with this,then again, maybe not > gary- worst thing you can do is tease a guy who is mad. > > joe- go to it man, if you have an opinion you should express it, nobody should tell you, you can't. If a company has a problem hearing a customers truth(you might think they would learn what we want from it and fix the problems,then they could say so on Histonet and all would benefit.) I don't think any company is problem free and they seem to watch our conversations for product needs, and to make money from us, so if you want the cash then deal with it all!!!!!! my perspective only. detection kit 400.00 > technical problems 1,000000's > able to express ones opinion on Histonet---priceless > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jnocito <@t> satx.rr.com Thu Aug 12 16:47:33 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this, then again, maybe not References: <1CEDD130.613A9B5A.0A1F969F@aol.com> Message-ID: <028b01c480b5$f9c771d0$5d6bce44@yourxhtr8hvc4p> see, that's just it, I laughed at Gary's response, couldn't help it. Believe it or not, I do have a sense of humor, alibit a little off the wall, but to know me is to love me. Joe ----- Original Message ----- From: To: "Gary Gill" ; "'Joe Nocito'" ; "Histonet" Cc: Sent: Thursday, August 12, 2004 4:03 PM Subject: RE: [Histonet] You may want to hit the delete button with this,then again, maybe not > gary- worst thing you can do is tease a guy who is mad. > > joe- go to it man, if you have an opinion you should express it, nobody should tell you, you can't. If a company has a problem hearing a customers truth(you might think they would learn what we want from it and fix the problems,then they could say so on Histonet and all would benefit.) I don't think any company is problem free and they seem to watch our conversations for product needs, and to make money from us, so if you want the cash then deal with it all!!!!!! my perspective only. detection kit 400.00 > technical problems 1,000000's > able to express ones opinion on Histonet---priceless > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From RCazares <@t> schosp.org Thu Aug 12 16:53:06 2004 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] ROUND OF APPLAUSE FOR JOE! Message-ID: <229A3566B9F0D311826E00D0B7441D7907E0B0BF@swedish_nt1.schosp.org> APPLAUSE! APPLAUSE! APPLAUSE! APPLAUSE! APPLAUSE! APPLAUSE! APPLAUSE! APPLAUSE! Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ************************* From pathrm35 <@t> adelphia.net Thu Aug 12 16:58:06 2004 From: pathrm35 <@t> adelphia.net (Ron Martin) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] proper disposal of specimen transpot biohazard bags References: <0556BE8AC5551E4E8AF6BB9E42509BA2D94B7D@usca0082k08.labvision.apogent.com> Message-ID: <002001c480b7$73812330$5dedeb44@Pathrm35> Thanks. Does anyone know if it is illegal (federal or state laws)or against OSHA regs to dispose of biohazard bags in regular trash? I'm in Florida so I would like to hear from some Florida techs if possible. ----- Original Message ----- From: "Morken, Tim - Labvision" To: Sent: Thursday, August 12, 2004 4:17 PM Subject: RE: [Histonet] proper disposal of specimen transpot biohazard bags > Well... When working at a nameless lab many years ago we had a call from a > local landfill that freaked when they found an empty biohazard bag in a load > of our "regular" trash, so I don't think they like that kind of thing. > > Tim Morken > > > -----Original Message----- > From: Ron Martin [mailto:pathrm35@adelphia.net] > Sent: Thursday, August 12, 2004 2:01 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] proper disposal of specimen transpot biohazard bags > > > Fellow techs, > What is the proper disposal means for the specimen transport biohazard bags? > The bags we use say "stat" and "biohazard" on them. I always thought that > they were to be discarded in a larger biohazard bag with the other > contaminated material. My manager says that they are to be discarded in the > regular trash unless formalin or other fluid leaks in them. What is everyone > else doing? Thanks, Ron Martin > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Thu Aug 12 17:06:18 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this,th en again, maybe not Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070466@HALIBUT.dcla.com> Off the wall is good, in the drink is not. Gary Gill -----Original Message----- From: Joe Nocito [mailto:jnocito@satx.rr.com] Sent: Thursday, August 12, 2004 4:48 PM To: DDittus787@aol.com; Gary Gill; 'Joe Nocito'; Histonet Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] You may want to hit the delete button with this,then again, maybe not see, that's just it, I laughed at Gary's response, couldn't help it. Believe it or not, I do have a sense of humor, alibit a little off the wall, but to know me is to love me. Joe ----- Original Message ----- From: To: "Gary Gill" ; "'Joe Nocito'" ; "Histonet" Cc: Sent: Thursday, August 12, 2004 4:03 PM Subject: RE: [Histonet] You may want to hit the delete button with this,then again, maybe not > gary- worst thing you can do is tease a guy who is mad. > > joe- go to it man, if you have an opinion you should express it, > nobody should tell you, you can't. If a company has a problem hearing a customers truth(you might think they would learn what we want from it and fix the problems,then they could say so on Histonet and all would benefit.) I don't think any company is problem free and they seem to watch our conversations for product needs, and to make money from us, so if you want the cash then deal with it all!!!!!! my perspective only. detection kit 400.00 > technical problems 1,000000's > able to express ones opinion on Histonet---priceless > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From garygill <@t> dcla.com Thu Aug 12 17:06:18 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this,th en again, maybe not Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070466@HALIBUT.dcla.com> Off the wall is good, in the drink is not. Gary Gill -----Original Message----- From: Joe Nocito [mailto:jnocito@satx.rr.com] Sent: Thursday, August 12, 2004 4:48 PM To: DDittus787@aol.com; Gary Gill; 'Joe Nocito'; Histonet Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] You may want to hit the delete button with this,then again, maybe not see, that's just it, I laughed at Gary's response, couldn't help it. Believe it or not, I do have a sense of humor, alibit a little off the wall, but to know me is to love me. Joe ----- Original Message ----- From: To: "Gary Gill" ; "'Joe Nocito'" ; "Histonet" Cc: Sent: Thursday, August 12, 2004 4:03 PM Subject: RE: [Histonet] You may want to hit the delete button with this,then again, maybe not > gary- worst thing you can do is tease a guy who is mad. > > joe- go to it man, if you have an opinion you should express it, > nobody should tell you, you can't. If a company has a problem hearing a customers truth(you might think they would learn what we want from it and fix the problems,then they could say so on Histonet and all would benefit.) I don't think any company is problem free and they seem to watch our conversations for product needs, and to make money from us, so if you want the cash then deal with it all!!!!!! my perspective only. detection kit 400.00 > technical problems 1,000000's > able to express ones opinion on Histonet---priceless > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ihc <@t> unipathllc.com Thu Aug 12 17:14:13 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Mallory's PTAH In-Reply-To: <000101c480b1$f5bb8350$034dbad0@hppav> Message-ID: <000001c480b9$b3b25350$4500a8c0@unipath02> George, Sorry, I didn't realize my full address wasn't on there. It is: bjackson@unipathllc.com. I really appreciate your help. I'll let you know how it turns out. -----Original Message----- From: George Cole [mailto:georgecole@ev1.net] Sent: Thursday, August 12, 2004 3:19 PM To: 'UniPath IHC' Subject: RE: [Histonet] Mallory's PTAH Brianna; This second message must have been to someone else----the procedures I send you will get your sections. Brianna, there is nothing keeping you from fixing the sections after they are cut. But I urge you to look at and try the methods I will send you. They outdid in appearance and utility anything in the standard procedure!!! georgecole@ev1.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of UniPath IHC Sent: Thursday, August 12, 2004 11:13 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mallory's PTAH I actually already tried to use formalin-fixed muscle. You're exactly right, it sectioned great, but I could never get any decent staining. I tried post-fixing for 24 hrs. in Zenker's vs. post-fixing for 3 Hrs in Zenker's at 56 degrees. I tried all three protocols in Sheehan and the protocol in Carson. I'm at a loss for what to do next if I can't make the Zenker fixed muscle work. At least I've learned a lot about the stain. :-) Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Aug 12 17:24:44 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Fluorescence and photobleaching discussion In-Reply-To: <200408122122.i7CLMENs413896@mail3.msu.montana.edu> References: <200408122122.i7CLMENs413896@mail3.msu.montana.edu> Message-ID: <6.0.0.22.1.20040812161830.01af7a28@gemini.msu.montana.edu> Teri et al, The same bubble problem/media retraction problem occurs with Molecular Probes Prolong Gold ready to use and that is why they suggest sealing edges of coverslip for long term storage. Prolong Gold is thicker in consistency but be sure to seal down edges. I have this occur with AquaMount, an all purpose aqueous mounting media with AEC chromogen - edges get sealed with this media too. Gayle Callis At 03:17 PM 8/12/2004, you wrote: >Gayle writes: **Vector has VectaShield Hard Set that is supposed to work >also. It is >nice to have an aqueous mounting media for fluorescent work that sets up > >hard!! > >I have used this product and I don't like it. Initially I did because >it is thin in consistency and spreads well. However, with time, thick >specimens (12 microns) developed air pockets in some of them. Having >learned this, I used more on the next batch of slides, and ended up with >the same problem. Also, they state that you can easily remount if >necessary by soaking in water to remove the coverslip. It took 3 days >before the coverslip was loose enough to remove, and remounting wasn't >really satisfactory for the coverslips that had cultured cells on it. >Some of the cells adhered to the glass slide. > >Others in our facility use this stuff and love it. > > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, Missouri 64110 >tjj@stowers-institute.org > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Thu Aug 12 18:57:18 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this,th en again, maybe not Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E23A@simba.kids> Did you mention drink? I would love one (or 2, or 3) Ah, what were we talking about? Olympics are on. My diploma in couch potatoism will be put to good use. Hope everyone has a good 2 weeks Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Friday, 13 August 2004 8:06 AM To: 'Joe Nocito'; DDittus787@aol.com; Gary Gill; 'Joe Nocito'; Histonet Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] You may want to hit the delete button with this,th en again, maybe not Off the wall is good, in the drink is not. Gary Gill -----Original Message----- From: Joe Nocito [mailto:jnocito@satx.rr.com] Sent: Thursday, August 12, 2004 4:48 PM To: DDittus787@aol.com; Gary Gill; 'Joe Nocito'; Histonet Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] You may want to hit the delete button with this,then again, maybe not see, that's just it, I laughed at Gary's response, couldn't help it. Believe it or not, I do have a sense of humor, alibit a little off the wall, but to know me is to love me. Joe ----- Original Message ----- From: To: "Gary Gill" ; "'Joe Nocito'" ; "Histonet" Cc: Sent: Thursday, August 12, 2004 4:03 PM Subject: RE: [Histonet] You may want to hit the delete button with this,then again, maybe not > gary- worst thing you can do is tease a guy who is mad. > > joe- go to it man, if you have an opinion you should express it, > nobody should tell you, you can't. If a company has a problem hearing a customers truth(you might think they would learn what we want from it and fix the problems,then they could say so on Histonet and all would benefit.) I don't think any company is problem free and they seem to watch our conversations for product needs, and to make money from us, so if you want the cash then deal with it all!!!!!! my perspective only. detection kit 400.00 > technical problems 1,000000's > able to express ones opinion on Histonet---priceless > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Thu Aug 12 18:57:18 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this,th en again, maybe not Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E23A@simba.kids> Did you mention drink? I would love one (or 2, or 3) Ah, what were we talking about? Olympics are on. My diploma in couch potatoism will be put to good use. Hope everyone has a good 2 weeks Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Friday, 13 August 2004 8:06 AM To: 'Joe Nocito'; DDittus787@aol.com; Gary Gill; 'Joe Nocito'; Histonet Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] You may want to hit the delete button with this,th en again, maybe not Off the wall is good, in the drink is not. Gary Gill -----Original Message----- From: Joe Nocito [mailto:jnocito@satx.rr.com] Sent: Thursday, August 12, 2004 4:48 PM To: DDittus787@aol.com; Gary Gill; 'Joe Nocito'; Histonet Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] You may want to hit the delete button with this,then again, maybe not see, that's just it, I laughed at Gary's response, couldn't help it. Believe it or not, I do have a sense of humor, alibit a little off the wall, but to know me is to love me. Joe ----- Original Message ----- From: To: "Gary Gill" ; "'Joe Nocito'" ; "Histonet" Cc: Sent: Thursday, August 12, 2004 4:03 PM Subject: RE: [Histonet] You may want to hit the delete button with this,then again, maybe not > gary- worst thing you can do is tease a guy who is mad. > > joe- go to it man, if you have an opinion you should express it, > nobody should tell you, you can't. If a company has a problem hearing a customers truth(you might think they would learn what we want from it and fix the problems,then they could say so on Histonet and all would benefit.) I don't think any company is problem free and they seem to watch our conversations for product needs, and to make money from us, so if you want the cash then deal with it all!!!!!! my perspective only. detection kit 400.00 > technical problems 1,000000's > able to express ones opinion on Histonet---priceless > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From tinayates <@t> comcast.net Thu Aug 12 20:40:55 2004 From: tinayates <@t> comcast.net (Comcast Mail) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] SEEKING HISTOLOGIST Message-ID: <000c01c480d6$93f8cf90$abdb1418@DFC4J821> Salem Hospital is a 454-bed facility located in the beautiful Willamette Valley of western Oregon. Salem Hospital Regional Laboratory, Advance Magazine's 2004 Lab of the Year, is currently seeking an individual for placement as a Histologist HT/HTL(ASCP). This opening is full-time and includes participation in both weekend and IHC rotations. Our busy and varied Surgical Pathology environment requires candidates who possess skilled experience in tissue processing, embedding, microtomy, special stains and automated immunohistochemistry. Our values of customer service require candidates who possess the ability to fully integrate into our culture of technical excellence in a team-oriented environment. Sign-on bonus and relocation assistance available. Please apply online at www.salemhospital.org or call 1.800.825.5199 for more information. From Jason.PALMER <@t> svhm.org.au Thu Aug 12 21:13:10 2004 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] RE: replies to immunostaining on technovit 8100 Message-ID: Thanks Pam Marcum and Gayle (especially) for your thoughtful responses. Perhaps I should have been clearer as to why we want to use this rather than paraffin - the tissues we collect are grown on a PLGA (a polymer of lactic and glycolic acids) scaffold, which melts / distorts at any temp above 45 degrees c, not to mention in alcohols,acetone, xylene and the like. Hence our desire to avoid paraffin. Technovit 8100 polymerises at zero degrees, so no heat needed at all, and we use inert dehydration to remove excess water from the PLGA / tissues. And, supposedly immunohistochemistry compatible.... And yes, I got the mouse and rat confused with the antibody - mouse anti rat ED1 it is! Cheers, Jason Palmer Bernard O'Brien Institute of Microsurgery Melbourne, Australia -------------- next part -------------- Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. From sjones <@t> cvm.tamu.edu Thu Aug 12 22:24:27 2004 From: sjones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] You may want to hit the delete button with this,then again, maybe not Message-ID: Hi Joe, This company sounds like the poster child for how to do poor customer service. I took an excellent class at the last NSH Convention on customer service; maybe their customer service personel could sign up for the class instead of harassing you. Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Joe Nocito" 08/12/04 3:34 PM >>> Afternoon Netter's, Remember about a week or two the topic was getting "flamed"? Well, here's a new one. Sometime last year, I made several postings about a certain company and their product that I used and it did not work for me. I wrote them a letter and returned all the merchandise back to them saying I was not happy with their product. I waited several months and never got a response, so I named the company on the Histonet. Since then, I have been throwing away their flyers, catalogs and any other correspondence I receive. Obviously, they are doing fine without my business and I am doing fine without their products. Today, I receive a letter from the company with a copy of my Histonet postings from over a year ago, telling me that this has been in customer service for about a year (hmmmmmmmm, I'm trying to find out where the customer service is in all of this. (One year, okay, I'll get back to that), that their customer service has tried several times to contact me but to no avail. (Hmmmmm, one way would have been to respond to my email address, but I'll come back to this also). So after all this time, they want to do business with me. (So close to the NSH meeting, hmmmmmmmmm, another point to ponder). My big beef is that these companies read this forum and try to use strong arm tactics on people who post negative comments about them or their products. I have many friends that are sales reps in this field (I'm talking like come over my house and eat type of friends) and I love these people. I must reiterate that this forum was set up to exchange ideas, theories and experiences, both positive and negative. If this was such a "hot item", point #1, why did it take a whole year to get a response when they had my company and address on file? Is this how customer service works? I don't think so!! Point #2 If you are a vendor and don't like a particular posting, why don't you reply back to that person via email. Point #3 don't wait until it's almost time for the NSH annual S/C to fix a problem, it's probably too late by then. Point #4 If you haven't had a purchase order or have not corresponded with a customer for over a year, chances are that customer does not want to do business with you. Let lying dogs lie and don't open old scars. All you did was get me going on a tangent. I know, there goes Nocito again, but I refuse to be treated like this from any one. Like they say in the movies, "Don't call me, I'll call you". Okay, I've said my piece now let the FLAMING BEGIN!!!!!!!!! As always, the opinions stated by this author in no way reflects the opinions of his employer, subordinates, friends or relatives. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histophilhuff <@t> yahoo.com Thu Aug 12 23:02:38 2004 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Bubbles in Thick Sections - Suggestions? Message-ID: <20040813040238.8280.qmail@web50308.mail.yahoo.com> We cut sections of tissue between 20 um and 30 um thick to view blood vessels after being stained with CD31. We also section tissue at thicknesses of 12 to 16 um for our H+E stains. We sometimes get bubbles when we coverslip with non-aqueous mounting media and hate to see them in well-stained slides. Does anyone have any suggestions for not getting bubbles in thick sections of tissue? After the last xylene step we allow the sections to dry to ensure that the tissues are not wet and we are currently using CureMount II, a non-aqueous UV-cured mounting media from Instrumedics. Help me destroy the bubble monster! Please.... Thanks in advance, Phil --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From pathrm35 <@t> adelphia.net Fri Aug 13 04:27:14 2004 From: pathrm35 <@t> adelphia.net (Ron Martin) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: <002701c48117$b8cbc750$5dedeb44@Pathrm35> Fellow techs, I am in a difficult situation and need some serious advice. I recently (4 months ago) accepted a technical supervisor position in a dermatology lab. I went from a bench tech at my old job to this position. I also walked away from a raise at my old position so I could step into a supervision position. I took the new position because I was told by my manager that she would teach me some supervision, management and financial skills that I currently do not have as well as the growth potential of the company.This position is not what I was told it would be. Part of my "duties" include emptying the trash, clean the bathroom (not happening) and taking her personal and professional calls. The question on ethics is the high volume of mistakes made by our technician, our offices and also by my manager herself. I caught a mistake made by our tech a few weeks ago.She put the wrong tissue on the wrong slides as she inverted the two cases. I caught her mistake before it went out. One time she assigned the same number to two different cases. She then sent the correct case out for a consult (which wasn't needed) and put the wrong patient name on the slide. Every other day there is something different.My manager will not terminate her as she knows I am seeking employment elsewhere and she cannot afford to lose a tech. One day my manager gave three cases the same accession number.She caught her mistake on one of the cases but I still ended up with two cases with the same number. When I first started there my manager had a case in which there " was no tissue in the container". She said that she notified the office about the situation however up to six weeks later the office was still calling looking for results.At that point she wrote up an incident report and dated it six weeks prior to coincide with the surgery date. Our offices are not any better. One case came in with the wrong patient information. It took about 7 weeks for the office to realize that they sent the wrong patient information with the biopsy as the names were close in spelling. The offices continually send mistakes with incorrect spelling of patients names and incorrect anatomical locations. I have tired to document everything but there are too many mistakes and I don't have enough time or energy to keep up with them.My manager wants us to do our own "internal quality control". My interpretation of this is that she doesn't want our physician and risk manager to know of these mistakes. Are these becoming common problems or is it just my situation? I want to emphasize that I hope I am not being unethical myself for revealing this information but I really need some advice and support.I have very high standards and they are not being met in this current situation. I am currently seeking a new position but I need employment and cannot resign until another position becomes available. I would like to stay in Florida and if anyone knows of any positions please inform me as I am at my wits end. Thanks in advance. Ron From lpwenk <@t> sbcglobal.net Fri Aug 13 03:59:20 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Mallory's PTAH References: <000001c48098$00735800$4500a8c0@unipath02> Message-ID: <009301c48113$d3e6da60$d126d445@domainnotset.invalid> Here's what worked for us - Out of the old Ann Preece book. Fix as usual in formalin, and process as you would for similar size tissue. Section 4-5 um (I always vote for thinner, when working on registry material) and place on a subbed slide. (Charged, poly-L-lysine, etc.) Helps to hold it on through the rest of the steps. Dry in 60 degree oven for about an hour (helps to hold the tissue on the slide during the rest of the steps). After deparaffinization and running down to water, post-fix in Bouin for 1 hour in a 60 degree C. oven or waterbath. Then wash in slowly running cold water for about 10 minutes, or until all the yellow is gone. (Yes, just like post-fixing to do a trichrome.) Then, stain as usual with the PTAH. Should work great, nice intense colors with very little splotching, and sectioning should be your usual formalin quality. No mercury disposal to worry about. This is assuming, of course, that the PTAH solution is still good and the procedure was followed correctly. Now - as far as any PTAH questions on the written/computer test - the correct answer to fixation/post-fixation for PTAH is "Zenker". As that's what is written in all the rest of the histo books. Hope that helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "UniPath IHC" To: Sent: Thursday, August 12, 2004 2:13 PM Subject: [Histonet] Mallory's PTAH I actually already tried to use formalin-fixed muscle. You're exactly right, it sectioned great, but I could never get any decent staining. I tried post-fixing for 24 hrs. in Zenker's vs. post-fixing for 3 Hrs in Zenker's at 56 degrees. I tried all three protocols in Sheehan and the protocol in Carson. I'm at a loss for what to do next if I can't make the Zenker fixed muscle work. At least I've learned a lot about the stain. :-) Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dobbin <@t> upei.ca Fri Aug 13 07:18:42 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics In-Reply-To: <002701c48117$b8cbc750$5dedeb44@Pathrm35> Message-ID: <411C8771.9200.C2635@localhost> Ron, You have Risk Managers? Meet with them right away! Today, not tommorrow! Unless the whole system there is corrupt, they will take care of you (job security) and you will rest easier knowing someone else is helping you "manage" the risk!! You owe it to the patients to make this situation right. Very serious consequences are inevidable in that current situation. Good luck. And keep us posted on new developments. Greg From: "Ron Martin" To: Date sent: Fri, 13 Aug 2004 04:27:14 -0500 Subject: [Histonet] a question on ethics > Fellow techs, > I am in a difficult situation and need some serious advice. I recently (4 months ago) accepted a technical supervisor position in a dermatology lab. I went from a bench tech at my old job to this position. I also walked away from a raise at my old position so I could step into a supervision position. I took the new position because I was told by my manager that she would teach me some supervision, management and financial skills that I currently do not have as well as the growth potential of the company.This position is not what I was told it would be. Part of my "duties" include emptying the trash, clean the bathroom (not happening) and taking her personal and professional calls. The question on ethics is the high volume of mistakes made by our technician, our offices and also by my manager herself. > I caught a mistake made by our tech a few weeks ago.She put the wrong tissue on the wrong slides as she inverted the two cases. I caught her mistake before it went out. One time she assigned the same number to two different cases. She then sent the correct case out for a consult (which wasn't needed) and put the wrong patient name on the slide. Every other day there is something different.My manager will not terminate her as she knows I am seeking employment elsewhere and she cannot afford to lose a tech. > One day my manager gave three cases the same accession number.She caught her mistake on one of the cases but I still ended up with two cases with the same number. When I first started there my manager had a case in which there " was no tissue in the container". She said that she notified the office about the situation however up to six weeks later the office was still calling looking for results.At that point she wrote up an incident report and dated it six weeks prior to coincide with the surgery date. > Our offices are not any better. One case came in with the wrong patient information. It took about 7 weeks for the office to realize that they sent the wrong patient information with the biopsy as the names were close in spelling. The offices continually send mistakes with incorrect spelling of patients names and incorrect anatomical locations. > I have tired to document everything but there are too many mistakes and I don't have enough time or energy to keep up with them.My manager wants us to do our own "internal quality control". My interpretation of this is that she doesn't want our physician and risk manager to know of these mistakes. Are these becoming common problems or is it just my situation? > I want to emphasize that I hope I am not being unethical myself for revealing this information but I really need some advice and support.I have very high standards and they are not being met in this current situation. I am currently seeking a new position but I need employment and cannot resign until another position becomes available. I would like to stay in Florida and if anyone knows of any positions please inform me as I am at my wits end. Thanks in advance. > > Ron > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Pathology Lab Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Making a living is getting; making a life is giving. From Jackie.O'Connor <@t> abbott.com Fri Aug 13 07:30:02 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Bubbles in Thick Sections - Suggestions? Message-ID: I must have missed something, but why are you cutting sections 20-30 um to look at CD31? Sections of what? Curious in Abbott Park Phillip Huff Sent by: histonet-bounces@lists.utsouthwestern.edu 08/12/2004 11:02 PM To: Histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Bubbles in Thick Sections - Suggestions? We cut sections of tissue between 20 um and 30 um thick to view blood vessels after being stained with CD31. We also section tissue at thicknesses of 12 to 16 um for our H+E stains. We sometimes get bubbles when we coverslip with non-aqueous mounting media and hate to see them in well-stained slides. Does anyone have any suggestions for not getting bubbles in thick sections of tissue? After the last xylene step we allow the sections to dry to ensure that the tissues are not wet and we are currently using CureMount II, a non-aqueous UV-cured mounting media from Instrumedics. Help me destroy the bubble monster! Please.... Thanks in advance, Phil --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri Aug 13 07:32:18 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: Run Ron Run. "Ron Martin" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 04:27 AM To: cc: Subject: [Histonet] a question on ethics Fellow techs, I am in a difficult situation and need some serious advice. I recently (4 months ago) accepted a technical supervisor position in a dermatology lab. I went from a bench tech at my old job to this position. I also walked away from a raise at my old position so I could step into a supervision position. I took the new position because I was told by my manager that she would teach me some supervision, management and financial skills that I currently do not have as well as the growth potential of the company.This position is not what I was told it would be. Part of my "duties" include emptying the trash, clean the bathroom (not happening) and taking her personal and professional calls. The question on ethics is the high volume of mistakes made by our technician, our offices and also by my manager herself. I caught a mistake made by our tech a few weeks ago.She put the wrong tissue on the wrong slides as she inverted the two cases. I caught her mistake before it went out. One time she assigned the same number to two different cases. She then sent the correct case out for a consult (which wasn't needed) and put the wrong patient name on the slide. Every other day there is something different.My manager will not terminate her as she knows I am seeking employment elsewhere and she cannot afford to lose a tech. One day my manager gave three cases the same accession number.She caught her mistake on one of the cases but I still ended up with two cases with the same number. When I first started there my manager had a case in which there " was no tissue in the container". She said that she notified the office about the situation however up to six weeks later the office was still calling looking for results.At that point she wrote up an incident report and dated it six weeks prior to coincide with the surgery date. Our offices are not any better. One case came in with the wrong patient information. It took about 7 weeks for the office to realize that they sent the wrong patient information with the biopsy as the names were close in spelling. The offices continually send mistakes with incorrect spelling of patients names and incorrect anatomical locations. I have tired to document everything but there are too many mistakes and I don't have enough time or energy to keep up with them.My manager wants us to do our own "internal quality control". My interpretation of this is that she doesn't want our physician and risk manager to know of these mistakes. Are these becoming common problems or is it just my situation? I want to emphasize that I hope I am not being unethical myself for revealing this information but I really need some advice and support.I have very high standards and they are not being met in this current situation. I am currently seeking a new position but I need employment and cannot resign until another position becomes available. I would like to stay in Florida and if anyone knows of any positions please inform me as I am at my wits end. Thanks in advance. Ron _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jmahoney <@t> alegent.org Fri Aug 13 07:41:27 2004 From: jmahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: Ron, I'm sorry for your situation. It seems to me that something you must do immediately, to protect your patients and yourself, is to do exactly what your manager is suggesting. Get an internal quality program into place. I'm assuming as supervisor you have that authority. One suggestion is to have a person check the slides against the blocks before they go out to the Pathologists. Start a log sheet or some kind of documentation to track the errors. Decide on an acceptable number of errors for your techs. (hopefully less than .1%). I would do this before I went to your administration or any other source. Explain to your techs that this is for the patient and for them. Everyone is liable. Another thing you can do is to look at the process of each tech cutting slides. If possible, standardize the way they check the numbers and transcribe them. Include in this standardization a minute to inspect the slide against the block before they put it in a staining rack. I applaud your dedication to quality. Do what you can now. Good Luck. Jan Mahoney Janice Mahoney Histology/Cytology Coordinator Alegent Health Laboratory Phone (402)717-2889 Fax (402)717-5231 "There's NO excuse for Domestic Violence" Crisis Lines: Omaha YWCA 402-345-7273 Sarpy County 1-800-523-3666 Council Bluffs 1-712-328-0266 NE's Statewide 1-800-876-6238 >>> "Greg Dobbin" 08/13/2004 9:18:42 AM >>> Ron, You have Risk Managers? Meet with them right away! Today, not tommorrow! Unless the whole system there is corrupt, they will take care of you (job security) and you will rest easier knowing someone else is helping you "manage" the risk!! You owe it to the patients to make this situation right. Very serious consequences are inevidable in that current situation. Good luck. And keep us posted on new developments. Greg From: "Ron Martin" To: Date sent: Fri, 13 Aug 2004 04:27:14 -0500 Subject: [Histonet] a question on ethics > Fellow techs, > I am in a difficult situation and need some serious advice. I recently (4 months ago) accepted a technical supervisor position in a dermatology lab. I went from a bench tech at my old job to this position. I also walked away from a raise at my old position so I could step into a supervision position. I took the new position because I was told by my manager that she would teach me some supervision, management and financial skills that I currently do not have as well as the growth potential of the company.This position is not what I was told it would be. Part of my "duties" include emptying the trash, clean the bathroom (not happening) and taking her personal and professional calls. The question on ethics is the high volume of mistakes made by our technician, our offices and also by my manager herself. > I caught a mistake made by our tech a few weeks ago.She put the wrong tissue on the wrong slides as she inverted the two cases. I caught her mistake before it went out. One time she assigned the same number to two different cases. She then sent the correct case out for a consult (which wasn't needed) and put the wrong patient name on the slide. Every other day there is something different.My manager will not terminate her as she knows I am seeking employment elsewhere and she cannot afford to lose a tech. > One day my manager gave three cases the same accession number.She caught her mistake on one of the cases but I still ended up with two cases with the same number. When I first started there my manager had a case in which there " was no tissue in the container". She said that she notified the office about the situation however up to six weeks later the office was still calling looking for results.At that point she wrote up an incident report and dated it six weeks prior to coincide with the surgery date. > Our offices are not any better. One case came in with the wrong patient information. It took about 7 weeks for the office to realize that they sent the wrong patient information with the biopsy as the names were close in spelling. The offices continually send mistakes with incorrect spelling of patients names and incorrect anatomical locations. > I have tired to document everything but there are too many mistakes and I don't have enough time or energy to keep up with them.My manager wants us to do our own "internal quality control". My interpretation of this is that she doesn't want our physician and risk manager to know of these mistakes. Are these becoming common problems or is it just my situation? > I want to emphasize that I hope I am not being unethical myself for revealing this information but I really need some advice and support.I have very high standards and they are not being met in this current situation. I am currently seeking a new position but I need employment and cannot resign until another position becomes available. I would like to stay in Florida and if anyone knows of any positions please inform me as I am at my wits end. Thanks in advance. > > Ron > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Pathology Lab Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Making a living is getting; making a life is giving. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Fri Aug 13 07:40:53 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: Yes, Do run Ron run Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: 13 August 2004 13:32 To: Ron Martin Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] a question on ethics Run Ron Run. "Ron Martin" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 04:27 AM To: cc: Subject: [Histonet] a question on ethics Fellow techs, I am in a difficult situation and need some serious advice. I recently (4 months ago) accepted a technical supervisor position in a dermatology lab. I went from a bench tech at my old job to this position. I also walked away from a raise at my old position so I could step into a supervision position. I took the new position because I was told by my manager that she would teach me some supervision, management and financial skills that I currently do not have as well as the growth potential of the company.This position is not what I was told it would be. Part of my "duties" include emptying the trash, clean the bathroom (not happening) and taking her personal and professional calls. The question on ethics is the high volume of mistakes made by our technician, our offices and also by my manager herself. I caught a mistake made by our tech a few weeks ago.She put the wrong tissue on the wrong slides as she inverted the two cases. I caught her mistake before it went out. One time she assigned the same number to two different cases. She then sent the correct case out for a consult (which wasn't needed) and put the wrong patient name on the slide. Every other day there is something different.My manager will not terminate her as she knows I am seeking employment elsewhere and she cannot afford to lose a tech. One day my manager gave three cases the same accession number.She caught her mistake on one of the cases but I still ended up with two cases with the same number. When I first started there my manager had a case in which there " was no tissue in the container". She said that she notified the office about the situation however up to six weeks later the office was still calling looking for results.At that point she wrote up an incident report and dated it six weeks prior to coincide with the surgery date. Our offices are not any better. One case came in with the wrong patient information. It took about 7 weeks for the office to realize that they sent the wrong patient information with the biopsy as the names were close in spelling. The offices continually send mistakes with incorrect spelling of patients names and incorrect anatomical locations. I have tired to document everything but there are too many mistakes and I don't have enough time or energy to keep up with them.My manager wants us to do our own "internal quality control". My interpretation of this is that she doesn't want our physician and risk manager to know of these mistakes. Are these becoming common problems or is it just my situation? I want to emphasize that I hope I am not being unethical myself for revealing this information but I really need some advice and support.I have very high standards and they are not being met in this current situation. I am currently seeking a new position but I need employment and cannot resign until another position becomes available. I would like to stay in Florida and if anyone knows of any positions please inform me as I am at my wits end. Thanks in advance. Ron _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.Eyres <@t> sanofi-synthelabo.com Fri Aug 13 07:56:12 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: Ron, Don't run Ron, Ron, don't run Ron (there's a song that goes like that... something about meeting someone on a Monday). Seriously, though. I agree with a previous respondent that you need to tell someone with lots of clout to sort make sure the mess is sorted out. From the patient's point of view, the very fact you've raised the issue means that you're the best person there to do the sorting out. The person with the clout, will act as your shield, though a bit of Kelvar might come in handy!! Best of luck. Steve "Marshall Terry Dr, Consultant Histopathologist" To: > cc: histonet@lists.utsouthwestern.edu Sent by: Subject: RE: [Histonet] a question on ethics histonet-bounces@lists.utsouth western.edu 13/08/2004 13:40 Yes, Do run Ron run Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: 13 August 2004 13:32 To: Ron Martin Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] a question on ethics Run Ron Run. "Ron Martin" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 04:27 AM To: cc: Subject: [Histonet] a question on ethics Fellow techs, I am in a difficult situation and need some serious advice. I recently (4 months ago) accepted a technical supervisor position in a dermatology lab. I went from a bench tech at my old job to this position. I also walked away from a raise at my old position so I could step into a supervision position. I took the new position because I was told by my manager that she would teach me some supervision, management and financial skills that I currently do not have as well as the growth potential of the company.This position is not what I was told it would be. Part of my "duties" include emptying the trash, clean the bathroom (not happening) and taking her personal and professional calls. The question on ethics is the high volume of mistakes made by our technician, our offices and also by my manager herself. I caught a mistake made by our tech a few weeks ago.She put the wrong tissue on the wrong slides as she inverted the two cases. I caught her mistake before it went out. One time she assigned the same number to two different cases. She then sent the correct case out for a consult (which wasn't needed) and put the wrong patient name on the slide. Every other day there is something different.My manager will not terminate her as she knows I am seeking employment elsewhere and she cannot afford to lose a tech. One day my manager gave three cases the same accession number.She caught her mistake on one of the cases but I still ended up with two cases with the same number. When I first started there my manager had a case in which there " was no tissue in the container". She said that she notified the office about the situation however up to six weeks later the office was still calling looking for results.At that point she wrote up an incident report and dated it six weeks prior to coincide with the surgery date. Our offices are not any better. One case came in with the wrong patient information. It took about 7 weeks for the office to realize that they sent the wrong patient information with the biopsy as the names were close in spelling. The offices continually send mistakes with incorrect spelling of patients names and incorrect anatomical locations. I have tired to document everything but there are too many mistakes and I don't have enough time or energy to keep up with them.My manager wants us to do our own "internal quality control". My interpretation of this is that she doesn't want our physician and risk manager to know of these mistakes. Are these becoming common problems or is it just my situation? I want to emphasize that I hope I am not being unethical myself for revealing this information but I really need some advice and support.I have very high standards and they are not being met in this current situation. I am currently seeking a new position but I need employment and cannot resign until another position becomes available. I would like to stay in Florida and if anyone knows of any positions please inform me as I am at my wits end. Thanks in advance. Ron _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- From JNocito <@t> Pathreflab.com Fri Aug 13 08:00:22 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics In-Reply-To: <002701c48117$b8cbc750$5dedeb44@Pathrm35> Message-ID: Ron, I sympathize greatly with you. You really need to take all your documentation to Risk Management today. Have you tried talking to your manager's supervisor? If you have and nothing changed, I would go to that person and tell them you are going to R.M. and that you were just telling them as a courtesy and don't let them talk you out of going to R.M. The reason I say this is no one likes to be blind-sided and then they can't come back and say you should have let them know. Prior military experience tells me to go through the management chain. In this situation, I would have all my ducks lines up so there could be no back-lash on you. You have the right and responsibility to be the patient's advocate. There are too many horror stories about mixing specimens up. And I would definitely start looking for another job. As always, the opinions of this author do not reflect the opinions of his employer. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ron Martin Sent: Friday, August 13, 2004 4:27 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] a question on ethics Fellow techs, I am in a difficult situation and need some serious advice. I recently (4 months ago) accepted a technical supervisor position in a dermatology lab. I went from a bench tech at my old job to this position. I also walked away from a raise at my old position so I could step into a supervision position. I took the new position because I was told by my manager that she would teach me some supervision, management and financial skills that I currently do not have as well as the growth potential of the company.This position is not what I was told it would be. Part of my "duties" include emptying the trash, clean the bathroom (not happening) and taking her personal and professional calls. The question on ethics is the high volume of mistakes made by our technician, our offices and also by my manager herself. I caught a mistake made by our tech a few weeks ago.She put the wrong tissue on the wrong slides as she inverted the two cases. I caught her mistake before it went out. One time she assigned the same number to two different cases. She then sent the correct case out for a consult (which wasn't needed) and put the wrong patient name on the slide. Every other day there is something different.My manager will not terminate her as she knows I am seeking employment elsewhere and she cannot afford to lose a tech. One day my manager gave three cases the same accession number.She caught her mistake on one of the cases but I still ended up with two cases with the same number. When I first started there my manager had a case in which there " was no tissue in the container". She said that she notified the office about the situation however up to six weeks later the office was still calling looking for results.At that point she wrote up an incident report and dated it six weeks prior to coincide with the surgery date. Our offices are not any better. One case came in with the wrong patient information. It took about 7 weeks for the office to realize that they sent the wrong patient information with the biopsy as the names were close in spelling. The offices continually send mistakes with incorrect spelling of patients names and incorrect anatomical locations. I have tired to document everything but there are too many mistakes and I don't have enough time or energy to keep up with them.My manager wants us to do our own "internal quality control". My interpretation of this is that she doesn't want our physician and risk manager to know of these mistakes. Are these becoming common problems or is it just my situation? I want to emphasize that I hope I am not being unethical myself for revealing this information but I really need some advice and support.I have very high standards and they are not being met in this current situation. I am currently seeking a new position but I need employment and cannot resign until another position becomes available. I would like to stay in Florida and if anyone knows of any positions please inform me as I am at my wits end. Thanks in advance. Ron _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Aug 13 08:03:16 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A62E6@sjhaexc02.sjha.org> Ron, I feel your pain!! I'm not sure what your resources are. But if you have a risk manager, I would first of all ask your manager to meet with you and the risk manager. If she refused, I'd let her know that I intended to request a meeting. I would also involve the pathologists as they are at risk for malpractice. The consensus of management gurus would be that processes cause most errors - not people. JCAHO now requires a "Root Cause Analysis" of serious error, to help determine what processes could be put into place to eliminate errors. I would review the error with this in mind to see what could be put into place to prevent errors and to catch them when they occur. One suggestion would be to ink the tissue - not just different colored cassettes or slides. There are kits of dyes in neat little dispensers that you would rotate through all the skins that would be dictated in the gross to help identify the specimen. That way you would know that Patient A was red, B was blue, There would be very little one could do to separate two skins of like size, other than something like this, I would think. Didn't intend to write a book. Good luck! Joyce Have a good weekend everyone! Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Charles.Embrey <@t> carle.com Fri Aug 13 08:28:22 2004 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] proper disposal of specimen transpot biohazard bag s Message-ID: Here in Illinois it is illegal to dispose of anything with a "biohazard" symbol on it in the regular trash. I have also felt that it was against federal OSHA laws. Charles Embrey Urbana IL -----Original Message----- From: Ron Martin [mailto:pathrm35@adelphia.net] Sent: Thursday, August 12, 2004 4:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] proper disposal of specimen transpot biohazard bags Fellow techs, What is the proper disposal means for the specimen transport biohazard bags? The bags we use say "stat" and "biohazard" on them. I always thought that they were to be discarded in a larger biohazard bag with the other contaminated material. My manager says that they are to be discarded in the regular trash unless formalin or other fluid leaks in them. What is everyone else doing? Thanks, Ron Martin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Fri Aug 13 08:28:03 2004 From: mprice26 <@t> juno.com (Marsha R Price) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: <20040813.082804.2416.0.mprice26@juno.com> Ron, You need to have an Quality Assurance Plan in place. That is an CAP checklist question. I have a manual with logs etc. that you can have a copy of. It can be customized for your lab. You can turn this lab around with a well defined QA Program. QA is not an option it is a must/requirement if you are a CLIA or CAP certified lab. Let me know if you would like a copy of the manual. It was done on Word. So I can send to you electronically and you can make changes to it. Marsha Price On Fri, 13 Aug 2004 04:27:14 -0500 "Ron Martin" writes: > Fellow techs, > I am in a difficult situation and need some serious advice. I > recently (4 months ago) accepted a technical supervisor position in > a dermatology lab. I went from a bench tech at my old job to this > position. I also walked away from a raise at my old position so I > could step into a supervision position. I took the new position > because I was told by my manager that she would teach me some > supervision, management and financial skills that I currently do not > have as well as the growth potential of the company.This position is > not what I was told it would be. Part of my "duties" include > emptying the trash, clean the bathroom (not happening) and taking > her personal and professional calls. The question on ethics is the > high volume of mistakes made by our technician, our offices and also > by my manager herself. > I caught a mistake made by our tech a few weeks ago.She put the > wrong tissue on the wrong slides as she inverted the two cases. I > caught her mistake before it went out. One time she assigned the > same number to two different cases. She then sent the correct case > out for a consult (which wasn't needed) and put the wrong patient > name on the slide. Every other day there is something different.My > manager will not terminate her as she knows I am seeking employment > elsewhere and she cannot afford to lose a tech. > One day my manager gave three cases the same accession number.She > caught her mistake on one of the cases but I still ended up with two > cases with the same number. When I first started there my manager > had a case in which there " was no tissue in the container". She > said that she notified the office about the situation however up to > six weeks later the office was still calling looking for results.At > that point she wrote up an incident report and dated it six weeks > prior to coincide with the surgery date. > Our offices are not any better. One case came in with the wrong > patient information. It took about 7 weeks for the office to realize > that they sent the wrong patient information with the biopsy as the > names were close in spelling. The offices continually send mistakes > with incorrect spelling of patients names and incorrect anatomical > locations. > I have tired to document everything but there are too many mistakes > and I don't have enough time or energy to keep up with them.My > manager wants us to do our own "internal quality control". My > interpretation of this is that she doesn't want our physician and > risk manager to know of these mistakes. Are these becoming common > problems or is it just my situation? > I want to emphasize that I hope I am not being unethical myself for > revealing this information but I really need some advice and > support.I have very high standards and they are not being met in > this current situation. I am currently seeking a new position but I > need employment and cannot resign until another position becomes > available. I would like to stay in Florida and if anyone knows of > any positions please inform me as I am at my wits end. Thanks in > advance. > > Ron > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From mprice26 <@t> juno.com Fri Aug 13 08:33:45 2004 From: mprice26 <@t> juno.com (Marsha R Price) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: <20040813.083346.2416.1.mprice26@juno.com> Ron, Also, I would like to add that when specimens are mislabeled they need to be documented as well as what was done to correct the situation. This needs to be documented as part of your QA plan and discussed at monthly or quarterly meetings. Put in a file and records are to be retained for two years. Marsha Price On Fri, 13 Aug 2004 04:27:14 -0500 "Ron Martin" writes: > Fellow techs, > I am in a difficult situation and need some serious advice. I > recently (4 months ago) accepted a technical supervisor position in > a dermatology lab. I went from a bench tech at my old job to this > position. I also walked away from a raise at my old position so I > could step into a supervision position. I took the new position > because I was told by my manager that she would teach me some > supervision, management and financial skills that I currently do not > have as well as the growth potential of the company.This position is > not what I was told it would be. Part of my "duties" include > emptying the trash, clean the bathroom (not happening) and taking > her personal and professional calls. The question on ethics is the > high volume of mistakes made by our technician, our offices and also > by my manager herself. > I caught a mistake made by our tech a few weeks ago.She put the > wrong tissue on the wrong slides as she inverted the two cases. I > caught her mistake before it went out. One time she assigned the > same number to two different cases. She then sent the correct case > out for a consult (which wasn't needed) and put the wrong patient > name on the slide. Every other day there is something different.My > manager will not terminate her as she knows I am seeking employment > elsewhere and she cannot afford to lose a tech. > One day my manager gave three cases the same accession number.She > caught her mistake on one of the cases but I still ended up with two > cases with the same number. When I first started there my manager > had a case in which there " was no tissue in the container". She > said that she notified the office about the situation however up to > six weeks later the office was still calling looking for results.At > that point she wrote up an incident report and dated it six weeks > prior to coincide with the surgery date. > Our offices are not any better. One case came in with the wrong > patient information. It took about 7 weeks for the office to realize > that they sent the wrong patient information with the biopsy as the > names were close in spelling. The offices continually send mistakes > with incorrect spelling of patients names and incorrect anatomical > locations. > I have tired to document everything but there are too many mistakes > and I don't have enough time or energy to keep up with them.My > manager wants us to do our own "internal quality control". My > interpretation of this is that she doesn't want our physician and > risk manager to know of these mistakes. Are these becoming common > problems or is it just my situation? > I want to emphasize that I hope I am not being unethical myself for > revealing this information but I really need some advice and > support.I have very high standards and they are not being met in > this current situation. I am currently seeking a new position but I > need employment and cannot resign until another position becomes > available. I would like to stay in Florida and if anyone knows of > any positions please inform me as I am at my wits end. Thanks in > advance. > > Ron > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From garygill <@t> dcla.com Fri Aug 13 08:36:18 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070467@HALIBUT.dcla.com> Current CLIA has replaced quality assurance with quality assessment as the preferred term. Gary Gill -----Original Message----- From: Marsha R Price [mailto:mprice26@juno.com] Sent: Friday, August 13, 2004 8:28 AM To: pathrm35@adelphia.net Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] a question on ethics Ron, You need to have an Quality Assurance Plan in place. That is an CAP checklist question. I have a manual with logs etc. that you can have a copy of. It can be customized for your lab. You can turn this lab around with a well defined QA Program. QA is not an option it is a must/requirement if you are a CLIA or CAP certified lab. Let me know if you would like a copy of the manual. It was done on Word. So I can send to you electronically and you can make changes to it. Marsha Price On Fri, 13 Aug 2004 04:27:14 -0500 "Ron Martin" writes: > Fellow techs, > I am in a difficult situation and need some serious advice. I > recently (4 months ago) accepted a technical supervisor position in > a dermatology lab. I went from a bench tech at my old job to this > position. I also walked away from a raise at my old position so I > could step into a supervision position. I took the new position > because I was told by my manager that she would teach me some > supervision, management and financial skills that I currently do not > have as well as the growth potential of the company.This position is > not what I was told it would be. Part of my "duties" include > emptying the trash, clean the bathroom (not happening) and taking > her personal and professional calls. The question on ethics is the > high volume of mistakes made by our technician, our offices and also > by my manager herself. > I caught a mistake made by our tech a few weeks ago.She put the > wrong tissue on the wrong slides as she inverted the two cases. I > caught her mistake before it went out. One time she assigned the > same number to two different cases. She then sent the correct case > out for a consult (which wasn't needed) and put the wrong patient > name on the slide. Every other day there is something different.My > manager will not terminate her as she knows I am seeking employment > elsewhere and she cannot afford to lose a tech. > One day my manager gave three cases the same accession number.She > caught her mistake on one of the cases but I still ended up with two > cases with the same number. When I first started there my manager > had a case in which there " was no tissue in the container". She > said that she notified the office about the situation however up to > six weeks later the office was still calling looking for results.At > that point she wrote up an incident report and dated it six weeks > prior to coincide with the surgery date. > Our offices are not any better. One case came in with the wrong > patient information. It took about 7 weeks for the office to realize > that they sent the wrong patient information with the biopsy as the > names were close in spelling. The offices continually send mistakes > with incorrect spelling of patients names and incorrect anatomical > locations. > I have tired to document everything but there are too many mistakes > and I don't have enough time or energy to keep up with them.My > manager wants us to do our own "internal quality control". My > interpretation of this is that she doesn't want our physician and > risk manager to know of these mistakes. Are these becoming common > problems or is it just my situation? > I want to emphasize that I hope I am not being unethical myself for > revealing this information but I really need some advice and > support.I have very high standards and they are not being met in > this current situation. I am currently seeking a new position but I > need employment and cannot resign until another position becomes > available. I would like to stay in Florida and if anyone knows of > any positions please inform me as I am at my wits end. Thanks in > advance. > > Ron > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jengirl1014 <@t> yahoo.com Fri Aug 13 08:36:53 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] RE: You may want to hit the delete button with this, then again maybe not Message-ID: <20040813133653.72258.qmail@web60607.mail.yahoo.com> Way to go Joe! This is a free country, we can talk about whpever and whatever we want. If it took them this long to get a hold of you, then Good Riddance!!! They don't need you! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From aostrand <@t> seattlecca.org Fri Aug 13 08:38:09 2004 From: aostrand <@t> seattlecca.org (Ostrander, Anita B) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B94801D309CB@wala01.seattlecca.org> Ron, If your manager absolutely refuses to set up various QC checks to help prevent errors, is hiding errors, and is unwilling to do anything to improve the quality of the work at your institution I would suggest the following. 1. I would ask for one more meeting with her where you presented all your concerns to her in writing. 2. I would present her with several possible solutions that you could implement for resolving each of the serious problems at your institution. 3. If she refuses to listen to you and continues to ignore the seriousness of this situation you have no choice but to go to the next level of management with your concerns and all of your written documentation. If that does not work, you need to continue to go up the ladder until someone is willing to listen to you and work with you and your manager to start resolving these problems. As a supervisor and as a Medical Professional you owe that to the patients that are utilizing your institution. I am really sorry that there are still Histology Mangers/Supervisors out there that are more concerned with working to cover up errors than spending that time in working to find ways to prevent them in the first place. I was involved in a similar situation many, many years ago when I was a newly minted Tech. Unfortunately, because I was so "new" nobody wanted to listen to my concerns about serious errors that were being covered up and hidden from management. You are definitely going to have some hard times ahead of you but I am sure you are going to have the support from everyone on the Histonet! Anita -----Original Message----- From: Ron Martin [mailto:pathrm35@adelphia.net] Sent: Friday, August 13, 2004 2:27 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] a question on ethics Fellow techs, I am in a difficult situation and need some serious advice. I recently (4 months ago) accepted a technical supervisor position in a dermatology lab. I went from a bench tech at my old job to this position. I also walked away from a raise at my old position so I could step into a supervision position. I took the new position because I was told by my manager that she would teach me some supervision, management and financial skills that I currently do not have as well as the growth potential of the company.This position is not what I was told it would be. Part of my "duties" include emptying the trash, clean the bathroom (not happening) and taking her personal and professional calls. The question on ethics is the high volume of mistakes made by our technician, our offices and also by my manager herself. I caught a mistake made by our tech a few weeks ago.She put the wrong tissue on the wrong slides as she inverted the two cases. I caught her mistake before it went out. One time she assigned the same number to two different cases. She then sent the correct case out for a consult (which wasn't needed) and put the wrong patient name on the slide. Every other day there is something different.My manager will not terminate her as she knows I am seeking employment elsewhere and she cannot afford to lose a tech. One day my manager gave three cases the same accession number.She caught her mistake on one of the cases but I still ended up with two cases with the same number. When I first started there my manager had a case in which there " was no tissue in the container". She said that she notified the office about the situation however up to six weeks later the office was still calling looking for results.At that point she wrote up an incident report and dated it six weeks prior to coincide with the surgery date. Our offices are not any better. One case came in with the wrong patient information. It took about 7 weeks for the office to realize that they sent the wrong patient information with the biopsy as the names were close in spelling. The offices continually send mistakes with incorrect spelling of patients names and incorrect anatomical locations. I have tired to document everything but there are too many mistakes and I don't have enough time or energy to keep up with them.My manager wants us to do our own "internal quality control". My interpretation of this is that she doesn't want our physician and risk manager to know of these mistakes. Are these becoming common problems or is it just my situation? I want to emphasize that I hope I am not being unethical myself for revealing this information but I really need some advice and support.I have very high standards and they are not being met in this current situation. I am currently seeking a new position but I need employment and cannot resign until another position becomes available. I would like to stay in Florida and if anyone knows of any positions please inform me as I am at my wits end. Thanks in advance. Ron _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From JWEEMS <@t> sjha.org Fri Aug 13 08:38:09 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A62E8@sjhaexc02.sjha.org> We call it "Performance Improvement". Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gary Gill Sent: Friday, August 13, 2004 9:36 AM To: 'Marsha R Price'; pathrm35@adelphia.net Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] a question on ethics Current CLIA has replaced quality assurance with quality assessment as the preferred term. Gary Gill -----Original Message----- From: Marsha R Price [mailto:mprice26@juno.com] Sent: Friday, August 13, 2004 8:28 AM To: pathrm35@adelphia.net Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] a question on ethics Ron, You need to have an Quality Assurance Plan in place. That is an CAP checklist question. I have a manual with logs etc. that you can have a copy of. It can be customized for your lab. You can turn this lab around with a well defined QA Program. QA is not an option it is a must/requirement if you are a CLIA or CAP certified lab. Let me know if you would like a copy of the manual. It was done on Word. So I can send to you electronically and you can make changes to it. Marsha Price On Fri, 13 Aug 2004 04:27:14 -0500 "Ron Martin" writes: > Fellow techs, > I am in a difficult situation and need some serious advice. I > recently (4 months ago) accepted a technical supervisor position in > a dermatology lab. I went from a bench tech at my old job to this > position. I also walked away from a raise at my old position so I > could step into a supervision position. I took the new position > because I was told by my manager that she would teach me some > supervision, management and financial skills that I currently do not > have as well as the growth potential of the company.This position is > not what I was told it would be. Part of my "duties" include > emptying the trash, clean the bathroom (not happening) and taking > her personal and professional calls. The question on ethics is the > high volume of mistakes made by our technician, our offices and also > by my manager herself. > I caught a mistake made by our tech a few weeks ago.She put the > wrong tissue on the wrong slides as she inverted the two cases. I > caught her mistake before it went out. One time she assigned the > same number to two different cases. She then sent the correct case > out for a consult (which wasn't needed) and put the wrong patient > name on the slide. Every other day there is something different.My > manager will not terminate her as she knows I am seeking employment > elsewhere and she cannot afford to lose a tech. > One day my manager gave three cases the same accession number.She > caught her mistake on one of the cases but I still ended up with two > cases with the same number. When I first started there my manager > had a case in which there " was no tissue in the container". She > said that she notified the office about the situation however up to > six weeks later the office was still calling looking for results.At > that point she wrote up an incident report and dated it six weeks > prior to coincide with the surgery date. > Our offices are not any better. One case came in with the wrong > patient information. It took about 7 weeks for the office to realize > that they sent the wrong patient information with the biopsy as the > names were close in spelling. The offices continually send mistakes > with incorrect spelling of patients names and incorrect anatomical > locations. > I have tired to document everything but there are too many mistakes > and I don't have enough time or energy to keep up with them.My > manager wants us to do our own "internal quality control". My > interpretation of this is that she doesn't want our physician and > risk manager to know of these mistakes. Are these becoming common > problems or is it just my situation? > I want to emphasize that I hope I am not being unethical myself for > revealing this information but I really need some advice and > support.I have very high standards and they are not being met in > this current situation. I am currently seeking a new position but I > need employment and cannot resign until another position becomes > available. I would like to stay in Florida and if anyone knows of > any positions please inform me as I am at my wits end. Thanks in > advance. > > Ron > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From kmerriam2003 <@t> yahoo.com Fri Aug 13 09:16:03 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Image analysis and slide scanning machine Message-ID: <20040813141603.13492.qmail@web52505.mail.yahoo.com> Hello everyone, Years ago, I heard of a machine that you could load an entire slide into, scan the slide and then do image analysis on the tissue. I have no idea who made this machine or even how to search for such a thing on the internet (or the histonet archives). If anyone has the name of this company, can someone please let me know. I am currently doing measurements of pancreatic islets as well as measurements of whole mouse pancreas'. I am needing to take 4-6 pictures per pancreas in order to do this, and it is very time consuming (these studies have up to 90 animals in them). I need a quicker and easier way to do this! Help! Kim Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From garygill <@t> dcla.com Fri Aug 13 09:32:06 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Image analysis and slide scanning machine Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070470@HALIBUT.dcla.com> CAS-200 Workstation: http://www.bacuslabs.com/indexcas200.html. There have been others, though most have died. Gary Gill -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: Friday, August 13, 2004 9:16 AM To: Histonet Subject: [Histonet] Image analysis and slide scanning machine Hello everyone, Years ago, I heard of a machine that you could load an entire slide into, scan the slide and then do image analysis on the tissue. I have no idea who made this machine or even how to search for such a thing on the internet (or the histonet archives). If anyone has the name of this company, can someone please let me know. I am currently doing measurements of pancreatic islets as well as measurements of whole mouse pancreas'. I am needing to take 4-6 pictures per pancreas in order to do this, and it is very time consuming (these studies have up to 90 animals in them). I need a quicker and easier way to do this! Help! Kim Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rbarnhart <@t> summithealth.org Fri Aug 13 09:38:39 2004 From: rbarnhart <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: Ron, I feel for you. I thought the mistakes in my lab were bad but nothing compared to yours. We just have some secretaries that have more important things to do then their jobs. I agree with everyone else talk to your manager one last time and keep going up the ladder until someone listen and does something. We are medical professionals and the mistakes you mentioned are totally unacceptable. Why are the pathologist not in an up roar? I know if our pathologist would get slides mislabeled, find out of slides being mislabeled or any of the mistakes in your lab he would not take it lightly and would be taking action himself. Marsha Can you email me your QA plan manual. I have been working on this in my lab and have some items in place but I would love to see your ideas. Thanks Becky >>> Marsha R Price 8/13/2004 9:28:03 AM >>> Ron, You need to have an Quality Assurance Plan in place. That is an CAP checklist question. I have a manual with logs etc. that you can have a copy of. It can be customized for your lab. You can turn this lab around with a well defined QA Program. QA is not an option it is a must/requirement if you are a CLIA or CAP certified lab. Let me know if you would like a copy of the manual. It was done on Word. So I can send to you electronically and you can make changes to it. Marsha Price On Fri, 13 Aug 2004 04:27:14 -0500 "Ron Martin" writes: > Fellow techs, > I am in a difficult situation and need some serious advice. I > recently (4 months ago) accepted a technical supervisor position in > a dermatology lab. I went from a bench tech at my old job to this > position. I also walked away from a raise at my old position so I > could step into a supervision position. I took the new position > because I was told by my manager that she would teach me some > supervision, management and financial skills that I currently do not > have as well as the growth potential of the company.This position is > not what I was told it would be. Part of my "duties" include > emptying the trash, clean the bathroom (not happening) and taking > her personal and professional calls. The question on ethics is the > high volume of mistakes made by our technician, our offices and also > by my manager herself. > I caught a mistake made by our tech a few weeks ago.She put the > wrong tissue on the wrong slides as she inverted the two cases. I > caught her mistake before it went out. One time she assigned the > same number to two different cases. She then sent the correct case > out for a consult (which wasn't needed) and put the wrong patient > name on the slide. Every other day there is something different.My > manager will not terminate her as she knows I am seeking employment > elsewhere and she cannot afford to lose a tech. > One day my manager gave three cases the same accession number.She > caught her mistake on one of the cases but I still ended up with two > cases with the same number. When I first started there my manager > had a case in which there " was no tissue in the container". She > said that she notified the office about the situation however up to > six weeks later the office was still calling looking for results.At > that point she wrote up an incident report and dated it six weeks > prior to coincide with the surgery date. > Our offices are not any better. One case came in with the wrong > patient information. It took about 7 weeks for the office to realize > that they sent the wrong patient information with the biopsy as the > names were close in spelling. The offices continually send mistakes > with incorrect spelling of patients names and incorrect anatomical > locations. > I have tired to document everything but there are too many mistakes > and I don't have enough time or energy to keep up with them.My > manager wants us to do our own "internal quality control". My > interpretation of this is that she doesn't want our physician and > risk manager to know of these mistakes. Are these becoming common > problems or is it just my situation? > I want to emphasize that I hope I am not being unethical myself for > revealing this information but I really need some advice and > support.I have very high standards and they are not being met in > this current situation. I am currently seeking a new position but I > need employment and cannot resign until another position becomes > available. I would like to stay in Florida and if anyone knows of > any positions please inform me as I am at my wits end. Thanks in > advance. > > Ron > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Fri Aug 13 09:44:10 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070471@HALIBUT.dcla.com> Read about the fiasco at Maryland General Hospital, which appears to parallel your situation: http://www.hhs.gov/asl/testify/t040518.html Get thee to your organization's compliance officer. The behaviors you described put everyone at risk. Your organization's compliance policy should include no retribution for those who report compliance problems in good faith. Gary Gill Corporate Compliance Officer -----Original Message----- From: Rebecca Barnhart [mailto:rbarnhart@summithealth.org] Sent: Friday, August 13, 2004 9:39 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] a question on ethics Ron, I feel for you. I thought the mistakes in my lab were bad but nothing compared to yours. We just have some secretaries that have more important things to do then their jobs. I agree with everyone else talk to your manager one last time and keep going up the ladder until someone listen and does something. We are medical professionals and the mistakes you mentioned are totally unacceptable. Why are the pathologist not in an up roar? I know if our pathologist would get slides mislabeled, find out of slides being mislabeled or any of the mistakes in your lab he would not take it lightly and would be taking action himself. Marsha Can you email me your QA plan manual. I have been working on this in my lab and have some items in place but I would love to see your ideas. Thanks Becky >>> Marsha R Price 8/13/2004 9:28:03 AM >>> Ron, You need to have an Quality Assurance Plan in place. That is an CAP checklist question. I have a manual with logs etc. that you can have a copy of. It can be customized for your lab. You can turn this lab around with a well defined QA Program. QA is not an option it is a must/requirement if you are a CLIA or CAP certified lab. Let me know if you would like a copy of the manual. It was done on Word. So I can send to you electronically and you can make changes to it. Marsha Price On Fri, 13 Aug 2004 04:27:14 -0500 "Ron Martin" writes: > Fellow techs, > I am in a difficult situation and need some serious advice. I > recently (4 months ago) accepted a technical supervisor position in > a dermatology lab. I went from a bench tech at my old job to this > position. I also walked away from a raise at my old position so I > could step into a supervision position. I took the new position > because I was told by my manager that she would teach me some > supervision, management and financial skills that I currently do not > have as well as the growth potential of the company.This position is > not what I was told it would be. Part of my "duties" include > emptying the trash, clean the bathroom (not happening) and taking > her personal and professional calls. The question on ethics is the > high volume of mistakes made by our technician, our offices and also > by my manager herself. > I caught a mistake made by our tech a few weeks ago.She put the > wrong tissue on the wrong slides as she inverted the two cases. I > caught her mistake before it went out. One time she assigned the > same number to two different cases. She then sent the correct case > out for a consult (which wasn't needed) and put the wrong patient > name on the slide. Every other day there is something different.My > manager will not terminate her as she knows I am seeking employment > elsewhere and she cannot afford to lose a tech. > One day my manager gave three cases the same accession number.She > caught her mistake on one of the cases but I still ended up with two > cases with the same number. When I first started there my manager > had a case in which there " was no tissue in the container". She > said that she notified the office about the situation however up to > six weeks later the office was still calling looking for results.At > that point she wrote up an incident report and dated it six weeks > prior to coincide with the surgery date. > Our offices are not any better. One case came in with the wrong > patient information. It took about 7 weeks for the office to realize > that they sent the wrong patient information with the biopsy as the > names were close in spelling. The offices continually send mistakes > with incorrect spelling of patients names and incorrect anatomical > locations. > I have tired to document everything but there are too many mistakes > and I don't have enough time or energy to keep up with them.My > manager wants us to do our own "internal quality control". My > interpretation of this is that she doesn't want our physician and > risk manager to know of these mistakes. Are these becoming common > problems or is it just my situation? > I want to emphasize that I hope I am not being unethical myself for > revealing this information but I really need some advice and > support.I have very high standards and they are not being met in > this current situation. I am currently seeking a new position but I > need employment and cannot resign until another position becomes > available. I would like to stay in Florida and if anyone knows of > any positions please inform me as I am at my wits end. Thanks in > advance. > > Ron > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From John.Auld <@t> whnt.nhs.uk Fri Aug 13 09:55:45 2004 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Re: a question of ethics Message-ID: Ron It sounds like you are not in a position to run - so don't! Go to your manager and say that QCing is required. Give reasons, suggestions (firmly but politely), see what response is. If not favourable or nothing happens SOON, go to next up the chain of command and Risk Management. If nothing happens there then LEAVE it's not the sort of place that a professional should be working in! Who knows maybe if you're real luckly you could end up as the manager:>) Also involve your pathologists - it's their backside in court potentially. Good luck, be brave John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral Internal extn 2560 External Tel 0151 604 7025 Subject: [Histonet] a question on ethics Fellow techs, I am in a difficult situation and need some serious advice. I recently (4 months ago) accepted a technical supervisor position in a dermatology lab. I went from a bench tech at my old job to this position. I also walked away from a raise at my old position so I could step into a supervision position. I took the new position because I was told by my manager that she would teach me some supervision, management and financial skills that I currently do not have as well as the growth potential of the company.This position is not what I was told it would be. Part of my "duties" include emptying the trash, clean the bathroom (not happening) and taking her personal and professional calls. The question on ethics is the high volume of mistakes made by our technician, our offices and also by my manager herself. I caught a mistake made by our tech a few weeks ago.She put the wrong tissue on the wrong slides as she inverted the two cases. I caught her mistake before it went out. One time she assigned the same number to two different cases. She then sent the correct case out for a consult (which wasn't needed) and put the wrong patient name on the slide. Every other day there is something different.My manager will not terminate her as she knows I am seeking employment elsewhere and she cannot afford to lose a tech. One day my manager gave three cases the same accession number.She caught her mistake on one of the cases but I still ended up with two cases with the same number. When I first started there my manager had a case in which there " was no tissue in the container". She said that she notified the office about the situation however up to six weeks later the office was still calling looking for results.At that point she wrote up an incident report and dated it six weeks prior to coincide with the surgery date. Our offices are not any better. One case came in with the wrong patient information. It took about 7 weeks for the office to realize that they sent the wrong patient information with the biopsy as the names were close in spelling. The offices continually send mistakes with incorrect spelling of patients names and incorrect anatomical locations. I have tired to document everything but there are too many mistakes and I don't have enough time or energy to keep up with them.My manager wants us to do our own "internal quality control". My interpretation of this is that she doesn't want our physician and risk manager to know of these mistakes. Are these becoming common problems or is it just my situation? I want to emphasize that I hope I am not being unethical myself for revealing this information but I really need some advice and support.I have very high standards and they are not being met in this current situation. I am currently seeking a new position but I need employment and cannot resign until another position becomes available. I would like to stay in Florida and if anyone knows of any positions please inform me as I am at my wits end. Thanks in advance. Ron From al.floyd <@t> juno.com Fri Aug 13 09:49:07 2004 From: al.floyd <@t> juno.com (Alton D. Floyd) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Image analysis and slide scanning machine Message-ID: <20040813.104911.-581867.2.al.floyd@juno.com> Hello Kim, There are at least three companies which are developing instruments for creating the "virtual slide" or "electronic slide". They are MicroBrightfield, Interscope and Aperio. Each uses somewhat different technology, and each generates extremely large files (hundreds of megabytes). One you have such a large file, it very time consuming to perform image analysis, unless you chop the file up into more manageable pieces. It can be done, but at the current state of technology development, it may not be the time saver you are looking for. You might also look at the offerings from Chromavision. This company also offers a scanning microscope, that can find objects at low magnification (assuming a specific stain), and then return to that area of the slide and grab a higher resolution image. Similar capability can be found in the Ariol offering from Applied Image. It would also be possible to create a system to do this using a general image analysis program such as ImagePro, Northern Eclipse, Aphelion, or others, but this would need a considerable amount of customization. Al Floyd Alton D. Floyd, Ph.D. 23126 South Shore Drive Edwardsburg, MI 49112 (269) 699-7182 phone & fax From Jackie.O'Connor <@t> abbott.com Fri Aug 13 09:50:01 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Image analysis and slide scanning machine Message-ID: Chromavision does this, as well as the Zeiss Mosaix program. I use the Mosaix to photograph the entire slide by taking a series of individual shots, (the program does this automatically, moving the stage to your set coordinates) and putting them back together like mosaic tiles. The photo has impeccable clarity - you'd never know it wasn't one whole photograph. I do image analysis for IHC (apoptosis, proliferation, microvessel density) as well as measuring areas of tumor growth, necrosis, etc.- it's a great system - I'm impressed. Jackie O'Connor Abbott Labs Gary Gill Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 09:32 AM To: "'Kim Merriam'" , Histonet cc: Subject: RE: [Histonet] Image analysis and slide scanning machine CAS-200 Workstation: http://www.bacuslabs.com/indexcas200.html. There have been others, though most have died. Gary Gill -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: Friday, August 13, 2004 9:16 AM To: Histonet Subject: [Histonet] Image analysis and slide scanning machine Hello everyone, Years ago, I heard of a machine that you could load an entire slide into, scan the slide and then do image analysis on the tissue. I have no idea who made this machine or even how to search for such a thing on the internet (or the histonet archives). If anyone has the name of this company, can someone please let me know. I am currently doing measurements of pancreatic islets as well as measurements of whole mouse pancreas'. I am needing to take 4-6 pictures per pancreas in order to do this, and it is very time consuming (these studies have up to 90 animals in them). I need a quicker and easier way to do this! Help! Kim Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Fri Aug 13 10:14:34 2004 From: mprice26 <@t> juno.com (Marsha R Price) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: <20040813.101434.2328.0.mprice26@juno.com> Actually, the hospital where I worked for the last year called it "Performance Improvement" also. I do not think they care much about what you call it as long as you have a well defined and documented plan in place. Marsha On Fri, 13 Aug 2004 08:36:18 -0500 Gary Gill writes: > Current CLIA has replaced quality assurance with quality assessment > as the > preferred term. > > Gary Gill > > -----Original Message----- > From: Marsha R Price [mailto:mprice26@juno.com] > Sent: Friday, August 13, 2004 8:28 AM > To: pathrm35@adelphia.net > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] a question on ethics > > > Ron, > You need to have an Quality Assurance Plan in place. That is an CAP > checklist question. I have a manual with logs etc. that you can have > a copy > of. It can be customized for your lab. You can turn this lab around > with a > well defined QA Program. QA is not an option it is a > must/requirement if you > are a CLIA or CAP certified lab. > > Let me know if you would like a copy of the manual. It was done on > Word. So > I can send to you electronically and you can make changes to it. > > Marsha Price > > On Fri, 13 Aug 2004 04:27:14 -0500 "Ron Martin" > > writes: > > Fellow techs, > > I am in a difficult situation and need some serious advice. I > > recently (4 months ago) accepted a technical supervisor position > in > > a dermatology lab. I went from a bench tech at my old job to this > > > position. I also walked away from a raise at my old position so I > > > could step into a supervision position. I took the new position > > because I was told by my manager that she would teach me some > > supervision, management and financial skills that I currently do > not > > have as well as the growth potential of the company.This position > is > > not what I was told it would be. Part of my "duties" include > > emptying the trash, clean the bathroom (not happening) and taking > > > her personal and professional calls. The question on ethics is the > > > high volume of mistakes made by our technician, our offices and > also > > by my manager herself. > > I caught a mistake made by our tech a few weeks ago.She put the > > wrong tissue on the wrong slides as she inverted the two cases. I > > caught her mistake before it went out. One time she assigned the > > same number to two different cases. She then sent the correct case > > > out for a consult (which wasn't needed) and put the wrong patient > > > name on the slide. Every other day there is something different.My > > > manager will not terminate her as she knows I am seeking > employment > > elsewhere and she cannot afford to lose a tech. > > One day my manager gave three cases the same accession number.She > > > caught her mistake on one of the cases but I still ended up with > two > > cases with the same number. When I first started there my manager > > > had a case in which there " was no tissue in the container". She > > said that she notified the office about the situation however up > to > > six weeks later the office was still calling looking for > results.At > > that point she wrote up an incident report and dated it six weeks > > > prior to coincide with the surgery date. > > Our offices are not any better. One case came in with the wrong > > patient information. It took about 7 weeks for the office to > realize > > that they sent the wrong patient information with the biopsy as > the > > names were close in spelling. The offices continually send > mistakes > > with incorrect spelling of patients names and incorrect anatomical > > > locations. > > I have tired to document everything but there are too many > mistakes > > and I don't have enough time or energy to keep up with them.My > > manager wants us to do our own "internal quality control". My > > interpretation of this is that she doesn't want our physician and > > > risk manager to know of these mistakes. Are these becoming common > > > problems or is it just my situation? > > I want to emphasize that I hope I am not being unethical myself > for > > revealing this information but I really need some advice and > > support.I have very high standards and they are not being met in > > this current situation. I am currently seeking a new position but > I > > need employment and cannot resign until another position becomes > > available. I would like to stay in Florida and if anyone knows of > > > any positions please inform me as I am at my wits end. Thanks in > > advance. > > > > Ron > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ________________________________________________________________ > The best thing to hit the Internet in years - Juno SpeedBand! Surf > the Web > up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to > sign up > today! > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From STEGTM <@t> samcstl.org Fri Aug 13 10:26:10 2004 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Staining smears Message-ID: A question, if you will. Does anyone out there know of a method to stain peripheral and bone marrow smears on an automated stainer (we recently obtained a Ventana Nexes). I'm referrring to the routine iron stains we usually run on several smears, along with the paraffin-embedded marrow clot slides. Right now, I'm running the paraffin sections on the stainer, and doing the smears by hand for fear of losing the smears on the machine. Any ideas? Peace, Terre From garygill <@t> dcla.com Fri Aug 13 10:43:18 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] a question on ethics Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070474@HALIBUT.dcla.com> True, but it's helpful to use the same language as that used by the regulatory folks. Regardless, make certain everyone understands the difference between QC and QA and puts various activities in the right bin: QUALITY CONTROL AND QUALITY ASSESSMENT Quality control activities look forward. They define the product's quality, imparting to it the credibility needed for its intended purpose. QC activities are the result of planning and are applied prospectively to everything that contributes to the final product, thereby impacting the outcome. QC activities are deterministic (i.e., lead to expected results when followed). Quality assessment activities look backward. They measure the degree to which desired outcomes are successful (i.e., their impact). QA activities, therefore, retrospectively sample outcomes. The findings modify the processes that contribute to the final product (e.g., did the patient have cancer as reported; if not, why?). As a practical matter quality assessment activities are probabilistic (i.e., have attendant uncertainty relative to reliability), as it not possible to review all product outcomes. DIFFERENTIAL FEATURES OF QUALITY CONTROL AND ASSESSMENT The following table is intended to help distinguish whether an activity is one of quality control or quality assessment: Differential Feature Quality Control Quality Assessment Purpose Defines Quality Measures Success Timing Prospective Retrospective Application All Processes Sample Outcomes Impact Outcomes Processes Nature Deterministic Probabilistic Gary Gill -----Original Message----- From: Marsha R Price [mailto:mprice26@juno.com] Sent: Friday, August 13, 2004 10:15 AM To: garygill@dcla.com Cc: pathrm35@adelphia.net; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] a question on ethics Actually, the hospital where I worked for the last year called it "Performance Improvement" also. I do not think they care much about what you call it as long as you have a well defined and documented plan in place. Marsha On Fri, 13 Aug 2004 08:36:18 -0500 Gary Gill writes: > Current CLIA has replaced quality assurance with quality assessment > as the > preferred term. > > Gary Gill > > -----Original Message----- > From: Marsha R Price [mailto:mprice26@juno.com] > Sent: Friday, August 13, 2004 8:28 AM > To: pathrm35@adelphia.net > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] a question on ethics > > > Ron, > You need to have an Quality Assurance Plan in place. That is an CAP > checklist question. I have a manual with logs etc. that you can have a > copy of. It can be customized for your lab. You can turn this lab > around with a > well defined QA Program. QA is not an option it is a > must/requirement if you > are a CLIA or CAP certified lab. > > Let me know if you would like a copy of the manual. It was done on > Word. So > I can send to you electronically and you can make changes to it. > > Marsha Price > > On Fri, 13 Aug 2004 04:27:14 -0500 "Ron Martin" > > writes: > > Fellow techs, > > I am in a difficult situation and need some serious advice. I > > recently (4 months ago) accepted a technical supervisor position > in > > a dermatology lab. I went from a bench tech at my old job to this > > > position. I also walked away from a raise at my old position so I > > > could step into a supervision position. I took the new position > > because I was told by my manager that she would teach me some > > supervision, management and financial skills that I currently do > not > > have as well as the growth potential of the company.This position > is > > not what I was told it would be. Part of my "duties" include > > emptying the trash, clean the bathroom (not happening) and taking > > > her personal and professional calls. The question on ethics is the > > > high volume of mistakes made by our technician, our offices and > also > > by my manager herself. > > I caught a mistake made by our tech a few weeks ago.She put the > > wrong tissue on the wrong slides as she inverted the two cases. I > > caught her mistake before it went out. One time she assigned the > > same number to two different cases. She then sent the correct case > > > out for a consult (which wasn't needed) and put the wrong patient > > > name on the slide. Every other day there is something different.My > > > manager will not terminate her as she knows I am seeking > employment > > elsewhere and she cannot afford to lose a tech. > > One day my manager gave three cases the same accession number.She > > > caught her mistake on one of the cases but I still ended up with > two > > cases with the same number. When I first started there my manager > > > had a case in which there " was no tissue in the container". She > > said that she notified the office about the situation however up > to > > six weeks later the office was still calling looking for > results.At > > that point she wrote up an incident report and dated it six weeks > > > prior to coincide with the surgery date. > > Our offices are not any better. One case came in with the wrong > > patient information. It took about 7 weeks for the office to > realize > > that they sent the wrong patient information with the biopsy as > the > > names were close in spelling. The offices continually send > mistakes > > with incorrect spelling of patients names and incorrect anatomical > > > locations. > > I have tired to document everything but there are too many > mistakes > > and I don't have enough time or energy to keep up with them.My > > manager wants us to do our own "internal quality control". My > > interpretation of this is that she doesn't want our physician and > > > risk manager to know of these mistakes. Are these becoming common > > > problems or is it just my situation? > > I want to emphasize that I hope I am not being unethical myself > for > > revealing this information but I really need some advice and > > support.I have very high standards and they are not being met in > > this current situation. I am currently seeking a new position but > I > > need employment and cannot resign until another position becomes > > available. I would like to stay in Florida and if anyone knows of > > > any positions please inform me as I am at my wits end. Thanks in > > advance. > > > > Ron > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ________________________________________________________________ > The best thing to hit the Internet in years - Juno SpeedBand! Surf > the Web > up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to > sign up > today! > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From laurie.colbert <@t> huntingtonhospital.com Fri Aug 13 10:59:00 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Staining smears Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5C001@EXCHANGE1.huntingtonhospital.com> We have the Ventana Nexes and we occasionally run special stains on smears. We fix the smears in 95% alcohol and rinse before staining. Laurie Colbert -----Original Message----- From: Therersa Stegall [mailto:STEGTM@samcstl.org] Sent: Friday, August 13, 2004 8:26 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Staining smears A question, if you will. Does anyone out there know of a method to stain peripheral and bone marrow smears on an automated stainer (we recently obtained a Ventana Nexes). I'm referrring to the routine iron stains we usually run on several smears, along with the paraffin-embedded marrow clot slides. Right now, I'm running the paraffin sections on the stainer, and doing the smears by hand for fear of losing the smears on the machine. Any ideas? Peace, Terre _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PWebster <@t> hei.org Fri Aug 13 11:09:40 2004 From: PWebster <@t> hei.org (Webster, Paul) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] RE: replies to immunostaining on technovit 8100 Message-ID: <4E8C1F1E4E8FA748B5487C50C91695F0862FA4@heimail.hei.org> There is a new resin that has just bee released called Technovit 9100 New. I have no experience with this resin but in addition to polymerizing at low temperatures, it is also possible to remove the resin from tissue sections using 2-methoxyethyl acetate. There is a paper describing its use in European Cells and Materials 2003 vol 6 pages 57-71. Authors are Yang, R., CM Davies, CW Archer and RG Richards. Any comments from people who may have used this resin would be of interest to me. Regards, Paul Webster. Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 phone (213) 273 8026 fax (213) 413 6739 email: pwebster@hei.org > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of PALMER Jason (SVHM) > Sent: Thursday, August 12, 2004 7:13 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: replies to immunostaining on technovit 8100 > > <><> > > > > > Thanks Pam Marcum and Gayle (especially) for your thoughtful responses. > > Perhaps I should have been clearer as to why we want to use this rather than paraffin - the tissues we collect are grown on a PLGA (a polymer of lactic and glycolic acids) scaffold, which melts / distorts at any temp above 45 degrees c, not to mention in alcohols,acetone, xylene and the like. Hence our desire to avoid paraffin. Technovit 8100 polymerises at zero degrees, so no heat needed at all, and we use inert dehydration to remove excess water from the PLGA / tissues. And, supposedly immunohistochemistry compatible.... > > And yes, I got the mouse and rat confused with the antibody - mouse anti rat ED1 it is! > > Cheers, > > Jason Palmer > Bernard O'Brien Institute of Microsurgery > Melbourne, Australia > > > > From LPrestridge <@t> seton.org Fri Aug 13 11:10:42 2004 From: LPrestridge <@t> seton.org (Prestridge, Linda) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] biopsies swelling? Message-ID: Dear Techs, Our lab is noticing that our small bx.'s ie gastric mainly . After they have been sectioned, and we need to go back for add't sections for s.stains, recuts, IHC- the tissue has swelled up, and is white. The Pathologists haven't noticed anything. We have formalin, penfix, 80%, 95%,100% on the VIP--so we know they are getting dehydrated. We don't leave them on the iceplate to cause water swelling. Any ideas. Thanks Linda- Seton Hosp. Austin,Tx. From Barbara_Lentz <@t> dahlchase.com Fri Aug 13 11:30:05 2004 From: Barbara_Lentz <@t> dahlchase.com (Barbara Lentz) Date: Fri Sep 16 15:23:53 2005 Subject: [Histonet] Staining smears Message-ID: We have been running our smears on the Artisan stainer. However, we found the Wash solution was hemolyzing the RBC's. We then started fixing the smears with Methanol prior to staining. When we brought the Ventana on board for testing, we did the same thing. We were getting fine staining on both machines. Barb >>> "Therersa Stegall" 08/13/04 11:26AM >>> A question, if you will. Does anyone out there know of a method to stain peripheral and bone marrow smears on an automated stainer (we recently obtained a Ventana Nexes). I'm referrring to the routine iron stains we usually run on several smears, along with the paraffin-embedded marrow clot slides. Right now, I'm running the paraffin sections on the stainer, and doing the smears by hand for fear of losing the smears on the machine. Any ideas? Peace, Terre _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Aug 13 11:41:59 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Bubbles in Thick Sections - Suggestions? In-Reply-To: <20040813040238.8280.qmail@web50308.mail.yahoo.com> References: <20040813040238.8280.qmail@web50308.mail.yahoo.com> Message-ID: <6.0.0.22.1.20040813103353.01b47128@gemini.msu.montana.edu> Are your sections tape transferred? I think you need the xylene or clearant to allow the flow of mounting media into and around all spaces of the thicker sections. I would go back to using a permanent mounting media and mount the coverslip on sections just coming out of your clearant. Less air is trapped if the solvent is there, although you can add bubbles, obviously. Make sure the coverslip is lowered onto a generous drop of mounting media so thicker section fills with media, do not overly squeege the excess from under coverslip - you can even put the media on top of the section to help fill spaces in clearant wetted section. If you coverslip air dried sections, thin out a permanent mounting media so it flows easily while coverslip is being mounted, you can also add a tiny drop of clearant to edge of mounted coverslip, it will flow under and join media, things fill in nicely. Good luck At 10:02 PM 8/12/2004, you wrote: >We cut sections of tissue between 20 um and 30 um thick to view blood >vessels after being stained with CD31. We also section tissue at >thicknesses of 12 to 16 um for our H+E stains. > >We sometimes get bubbles when we coverslip with non-aqueous mounting media >and hate to see them in well-stained slides. Does anyone have any >suggestions for not getting bubbles in thick sections of tissue? After the >last xylene step we allow the sections to dry to ensure that the tissues >are not wet and we are currently using CureMount II, a non-aqueous >UV-cured mounting media from Instrumedics. > >Help me destroy the bubble monster! Please.... > >Thanks in advance, > >Phil > > >--------------------------------- >Do you Yahoo!? >Yahoo! Mail Address AutoComplete - You start. We finish. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From pathrm35 <@t> adelphia.net Fri Aug 13 12:33:44 2004 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] a question on ethics Message-ID: <20040813173343.CJSU24693.mta13.adelphia.net@mail.adelphia.net> I sent copies of incident reports to RM but my manager wants us to do our own internal quality control. How are things suppose to get fixed that way? My physician doesn't like to deal with these situations and refers me to my manager. I'm running in circles!!! Ron > > From: "pam marcum" > Date: 2004/08/13 Fri AM 09:11:34 EDT > To: , , "Joe Nocito" > Subject: RE: [Histonet] a question on ethics > > From pathrm35 <@t> adelphia.net Fri Aug 13 12:34:04 2004 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] a question on ethics Message-ID: <20040813173404.CJXJ24693.mta13.adelphia.net@mail.adelphia.net> I sent copies of incident reports to RM but my manager wants us to do our own internal quality control. How are things suppose to get fixed that way? My physician doesn't like to deal with these situations and refers me to my manager. I'm running in circles!!! Ron > > From: "pam marcum" > Date: 2004/08/13 Fri AM 09:11:34 EDT > To: , , "Joe Nocito" > Subject: RE: [Histonet] a question on ethics > > From garygill <@t> dcla.com Fri Aug 13 12:39:01 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] a question on ethics Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070483@HALIBUT.dcla.com> Let your physician know that the person in charge is ultimately responsible that the lab is run by the book. There are right ways and wrong ways to do things. Wrong is indefensible. Among possible penalties, CMS can shut down the lab. Surely your boss doesn't want to see the lab's name in the local newspaper headlines. Gary Gill -----Original Message----- From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] Sent: Friday, August 13, 2004 12:34 PM To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito Cc: histonet@lists.utsouthwestern.edu Subject: Re: RE: [Histonet] a question on ethics I sent copies of incident reports to RM but my manager wants us to do our own internal quality control. How are things suppose to get fixed that way? My physician doesn't like to deal with these situations and refers me to my manager. I'm running in circles!!! Ron > > From: "pam marcum" > Date: 2004/08/13 Fri AM 09:11:34 EDT > To: , , > "Joe Nocito" > Subject: RE: [Histonet] a question on ethics > > From WesterM <@t> MedImmune.com Fri Aug 13 12:46:31 2004 From: WesterM <@t> MedImmune.com (Wester, Martha) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Image analysis and slide scanning machine Message-ID: <83899F0EC7671543B305FB5694024DE605F88416@medimmune4.medimmune.com> Hi Kim We just purchased a Chromavision system and our pathologist loves it! http://www.chromavision.com/ Martha Wester westem@medimmune.com Associate Scientist MedImmune, Inc. Gaithersburg, MD 20878 -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: Friday, August 13, 2004 9:16 AM To: Histonet Subject: [Histonet] Image analysis and slide scanning machine Hello everyone, Years ago, I heard of a machine that you could load an entire slide into, scan the slide and then do image analysis on the tissue. I have no idea who made this machine or even how to search for such a thing on the internet (or the histonet archives). If anyone has the name of this company, can someone please let me know. I am currently doing measurements of pancreatic islets as well as measurements of whole mouse pancreas'. I am needing to take 4-6 pictures per pancreas in order to do this, and it is very time consuming (these studies have up to 90 animals in them). I need a quicker and easier way to do this! Help! Kim Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From pathrm35 <@t> adelphia.net Fri Aug 13 12:47:06 2004 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] a question on ethics Message-ID: <20040813174706.CQCA24693.mta13.adelphia.net@mail.adelphia.net> There is also the issue that this position is not what I was told it would be. I also didn't expect to have so many issues such as a tech that can't work up to my standards. Also I keep asking myself if it is worth it esp. when this isn't turning out to be the job I expected it to be. Lastly, how can I work for someone I can't respect or trust? Any thought on that? > > From: Gary Gill > Date: 2004/08/13 Fri PM 01:39:01 EDT > To: "'pathrm35@adelphia.net'" , pam marcum > , histonet@lists.utsouthwestern.edu, Joe Nocito > > CC: histonet@lists.utsouthwestern.edu > Subject: RE: RE: [Histonet] a question on ethics > > Let your physician know that the person in charge is ultimately responsible > that the lab is run by the book. There are right ways and wrong ways to do > things. Wrong is indefensible. Among possible penalties, CMS can shut down > the lab. Surely your boss doesn't want to see the lab's name in the local > newspaper headlines. > > Gary Gill > > -----Original Message----- > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > Sent: Friday, August 13, 2004 12:34 PM > To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: RE: [Histonet] a question on ethics > > > I sent copies of incident reports to RM but my manager wants us to do our > own internal quality control. How are things suppose to get fixed that way? > My physician doesn't like to deal with these situations and refers me to my > manager. I'm running in circles!!! Ron > > > > From: "pam marcum" > > Date: 2004/08/13 Fri AM 09:11:34 EDT > > To: , , > > "Joe Nocito" > > Subject: RE: [Histonet] a question on ethics > > > > > From andrae <@t> u.washington.edu Fri Aug 13 13:14:55 2004 From: andrae <@t> u.washington.edu (A. Erickson) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] a question on ethics In-Reply-To: <20040813174706.CQCA24693.mta13.adelphia.net@mail.adelphia.net> References: <20040813174706.CQCA24693.mta13.adelphia.net@mail.adelphia.net> Message-ID: It sounds to me, from the thread of your post(s) that you have already made the decision to leave, just not implemented it! Good luck! Andra Erickson, HT, Research Technologist On Fri, 13 Aug 2004 pathrm35@adelphia.net wrote: > There is also the issue that this position is not what I was told it would be. I also didn't expect to have so many issues such as a tech that can't work up to my standards. Also I keep asking myself if it is worth it esp. when this isn't turning out to be the job I expected it to be. Lastly, how can I work for someone I can't respect or trust? Any thought on that? > > > > From: Gary Gill > > Date: 2004/08/13 Fri PM 01:39:01 EDT > > To: "'pathrm35@adelphia.net'" , pam marcum > > , histonet@lists.utsouthwestern.edu, Joe Nocito > > > > CC: histonet@lists.utsouthwestern.edu > > Subject: RE: RE: [Histonet] a question on ethics > > > > Let your physician know that the person in charge is ultimately responsible > > that the lab is run by the book. There are right ways and wrong ways to do > > things. Wrong is indefensible. Among possible penalties, CMS can shut down > > the lab. Surely your boss doesn't want to see the lab's name in the local > > newspaper headlines. > > > > Gary Gill > > > > -----Original Message----- > > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > > Sent: Friday, August 13, 2004 12:34 PM > > To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > > Cc: histonet@lists.utsouthwestern.edu > > Subject: Re: RE: [Histonet] a question on ethics > > > > > > I sent copies of incident reports to RM but my manager wants us to do our > > own internal quality control. How are things suppose to get fixed that way? > > My physician doesn't like to deal with these situations and refers me to my > > manager. I'm running in circles!!! Ron > > > > > > From: "pam marcum" > > > Date: 2004/08/13 Fri AM 09:11:34 EDT > > > To: , , > > > "Joe Nocito" > > > Subject: RE: [Histonet] a question on ethics > > > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From BlazekL <@t> childrensdayton.org Fri Aug 13 13:15:19 2004 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] a question on ethics Message-ID: Seems to me you have already shut down and have made up your mind that you are done and don't want to accept some of the great ideas that have been offered today. If your goal is to solve the problems you have encountered then the histonet has offered some wonderful help. >>> 8/13/2004 1:47:06 PM >>> There is also the issue that this position is not what I was told it would be. I also didn't expect to have so many issues such as a tech that can't work up to my standards. Also I keep asking myself if it is worth it esp. when this isn't turning out to be the job I expected it to be. Lastly, how can I work for someone I can't respect or trust? Any thought on that? > > From: Gary Gill > Date: 2004/08/13 Fri PM 01:39:01 EDT > To: "'pathrm35@adelphia.net'" , pam marcum > , histonet@lists.utsouthwestern.edu, Joe Nocito > > CC: histonet@lists.utsouthwestern.edu > Subject: RE: RE: [Histonet] a question on ethics > > Let your physician know that the person in charge is ultimately responsible > that the lab is run by the book. There are right ways and wrong ways to do > things. Wrong is indefensible. Among possible penalties, CMS can shut down > the lab. Surely your boss doesn't want to see the lab's name in the local > newspaper headlines. > > Gary Gill > > -----Original Message----- > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > Sent: Friday, August 13, 2004 12:34 PM > To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: RE: [Histonet] a question on ethics > > > I sent copies of incident reports to RM but my manager wants us to do our > own internal quality control. How are things suppose to get fixed that way? > My physician doesn't like to deal with these situations and refers me to my > manager. I'm running in circles!!! Ron > > > > From: "pam marcum" > > Date: 2004/08/13 Fri AM 09:11:34 EDT > > To: , , > > "Joe Nocito" > > Subject: RE: [Histonet] a question on ethics > > > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pathrm35 <@t> adelphia.net Fri Aug 13 13:35:09 2004 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] a question on ethics Message-ID: <20040813183509.TGEP9204.mta10.adelphia.net@mail.adelphia.net> I am willing to accept any and all ideas for a resolution. I value everyones opinion on histonet, that's why I posted my situation. > > From: "Linda Blazek" > Date: 2004/08/13 Fri PM 02:15:19 EDT > To: pathrm35@adelphia.net, mucram11@comcast.net, garygill@dcla.com, > jnocito@pathreflab.com > CC: histonet@lists.utsouthwestern.edu > Subject: RE: RE: [Histonet] a question on ethics > > Seems to me you have already shut down and have made up your mind that > you are done and don't want to accept some of the great ideas that have > been offered today. If your goal is to solve the problems you have > encountered then the histonet has offered some wonderful help. > > > >>> 8/13/2004 1:47:06 PM >>> > > There is also the issue that this position is not what I was told it > would be. I also didn't expect to have so many issues such as a tech > that can't work up to my standards. Also I keep asking myself if it is > worth it esp. when this isn't turning out to be the job I expected it to > be. Lastly, how can I work for someone I can't respect or trust? Any > thought on that? > > > > From: Gary Gill > > Date: 2004/08/13 Fri PM 01:39:01 EDT > > To: "'pathrm35@adelphia.net'" , pam marcum > > , histonet@lists.utsouthwestern.edu, Joe > Nocito > > > > CC: histonet@lists.utsouthwestern.edu > > Subject: RE: RE: [Histonet] a question on ethics > > > > Let your physician know that the person in charge is ultimately > responsible > > that the lab is run by the book. There are right ways and wrong ways > to do > > things. Wrong is indefensible. Among possible penalties, CMS can > shut down > > the lab. Surely your boss doesn't want to see the lab's name in the > local > > newspaper headlines. > > > > Gary Gill > > > > -----Original Message----- > > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > > Sent: Friday, August 13, 2004 12:34 PM > > To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > > Cc: histonet@lists.utsouthwestern.edu > > Subject: Re: RE: [Histonet] a question on ethics > > > > > > I sent copies of incident reports to RM but my manager wants us to do > our > > own internal quality control. How are things suppose to get fixed > that way? > > My physician doesn't like to deal with these situations and refers me > to my > > manager. I'm running in circles!!! Ron > > > > > > From: "pam marcum" > > > Date: 2004/08/13 Fri AM 09:11:34 EDT > > > To: , , > > > > "Joe Nocito" > > > Subject: RE: [Histonet] a question on ethics > > > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From akbitting <@t> geisinger.edu Fri Aug 13 14:05:21 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Diastase dilemma Message-ID: Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From JNocito <@t> Pathreflab.com Fri Aug 13 14:27:53 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Diastase dilemma In-Reply-To: Message-ID: Angela, It could be that the patient section contains mucin, which will not digest out. If this is not the case, then I suggest trying to digest the slides on a slide rack or something to lay the slides on a horizontal surface. Just my 21/2 cents worth. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Angela Bitting Sent: Friday, August 13, 2004 2:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Diastase dilemma Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri Aug 13 14:31:07 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Diastase dilemma Message-ID: Did you stop to think that your patient just might be positive for glycogen? "Angela Bitting" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 02:05 PM To: cc: Subject: [Histonet] Diastase dilemma Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dmikita <@t> wmcnet.org Fri Aug 13 14:44:05 2004 From: dmikita <@t> wmcnet.org (Daryl Mikita) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] pHing of eosin Message-ID: Hello, We are looking at pHing our eosin to the correct pH, instead of just using it straight from the bottle. What should we use to adjust the pH? Should it be 1N sodium hydroxide or 1N hydrochloric acid? Thanks Daryl From dmikita <@t> wmcnet.org Fri Aug 13 14:47:19 2004 From: dmikita <@t> wmcnet.org (Daryl Mikita) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Another question Message-ID: Hello, We were looking at our HQIP and we were wondering where in the H&E staining procedure, people use heat? We could not think of any place except for drying the slides. Thanks Daryl From garygill <@t> dcla.com Fri Aug 13 15:07:51 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] pHing of eosin Message-ID: <0B11620AEE5EFB4FA7A7558339972A8307048A@HALIBUT.dcla.com> Suggest using glacial acetic acid: 5 mL per liter of eosin. Be prepared for a color explosion. Gary Gill -----Original Message----- From: Daryl Mikita [mailto:dmikita@wmcnet.org] Sent: Friday, August 13, 2004 2:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pHing of eosin Hello, We are looking at pHing our eosin to the correct pH, instead of just using it straight from the bottle. What should we use to adjust the pH? Should it be 1N sodium hydroxide or 1N hydrochloric acid? Thanks Daryl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From amosbrooks <@t> earthlink.net Fri Aug 13 15:11:47 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] a question on ethics In-Reply-To: <200408130629.1bVC7Y1mH3NZFjV0@sparrow> References: <200408130629.1bVC7Y1mH3NZFjV0@sparrow> Message-ID: <6.0.0.22.0.20040813154852.01be9008@pop.earthlink.net> Ron, When a ship is hitting a reef there is 2 options. First option: Bail out, hit the life rafts and get out of there before you go down with the ship. Second option is to get the ship off the reef and repair the damage. This sounds like a problematic situation and you may be in a good position to make drastic improvements to the system. The question you need to decide is which course you intend to take, and stick with it. If you decide to leave, do it cautiously as these problems could still be pinned on you as the guy that came in caused problems and left. (I know that's not the way it is but people often shift blame for problems on people that can't defend themselves). You still should mention in an exit interview (assuming you get one) about the issues, so that someone else can fix them when you are gone, and so it is documented that you were aware of the situation and not the cause. If you decide to fix the ship, expect some rough waters. Very often when you point out errors to people they start looking for things to complain about with you. You should proceed with caution and document, document, document! The root cause survey that was previously mentioned sounds like a good plan. Often people make errors that they can prevent just by a simple modification of their procedure. Establishing redundancy is a good way to prevent most of the errors you have described. For example, picking a block and a slide up and comparing them before they go into the microtome, then again after the slide is cut and finally comparing the block to the slides before sending them out. Time consuming, yes, but it is hard to make the same mistake 3 times. Similar redundancies can be established in most other situations too. Turn around time will probably suffer a bit, but if it keeps you from getting sued it's worth it. Good luck which ever way you choose, Amos Brooks At 09:29 AM 8/13/2004, you wrote: >Message: 9 >Date: Fri, 13 Aug 2004 04:27:14 -0500 >From: "Ron Martin" >Subject: [Histonet] a question on ethics >To: >Message-ID: <002701c48117$b8cbc750$5dedeb44@Pathrm35> >Content-Type: text/plain; charset="iso-8859-1" > >Fellow techs, >I am in a difficult situation and need some serious advice. I recently (4 >months ago) accepted a technical supervisor position in a dermatology lab. >I went from a bench tech at my old job to this position. I also walked >away from a raise at my old position so I could step into a supervision >position. I took the new position because I was told by my manager that >she would teach me some supervision, management and financial skills that >I currently do not have as well as the growth potential of the >company.This position is not what I was told it would be. Part of my >"duties" include emptying the trash, clean the bathroom (not happening) >and taking her personal and professional calls. The question on ethics is >the high volume of mistakes made by our technician, our offices and also >by my manager herself. >I caught a mistake made by our tech a few weeks ago.She put the wrong >tissue on the wrong slides as she inverted the two cases. I caught her >mistake before it went out. One time she assigned the same number to two >different cases. She then sent the correct case out for a consult (which >wasn't needed) and put the wrong patient name on the slide. Every other >day there is something different.My manager will not terminate her as she >knows I am seeking employment elsewhere and she cannot afford to lose a tech. >One day my manager gave three cases the same accession number.She caught >her mistake on one of the cases but I still ended up with two cases with >the same number. When I first started there my manager had a case in which >there " was no tissue in the container". She said that she notified the >office about the situation however up to six weeks later the office was >still calling looking for results.At that point she wrote up an incident >report and dated it six weeks prior to coincide with the surgery date. >Our offices are not any better. One case came in with the wrong patient >information. It took about 7 weeks for the office to realize that they >sent the wrong patient information with the biopsy as the names were close >in spelling. The offices continually send mistakes with incorrect >spelling of patients names and incorrect anatomical locations. > I have tired to document everything but there are too many mistakes and > I don't have enough time or energy to keep up with them.My manager wants > us to do our own "internal quality control". My interpretation of this is > that she doesn't want our physician and risk manager to know of these > mistakes. Are these becoming common problems or is it just my situation? >I want to emphasize that I hope I am not being unethical myself for >revealing this information but I really need some advice and support.I >have very high standards and they are not being met in this current >situation. I am currently seeking a new position but I need employment and >cannot resign until another position becomes available. I would like to >stay in Florida and if anyone knows of any positions please inform me as I >am at my wits end. Thanks in advance. > >Ron From juan.gutierrez <@t> christushealth.org Fri Aug 13 15:12:37 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] You may want to hit the delete button with this, then again, maybe not Message-ID: Hey at least they tried to talk to you(a year later)but they tried YOU! Like I told you on my last e-mail my behind is about two pounds lighter after my boss bit some of it off because of my comments on the histonet about a certain German company who should remain nameless. Apparently my speaking the truth, the whole truth, and nothing but the truth on this forum made them a little mad. If they made good products to begin with, we wouldn't be having this conversation. Have anybody noticed how many pages of the phonebook are dedicated to lawyers? There is about twice the number of lawyers compared to doctors. Aint that scary? I wonder when lawyer season is going to open, heck there's more of them than there is deer. Just kidding. I know one good lawyer, one. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My views are my own and do not reflect those of my employer. Long live free speech! "We have nothing to fear but fear itself". -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Thursday, August 12, 2004 3:35 PM To: Histonet Subject: [Histonet] You may want to hit the delete button with this,then again, maybe not Afternoon Netter's, Remember about a week or two the topic was getting "flamed"? Well, here's a new one. Sometime last year, I made several postings about a certain company and their product that I used and it did not work for me. I wrote them a letter and returned all the merchandise back to them saying I was not happy with their product. I waited several months and never got a response, so I named the company on the Histonet. Since then, I have been throwing away their flyers, catalogs and any other correspondence I receive. Obviously, they are doing fine without my business and I am doing fine without their products. Today, I receive a letter from the company with a copy of my Histonet postings from over a year ago, telling me that this has been in customer service for about a year (hmmmmmmmm, I'm trying to find out where the customer service is in all of this. (One year, okay, I'll get back to that), that their customer service has tried several times to contact me but to no avail. (Hmmmmm, one way would have been to respond to my email address, but I'll come back to this also). So after all this time, they want to do business with me. (So close to the NSH meeting, hmmmmmmmmm, another point to ponder). My big beef is that these companies read this forum and try to use strong arm tactics on people who post negative comments about them or their products. I have many friends that are sales reps in this field (I'm talking like come over my house and eat type of friends) and I love these people. I must reiterate that this forum was set up to exchange ideas, theories and experiences, both positive and negative. If this was such a "hot item", point #1, why did it take a whole year to get a response when they had my company and address on file? Is this how customer service works? I don't think so!! Point #2 If you are a vendor and don't like a particular posting, why don't you reply back to that person via email. Point #3 don't wait until it's almost time for the NSH annual S/C to fix a problem, it's probably too late by then. Point #4 If you haven't had a purchase order or have not corresponded with a customer for over a year, chances are that customer does not want to do business with you. Let lying dogs lie and don't open old scars. All you did was get me going on a tangent. I know, there goes Nocito again, but I refuse to be treated like this from any one. Like they say in the movies, "Don't call me, I'll call you". Okay, I've said my piece now let the FLAMING BEGIN!!!!!!!!! As always, the opinions stated by this author in no way reflects the opinions of his employer, subordinates, friends or relatives. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri Aug 13 15:12:07 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Diastase dilemma Message-ID: Once I read my own answer, I saw how stupid it is. My apologies. It's been a long week. Think before you type, Jack . . .think..... Jackie.O'Connor@abbott.com Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 02:31 PM To: "Angela Bitting" cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Diastase dilemma Did you stop to think that your patient just might be positive for glycogen? "Angela Bitting" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 02:05 PM To: cc: Subject: [Histonet] Diastase dilemma Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Aug 13 15:15:00 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] pHing of eosin In-Reply-To: References: Message-ID: <6.0.0.22.1.20040813141110.01b227a8@gemini.msu.montana.edu> If you are buying eosin Y, already made up, check the MSDS, and the pH, it may be adjusted correctly. I know that this is the case for Richard Allan Eosin Y solution. If you are making up eosin y from the dry dye, then the recipe is (Hrapchak and Sheehan) eosin y 0.5g 95% ethanol 50 ml glacial acetic acid 1 drop If you are adjusting already made up, use acetic acid. At 01:44 PM 8/13/2004, you wrote: >Hello, > >We are looking at pHing our eosin to the correct pH, instead of just using >it straight from the bottle. What should we use to adjust the pH? Should >it be 1N sodium hydroxide or 1N hydrochloric acid? > >Thanks >Daryl > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From terribraud <@t> msn.com Fri Aug 13 15:15:48 2004 From: terribraud <@t> msn.com (TERRI BRAUD) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] RE: a Question of Ethics Message-ID: I can certainly understand the frustration of working in new management position that does not offer the training and responsibilities to go with it. And while I support going over a manager's head when patient care is impacted, you also must ask yourself if the activity is being tolerated by the pathologist(s)? If so, then look for another job, because you MUST have the support of your medical staff. Beyond that, I would encourage you to look for management training from outside sources, such as the program offered at the NSH convention, on-line courses in Quality Improvement and Laboratory Management, subscription to the Manager's edition of Advance, etc. These will help give you the educational tools to get started, but they in no way will make you an effective leader. Speaking from 15 years of management experience, to be an effective leader you have to be out in front, not someone who just points the way. As for any job, this often means "paying dues" by performing "other" duties until you are accepted as part of the team. If this means you have to clean bathrooms or empty waste to show you are part of a team, then do it cheerfully for a time. If a team sees your willingness to perform the "nasty" tasks, then eventually they will be more willing to take on those tasks, and others, to free your time for management duties. Anyone can point to problems within any system, but the true leader is the one that offers concrete solutions and suggestions and is willing to go the extra miles to put them into place. As we all know, Histology is a manual labor intensive process, prone to identity errors. As a leader, you must provide your employees with a system that has enough system checks and double checks that limit their ability to make a mistake, or catch it before it impacts patient care. Then, when mistakes are made, use a carefully constructed system to document and to review the system process. Maybe these "mistake-prone" employees have suffered the same frustrations that you have, even to the point of not caring anymore. Have they been given the tools, the training, the education needed to perform their jobs accurately and with positive feedback? If you, as a new manager, are not being helped, how much do you think is trickling down to their level? I would guess very little. This lab may or may not be the place to learn to be a supervisor, but it sounds like a place that is "ripe" for the picking. Anyone can step into a smooth running lab, but having the opportunity to make a real difference in a poorly functioning lab is not as easy. Laboratory administrators often come from a clinical background with very little AP experience and usually welcome the Histology Supervisor that comes in with fresh ideas to improve a lab's performance. Study process improvement. Offer well thought out, budget neutral suggestions for process improvement. And remember, being a supervisor means that your most important skill should be in "people management". This means coming up with ways to inspire and empower your employees to do a better job! Good luck with your quest, where ever it takes you. Sincerely, Terri L. Braud, HT(ASCP) Surgical Pathology Manager University of Virginia Health Systems Charlottesville, VA 22908 From RossS <@t> BaylorHealth.edu Fri Aug 13 15:30:12 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Diastase dilemma Message-ID: Are you running a plain PAS on the patient as well? I was always taught to always run a PAS and PASD even if the Dr only wanted a PASD. It serves as a control. If you are doing that, and know there is no diference digested vs undigested in the patient then I don't know what to say. I would normally look to something in the tissue that is causing this. If you were still using spit to digest I would blame it on you eating sugar before spitting on the slide, or not drinking enough coffee. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Friday, August 13, 2004 2:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Diastase dilemma Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From lindas <@t> awesomenet.net Fri Aug 13 15:46:36 2004 From: lindas <@t> awesomenet.net (Linda Davis) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Re: Smears on the Ventana Message-ID: <007c01c48176$bfbe2490$05a16243@D6JLZ851> Terre, We fix the smear in 95% alcohol, rinse in water, and place on the Ventana to run along with the paraffin sections. Linda Davis, HT (ASCP) Histology Supervisor Rio Grande Regional Hospital McAllen, TX. From Charles.Embrey <@t> carle.com Fri Aug 13 16:29:01 2004 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Diastase dilemma Message-ID: Did you ever stop to think.... that if the patient tissue were positive for Glycogen then the digestion would remove it and it would show no PAS staining? The positive reaction on could be mucin or acid mucosubstances. Charles Embrey -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Friday, August 13, 2004 2:31 PM To: Angela Bitting Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Diastase dilemma Did you stop to think that your patient just might be positive for glycogen? "Angela Bitting" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 02:05 PM To: cc: Subject: [Histonet] Diastase dilemma Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ynwang <@t> u.washington.edu Fri Aug 13 16:35:07 2004 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] MMP-1 immuno labelling of placenta Message-ID: Hello, Has anyone had any experience immuno labelling placenta tissue with anti-MMP1? We have tried this (cryosections with ABC kit with DAB) but are getting alot of (what we assume to be) background and nothing that looks like images we have seen on-line. Seeing as we have not had any experience with placenta tissue we are questioning if we are took the tissue from the correct place from the placenta (we were given a whole one which we took random samples from). Thank you for your help Yak-Nam Wang Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 From Sue.Kapoor <@t> uhsi.org Fri Aug 13 16:40:44 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] RE: a Question of Ethics Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F563D@khmcexch.uhsi.org> As for any job, this often means "paying dues" by performing "other" duties until you are accepted as part of the team. If this means you have to clean bathrooms or empty waste to show you are part of a team, then do it cheerfully for a time. -I'm sorry, but as a Histology Supervisor, cleaning bathrooms or emptying waste does NOT show you are a "part of the team"...it shows that management has no respect for your position. Why isn't the housekeeping dept. doing these duties? Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From dmccaig <@t> ckha.on.ca Fri Aug 13 17:03:58 2004 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] acid washed glassware Message-ID: <3E5A3F039F0BD8118B4700C00D002024043250@CKHA9> What concentration and what acid do you use to clean glassware prior to silver impregnation staining. Do you simply rinse it or is it soaked for a specific period of time? Air or oven dried? Diana McCaig, R.T. Charge Tech, Histology Chatham Kent Health Alliance 519-352-6401 (6604) From ploykasek <@t> phenopath.com Fri Aug 13 17:27:44 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] B+ fixative Message-ID: Hi all. We have a new client that uses B+ fixative. I am totally unfamiliar with this fixative. I'd appreciate any info on it. My main question concerns IHC on this tissue. We've done a CD20 with our standard heat pretreatment & standard titer, and gotten a negative result - even internal controls are negative. I'd really appreciate some help with this. Thanks. Patti Loykasek PhenoPath Laboratories Seattle, WA From JColCLEFA <@t> aol.com Fri Aug 13 17:52:37 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Prostate IHC, storing IF's Message-ID: We don't do Thymidine Synthetase but we do 34bE12, and AMACR (P504s) we are also going to be adding Ki67 and possibly p63. there is a mountant (aqeuos for IF) called Gelvatol (google search for the recepie) which keeps refrigerated IF slides for many weeks, if held in an airtight plastic box in the dark @ 2-8C. we've noticed no loss of signal, virtually no change for at least 2 weeks, sometimes longer From TJasper <@t> smdc.org Fri Aug 13 18:18:30 2004 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:23:54 2005 Subject: FW: RE: [Histonet] a question on ethics Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E44F77@harrier> Thomas Jasper HT(ASCP) BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org > -----Original Message----- > From: Jasper, Thomas G. > Sent: Friday, August 13, 2004 3:47 PM > To: 'pathrm35@adelphia.net' > Subject: RE: RE: [Histonet] a question on ethics > > Hey Ron, > > Have just followed your thread and hopefully I am understanding this > correctly. First of all the suggestions made to you by everyone on the > histonet are right, any or all of them should be implemented. Sounds to > me like you are running into a brick wall with a manager (you directly > report to) that is incompetent. Therefore, it is extremely difficult for > you to put these suggestions into place. Secondly, you were deceived into > exactly what the position was (is). This only complicates any effort on > your part to remedy a dire situation. > I will reiterate what others have said, something must be done as patients > are at risk. This "something" can only be done once the proper authority > at your institution deems it so. Whether this comes about from a sentinel > event (a death), some sort of legal salvo, action from an accrediting > agency or any combination of the previously mentioned, I am quite certain > it will happen. Once it happens it could be very ugly. > You said your physician (pathologist?) doesn't like to deal with these > situations. Hopefully your physician is more comfortable in a court of > law than he/she is in dealing with your institution's issues. You must go > to your MD as someone stated previously he/she is the one on the line. > Internal QC, QI, QA, PI or whatever your manager wants to call it sounds > bogus to me. I doubt your manager understands what a quality concept is. > How can you apply something you don't understand? > You may also want to remind your manager, risk management people and > physician about a little incident that took place up here in Minnesota not > too long ago. Two breast cases were mixed up resulting in surgery for a > patient (that was not required) and a missed diagnosis on another (that > was required). This was NOT the fault of the technical staff involved as > the pathologist mixed the cases up. Nevertheless, this was a wake-up call > to MOST path labs in the country. You are setting yourself up for, or > already have set yourself up for something along the same lines. > You may also want to remind the people senior to you that any one of them > could be the "patient". How would they feel about that? Also which > agency inspects your lab? CAP? (I doubt it) JCAHO? (seems unlikely). No > rap on you Ron but situations like yours are the whole reason responsible > inspection and auditing takes place. What goes on at your place should > NOT be happening. Fact is stranger than fiction I guess. > Finally, if I were you I would leave. Not to bail out on patients, but > this thing is bigger than you. The Boy Scouts of America didn't capture > Saddam Hussein. That's an extreme comparison but I think it makes the > point. I believe you were set up for this whole debacle, you were > probably viewed as a babe in the woods and ripe for the picking. Frankly, > you don't owe these deceptive jackdaws a thing. The only thing you can > really do, is defer to higher authority and hang in there until you can > get away from these toxic people. Good luck! > > > Thomas Jasper HT(ASCP)BAS > Anatomic Pathology Coordinator > SMDC Clinical Laboratory > Duluth, MN > tjasper@smdc.org > > > -----Original Message----- > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > Sent: Friday, August 13, 2004 12:47 PM > To: Gary Gill; pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: RE: [Histonet] a question on ethics > > > There is also the issue that this position is not what I was told it would > be. I also didn't expect to have so many issues such as a tech that can't > work up to my standards. Also I keep asking myself if it is worth it esp. > when this isn't turning out to be the job I expected it to be. Lastly, how > can I work for someone I can't respect or trust? Any thought on that? > > > > From: Gary Gill > > Date: 2004/08/13 Fri PM 01:39:01 EDT > > To: "'pathrm35@adelphia.net'" , pam marcum > > , histonet@lists.utsouthwestern.edu, Joe > Nocito > > > > CC: histonet@lists.utsouthwestern.edu > > Subject: RE: RE: [Histonet] a question on ethics > > > > Let your physician know that the person in charge is ultimately > responsible > > that the lab is run by the book. There are right ways and wrong ways to > do > > things. Wrong is indefensible. Among possible penalties, CMS can shut > down > > the lab. Surely your boss doesn't want to see the lab's name in the > local > > newspaper headlines. > > > > Gary Gill > > > > -----Original Message----- > > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > > Sent: Friday, August 13, 2004 12:34 PM > > To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > > Cc: histonet@lists.utsouthwestern.edu > > Subject: Re: RE: [Histonet] a question on ethics > > > > > > I sent copies of incident reports to RM but my manager wants us to do > our > > own internal quality control. How are things suppose to get fixed that > way? > > My physician doesn't like to deal with these situations and refers me to > my > > manager. I'm running in circles!!! Ron > > > > > > From: "pam marcum" > > > Date: 2004/08/13 Fri AM 09:11:34 EDT > > > To: , , > > > "Joe Nocito" > > > Subject: RE: [Histonet] a question on ethics > > > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Gervaip <@t> aol.com Fri Aug 13 18:28:50 2004 From: Gervaip <@t> aol.com (Gervaip@aol.com) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] proper disposal of specimen transpot biohazard bags Message-ID: <79.30ed33ba.2e4ea8b2@aol.com> I just recently attended an OSHA workshop and the question came up about the disposal of gloves and the transport specimen bags. If they are not visibly soiled, regular trash is all right. If you think the gloves might be contaminated, throw them in the Biohazard can. The cost of disposing of the Biohazard waste is determined normally by the weight. So the unnecessary disposal in a Biohazard waste container will just drive up operating cost. Pearl Gervais, New Orleans From lpwenk <@t> sbcglobal.net Fri Aug 13 19:14:02 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] acid washed glassware References: <3E5A3F039F0BD8118B4700C00D002024043250@CKHA9> Message-ID: <001601c48193$9b32f480$18e2d445@domainnotset.invalid> "Acid-cleaned", now a days, can mean cleaned with a commercially available product, several varieties, from several companies (maybe they can identify themselves and their products to you). Or, it could mean the original acid, known as "aqua regia". (more about this one in a moment) If a glassware is not washed well after being used with another stain, a chemical could be left behind on the glassware, such as dye or a metal salt (ferric chloride for example). Or if the glassware is not rinsed well after washing, soap would be left behind. Any of these contaminants that are left behind could react with the silver stain. The silver solution will then precipitate out, all over your tissue, slide, etc.. The obvious solution is to scrub really well in all the nooks and crannies and rinse lots of time with d. water. But sometimes even that doesn't get all the contaminants To really make certain that all the unwanted left-behind dyes, soap, etc. are removed, then a strong acid can be used to dissolve away the contaminants. The original acid cleaner is "aqua regia", or "royal water". This was the only solution that could dissolve gold and platinum, the metals that only really rich people (i.e., royalty) could afford. Aqua regia is made with 1 part conc. nitric acid to 3 parts conc. hydrochloric acid. (e.g., 50 mL nitric acid + 150 mL hydrochloric acid) Can be stored at room temperature in a clear bottle. Can be reused until it starts taking too long to clean, or turns really dark yellow/orange (it's light yellowish when first combined). Pour the aqua regia in the glassware and let it set in the glassware for 1-5 minutes. Pour the aqua regia back into the original bottle. Slowly add d. water to the glassware (I know, water into acid is a bad idea, that's why I said slowly). There will be a little white wispy "smoke". Dump out the d. water, and do several more rinses with d. water. Air drying is fine, but if you're in a hurry, oven drying would be OK too. Acid precautions apply = goggle, heavy nitrile-type gloves, store in acid cabinet, pour away from your face, etc. Think beyond acid-clean glassware. Ever get a bottle that you just can't get the dye or chemical out? No matter how hard you scrub or how long you let it soak or how much bleach you put it? This will definitely get it out. I keep a bottle of aqua regia in the lab for this purpose. Works great. Sorry for being a little wordy on this answer. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Diana McCaig" To: Sent: Friday, August 13, 2004 6:03 PM Subject: [Histonet] acid washed glassware > What concentration and what acid do you use to clean glassware prior to > silver impregnation staining. Do you simply rinse it or is it soaked for a > specific period of time? Air or oven dried? > Diana McCaig, R.T. > Charge Tech, Histology > Chatham Kent Health Alliance > 519-352-6401 (6604) > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From terribraud <@t> msn.com Fri Aug 13 19:40:32 2004 From: terribraud <@t> msn.com (TERRI BRAUD) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] RE: emptying waste and cleaning bathrooms Message-ID: -I'm sorry, but as a Histology Supervisor, cleaning bathrooms or emptying waste does NOT show you are a "part of the team"...it shows that management has no respect for your position. Why isn't the housekeeping dept. doing these duties? Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 Lets agree to disagree, ok? Some of us may not have the luxury of a well staffed housekeeping department, and I know that my equally overworked housekeeping team members appreciate an occasional helping hand. Notice I said occassional! In return, I know that the 2 regular housekeepers that attend to my department take extra pains to ensure that my department sparkles and that they willingly offer to help me with the occasional task that is outside of their duties. True teamwork, for me, does not stop at the department door. Terri Braud From TJJ <@t> Stowers-Institute.org Fri Aug 13 21:00:12 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Re: Diastase dilemma Message-ID: This doesn't solve your dilemma (sorry) but I learned that the best control to use for PASD is cervix with endocervix. The glycogen in the epithelium will digest, and the mucin in the glands will not. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research Kansas City, MO From histo20 <@t> hotmail.com Fri Aug 13 21:06:05 2004 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] RE: Artisan Stainer Message-ID: We have had the Artisan for nearly a year now. We had had some problems with it initially, due to lack of information and training. We found you had to initialize between each run, the refrigerated packs, especially the silvers HAD to come to room temperture, otherwise there would be a precipitate on the slide. We also learned that when filling the bulk fluids, you had to decrease the volume by 10%. So if the reagent bottle was actually 950 ml, then you would tell it 900. What we thought was precipitate was actually reagents not being rinsed off the slide because of an empty bulk fluid bottle. We also needed a power back-up and surge protector. Those seemed to be the end of our problems. The trichrome stain needs to be by itself since there is something wrong with the packs. We were told that the packs squirt and that manufacturing is working on the problem. Our turn around time for specials has improved and I also lost a tech with this machine (someone not being replaced). Stains are beautiful and consistent now. Paula Wilder St. Joseph Medical Center Baltimore, MD. 21204 _________________________________________________________________ On the road to retirement? Check out MSN Life Events for advice on how to get there! http://lifeevents.msn.com/category.aspx?cid=Retirement From JQB7 <@t> CDC.GOV Sat Aug 14 09:31:52 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] acid washed glassware Message-ID: We use 20% nitric acid in distilled water. We let it sit in the glassware for at least 30 minutes then pour it back into the container (we re-use the solution for 3 months) and then thoroughly rinse the glassware with distilled water. We clean the glassware just before use and we primarily use acid cleaned glassware for silver procedures. We routinely have our glassware cleaned by our glassware department which uses strong detergents and high temperatures so the glassware is cleaned very well but we still acid clean for our silver procedures. Jeanine Bartlett CDC -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Diana McCaig Sent: Fri 8/13/2004 6:03 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] acid washed glassware What concentration and what acid do you use to clean glassware prior to silver impregnation staining. Do you simply rinse it or is it soaked for a specific period of time? Air or oven dried? Diana McCaig, R.T. Charge Tech, Histology Chatham Kent Health Alliance 519-352-6401 (6604) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Sat Aug 14 10:12:57 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] canada balsam Message-ID: <000a01c48211$2e96dbb0$eeeea8c0@server> Hi all, A little question by interest. Was it canada balsam where you have to margin the coverslip with paraffin or nail polish to avoid drying out? Or was this only necessary with aqueous medias? Gudrun From froyer <@t> bitstream.net Sat Aug 14 12:08:48 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] RE: emptying waste and cleaning bathrooms In-Reply-To: References: Message-ID: <411E4720.20700@bitstream.net> Well said Terri. Many of the Techs that I used to work with looked down on the hospital housekeeping and maintenance staff. When our in-lab glassware dishwasher was on extended medical leave for major surgery, they even resented having to load the dishwasher themselves. I felt differently. Without these "ancillary staff" we "professionals" would not be able to perform our honorable duties of "saving lives and stamping out disease". I made a point to get to know our housekeeping & maintenance people... learned their names and never failed to say "hello" to them or share a joke when we passed in the hall. If a waste basket was overflowing, I would dump it myself. If I was asked to take a shift in the dishwashing room, I did it gladly. As a result, whenever I needed a mess (that I created) cleaned up, or something fixed in my department, help was a quick phone call away. They would respond in seconds. My co-supervisors could never figure out why I got such good service from "these people" when it would take them days to respond to one of their requests. A smile (and a simple hello) is something nice to see and it doesn't cost a cent.... ~ Ford Ford M. Royer, MT(ASCP) Minneapolis, MN TERRI BRAUD wrote: > -I'm sorry, but as a Histology Supervisor, cleaning bathrooms or emptying > waste does NOT show you are a "part of the team"...it shows that > management > has no respect for your position. Why isn't the housekeeping dept. doing > these duties? > Sue Kapoor, HT (ASCP) > Histology Coordinator > Kenosha Medical Center > Kenosha, WI > 262-653-5570 > > Lets agree to disagree, ok? > Some of us may not have the luxury of a well staffed housekeeping > department, and I know that my equally overworked housekeeping team > members appreciate an occasional helping hand. Notice I said > occassional! In return, I know that the 2 regular housekeepers that > attend to my department take extra pains to ensure that my department > sparkles and that they willingly offer to help me with the occasional > task that is outside of their duties. True teamwork, for me, does not > stop at the department door. > Terri Braud > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From amosbrooks <@t> earthlink.net Sat Aug 14 20:36:43 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] proper disposal of specimen transport biohazard bags In-Reply-To: <200408141007.1bW20A6QN3NZFmQ0@tanager.mail.pas.earthlink.n et> References: <200408141007.1bW20A6QN3NZFmQ0@tanager.mail.pas.earthlink.net> Message-ID: <6.0.0.22.0.20040814212531.01c09cc8@pop.earthlink.net> Attention ... please turn on your sarcasm filter! Those with sarcasm sensitivity please hit delete now! Because we all know we can see hazardous microbes on surfaces of gloves. Let's listen in on this conversation in a hypothetical lab. Are those gloves contaminated with hepatitis? Gosh, I don't see any thing on them... they must be clean. ... Is that a coffee in your hand? Get out of the lab with it hepatis could be in the air and land in your cup! OSHA is so stupid... Ever heard of universal precautions? OK nonsense rant over you may now return to your regularly scheduled discussion Amos Brooks At 01:07 PM 8/14/2004, you wrote: >Message: 4 >Date: Fri, 13 Aug 2004 19:28:50 EDT >From: Gervaip@aol.com >Subject: Re: [Histonet] proper disposal of specimen transpot biohazard > bags >To: pathrm35@adelphia.net, histonet@pathology.swmed.edu >Message-ID: <79.30ed33ba.2e4ea8b2@aol.com> >Content-Type: text/plain; charset="US-ASCII" > >I just recently attended an OSHA workshop and the question came up about the >disposal of gloves and the transport specimen bags. If they are not visibly >soiled, regular trash is all right. If you think the gloves might be >contaminated, throw them in the Biohazard can. > >The cost of disposing of the Biohazard waste is determined normally by the >weight. So the unnecessary disposal in a Biohazard waste container will just >drive up operating cost. > > >Pearl Gervais, New Orleans From leahcox27 <@t> yahoo.com Sat Aug 14 21:14:20 2004 From: leahcox27 <@t> yahoo.com (Leah Cox) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] test Message-ID: <20040815021420.54891.qmail@web50206.mail.yahoo.com> This is a test --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - Send 10MB messages! From lpwenk <@t> sbcglobal.net Sat Aug 14 21:43:31 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] canada balsam References: <000a01c48211$2e96dbb0$eeeea8c0@server> Message-ID: <001b01c48271$a84620a0$2530d445@domainnotset.invalid> We ring our aqueous mounting media with finger nail polish. Aqueous mounting media does not really dry out and harden, and keeps oozing out from under the coverslip and the coverslip keeps moving, so was always making a mess of the slide, especially for filing. The aqueous mounting media will eventually dry out, but then all you are left with is dried sugar under the coverslip(terrible refractive index at that point (tongue-in-cheek)). The finger nail polish kept the aq. mounting media from oozing and slowed down the drying out of the aq. mounting media to sugar. On the other hand, Canadian balsam is basically pine tree sap diluted with xylene or toluene. Stays runny and sticky for a long time. I remember placing the slides in the 60 degree C. oven for 1 week after coverslipping, to solidify the mounting media enough for us to file the slides. I suppose it is possible that someone could have ringed the coverslip with finger nail polish after coverslipping, just to prevent the slight oozing and the coverslip from moving around. I had never thought to do it, but then we were coverslipping 300 slides a day in the lab at the time, and ringing is not time efficient for that quantity. Thank goodness for the modern day mounting medias with their faster drying times (30 minutes to non-stickiness) and their non-acidic pH so the stains don't fade with time (which is why we have to restain old slides of H&E that were originally mounted with Canadian balsam). Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Gudrun Lang" To: "Histonetliste" Sent: Saturday, August 14, 2004 11:12 AM Subject: [Histonet] canada balsam Hi all, A little question by interest. Was it canada balsam where you have to margin the coverslip with paraffin or nail polish to avoid drying out? Or was this only necessary with aqueous medias? Gudrun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From WWmn916 <@t> aol.com Sat Aug 14 22:49:27 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Iron stain question Message-ID: Hello again histonetters, Can anyone give me any insight as to why some smear slides stained for iron would not stains positive as it was expected they would? Doctor say's the slides should have stained positive for iron because the patient has leukemia. The control stained positive as it should have. Staining tech is sure all her slides were exposed to solution. Thanks for any help. Deb King, HT(ASCP) Sacramento, CA From chesarato <@t> hotmail.com Sat Aug 14 23:02:43 2004 From: chesarato <@t> hotmail.com (=?iso-8859-1?Q?C=E9sar_Romero?=) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] S100 A6 Message-ID: Dear People from Histonet I would like to know if somebody has any experience with this Antibody ( S100A6 ) from Dako Corporation. It is suposed to stain Spitz Nevus and not Common Nevus or Melanomas. It also stains Neurothekeomas. I have tried it but in my cases the antibody stained everything ( anything ). Thank you in advance. Dr C?sar Romero Pathologist chesarato@hotmail.com From lpwenk <@t> sbcglobal.net Sun Aug 15 06:39:41 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Iron stain question References: Message-ID: <001001c482bc$8ee129c0$9936d445@domainnotset.invalid> Usually, it's the bone core that is negative due to the decalcification, while the clot and the smear are positive. Couple off the wall thoughts. Any chance that the smear slide was in the same coplin jar slot as the control, so that the smear was stuck against the back of the previous slide? That would prevent the ferrocyanide solution from "touching" the smear. If this is possible, remove the coverslip, remove the mounting media with xylene (or whatever you use), run the slide back down through alcohols to water, and restain. Any chance that the smear was fixed in something containing an acid, like acetic acid? That might remove the iron. No way to correct this problem. Did they do a bone core and/or clot? Were they negative? If so, then the person IS iron deficient, regardless of whether the person has leukemia or not. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Saturday, August 14, 2004 11:49 PM Subject: [Histonet] Iron stain question > Hello again histonetters, > > Can anyone give me any insight as to why some smear slides stained for iron > would not stains positive as it was expected they would? Doctor say's the > slides should have stained positive for iron because the patient has leukemia. > The control stained positive as it should have. Staining tech is sure all her > slides were exposed to solution. Thanks for any help. > > Deb King, HT(ASCP) > Sacramento, CA > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Sun Aug 15 11:48:26 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] You may want to hit the delete button with Message-ID: <003d01c482e7$afc29e00$c6949a51@home> "(I'm talking like come over my house and eat type of friends) and I love these people." I wonder if any of Joe's Salespeople friends were called Raoul ;-))))) Remember that Paul Bartel film? --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.726 / Virus Database: 481 - Release Date: 22/07/2004 From carl.hobbs <@t> kcl.ac.uk Sun Aug 15 12:01:39 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Image analysis and slide scanning machine Message-ID: <005201c482e9$886e5720$c6949a51@home> Jackie...interested to read that you have the Mosaix from Zeiss. Am about to take delivery of this software along with an Axioskop set up with the Apotome system......I understand that the Mosaix software has undergone a further revision. Be grateful for more info on your experiences with this eg; what software do you use for analysis once you've captured images? I got KS300....do you have a dedicated macro? Thanks Carl --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.726 / Virus Database: 481 - Release Date: 22/07/2004 From carl.hobbs <@t> kcl.ac.uk Sun Aug 15 12:28:14 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] RE: replies to immunostaining on technovit Message-ID: <005b01c482ed$3efa5630$c6949a51@home> Thanks, Paul. Very interesting! I have used Tech8200 for many years for morphological studies, very successfully, on many species albeit with very poor results for immuno work. Interestingly, for whole- mount fluorochrome, Bgal, DAB- stained specimens it is superb! I have tried to etch/ dissolve/ Ag- retrieve with many chemicals/protocols with inadequate results. If I lower the amount of cross-linker( getting a rather sticky block) the results are better, but never satisfactory/consistent. Any experience with LRWhite for immuno? It's Hydroxyethyl, rather than H.glycol methacrylate. I will read the article for which you kindly provided the link --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.726 / Virus Database: 481 - Release Date: 22/07/2004 From carl.hobbs <@t> kcl.ac.uk Sun Aug 15 12:46:03 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Mowiol aq mountant Message-ID: <007501c482ef$bc544350$c6949a51@home> Reading the posts re aq. mountants......I use a Mowiol - based, setting mountant for fluorochrome preps with great results. OK, under confocal conditions it fades upon examination but much slower than any other I've tried. Anyone else use this? Be grateful for comments. --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.726 / Virus Database: 481 - Release Date: 22/07/2004 From WWmn916 <@t> aol.com Sun Aug 15 14:12:52 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Iron stain question Message-ID: Hi Peggy, Thanks for your response. Very helpful. Tech is positive the slides were not stuck together. I had similar thoughts which is why the histonet is so wonderful. Deb King, HT(ASCP) Sacramento, CA From PWebster <@t> hei.org Sun Aug 15 16:27:21 2004 From: PWebster <@t> hei.org (Webster, Paul) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Mowiol aq mountant Message-ID: <4E8C1F1E4E8FA748B5487C50C91695F001280006@heimail.hei.org> Try adding a pinch of DABCO or PPI (see the Sigma Catalog for details) to the Moviol to reduce the fading. Paul Webster. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Carl Sent: Sun 8/15/2004 10:46 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Mowiol aq mountant Reading the posts re aq. mountants......I use a Mowiol - based, setting mountant for fluorochrome preps with great results. OK, under confocal conditions it fades upon examination but much slower than any other I've tried. Anyone else use this? Be grateful for comments. --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.726 / Virus Database: 481 - Release Date: 22/07/2004 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JColCLEFA <@t> aol.com Sun Aug 15 17:36:21 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] division of "LAB"or, iron stain Message-ID: <12f.48d9a1ba.2e513f65@aol.com> Division of labor--I am the Sr. IHC Tech for a large hospital corporation and my experience is that a division of labor functions best when the lines are defined. I offered temporary help assisting autopsies because the full time autopsy assistant would call in sick when bodies would be available to be autopsied (member of the team, right?) I was on call every weekend for 6 years. When I finally put my foot down and took a week off to go to the NSH convention, I was assured by my supervisor that the morgue would be covered by assistants from other sources. I offered to train them and my boss told me they were already fully trained . I was called back from the convention a day early because none of the people called in were fully trained and I was held accountable (bad annual review which specifically mentioned that one week) for the SNAFU even though my job description has no mention of autopsy responsibility. Also the housekeeping staff had the habit of forgetting to take out the trash in the morgue, so I decided to be a team player and toss the trash myself when they "forgot". When the noble housekeeping staff found out that I would do this they stopped taking out the trash altogether. I earned back their respect (their fear?) only after complaining via e-mail cc'd to their supervisor's manager, and threats to report the manager to the VP for creating a dangerous environment. Six years of trusting that people would do the right thing if given the chance did nothing but turn me into a Macheavellian. I don't care if the houskeeping staff likes me or not. They earned my disdain, and now my hospital is safer. Iron stain-- maybe it is negative. From AnthonyH <@t> chw.edu.au Sun Aug 15 18:21:04 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Mallory's PTAH Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E07F@simba.kids> I've found the Cherukian's modification of the PTAH stain to be hard to beat. It does not require Bouin's or Zenkers pre-treatment. Method as follows: Document Procedure: Phosphotungstic Acid Haematoxylin Pathnet Code: S PTAH M or G Principle: Cherukian's modification employs an eosin solution that stains the erythrocytes red and differentiates them from the blue fibrin. Fixation: 10% buffered formalin. Microtomy: paraffin sections at 5um. Controls: Use brain sections and section of muscle. A good stain will demonstrate the dendrites as blue where as in a bad stain they appear light grey to salmon in colour. Nuclei, fibrin, platelets and muscle will be blue, red cells and collagen appear red. Muscle striations should be well defined. Reagents: 1. Eosin: - Warning: Flammable - see MSDS Eosin Y, water soluble (CI 145380) 0.5g Distilled Water 10ml 80% Ethanol 190ml Working: 10ml stock and Before use add 50ul glacial acetic acid. 2. 1% Periodic Acid 3. PTAH solution Haematoxylin (CI 75290) 0.5g Phosphotungstic Acid 10g Distilled water 500ml Dissolve solid ingredients in separate portions of the water. Use gentle heat for Haematoxylin. Combine solutions when cool. Add 0.088g potassium permanganate to ripen. The stain is ready to use. Procedure: 1. Dewax and hydrate sections to 80% alcohol. 2. Place slides in eosin for 30 seconds. 3. Wash slides in distilled water for a few seconds. 4. Place slides in 1% periodic acid for 20 minutes. 5. Wash slides in water for 3 minutes. 6. Place slides in PTAH for 30-90 minutes in 60oC oven. Check from 30 minutes on. 7. Dehydrate, clear and mount. Results: Dendrites, nuclei, fibrin, platelets and muscle - blue Red blood cells and collagen - red. Notes: Reference: 1. Cherukian, C.J., Histologic. 8(4); 105, (1977). 2. Luna, L., Histologic. 5(2); 66, (1975). Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] Sent: Friday, 13 August 2004 6:59 PM To: UniPath IHC; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Mallory's PTAH Here's what worked for us - Out of the old Ann Preece book. Fix as usual in formalin, and process as you would for similar size tissue. Section 4-5 um (I always vote for thinner, when working on registry material) and place on a subbed slide. (Charged, poly-L-lysine, etc.) Helps to hold it on through the rest of the steps. Dry in 60 degree oven for about an hour (helps to hold the tissue on the slide during the rest of the steps). After deparaffinization and running down to water, post-fix in Bouin for 1 hour in a 60 degree C. oven or waterbath. Then wash in slowly running cold water for about 10 minutes, or until all the yellow is gone. (Yes, just like post-fixing to do a trichrome.) Then, stain as usual with the PTAH. Should work great, nice intense colors with very little splotching, and sectioning should be your usual formalin quality. No mercury disposal to worry about. This is assuming, of course, that the PTAH solution is still good and the procedure was followed correctly. Now - as far as any PTAH questions on the written/computer test - the correct answer to fixation/post-fixation for PTAH is "Zenker". As that's what is written in all the rest of the histo books. Hope that helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "UniPath IHC" To: Sent: Thursday, August 12, 2004 2:13 PM Subject: [Histonet] Mallory's PTAH I actually already tried to use formalin-fixed muscle. You're exactly right, it sectioned great, but I could never get any decent staining. I tried post-fixing for 24 hrs. in Zenker's vs. post-fixing for 3 Hrs in Zenker's at 56 degrees. I tried all three protocols in Sheehan and the protocol in Carson. I'm at a loss for what to do next if I can't make the Zenker fixed muscle work. At least I've learned a lot about the stain. :-) Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Sun Aug 15 18:27:54 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] pHing of eosin Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E080@simba.kids> We use 3% acetic acid Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Daryl Mikita [mailto:dmikita@wmcnet.org] Sent: Saturday, 14 August 2004 5:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pHing of eosin Hello, We are looking at pHing our eosin to the correct pH, instead of just using it straight from the bottle. What should we use to adjust the pH? Should it be 1N sodium hydroxide or 1N hydrochloric acid? Thanks Daryl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Sun Aug 15 18:38:33 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Iron stain question Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E081@simba.kids> What were the smears fixed in? If Carnoys or any fixative containing acid then iron will leach out. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Sunday, 15 August 2004 1:49 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Iron stain question Hello again histonetters, Can anyone give me any insight as to why some smear slides stained for iron would not stains positive as it was expected they would? Doctor say's the slides should have stained positive for iron because the patient has leukemia. The control stained positive as it should have. Staining tech is sure all her slides were exposed to solution. Thanks for any help. Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From info <@t> ihcworld.com Sun Aug 15 23:48:04 2004 From: info <@t> ihcworld.com (IHC World) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Immunohistochemistry Database In-Reply-To: <001b01c47dec$87d73b80$ae2dd445@domainnotset.invalid> Message-ID: Our protocol database has been updated recently to over 500 protocols. Hope this update will benefit your immunohistochemistry research needs. Here is the link: http://www.ihcworld.com Also everyone is welcome to join our discussion forum at http://www.ihcworld.com/forum to ask and answer questions about IHC and histology. Best regards, Richard IHC World - Online Information Center for Immmunohistochemistry. http://www.ihcworld.com From rentonlf <@t> bru.wits.ac.za Mon Aug 16 02:44:22 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] MMP-1 immuno labelling of placenta Message-ID: <1092642262.91c1a8c0rentonlf@bru.wits.ac.za> 1. Was the tissue fresh when sampled? autolysis can alter IHC dramatically 2.Placenta contains a lot of biotin...are you blocking adequately? Regards -----Original Message----- From: "Y. Wang" To: Histonet@lists.utsouthwestern.edu Date: Fri, 13 Aug 2004 14:35:07 -0700 (PDT) Subject: [Histonet] MMP-1 immuno labelling of placenta Hello, Has anyone had any experience immuno labelling placenta tissue with anti-MMP1? We have tried this (cryosections with ABC kit with DAB) but are getting alot of (what we assume to be) background and nothing that looks like images we have seen on-line. Seeing as we have not had any experience with placenta tissue we are questioning if we are took the tissue from the correct place from the placenta (we were given a whole one which we took random samples from). Thank you for your help Yak-Nam Wang Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From Barry.R.Rittman <@t> uth.tmc.edu Mon Aug 16 06:37:20 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:23:54 2005 Subject: FW: [Histonet] pHing of eosin Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635B04@UTHEVS3.mail.uthouston.edu> I believe that care should be used in universally adding specific amounts of acetic acid to acidify eosin solutions that differ. I asume that different batches of dye have different dye contents so that the 5% stain that one user prepares is a little different from that used by others. Each lab should prepare their eosin with this is mind. I usually add a low percentage of acetic acid to the eosin until a faint opalescence occurs. I have been told that this is the point at which the dye acid just starts to precipitate. For our eosin solutions this involves 5 drops or so of 2% acetic acid per 100 ml of 5% eosin. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tony Henwood Sent: Sun 8/15/2004 6:27 PM To: 'Daryl Mikita'; histonet@lists.utsouthwestern.edu Cc: Subject: RE: [Histonet] pHing of eosin We use 3% acetic acid Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Daryl Mikita [mailto:dmikita@wmcnet.org] Sent: Saturday, 14 August 2004 5:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pHing of eosin Hello, We are looking at pHing our eosin to the correct pH, instead of just using it straight from the bottle. What should we use to adjust the pH? Should it be 1N sodium hydroxide or 1N hydrochloric acid? Thanks Daryl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Mon Aug 16 07:06:34 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] a question on ethics In-Reply-To: <1DB04B57B04C5747B87C3B39F3E605AA02E44F77@harrier> Message-ID: After thinking about this all weekend, one thing that hasn't been mentioned and should be used as a last resort is CLIA. Every state has a government agency that overlooks the lab and healthcare. I worked in a hospital once that a disgruntled employee called in CLIA. We lost the lab manager and three section supervisors were put on probation. Not a happy situation, but the tech got what they wanted. The manager fired. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jasper, Thomas G. Sent: Friday, August 13, 2004 6:19 PM To: 'histonet@pathology.swmed.edu' Subject: FW: RE: [Histonet] a question on ethics Thomas Jasper HT(ASCP) BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org > -----Original Message----- > From: Jasper, Thomas G. > Sent: Friday, August 13, 2004 3:47 PM > To: 'pathrm35@adelphia.net' > Subject: RE: RE: [Histonet] a question on ethics > > Hey Ron, > > Have just followed your thread and hopefully I am understanding this > correctly. First of all the suggestions made to you by everyone on the > histonet are right, any or all of them should be implemented. Sounds to > me like you are running into a brick wall with a manager (you directly > report to) that is incompetent. Therefore, it is extremely difficult for > you to put these suggestions into place. Secondly, you were deceived into > exactly what the position was (is). This only complicates any effort on > your part to remedy a dire situation. > I will reiterate what others have said, something must be done as patients > are at risk. This "something" can only be done once the proper authority > at your institution deems it so. Whether this comes about from a sentinel > event (a death), some sort of legal salvo, action from an accrediting > agency or any combination of the previously mentioned, I am quite certain > it will happen. Once it happens it could be very ugly. > You said your physician (pathologist?) doesn't like to deal with these > situations. Hopefully your physician is more comfortable in a court of > law than he/she is in dealing with your institution's issues. You must go > to your MD as someone stated previously he/she is the one on the line. > Internal QC, QI, QA, PI or whatever your manager wants to call it sounds > bogus to me. I doubt your manager understands what a quality concept is. > How can you apply something you don't understand? > You may also want to remind your manager, risk management people and > physician about a little incident that took place up here in Minnesota not > too long ago. Two breast cases were mixed up resulting in surgery for a > patient (that was not required) and a missed diagnosis on another (that > was required). This was NOT the fault of the technical staff involved as > the pathologist mixed the cases up. Nevertheless, this was a wake-up call > to MOST path labs in the country. You are setting yourself up for, or > already have set yourself up for something along the same lines. > You may also want to remind the people senior to you that any one of them > could be the "patient". How would they feel about that? Also which > agency inspects your lab? CAP? (I doubt it) JCAHO? (seems unlikely). No > rap on you Ron but situations like yours are the whole reason responsible > inspection and auditing takes place. What goes on at your place should > NOT be happening. Fact is stranger than fiction I guess. > Finally, if I were you I would leave. Not to bail out on patients, but > this thing is bigger than you. The Boy Scouts of America didn't capture > Saddam Hussein. That's an extreme comparison but I think it makes the > point. I believe you were set up for this whole debacle, you were > probably viewed as a babe in the woods and ripe for the picking. Frankly, > you don't owe these deceptive jackdaws a thing. The only thing you can > really do, is defer to higher authority and hang in there until you can > get away from these toxic people. Good luck! > > > Thomas Jasper HT(ASCP)BAS > Anatomic Pathology Coordinator > SMDC Clinical Laboratory > Duluth, MN > tjasper@smdc.org > > > -----Original Message----- > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > Sent: Friday, August 13, 2004 12:47 PM > To: Gary Gill; pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: RE: [Histonet] a question on ethics > > > There is also the issue that this position is not what I was told it would > be. I also didn't expect to have so many issues such as a tech that can't > work up to my standards. Also I keep asking myself if it is worth it esp. > when this isn't turning out to be the job I expected it to be. Lastly, how > can I work for someone I can't respect or trust? Any thought on that? > > > > From: Gary Gill > > Date: 2004/08/13 Fri PM 01:39:01 EDT > > To: "'pathrm35@adelphia.net'" , pam marcum > > , histonet@lists.utsouthwestern.edu, Joe > Nocito > > > > CC: histonet@lists.utsouthwestern.edu > > Subject: RE: RE: [Histonet] a question on ethics > > > > Let your physician know that the person in charge is ultimately > responsible > > that the lab is run by the book. There are right ways and wrong ways to > do > > things. Wrong is indefensible. Among possible penalties, CMS can shut > down > > the lab. Surely your boss doesn't want to see the lab's name in the > local > > newspaper headlines. > > > > Gary Gill > > > > -----Original Message----- > > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > > Sent: Friday, August 13, 2004 12:34 PM > > To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > > Cc: histonet@lists.utsouthwestern.edu > > Subject: Re: RE: [Histonet] a question on ethics > > > > > > I sent copies of incident reports to RM but my manager wants us to do > our > > own internal quality control. How are things suppose to get fixed that > way? > > My physician doesn't like to deal with these situations and refers me to > my > > manager. I'm running in circles!!! Ron > > > > > > From: "pam marcum" > > > Date: 2004/08/13 Fri AM 09:11:34 EDT > > > To: , , > > > "Joe Nocito" > > > Subject: RE: [Histonet] a question on ethics > > > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> instrumedics.com Mon Aug 16 08:48:06 2004 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] bubbles Message-ID: <030801c48397$a8c6d5c0$6401a8c0@INSTRUMEDICS22> Phil, When you are using CureMount you MUST be very sure that before you place the slide under the UV light you have not trapped any bubbles in the coverslipping process. I believe that with care you will find that the bubbles will not be present in the cured mounting medium. Please let us know if this suggestion helps. Bernice From VanmetMJ <@t> pbrc.edu Mon Aug 16 09:27:44 2004 From: VanmetMJ <@t> pbrc.edu (Montina Van Meter) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] 5-HT 2C Receptor Message-ID: Fellow Histonetters, Does anyone know of a commercially available 5-HT 2C receptor that works well on rat CNS for immunofluorescence? The tissue has been fixed with 4% paraformaldehyde, and cut at 40 microns on a freezing microtome. Thank you in advance, Tina Van Meter Montina J. Van Meter Lab Coordinator Autonomic Neuroscience Pennington Biomedical Research Center Baton Rouge, LA 70808 225-603-0953 vanmetmj@pbrc.edu From juan.gutierrez <@t> christushealth.org Mon Aug 16 10:01:51 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Biohazard Bags Message-ID: Hello netters. A while back there was a posting about a regulation prohibiting lab personnel from reaching into a red bag to retrieve a specimen for grossing. Can someone send me a copy or reference for the regulation? We have checked with several hospitals in our area, including two military hospitals, and most people have not heard of such. Thanks for your help. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 (210)704-3180 Fax My views are my own and do not reflect those of my employer. Long live free speech! "We have nothing to fear but fear itself". From ynwang <@t> u.washington.edu Mon Aug 16 10:30:51 2004 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] MMP-1 immuno labelling of placenta In-Reply-To: <1092642262.91c1a8c0rentonlf@bru.wits.ac.za> References: <1092642262.91c1a8c0rentonlf@bru.wits.ac.za> Message-ID: Hello Louise, Thank you for your suggestions. 1) Seeing as we could not go to the farm to collect the tissue the tissue was kept on ice (~5 hours) while it was shipped to us and immediately embedded. Do you think this is a problem? 2)We have not done any biotin blocking for these samples. Thank you for suggesting that. We will try this next. Thanks again Yak-Nam Wang On Mon, 16 Aug 2004, renton louise mrs wrote: > 1. Was the tissue fresh when sampled? autolysis can alter IHC dramatically > 2.Placenta contains a lot of biotin...are you blocking adequately? > > Regards > > > > -----Original Message----- > From: "Y. Wang" > To: Histonet@lists.utsouthwestern.edu > Date: Fri, 13 Aug 2004 14:35:07 -0700 (PDT) > Subject: [Histonet] MMP-1 immuno labelling of placenta > > Hello, > > Has anyone had any experience immuno labelling placenta tissue with > anti-MMP1? We have tried this (cryosections with ABC kit with DAB) but are > getting alot of (what we assume to be) background and nothing that looks > like images we have seen on-line. Seeing as we have not had any experience > with placenta tissue we are questioning if we are took the tissue from the > correct place from the placenta (we were given a whole one which we took > random samples from). > > Thank you for your help > Yak-Nam Wang > > Senior Fellow > Department of Bioengineering > University of Washington > Box 357962 > Seattle, WA 98195 > > Tel.: (206)-221-5873 > Fax.: (206)-221-5874 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > .......so what IS the speed of dark? > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Susan.Walzer <@t> HCAHealthcare.com Mon Aug 16 10:43:54 2004 From: Susan.Walzer <@t> HCAHealthcare.com (Walzer Susan) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] a question on ethics Message-ID: <492A62ABF11DA240B063BC8278A88A34611080@orlex03.orl.medcity.net> I would definetly get the pathologists involved too, they are the ones signing off on these screw ups. -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Friday, August 13, 2004 8:32 AM To: Ron Martin Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] a question on ethics Run Ron Run. "Ron Martin" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 04:27 AM To: cc: Subject: [Histonet] a question on ethics Fellow techs, I am in a difficult situation and need some serious advice. I recently (4 months ago) accepted a technical supervisor position in a dermatology lab. I went from a bench tech at my old job to this position. I also walked away from a raise at my old position so I could step into a supervision position. I took the new position because I was told by my manager that she would teach me some supervision, management and financial skills that I currently do not have as well as the growth potential of the company.This position is not what I was told it would be. Part of my "duties" include emptying the trash, clean the bathroom (not happening) and taking her personal and professional calls. The question on ethics is the high volume of mistakes made by our technician, our offices and also by my manager herself. I caught a mistake made by our tech a few weeks ago.She put the wrong tissue on the wrong slides as she inverted the two cases. I caught her mistake before it went out. One time she assigned the same number to two different cases. She then sent the correct case out for a consult (which wasn't needed) and put the wrong patient name on the slide. Every other day there is something different.My manager will not terminate her as she knows I am seeking employment elsewhere and she cannot afford to lose a tech. One day my manager gave three cases the same accession number.She caught her mistake on one of the cases but I still ended up with two cases with the same number. When I first started there my manager had a case in which there " was no tissue in the container". She said that she notified the office about the situation however up to six weeks later the office was still calling looking for results.At that point she wrote up an incident report and dated it six weeks prior to coincide with the surgery date. Our offices are not any better. One case came in with the wrong patient information. It took about 7 weeks for the office to realize that they sent the wrong patient information with the biopsy as the names were close in spelling. The offices continually send mistakes with incorrect spelling of patients names and incorrect anatomical locations. I have tired to document everything but there are too many mistakes and I don't have enough time or energy to keep up with them.My manager wants us to do our own "internal quality control". My interpretation of this is that she doesn't want our physician and risk manager to know of these mistakes. Are these becoming common problems or is it just my situation? I want to emphasize that I hope I am not being unethical myself for revealing this information but I really need some advice and support.I have very high standards and they are not being met in this current situation. I am currently seeking a new position but I need employment and cannot resign until another position becomes available. I would like to stay in Florida and if anyone knows of any positions please inform me as I am at my wits end. Thanks in advance. Ron _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LPrestridge <@t> seton.org Mon Aug 16 11:00:02 2004 From: LPrestridge <@t> seton.org (Prestridge, Linda) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Substitute fixative for B5 Message-ID: We are trying to quit using B5. We are did study with B-Plus from BBc Chemical, IBF from Surgipath, Alcoholic Zinc from ANatech and AZF from Newcomers. Well the pathologists ruled it down to B-Plus and AZF from Newcomers. We then did battery of immuno's - Lymphoma panel. Most stained fair, and they narrowed it down to B-Plus. But...the problem is that I can't get the kappa and lambda (poly's) to stain. I tried AR and got some staining but very dirty. What are any of you using and have you had the same problem getting the kappa and lambda to stain??? Thanks, Linda From JKOBLER <@t> PARTNERS.ORG Mon Aug 16 11:11:28 2004 From: JKOBLER <@t> PARTNERS.ORG (Kobler, James) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Processing tissue engineering sample Message-ID: <57531340B9FDD611A8580008026158F1085F4BB3@phsexch26.mgh.harvard.edu> I would like advice about processing some experimental gels containing fibroblasts. The goal is to stain immunohistochemically for extracellular collagen in small gel samples (5mm diameter, 300 micron thick). It is desirable to cut the gels perpendicular to the surface in order to get multiple sections that can be stained with different antibodies. Cryosectioning would be preferable since dehydration would probably cause shrinkage of the gels. Questions: (1) should the gels be treated with a cryoperservative such as 30% sucrose before freeezing? (2) any tips for orienting the gel discs for nice cross-sections? (3) Is freezing in isopentane cooled with liquid nitrogen the best freezing method? Thanks very much, Jim Kobler, Ph.D. Mass General Hospital jkobler@partners.org 1-617-726-0212 From joshua71 <@t> gmx.de Mon Aug 16 11:23:47 2004 From: joshua71 <@t> gmx.de (Joshua Tree) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] (no subject) Message-ID: <25858.1092673427@www70.gmx.net> test -- NEU: WLAN-Router für 0,- EUR* - auch für DSL-Wechsler! GMX DSL = supergünstig & kabellos http://www.gmx.net/de/go/dsl From juan.gutierrez <@t> christushealth.org Mon Aug 16 11:35:35 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Substitute fixative for B5 Message-ID: Hi Linda, we are using B-Plus and doing our immunos on the Ventana system (Benchmarks). So far no problems with any of the common lymphocyte markers. We didn't even have to change our protocols. Good luck. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My views are my own and do not reflect those of my employer. Long live free speech! "We have nothing to fear but fear itself". -----Original Message----- From: Prestridge, Linda [mailto:LPrestridge@seton.org] Sent: Monday, August 16, 2004 11:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Substitute fixative for B5 We are trying to quit using B5. We are did study with B-Plus from BBc Chemical, IBF from Surgipath, Alcoholic Zinc from ANatech and AZF from Newcomers. Well the pathologists ruled it down to B-Plus and AZF from Newcomers. We then did battery of immuno's - Lymphoma panel. Most stained fair, and they narrowed it down to B-Plus. But...the problem is that I can't get the kappa and lambda (poly's) to stain. I tried AR and got some staining but very dirty. What are any of you using and have you had the same problem getting the kappa and lambda to stain??? Thanks, Linda _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Mon Aug 16 11:47:54 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Substitute fixative for B5 Message-ID: After eliminating B5 several years ago we experimented with several different fixatives. We ended up going back to formalin and, as long as the specimen is adequately fixed, the morphology has been excellent. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Prestridge, Linda" 08/16/04 12:00PM >>> We are trying to quit using B5. We are did study with B-Plus from BBc Chemical, IBF from Surgipath, Alcoholic Zinc from ANatech and AZF from Newcomers. Well the pathologists ruled it down to B-Plus and AZF from Newcomers. We then did battery of immuno's - Lymphoma panel. Most stained fair, and they narrowed it down to B-Plus. But...the problem is that I can't get the kappa and lambda (poly's) to stain. I tried AR and got some staining but very dirty. What are any of you using and have you had the same problem getting the kappa and lambda to stain??? Thanks, Linda _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cjohnston <@t> mdanderson.org Mon Aug 16 11:49:34 2004 From: cjohnston <@t> mdanderson.org (cjohnston@mdanderson.org) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Processing tissue engineering sample Message-ID: I have cut gels for IHC and maybe able to give you some information. Freezing in isopentane works for us. We do not use cryoprotection and have cut the gels to orient them. I would be glad to talk with you and send you some references if you would like. Carol M. Johnston HT(ASCP) M.D. Anderson Cancer Center 1515 Holcombe Blvd. Unit 443 Houston, Texas 77030 713-745-4625 "Kobler, James" To: Sent by: "'histonet@lists.utsouthwestern.edu'" histonet-bounces@lists.utsouth western.edu cc: 08/16/2004 11:11 AM Subject: [Histonet] Processing tissue engineering sample I would like advice about processing some experimental gels containing fibroblasts. The goal is to stain immunohistochemically for extracellular collagen in small gel samples (5mm diameter, 300 micron thick). It is desirable to cut the gels perpendicular to the surface in order to get multiple sections that can be stained with different antibodies. Cryosectioning would be preferable since dehydration would probably cause shrinkage of the gels. Questions: (1) should the gels be treated with a cryoperservative such as 30% sucrose before freeezing? (2) any tips for orienting the gel discs for nice cross-sections? (3) Is freezing in isopentane cooled with liquid nitrogen the best freezing method? Thanks very much, Jim Kobler, Ph.D. Mass General Hospital jkobler@partners.org 1-617-726-0212 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From weneng <@t> hotmail.com Mon Aug 16 12:32:15 2004 From: weneng <@t> hotmail.com (Wendy England) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] quantitative analysis for trichrome stain Message-ID: Dear histonetters, I did some trichrome stain on rat kidney sections. Now I was asked to quantify the collagen. I heard NIH-Image is a choice to do this. Could anybody give me more detail about this method? Many thanks! Wendy _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From carl.hobbs <@t> kcl.ac.uk Mon Aug 16 13:07:20 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] RE: Mowiol aq mountant Message-ID: <001301c483bb$dfa768b0$88412cd9@home> Thanks for reminder, Paul. I add DABCO and azide. Give enough retardation of fading to allow a good session on the confocal to get plenty of images before bleaching sets in. --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.726 / Virus Database: 481 - Release Date: 22/07/2004 From w.huang <@t> qmul.ac.uk Mon Aug 16 13:14:42 2004 From: w.huang <@t> qmul.ac.uk (wenlong) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] mouse monoclonal antibody for p-c-Jun Message-ID: <002b01c483bc$e71219f0$d24c258a@nigella> I recently bought a mouse monoclonal antibody against p-c-Jun from Santa Cruz Biotech (sc-822). I used this antibody for immunofluoresent staining of cryostat sections from injured rat spinal cord (4% PF perfusion of the rat). I got very strong non-specific labelling of the axons. Any kind advice of how to avoid such non-specific labelling? also anyone has a good protocol for this monoclonal antibody? Many thanks, Dr. WL Huang Neuroscience Centre Queen Mary University of London From HornHV <@t> archildrens.org Mon Aug 16 13:31:32 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] ?? about ferric chloride Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B920@EMAIL.archildrens.org> I ordered by mistake ferric chloride hexahydrate instead of plain old ferric chloride. Will the waters make any difference in staining? I can always just try it and see what happens, but if it doesn't work I'd be stuck with the open bottle of a chemical I can't use. Anybody know? Thanks. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From cwscouten <@t> myneurolab.com Mon Aug 16 13:36:43 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Processing tissue engineering sample Message-ID: For more convenient chilled isopentane, see the following link: http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=476401&catdesc=Histology+Equipment&CatThreeID=650&CatOneID=4&subcatdesc=Freezing+Devices&idsubcategory=187 Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 FAX 314 522 0377 cwscouten@myneurolab.com www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cjohnston@mdanderson.org Sent: Monday, August 16, 2004 11:50 AM To: Kobler, James Cc: 'histonet@lists.utsouthwestern.edu'; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Processing tissue engineering sample I have cut gels for IHC and maybe able to give you some information. Freezing in isopentane works for us. We do not use cryoprotection and have cut the gels to orient them. I would be glad to talk with you and send you some references if you would like. Carol M. Johnston HT(ASCP) M.D. Anderson Cancer Center 1515 Holcombe Blvd. Unit 443 Houston, Texas 77030 713-745-4625 "Kobler, James" To: Sent by: "'histonet@lists.utsouthwestern.edu'" histonet-bounces@lists.utsouth western.edu cc: 08/16/2004 11:11 AM Subject: [Histonet] Processing tissue engineering sample I would like advice about processing some experimental gels containing fibroblasts. The goal is to stain immunohistochemically for extracellular collagen in small gel samples (5mm diameter, 300 micron thick). It is desirable to cut the gels perpendicular to the surface in order to get multiple sections that can be stained with different antibodies. Cryosectioning would be preferable since dehydration would probably cause shrinkage of the gels. Questions: (1) should the gels be treated with a cryoperservative such as 30% sucrose before freeezing? (2) any tips for orienting the gel discs for nice cross-sections? (3) Is freezing in isopentane cooled with liquid nitrogen the best freezing method? Thanks very much, Jim Kobler, Ph.D. Mass General Hospital jkobler@partners.org 1-617-726-0212 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jmahoney <@t> alegent.org Mon Aug 16 13:57:06 2004 From: jmahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Special stains Message-ID: Can someone direct em to a good web page that shows pictures of special stains? I need it for a hsito student. Thanks, Jan Mahoney From ryaskovich <@t> dir.nidcr.nih.gov Mon Aug 16 14:31:39 2004 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Special stains Message-ID: <8F3AB322628548428A992EFB0E80F5D3A2F0EF@nihexchange8.nih.gov> Janice, Go to Google and click on images and type what you want. e.g. Histology brain slides etc. Have fun! Ruth Yaskovich National Institutes of Health National Intstitute of Dental and Crainiofacial Research Neuronal Gene Expression Section > ---------- > From: Janice A Mahoney > Sent: Monday, August 16, 2004 1:57 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Special stains > > Can someone direct em to a good web page that shows pictures of special > stains? I need it for a hsito student. > Thanks, > Jan Mahoney > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gu.lang <@t> gmx.at Mon Aug 16 14:35:19 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Special stains References: Message-ID: <008501c483c8$2a147c60$eeeea8c0@server> This is a very good site, but in German. http://alf3.urz.unibas.ch/pathopic/srch-f-thu.cfm Gudrun Lang ----- Original Message ----- From: "Janice A Mahoney" To: Sent: Monday, August 16, 2004 8:57 PM Subject: [Histonet] Special stains Can someone direct em to a good web page that shows pictures of special stains? I need it for a hsito student. Thanks, Jan Mahoney _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gsmith <@t> confocal.com Mon Aug 16 15:11:45 2004 From: gsmith <@t> confocal.com (Glenn Smith) Date: Fri Sep 16 15:23:54 2005 Subject: [Histonet] Image analysis and slide scanning machine (Wester, Martha) In-Reply-To: Message-ID: <00c101c483cd$45edc040$1f05a8c0@confocal.com> Hi Kim, We supply such an instrument, our TISSUEscope - combining confocal scanning laser microscopy and a wide field of view. Depending on your resolution requirements, the TISSUEscope can acquire a single image at up to 1 um resolution with no mosaic and no tiling of images. A standard slide (1" x 3") is loaded into the machine and the rest of operations is via software. We have another model which offers submicron resolution as well. Both brightfield and fluorescence detection is provided within the same instrument. Please feel free to contact me offline for more details. Also, we will have one of these instruments at the NSH meeting in Toronto - we'll be exhibiting in conjunction with Beecher Instruments. Regards Glenn Smith, P.Eng. 519.886.9013 x38 Biomedical Photometrics Inc/GeneFocus Widefield Confocal Scanning Instruments and Software -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: Friday, August 13, 2004 9:16 AM To: Histonet Subject: [Histonet] Image analysis and slide scanning machine Hello everyone, Years ago, I heard of a machine that you could load an entire slide into, scan the slide and then do image analysis on the tissue. I have no idea who made this machine or even how to search for such a thing on the internet (or the histonet archives). If anyone has the name of this company, can someone please let me know. I am currently doing measurements of pancreatic islets as well as measurements of whole mouse pancreas'. I am needing to take 4-6 pictures per pancreas in order to do this, and it is very time consuming (these studies have up to 90 animals in them). I need a quicker and easier way to do this! Help! Kim Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 5 Date: Fri, 13 Aug 2004 13:47:06 -0400 From: Subject: RE: RE: [Histonet] a question on ethics To: Gary Gill , pam marcum , , Joe Nocito Cc: histonet@lists.utsouthwestern.edu Message-ID: <20040813174706.CQCA24693.mta13.adelphia.net@mail.adelphia.net> Content-Type: text/plain; charset=ISO-8859-1 There is also the issue that this position is not what I was told it would be. I also didn't expect to have so many issues such as a tech that can't work up to my standards. Also I keep asking myself if it is worth it esp. when this isn't turning out to be the job I expected it to be. Lastly, how can I work for someone I can't respect or trust? Any thought on that? > > From: Gary Gill > Date: 2004/08/13 Fri PM 01:39:01 EDT > To: "'pathrm35@adelphia.net'" , pam marcum > , histonet@lists.utsouthwestern.edu, Joe Nocito > > CC: histonet@lists.utsouthwestern.edu > Subject: RE: RE: [Histonet] a question on ethics > > Let your physician know that the person in charge is ultimately > responsible that the lab is run by the book. There are right ways and > wrong ways to do things. Wrong is indefensible. Among possible > penalties, CMS can shut down the lab. Surely your boss doesn't want > to see the lab's name in the local newspaper headlines. > > Gary Gill > > -----Original Message----- > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > Sent: Friday, August 13, 2004 12:34 PM > To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: RE: [Histonet] a question on ethics > > > I sent copies of incident reports to RM but my manager wants us to do > our own internal quality control. How are things suppose to get fixed > that way? My physician doesn't like to deal with these situations and > refers me to my manager. I'm running in circles!!! Ron > > > > From: "pam marcum" > > Date: 2004/08/13 Fri AM 09:11:34 EDT > > To: , , > > "Joe Nocito" > > Subject: RE: [Histonet] a question on ethics > > > > > ------------------------------ Message: 6 Date: Fri, 13 Aug 2004 11:14:55 -0700 (PDT) From: "A. Erickson" Subject: RE: RE: [Histonet] a question on ethics To: pathrm35@adelphia.net Cc: histonet@lists.utsouthwestern.edu, Gary Gill , Joe Nocito Message-ID: Content-Type: TEXT/PLAIN; charset=US-ASCII It sounds to me, from the thread of your post(s) that you have already made the decision to leave, just not implemented it! Good luck! Andra Erickson, HT, Research Technologist On Fri, 13 Aug 2004 pathrm35@adelphia.net wrote: > There is also the issue that this position is not what I was told it > would be. I also didn't expect to have so many issues such as a tech that can't work up to my standards. Also I keep asking myself if it is worth it esp. when this isn't turning out to be the job I expected it to be. Lastly, how can I work for someone I can't respect or trust? Any thought on that? > > > > From: Gary Gill > > Date: 2004/08/13 Fri PM 01:39:01 EDT > > To: "'pathrm35@adelphia.net'" , pam marcum > > , histonet@lists.utsouthwestern.edu, Joe Nocito > > > > CC: histonet@lists.utsouthwestern.edu > > Subject: RE: RE: [Histonet] a question on ethics > > > > Let your physician know that the person in charge is ultimately > > responsible that the lab is run by the book. There are right ways > > and wrong ways to do things. Wrong is indefensible. Among possible > > penalties, CMS can shut down the lab. Surely your boss doesn't want > > to see the lab's name in the local newspaper headlines. > > > > Gary Gill > > > > -----Original Message----- > > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > > Sent: Friday, August 13, 2004 12:34 PM > > To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > > Cc: histonet@lists.utsouthwestern.edu > > Subject: Re: RE: [Histonet] a question on ethics > > > > > > I sent copies of incident reports to RM but my manager wants us to > > do our own internal quality control. How are things suppose to get > > fixed that way? My physician doesn't like to deal with these > > situations and refers me to my manager. I'm running in circles!!! > > Ron > > > > > > From: "pam marcum" > > > Date: 2004/08/13 Fri AM 09:11:34 EDT > > > To: , , > > > "Joe Nocito" > > > Subject: RE: [Histonet] a question on ethics > > > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 7 Date: Fri, 13 Aug 2004 14:15:19 -0400 From: "Linda Blazek" Subject: RE: RE: [Histonet] a question on ethics To: pathrm35@adelphia.net, mucram11@comcast.net, garygill@dcla.com, jnocito@pathreflab.com Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Seems to me you have already shut down and have made up your mind that you are done and don't want to accept some of the great ideas that have been offered today. If your goal is to solve the problems you have encountered then the histonet has offered some wonderful help. >>> 8/13/2004 1:47:06 PM >>> There is also the issue that this position is not what I was told it would be. I also didn't expect to have so many issues such as a tech that can't work up to my standards. Also I keep asking myself if it is worth it esp. when this isn't turning out to be the job I expected it to be. Lastly, how can I work for someone I can't respect or trust? Any thought on that? > > From: Gary Gill > Date: 2004/08/13 Fri PM 01:39:01 EDT > To: "'pathrm35@adelphia.net'" , pam marcum > , histonet@lists.utsouthwestern.edu, Joe Nocito > > CC: histonet@lists.utsouthwestern.edu > Subject: RE: RE: [Histonet] a question on ethics > > Let your physician know that the person in charge is ultimately responsible > that the lab is run by the book. There are right ways and wrong ways to do > things. Wrong is indefensible. Among possible penalties, CMS can shut down > the lab. Surely your boss doesn't want to see the lab's name in the local > newspaper headlines. > > Gary Gill > > -----Original Message----- > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > Sent: Friday, August 13, 2004 12:34 PM > To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: RE: [Histonet] a question on ethics > > > I sent copies of incident reports to RM but my manager wants us to do our > own internal quality control. How are things suppose to get fixed that way? > My physician doesn't like to deal with these situations and refers me to my > manager. I'm running in circles!!! Ron > > > > From: "pam marcum" > > Date: 2004/08/13 Fri AM 09:11:34 EDT > > To: , , > > "Joe Nocito" > > Subject: RE: [Histonet] a question on ethics > > > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Fri, 13 Aug 2004 14:35:09 -0400 From: Subject: RE: RE: [Histonet] a question on ethics To: "Linda Blazek" , , , Cc: histonet@lists.utsouthwestern.edu Message-ID: <20040813183509.TGEP9204.mta10.adelphia.net@mail.adelphia.net> Content-Type: text/plain; charset=ISO-8859-1 I am willing to accept any and all ideas for a resolution. I value everyones opinion on histonet, that's why I posted my situation. > > From: "Linda Blazek" > Date: 2004/08/13 Fri PM 02:15:19 EDT > To: pathrm35@adelphia.net, mucram11@comcast.net, garygill@dcla.com, > jnocito@pathreflab.com > CC: histonet@lists.utsouthwestern.edu > Subject: RE: RE: [Histonet] a question on ethics > > Seems to me you have already shut down and have made up your mind that > you are done and don't want to accept some of the great ideas that > have been offered today. If your goal is to solve the problems you > have encountered then the histonet has offered some wonderful help. > > > >>> 8/13/2004 1:47:06 PM >>> > > There is also the issue that this position is not what I was told it > would be. I also didn't expect to have so many issues such as a tech > that can't work up to my standards. Also I keep asking myself if it is > worth it esp. when this isn't turning out to be the job I expected it > to be. Lastly, how can I work for someone I can't respect or trust? > Any thought on that? > > > > From: Gary Gill > > Date: 2004/08/13 Fri PM 01:39:01 EDT > > To: "'pathrm35@adelphia.net'" , pam marcum > > , histonet@lists.utsouthwestern.edu, Joe > Nocito > > > > CC: histonet@lists.utsouthwestern.edu > > Subject: RE: RE: [Histonet] a question on ethics > > > > Let your physician know that the person in charge is ultimately > responsible > > that the lab is run by the book. There are right ways and wrong > > ways > to do > > things. Wrong is indefensible. Among possible penalties, CMS can > shut down > > the lab. Surely your boss doesn't want to see the lab's name in the > local > > newspaper headlines. > > > > Gary Gill > > > > -----Original Message----- > > From: pathrm35@adelphia.net [mailto:pathrm35@adelphia.net] > > Sent: Friday, August 13, 2004 12:34 PM > > To: pam marcum; histonet@lists.utsouthwestern.edu; Joe Nocito > > Cc: histonet@lists.utsouthwestern.edu > > Subject: Re: RE: [Histonet] a question on ethics > > > > > > I sent copies of incident reports to RM but my manager wants us to > > do > our > > own internal quality control. How are things suppose to get fixed > that way? > > My physician doesn't like to deal with these situations and refers > > me > to my > > manager. I'm running in circles!!! Ron > > > > > > From: "pam marcum" > > > Date: 2004/08/13 Fri AM 09:11:34 EDT > > > To: , , > > > > "Joe Nocito" > > > Subject: RE: [Histonet] a question on ethics > > > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ------------------------------ Message: 9 Date: Fri, 13 Aug 2004 15:05:21 -0400 From: "Angela Bitting" Subject: [Histonet] Diastase dilemma To: Message-ID: Content-Type: text/plain; charset=US-ASCII Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. ------------------------------ Message: 10 Date: Fri, 13 Aug 2004 14:27:53 -0500 From: "Joe Nocito" Subject: RE: [Histonet] Diastase dilemma To: "Angela Bitting" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Angela, It could be that the patient section contains mucin, which will not digest out. If this is not the case, then I suggest trying to digest the slides on a slide rack or something to lay the slides on a horizontal surface. Just my 21/2 cents worth. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Angela Bitting Sent: Friday, August 13, 2004 2:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Diastase dilemma Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Fri, 13 Aug 2004 14:31:07 -0500 From: Jackie.O'Connor@abbott.com Subject: Re: [Histonet] Diastase dilemma To: "Angela Bitting" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Did you stop to think that your patient just might be positive for glycogen? "Angela Bitting" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 02:05 PM To: cc: Subject: [Histonet] Diastase dilemma Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Fri, 13 Aug 2004 13:44:05 -0600 From: "Daryl Mikita" Subject: [Histonet] pHing of eosin To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hello, We are looking at pHing our eosin to the correct pH, instead of just using it straight from the bottle. What should we use to adjust the pH? Should it be 1N sodium hydroxide or 1N hydrochloric acid? Thanks Daryl ------------------------------ Message: 13 Date: Fri, 13 Aug 2004 13:47:19 -0600 From: "Daryl Mikita" Subject: [Histonet] Another question To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hello, We were looking at our HQIP and we were wondering where in the H&E staining procedure, people use heat? We could not think of any place except for drying the slides. Thanks Daryl ------------------------------ Message: 14 Date: Fri, 13 Aug 2004 15:07:51 -0500 From: Gary Gill Subject: RE: [Histonet] pHing of eosin To: 'Daryl Mikita' , histonet@lists.utsouthwestern.edu Message-ID: <0B11620AEE5EFB4FA7A7558339972A8307048A@HALIBUT.dcla.com> Content-Type: text/plain Suggest using glacial acetic acid: 5 mL per liter of eosin. Be prepared for a color explosion. Gary Gill -----Original Message----- From: Daryl Mikita [mailto:dmikita@wmcnet.org] Sent: Friday, August 13, 2004 2:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pHing of eosin Hello, We are looking at pHing our eosin to the correct pH, instead of just using it straight from the bottle. What should we use to adjust the pH? Should it be 1N sodium hydroxide or 1N hydrochloric acid? Thanks Daryl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Fri, 13 Aug 2004 16:11:47 -0400 From: Amos Brooks Subject: [Histonet] a question on ethics To: histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.0.20040813154852.01be9008@pop.earthlink.net> Content-Type: text/plain; charset="us-ascii"; format=flowed Ron, When a ship is hitting a reef there is 2 options. First option: Bail out, hit the life rafts and get out of there before you go down with the ship. Second option is to get the ship off the reef and repair the damage. This sounds like a problematic situation and you may be in a good position to make drastic improvements to the system. The question you need to decide is which course you intend to take, and stick with it. If you decide to leave, do it cautiously as these problems could still be pinned on you as the guy that came in caused problems and left. (I know that's not the way it is but people often shift blame for problems on people that can't defend themselves). You still should mention in an exit interview (assuming you get one) about the issues, so that someone else can fix them when you are gone, and so it is documented that you were aware of the situation and not the cause. If you decide to fix the ship, expect some rough waters. Very often when you point out errors to people they start looking for things to complain about with you. You should proceed with caution and document, document, document! The root cause survey that was previously mentioned sounds like a good plan. Often people make errors that they can prevent just by a simple modification of their procedure. Establishing redundancy is a good way to prevent most of the errors you have described. For example, picking a block and a slide up and comparing them before they go into the microtome, then again after the slide is cut and finally comparing the block to the slides before sending them out. Time consuming, yes, but it is hard to make the same mistake 3 times. Similar redundancies can be established in most other situations too. Turn around time will probably suffer a bit, but if it keeps you from getting sued it's worth it. Good luck which ever way you choose, Amos Brooks At 09:29 AM 8/13/2004, you wrote: >Message: 9 >Date: Fri, 13 Aug 2004 04:27:14 -0500 >From: "Ron Martin" >Subject: [Histonet] a question on ethics >To: >Message-ID: <002701c48117$b8cbc750$5dedeb44@Pathrm35> >Content-Type: text/plain; charset="iso-8859-1" > >Fellow techs, >I am in a difficult situation and need some serious advice. I recently >(4 >months ago) accepted a technical supervisor position in a dermatology lab. >I went from a bench tech at my old job to this position. I also walked >away from a raise at my old position so I could step into a supervision >position. I took the new position because I was told by my manager that >she would teach me some supervision, management and financial skills that >I currently do not have as well as the growth potential of the >company.This position is not what I was told it would be. Part of my >"duties" include emptying the trash, clean the bathroom (not happening) >and taking her personal and professional calls. The question on ethics is >the high volume of mistakes made by our technician, our offices and also >by my manager herself. >I caught a mistake made by our tech a few weeks ago.She put the wrong >tissue on the wrong slides as she inverted the two cases. I caught her >mistake before it went out. One time she assigned the same number to two >different cases. She then sent the correct case out for a consult (which >wasn't needed) and put the wrong patient name on the slide. Every other >day there is something different.My manager will not terminate her as she >knows I am seeking employment elsewhere and she cannot afford to lose a tech. >One day my manager gave three cases the same accession number.She caught >her mistake on one of the cases but I still ended up with two cases with >the same number. When I first started there my manager had a case in which >there " was no tissue in the container". She said that she notified the >office about the situation however up to six weeks later the office was >still calling looking for results.At that point she wrote up an incident >report and dated it six weeks prior to coincide with the surgery date. >Our offices are not any better. One case came in with the wrong patient >information. It took about 7 weeks for the office to realize that they >sent the wrong patient information with the biopsy as the names were close >in spelling. The offices continually send mistakes with incorrect >spelling of patients names and incorrect anatomical locations. > I have tired to document everything but there are too many mistakes and > I don't have enough time or energy to keep up with them.My manager wants > us to do our own "internal quality control". My interpretation of this is > that she doesn't want our physician and risk manager to know of these > mistakes. Are these becoming common problems or is it just my situation? >I want to emphasize that I hope I am not being unethical myself for >revealing this information but I really need some advice and support.I >have very high standards and they are not being met in this current >situation. I am currently seeking a new position but I need employment and >cannot resign until another position becomes available. I would like to >stay in Florida and if anyone knows of any positions please inform me as I >am at my wits end. Thanks in advance. > >Ron ------------------------------ Message: 16 Date: Fri, 13 Aug 2004 15:12:37 -0500 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] You may want to hit the delete button with this, then again, maybe not To: "Joe Nocito" , "Histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hey at least they tried to talk to you(a year later)but they tried YOU! Like I told you on my last e-mail my behind is about two pounds lighter after my boss bit some of it off because of my comments on the histonet about a certain German company who should remain nameless. Apparently my speaking the truth, the whole truth, and nothing but the truth on this forum made them a little mad. If they made good products to begin with, we wouldn't be having this conversation. Have anybody noticed how many pages of the phonebook are dedicated to lawyers? There is about twice the number of lawyers compared to doctors. Aint that scary? I wonder when lawyer season is going to open, heck there's more of them than there is deer. Just kidding. I know one good lawyer, one. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My views are my own and do not reflect those of my employer. Long live free speech! "We have nothing to fear but fear itself". -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Thursday, August 12, 2004 3:35 PM To: Histonet Subject: [Histonet] You may want to hit the delete button with this,then again, maybe not Afternoon Netter's, Remember about a week or two the topic was getting "flamed"? Well, here's a new one. Sometime last year, I made several postings about a certain company and their product that I used and it did not work for me. I wrote them a letter and returned all the merchandise back to them saying I was not happy with their product. I waited several months and never got a response, so I named the company on the Histonet. Since then, I have been throwing away their flyers, catalogs and any other correspondence I receive. Obviously, they are doing fine without my business and I am doing fine without their products. Today, I receive a letter from the company with a copy of my Histonet postings from over a year ago, telling me that this has been in customer service for about a year (hmmmmmmmm, I'm trying to find out where the customer service is in all of this. (One year, okay, I'll get back to that), that their customer service has tried several times to contact me but to no avail. (Hmmmmm, one way would have been to respond to my email address, but I'll come back to this also). So after all this time, they want to do business with me. (So close to the NSH meeting, hmmmmmmmmm, another point to ponder). My big beef is that these companies read this forum and try to use strong arm tactics on people who post negative comments about them or their products. I have many friends that are sales reps in this field (I'm talking like come over my house and eat type of friends) and I love these people. I must reiterate that this forum was set up to exchange ideas, theories and experiences, both positive and negative. If this was such a "hot item", point #1, why did it take a whole year to get a response when they had my company and address on file? Is this how customer service works? I don't think so!! Point #2 If you are a vendor and don't like a particular posting, why don't you reply back to that person via email. Point #3 don't wait until it's almost time for the NSH annual S/C to fix a problem, it's probably too late by then. Point #4 If you haven't had a purchase order or have not corresponded with a customer for over a year, chances are that customer does not want to do business with you. Let lying dogs lie and don't open old scars. All you did was get me going on a tangent. I know, there goes Nocito again, but I refuse to be treated like this from any one. Like they say in the movies, "Don't call me, I'll call you". Okay, I've said my piece now let the FLAMING BEGIN!!!!!!!!! As always, the opinions stated by this author in no way reflects the opinions of his employer, subordinates, friends or relatives. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Fri, 13 Aug 2004 15:12:07 -0500 From: Jackie.O'Connor@abbott.com Subject: Re: [Histonet] Diastase dilemma To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Once I read my own answer, I saw how stupid it is. My apologies. It's been a long week. Think before you type, Jack . . .think..... Jackie.O'Connor@abbott.com Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 02:31 PM To: "Angela Bitting" cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Diastase dilemma Did you stop to think that your patient just might be positive for glycogen? "Angela Bitting" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 02:05 PM To: cc: Subject: [Histonet] Diastase dilemma Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Fri, 13 Aug 2004 14:15:00 -0600 From: Gayle Callis Subject: Re: [Histonet] pHing of eosin To: "Daryl Mikita" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20040813141110.01b227a8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed If you are buying eosin Y, already made up, check the MSDS, and the pH, it may be adjusted correctly. I know that this is the case for Richard Allan Eosin Y solution. If you are making up eosin y from the dry dye, then the recipe is (Hrapchak and Sheehan) eosin y 0.5g 95% ethanol 50 ml glacial acetic acid 1 drop If you are adjusting already made up, use acetic acid. At 01:44 PM 8/13/2004, you wrote: >Hello, > >We are looking at pHing our eosin to the correct pH, instead of just >using >it straight from the bottle. What should we use to adjust the pH? Should >it be 1N sodium hydroxide or 1N hydrochloric acid? > >Thanks >Daryl > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 19 Date: Fri, 13 Aug 2004 16:15:48 -0400 From: "TERRI BRAUD" Subject: [Histonet] RE: a Question of Ethics To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed I can certainly understand the frustration of working in new management position that does not offer the training and responsibilities to go with it. And while I support going over a manager's head when patient care is impacted, you also must ask yourself if the activity is being tolerated by the pathologist(s)? If so, then look for another job, because you MUST have the support of your medical staff. Beyond that, I would encourage you to look for management training from outside sources, such as the program offered at the NSH convention, on-line courses in Quality Improvement and Laboratory Management, subscription to the Manager's edition of Advance, etc. These will help give you the educational tools to get started, but they in no way will make you an effective leader. Speaking from 15 years of management experience, to be an effective leader you have to be out in front, not someone who just points the way. As for any job, this often means "paying dues" by performing "other" duties until you are accepted as part of the team. If this means you have to clean bathrooms or empty waste to show you are part of a team, then do it cheerfully for a time. If a team sees your willingness to perform the "nasty" tasks, then eventually they will be more willing to take on those tasks, and others, to free your time for management duties. Anyone can point to problems within any system, but the true leader is the one that offers concrete solutions and suggestions and is willing to go the extra miles to put them into place. As we all know, Histology is a manual labor intensive process, prone to identity errors. As a leader, you must provide your employees with a system that has enough system checks and double checks that limit their ability to make a mistake, or catch it before it impacts patient care. Then, when mistakes are made, use a carefully constructed system to document and to review the system process. Maybe these "mistake-prone" employees have suffered the same frustrations that you have, even to the point of not caring anymore. Have they been given the tools, the training, the education needed to perform their jobs accurately and with positive feedback? If you, as a new manager, are not being helped, how much do you think is trickling down to their level? I would guess very little. This lab may or may not be the place to learn to be a supervisor, but it sounds like a place that is "ripe" for the picking. Anyone can step into a smooth running lab, but having the opportunity to make a real difference in a poorly functioning lab is not as easy. Laboratory administrators often come from a clinical background with very little AP experience and usually welcome the Histology Supervisor that comes in with fresh ideas to improve a lab's performance. Study process improvement. Offer well thought out, budget neutral suggestions for process improvement. And remember, being a supervisor means that your most important skill should be in "people management". This means coming up with ways to inspire and empower your employees to do a better job! Good luck with your quest, where ever it takes you. Sincerely, Terri L. Braud, HT(ASCP) Surgical Pathology Manager University of Virginia Health Systems Charlottesville, VA 22908 ------------------------------ Message: 20 Date: Fri, 13 Aug 2004 15:30:12 -0500 From: "Stapf, Ross" Subject: RE: [Histonet] Diastase dilemma To: "Angela Bitting" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Are you running a plain PAS on the patient as well? I was always taught to always run a PAS and PASD even if the Dr only wanted a PASD. It serves as a control. If you are doing that, and know there is no diference digested vs undigested in the patient then I don't know what to say. I would normally look to something in the tissue that is causing this. If you were still using spit to digest I would blame it on you eating sugar before spitting on the slide, or not drinking enough coffee. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Friday, August 13, 2004 2:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Diastase dilemma Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ------------------------------ Message: 21 Date: Fri, 13 Aug 2004 15:46:36 -0500 From: "Linda Davis" Subject: [Histonet] Re: Smears on the Ventana To: Message-ID: <007c01c48176$bfbe2490$05a16243@D6JLZ851> Content-Type: text/plain; charset="iso-8859-1" Terre, We fix the smear in 95% alcohol, rinse in water, and place on the Ventana to run along with the paraffin sections. Linda Davis, HT (ASCP) Histology Supervisor Rio Grande Regional Hospital McAllen, TX. ------------------------------ Message: 22 Date: Fri, 13 Aug 2004 16:29:01 -0500 From: "Charles.Embrey" Subject: RE: [Histonet] Diastase dilemma To: "'Jackie.O'Connor@abbott.com'" Cc: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Did you ever stop to think.... that if the patient tissue were positive for Glycogen then the digestion would remove it and it would show no PAS staining? The positive reaction on could be mucin or acid mucosubstances. Charles Embrey -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Friday, August 13, 2004 2:31 PM To: Angela Bitting Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Diastase dilemma Did you stop to think that your patient just might be positive for glycogen? "Angela Bitting" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/13/2004 02:05 PM To: cc: Subject: [Histonet] Diastase dilemma Bizarre things happen here all the time, but this is REALLY BIZARRE! We'll do a PAS w/ diastase and our control tissue will be digested but the patient tissue won't be. They are mounted on the same slide! The next day we will run the stain again and we tried turning the slide upside-down in the Coplin jar. Same problem. We mixed fresh diastase solution and tried again. Same problem. Tomorrow, we may run it again and it will work fine. What could be going on? I'm perplexed! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 23 Date: Fri, 13 Aug 2004 14:35:07 -0700 (PDT) From: "Y. Wang" Subject: [Histonet] MMP-1 immuno labelling of placenta To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: TEXT/PLAIN; charset=US-ASCII Hello, Has anyone had any experience immuno labelling placenta tissue with anti-MMP1? We have tried this (cryosections with ABC kit with DAB) but are getting alot of (what we assume to be) background and nothing that looks like images we have seen on-line. Seeing as we have not had any experience with placenta tissue we are questioning if we are took the tissue from the correct place from the placenta (we were given a whole one which we took random samples from). Thank you for your help Yak-Nam Wang Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 ------------------------------ Message: 24 Date: Fri, 13 Aug 2004 16:40:44 -0500 From: "Kapoor, Sue" Subject: RE: [Histonet] RE: a Question of Ethics To: histonet@lists.utsouthwestern.edu Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F563D@khmcexch.uhsi.org> Content-Type: text/plain; charset="iso-8859-1" As for any job, this often means "paying dues" by performing "other" duties until you are accepted as part of the team. If this means you have to clean bathrooms or empty waste to show you are part of a team, then do it cheerfully for a time. -I'm sorry, but as a Histology Supervisor, cleaning bathrooms or emptying waste does NOT show you are a "part of the team"...it shows that management has no respect for your position. Why isn't the housekeeping dept. doing these duties? Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 ------------------------------ Message: 25 Date: Fri, 13 Aug 2004 18:03:58 -0400 From: Diana McCaig Subject: [Histonet] acid washed glassware To: histonet@lists.utsouthwestern.edu Message-ID: <3E5A3F039F0BD8118B4700C00D002024043250@CKHA9> Content-Type: text/plain What concentration and what acid do you use to clean glassware prior to silver impregnation staining. Do you simply rinse it or is it soaked for a specific period of time? Air or oven dried? Diana McCaig, R.T. Charge Tech, Histology Chatham Kent Health Alliance 519-352-6401 (6604) ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 9, Issue 23 *************************************** From gentras <@t> vetmed.auburn.edu Mon Aug 16 15:38:44 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] FIZ Message-ID: <6.0.1.1.0.20040816153319.025ca3e0@mailhost.vetmed.auburn.edu> Hello, please will someone familiar with Film In Situ Zymography (FIZ) share with me either your recipe or source for the (not sure of the correct spelling) trichrolic, or trichloric, or trichlolic acid used to make the 0.3% Biebrich Scarlet ASAP? Thanks, Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From gsmith <@t> confocal.com Mon Aug 16 15:41:24 2004 From: gsmith <@t> confocal.com (Glenn Smith) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Image analysis and slide scanning machine (Wester, Martha) In-Reply-To: Message-ID: <00c501c483d1$6a4496e0$1f05a8c0@confocal.com> Hi Kim, We supply such an instrument, our TISSUEscope - combining confocal scanning laser microscopy and a wide field of view. Depending on your resolution requirements, the TISSUEscope can acquire a single image at up to 1 um resolution with no mosaic and no tiling of images. A standard slide (1" x 3") is loaded into the machine and the rest of operations is via software. We have another model which offers submicron resolution as well. Both brightfield and fluorescence detection is provided within the same instrument. Please feel free to contact me offline for more details. Also, we will have one of these instruments at the NSH meeting in Toronto - we'll be exhibiting in conjunction with Beecher Instruments. Regards Glenn Smith, P.Eng. 519.886.9013 x38 Biomedical Photometrics Inc/GeneFocus Widefield Confocal Scanning Instruments and Software -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: Friday, August 13, 2004 9:16 AM To: Histonet Subject: [Histonet] Image analysis and slide scanning machine Hello everyone, Years ago, I heard of a machine that you could load an entire slide into, scan the slide and then do image analysis on the tissue. I have no idea who made this machine or even how to search for such a thing on the internet (or the histonet archives). If anyone has the name of this company, can someone please let me know. I am currently doing measurements of pancreatic islets as well as measurements of whole mouse pancreas'. I am needing to take 4-6 pictures per pancreas in order to do this, and it is very time consuming (these studies have up to 90 animals in them). I need a quicker and easier way to do this! Help! Kim Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From lpwenk <@t> sbcglobal.net Mon Aug 16 19:13:27 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] ?? about ferric chloride References: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B920@EMAIL.archildrens.org> Message-ID: <004d01c483ef$05b6b960$f5e3d445@domainnotset.invalid> I think the answer is yes, but we have to do a little chemistry math. The formula for ferric chloride anhydrous is FeCl3 (1 iron, 3 chlorines) The formula for ferric chloride hexahydrate is FeCl3*6H2O (1 iron, 3 chlorines, and 6 waters) The molecular weight (MW) of FeCl3 anhydrous is 162. When 6 waters is added on, the MW of water is 6 x 18 = 108. So the molecular weight of FeCl3*6H2O is 162 + 108 = 270. So, out of the 270 MW of the hexahydrate, 108 is water, or about 40% of the weight (108/240). Hopefully, it would follow that, if you were to weigh out 10 g of the anhydrous, all 10 grams would be ferric chloride. However, if you weighed out 10 g of the hexahydrate, 40% would be water and only 60% would be ferric chloride, or about 6 grams. So to get the hexahydrate to equal anhydrous (for the ferric chloride), you would have take the amount you need and multiply it by 270/162 = 1.67. Or, if 10 g of anhydrous is needed, then multiply the 10 g x 1.67 = 16.7 g of ferric chloride hexahydrate would equal 10 g of ferric chloride anhydrous. Try that, and let us know how it works. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI ----- Original Message ----- From: "Horn, Hazel V" To: Sent: Monday, August 16, 2004 2:31 PM Subject: [Histonet] ?? about ferric chloride I ordered by mistake ferric chloride hexahydrate instead of plain old ferric chloride. Will the waters make any difference in staining? I can always just try it and see what happens, but if it doesn't work I'd be stuck with the open bottle of a chemical I can't use. Anybody know? Thanks. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 ---------------------------------------------------------------------------- -- The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eugenio.pittioni <@t> uniud.it Tue Aug 17 02:24:26 2004 From: eugenio.pittioni <@t> uniud.it (Eugenio PITTIONI) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Special stains In-Reply-To: <8F3AB322628548428A992EFB0E80F5D3A2F0EF@nihexchange8.nih.gov> References: <8F3AB322628548428A992EFB0E80F5D3A2F0EF@nihexchange8.nih.gov> Message-ID: <200408170924.26922.eugenio.pittioni@uniud.it> > > From: Janice A Mahoney > > > > Can someone direct em to a good web page that shows pictures of special > > stains? I need it for a hsito student. > > Thanks, > > Jan Mahoney > > Here you can find protocols and some nice pictures http://www.bris.ac.uk/Depts/PathAndMicro/CPL/histmeth.htm Bye Eugenio PITTIONI Veterinary Medicine University of UDINE ITALY From Janet.Bonner <@t> FLHOSP.ORG Tue Aug 17 07:33:53 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Iron stain question Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB400A@fh2k093.fhmis.net> Just a note on the side about the core being negative - this no longer has to be! BCC rapidcal immuno decals Bone Marrow cores without compromising the positivity of the iron (no- I don't work for them). We just discovered it and the immuno and iron results are beautiful. -----Original Message----- From: lpwenk@sbcglobal.net To: WWmn916@aol.com; histonet@pathology.swmed.edu Sent: 8/15/2004 7:39 AM Subject: Re: [Histonet] Iron stain question Usually, it's the bone core that is negative due to the decalcification, while the clot and the smear are positive. Couple off the wall thoughts. Any chance that the smear slide was in the same coplin jar slot as the control, so that the smear was stuck against the back of the previous slide? That would prevent the ferrocyanide solution from "touching" the smear. If this is possible, remove the coverslip, remove the mounting media with xylene (or whatever you use), run the slide back down through alcohols to water, and restain. Any chance that the smear was fixed in something containing an acid, like acetic acid? That might remove the iron. No way to correct this problem. Did they do a bone core and/or clot? Were they negative? If so, then the person IS iron deficient, regardless of whether the person has leukemia or not. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Saturday, August 14, 2004 11:49 PM Subject: [Histonet] Iron stain question > Hello again histonetters, > > Can anyone give me any insight as to why some smear slides stained for iron > would not stains positive as it was expected they would? Doctor say's the > slides should have stained positive for iron because the patient has leukemia. > The control stained positive as it should have. Staining tech is sure all her > slides were exposed to solution. Thanks for any help. > > Deb King, HT(ASCP) > Sacramento, CA > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From JWEEMS <@t> sjha.org Tue Aug 17 08:26:24 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Regarding the recent Biohazard Discussion Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A631E@sjhaexc02.sjha.org> Hi Everyone, There is an article in ADVANCE regarding this issue. Perhaps it shed light on the questions. (I haven't read it - just saw it!) j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From garygill <@t> dcla.com Tue Aug 17 09:49:43 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] pHing of eosin Message-ID: <0B11620AEE5EFB4FA7A7558339972A8307049A@HALIBUT.dcla.com> Re dye content variation in biological dyes certified by the Biological Stain Commission, eosin must contain at least 90% dye to be certified. The threshold used to be 80% until about 1996 or so. Dye content is printed on the labels of all certified dyes -- a notable exception being hematoxylin, as it is not a dye per se. The amount of dye that should be weighed out to produce the same total dye content (TDC) solution will vary, therefore. For example, to prepare 1 liter of 0.5% TDC eosin solution (raw dye content = 90% [0.9]), dissolve 5.55 gm eosin in 1 liter solvent (i.e., 5 gm ? 0.9 = 5.55 gm). If the raw dye content were 95%, one would weigh out 5.26 gm (i.e., 5 gm ? 0.95 = 5.26 gm). A difference of 0.29 gm will not make any visually appreciable difference. If one were using dyes with lower certification dye content thresholds, the differences between a corrected and uncorrected amount of dye could be visibly appreciable (e.g., light green SF yellowish = 65%, orange G = 80%). Gary Gill -----Original Message----- From: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: Monday, August 16, 2004 6:37 AM To: histonet@lists.utsouthwestern.edu Subject: FW: [Histonet] pHing of eosin I believe that care should be used in universally adding specific amounts of acetic acid to acidify eosin solutions that differ. I asume that different batches of dye have different dye contents so that the 5% stain that one user prepares is a little different from that used by others. Each lab should prepare their eosin with this is mind. I usually add a low percentage of acetic acid to the eosin until a faint opalescence occurs. I have been told that this is the point at which the dye acid just starts to precipitate. For our eosin solutions this involves 5 drops or so of 2% acetic acid per 100 ml of 5% eosin. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tony Henwood Sent: Sun 8/15/2004 6:27 PM To: 'Daryl Mikita'; histonet@lists.utsouthwestern.edu Cc: Subject: RE: [Histonet] pHing of eosin We use 3% acetic acid Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Daryl Mikita [mailto:dmikita@wmcnet.org] Sent: Saturday, 14 August 2004 5:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pHing of eosin Hello, We are looking at pHing our eosin to the correct pH, instead of just using it straight from the bottle. What should we use to adjust the pH? Should it be 1N sodium hydroxide or 1N hydrochloric acid? Thanks Daryl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Vivian.King <@t> CLS.ab.ca Tue Aug 17 10:45:19 2004 From: Vivian.King <@t> CLS.ab.ca (Vivian.King@CLS.ab.ca) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Muscle biopsies Message-ID: <30C050525B881C4AAFF41E6D16543E681927CF@mail3.cls.ab.ca> Does anyone out there get muscle biopsies sent to them from far away? (ie: a few hours transport time) How are they sent to you? (Fresh? or Frozen in proper orientation?) I am wondering if anyone has any idea how long a muscle bx can be held on ice before the diagnostic value of the tissue is compromised. Any information would be greatly appreciated. Vivian King Tech II - Neuropathology FMC Calgary, Alberta 403-944-1576 vivian.king@cls.ab.ca From dlbroo01 <@t> gwise.louisville.edu Tue Aug 17 10:50:59 2004 From: dlbroo01 <@t> gwise.louisville.edu (Donna L Brooks) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Re: Histonet Digest, Vol 9, Issue 27 Message-ID: Hello Dr. WL Huang, In regards to the mouse monoclonal antibody for p-c-Jun: Be sure to check if your secondary antibody is "rat adsorbed". I suspect that you may be experiencing a cross reactivity due to the species (rat/mouse) being so closely related. I have also done quite a bit of immunoflourescent and immunoperoxidase staining of rodent neural tissue from spinal cord injured rodents, and had this very same problem. Also, I would suggest checking the Ig groups. For example, IgG versus IgA. Did you use a protein blocking solution? If not you may want to try that as well. Be sure to watch the concentration of your antibodies and blocking serums/protein solutions. If the concentrations are too high (say 1:250 [high] vs 1:1000[lower]) you will get a high background. I suggest doing a titration method to see which concentrations give th From jmitchell <@t> neurology.wisc.edu Tue Aug 17 11:22:39 2004 From: jmitchell <@t> neurology.wisc.edu (Mitchell (Jean)) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Muscle biopsies Message-ID: There is an article in December 1998 Journal of Histotechnology "Delayed Processing of Muscle Biopsy Specimens: Does it Really Compromise Enzyme Histochemistry?" The paper addresses extended delay (up to 48 hours) in processing muscle biopsies. I can fax a copy if you would like. When muscle biopsies are transported to us, ideally we like to receive the tissue on regular ice (not dry ice) within 3 hours. At times it has taken up to 6 hours to receive samples. The staining & diagnostic value of the muscle tissue has remained intact in all cases. With extended transport time usually PAS staining is the most compromised due to glycogan leakage. But one can always note the difference in the section quality of a muscle biopsy that was frozen & sectioned on site vs a biopsy that was transported to us for processing. We tend to see more problems with muscle samples when they are frozen on site then transported to us on dry ice. Much more artifact with that method - but they are still diagnostic. Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vivian.King@CLS.ab.ca Sent: Tuesday, August 17, 2004 10:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Muscle biopsies Does anyone out there get muscle biopsies sent to them from far away? (ie: a few hours transport time) How are they sent to you? (Fresh? or Frozen in proper orientation?) I am wondering if anyone has any idea how long a muscle bx can be held on ice before the diagnostic value of the tissue is compromised. Any information would be greatly appreciated. Vivian King Tech II - Neuropathology FMC Calgary, Alberta 403-944-1576 vivian.king@cls.ab.ca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From failm <@t> musc.edu Tue Aug 17 11:19:44 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Muscle biopsies Message-ID: Vivian, We have outside clients wrap the fresh muscle in saline dampened gauze put it in a biohazard bag then place that bag in another containing ice. Most of the time the biopsies are sent by cab or by one of their security personnel. But they are sometimes shipped overnight on ice. We did run some tests on Autopsy muscle and saw only minimal loss of enzyme histochemistry after 72 hours of refrigerator storage. But we prefer than none go longer than 48 before snap freezing, and specify that the muscles must be received within 24 hours. Rena Fail >>> 08/17/04 11:45AM >>> Does anyone out there get muscle biopsies sent to them from far away? (ie: a few hours transport time) How are they sent to you? (Fresh? or Frozen in proper orientation?) I am wondering if anyone has any idea how long a muscle bx can be held on ice before the diagnostic value of the tissue is compromised. Any information would be greatly appreciated. Vivian King Tech II - Neuropathology FMC Calgary, Alberta 403-944-1576 vivian.king@cls.ab.ca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JQB7 <@t> CDC.GOV Tue Aug 17 11:34:41 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] silver question Message-ID: All: Has anyone experienced a problem with their Steiner's "fading" after a period of time? We had a slide that had been stained with the Steiner procedure and it was very positive, however when it was reviewed at a much later date it appeared negative. Another section was stained and it was positive. I know that leaving the sections for a long time in xylene before they are coverslipped can lead to fading but wouldn't this be apparent when the slides were originally viewed? Any suggestions? Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 From northma <@t> ohsu.edu Tue Aug 17 11:35:36 2004 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Muscle biopsies Message-ID: In our experience, if the specimen has been kept moist (not soaked in saline) and cold, the enzyme histochemisty stains still yield diagnostic results even after many hours following surgery. Some of our fresh specimens are not received until the day after surgery due to a faraway location. These have been refrigerated at the source hospital and then sent on ice, FedEx overnight. Mary North, HT(ASCP), HTL Neuromuscular Laboratory Oregon Health & Science University Portland, OR >>> 8/17/2004 8:45:19 AM >>> Does anyone out there get muscle biopsies sent to them from far away? (ie: a few hours transport time) How are they sent to you? (Fresh? or Frozen in proper orientation?) I am wondering if anyone has any idea how long a muscle bx can be held on ice before the diagnostic value of the tissue is compromised. Any information would be greatly appreciated. Vivian King Tech II - Neuropathology FMC Calgary, Alberta 403-944-1576 vivian.king@cls.ab.ca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kmerriam2003 <@t> yahoo.com Tue Aug 17 11:38:45 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Cutting fresh "squishy" rat brain Message-ID: <20040817163845.22190.qmail@web52505.mail.yahoo.com> Hello everyone, I need to cut sections of fresh rat brain, in certain stereotactic coordinates and then embed them in OCT. Does anyone have any tips as to how to do this without squashing the brain? We are trying to cut coronal sections of the brain to precisely expose a particular region, and we are using a fresh disposable microtome blade to do the cuts, but the brain is still very squishy when fresh and is very difficult to cut. We are doing LCM on these cryosections, so any type of fixation is out of the question. Kim Merriam Novartis Cambridge, MA Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Mail is new and improved - Check it out! From cormier <@t> MIT.EDU Tue Aug 17 11:47:58 2004 From: cormier <@t> MIT.EDU (Kathleen Cormier) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] T blue for mucin veronal acetate buffer Message-ID: <5.2.1.1.2.20040817123802.00af5b40@hesiod> Hello All! I am looking for a supplier for the veronal acetate buffer needed in the T blue for mucin (Theory and Practice ver 1980 pg 169-70) Since the buffer uses sodium barbiturate (alas! a controlled substance!) it would be much easier to get this buffer pre made. Any one know of anyone who sells this buffer or kit? Is there a similar stain that does not have sodium barbiturate? Is this a lost cause? This researcher is looking at gel membranes with glycosaminoglycans chains, and would like the T blue to stain the sulfated and nonsulfated mucopolysaccharides... Thanks! Kathy From gcallis <@t> montana.edu Tue Aug 17 11:53:23 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Cutting fresh "squishy" rat brain In-Reply-To: <20040817163845.22190.qmail@web52505.mail.yahoo.com> References: <20040817163845.22190.qmail@web52505.mail.yahoo.com> Message-ID: <6.0.0.22.1.20040817104743.01b1d888@gemini.msu.montana.edu> What about holding brain in one of the rat brain matrices holders. It allows holding the brain for precise slices, coronal orientation. www.Myneurolab.com has these, for coronal, sagittal, and different age and size of rats. You can cut as thin as 1 mm At 10:38 AM 8/17/2004, you wrote: >Hello everyone, > >I need to cut sections of fresh rat brain, in certain stereotactic >coordinates and then embed them in OCT. Does anyone have any tips as to >how to do this without squashing the brain? We are trying to cut coronal >sections of the brain to precisely expose a particular region, and we are >using a fresh disposable microtome blade to do the cuts, but the brain is >still very squishy when fresh and is very difficult to cut. We are doing >LCM on these cryosections, so any type of fixation is out of the question. > >Kim Merriam >Novartis >Cambridge, MA > > >Kim Merriam >Novartis >Cambridge, MA > >--------------------------------- >Do you Yahoo!? >Yahoo! Mail is new and improved - Check it out! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From kmerriam2003 <@t> yahoo.com Tue Aug 17 12:44:03 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Cutting fresh "squishy" rat brain In-Reply-To: <6.0.0.22.1.20040817104743.01b1d888@gemini.msu.montana.edu> Message-ID: <20040817174403.44128.qmail@web52505.mail.yahoo.com> Thanks so much for this information about brain matrices from myneurolab and ted pella. The histonet can be a lifesaver! Kim Gayle Callis wrote: What about holding brain in one of the rat brain matrices holders. It allows holding the brain for precise slices, coronal orientation. www.Myneurolab.com has these, for coronal, sagittal, and different age and size of rats. You can cut as thin as 1 mm At 10:38 AM 8/17/2004, you wrote: >Hello everyone, > >I need to cut sections of fresh rat brain, in certain stereotactic >coordinates and then embed them in OCT. Does anyone have any tips as to >how to do this without squashing the brain? We are trying to cut coronal >sections of the brain to precisely expose a particular region, and we are >using a fresh disposable microtome blade to do the cuts, but the brain is >still very squishy when fresh and is very difficult to cut. We are doing >LCM on these cryosections, so any type of fixation is out of the question. > >Kim Merriam >Novartis >Cambridge, MA > > >Kim Merriam >Novartis >Cambridge, MA > >--------------------------------- >Do you Yahoo!? >Yahoo! Mail is new and improved - Check it out! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. From chau0056 <@t> umn.edu Tue Aug 17 13:19:05 2004 From: chau0056 <@t> umn.edu (chau0056) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] antibodies for human and bovine collagen type 1 Message-ID: <200408171819.i7HIJ5S6025253@qix.software.umn.edu> I'm working with collagen matrixes, made of bovine type I dermal collagen. I am seeding the matrix with Human stromal fibroblast that secrete type I human collagen in to the matrix. My question is: are there primary antibodies specific to human and bovine collagen type 1? I have asked a few companies, and they are not sure about the cross reactivity between the two. I suspect that human and bovine collagen are too similar in structure for me to distinguish between the two with immunofluoresence confocal microscopy. Eric Chau Dept Mechanical Enginnering University of Minnesota From degaboh <@t> rice.edu Tue Aug 17 13:39:21 2004 From: degaboh <@t> rice.edu (Z. Patel) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Image analysis and slide scanning machine In-Reply-To: <20040817133438.168AD19897@fungible4.mail.rice.edu> Message-ID: <20040817183921.8B7221DB64@fungible10.mail.rice.edu> I think a machine called MicroTech 4000, or something along those lines, will scan an entire slide. Zarana From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: Friday, August 13, 2004 9:16 AM To: Histonet Subject: [Histonet] Image analysis and slide scanning machine Hello everyone, Years ago, I heard of a machine that you could load an entire slide into, scan the slide and then do image analysis on the tissue. I have no idea who made this machine or even how to search for such a thing on the internet (or the histonet archives). If anyone has the name of this company, can someone please let me know. I am currently doing measurements of pancreatic islets as well as measurements of whole mouse pancreas'. I am needing to take 4-6 pictures per pancreas in order to do this, and it is very time consuming (these studies have up to 90 animals in them). I need a quicker and easier way to do this! Help! Kim Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? From mbecker <@t> pathlabinc.com Tue Aug 17 13:48:05 2004 From: mbecker <@t> pathlabinc.com (Michele Becker) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Who stores the old blocks? Message-ID: Hello Histo-world! We have been processing small specimens for a physician in the area for several years. We send him the final H&E slide which he keeps. We have always stored the blocks on site and sent blocks on an as needed basis. Here is my question - Since he is closing his practice here and moving to another part of the country,what should happen with his block files? Apparently, he will be taking the slides with him. It makes sense for the block files to go with the slides. I just want to make sure it is the right thing to do. Thanks for your help. Michele From gcallis <@t> montana.edu Tue Aug 17 14:24:15 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Who stores the old blocks? In-Reply-To: References: Message-ID: <6.0.0.22.1.20040817131857.01aef338@gemini.msu.montana.edu> A similar thing happened to our lab, we did human renal biopsies for several years. All blocks and duplicate accession records were given to the physician. I suggets for future work like this, give them all blocks, unstained and stained slides are given to the physician for storage at the time you return a stained slide. You can help them set up a block storage unit - At 12:48 PM 8/17/2004, you wrote: >Hello Histo-world! > >We have been processing small specimens for a physician in the area for >several years. We send him the final H&E slide which he keeps. We have >always stored the blocks on site and sent blocks on an as needed basis. >Here is my question - Since he is closing his practice here and moving to >another part of the country,what should happen with his block files? >Apparently, he will be taking the slides with him. It makes sense for the >block files to go with the slides. I just want to make sure it is the right >thing to do. >Thanks for your help. > >Michele > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From nperson211 <@t> comcast.net Tue Aug 17 14:52:24 2004 From: nperson211 <@t> comcast.net (nperson211@comcast.net) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] tris HCl antigen retrieval Message-ID: <081720041952.8325.412261F80009A792000020852200762302CECECD02019C9D0A9F02@comcast.net> From nperson211 <@t> comcast.net Tue Aug 17 14:55:02 2004 From: nperson211 <@t> comcast.net (nperson211@comcast.net) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] tris-HCl antigen retrieval Message-ID: <081720041955.20554.41226295000E8D740000504A2200735446CECECD02019C9D0A9F02@comcast.net> Histonetters, Would any of you be so kind as to share any and all formulae for buffers utilizing Tris base and HCl for antigen retrieval, with a pH of 10.0? I have one formula but it is giving me uneven results. Thanks in advance, Nancy Lemke Hermelin Brain Tumor Center Henry Ford Hospital Detroit From cgfields <@t> lexhealth.org Tue Aug 17 15:39:00 2004 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Recycling Systems Message-ID: Hello Everyone, We are looking at reagent recycling systems at our hospital and would like to know the different companies techs are using. CBG Biotech came to talk with us today and I know of R&B but would like to know some of the pros and cons to some of these systems. I would really appreciate any information you can give us, good or bad. THX in advance Carole Fields Lexington Medical Center W.Columbia, SC _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From mcauliff <@t> umdnj.edu Tue Aug 17 19:29:14 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] T blue for mucin veronal acetate buffer In-Reply-To: <5.2.1.1.2.20040817123802.00af5b40@hesiod> References: <5.2.1.1.2.20040817123802.00af5b40@hesiod> Message-ID: <4122A2DA.2000300@umdnj.edu> Hi Kathy: You don't need veronal acetate buffer to get good tol. blue staining of GAGs, other acetate buffers (check the buffer tables at the back of most histotechnique books) will work just as well. Pick the pH you want and go for it. Metachromasia can sometimes be retained if you dehydrate in acetone rather than alcohol, then clear in fresh xylenes (w/o previous contamination w/ alcohols). Geoff Kathleen Cormier wrote: > Hello All! > > I am looking for a supplier for the veronal acetate buffer needed in > the T blue for mucin (Theory and Practice ver 1980 pg 169-70) Since > the buffer uses sodium barbiturate (alas! a controlled substance!) it > would be much easier to get this buffer pre made. Any one know of > anyone who sells this buffer or kit? Is there a similar stain that > does not have sodium barbiturate? Is this a lost cause? This > researcher is looking at gel membranes with glycosaminoglycans chains, > and would like the T blue to stain the sulfated and nonsulfated > mucopolysaccharides... > > Thanks! > > Kathy > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From histophilhuff <@t> yahoo.com Tue Aug 17 16:38:35 2004 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] tris-HCl antigen retrieval - Protocol Online - In-Reply-To: <081720041955.20554.41226295000E8D740000504A2200735446CECECD02019C9D0A9F02@comcast.net> Message-ID: <20040817213835.85354.qmail@web50307.mail.yahoo.com> Check out the following website. Scroll down for the relevant links to antigen retrieval techniques some of which involve tris buffer. http://www.protocol-online.org/prot/Immunology/Immunohistochemistry/Antigen_Retrieval/index.html The protocol online site has a very valuable selection of protocols. nperson211@comcast.net wrote: Histonetters, Would any of you be so kind as to share any and all formulae for buffers utilizing Tris base and HCl for antigen retrieval, with a pH of 10.0? I have one formula but it is giving me uneven results. Thanks in advance, Nancy Lemke Hermelin Brain Tumor Center Henry Ford Hospital Detroit _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! From AnthonyH <@t> chw.edu.au Tue Aug 17 17:25:04 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] silver question Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E08D@simba.kids> Check the mountant you are using. Some will fade Spirochete silver stains very quickly Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Bartlett, Jeanine [mailto:JQB7@CDC.GOV] Sent: Wednesday, 18 August 2004 2:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] silver question All: Has anyone experienced a problem with their Steiner's "fading" after a period of time? We had a slide that had been stained with the Steiner procedure and it was very positive, however when it was reviewed at a much later date it appeared negative. Another section was stained and it was positive. I know that leaving the sections for a long time in xylene before they are coverslipped can lead to fading but wouldn't this be apparent when the slides were originally viewed? Any suggestions? Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From tahseen <@t> brain.net.pk Tue Aug 17 18:18:50 2004 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] proposal for post graduate diploma program in histology Message-ID: <006801c484b0$8fc9e3e0$972bfea9@m7c0y4> Dear Histoneter, My medical director has given me a task to prepare a proposal for post graduate diploma program in histology. Would any one like to share with me in preparation of a comprehensive proposal for post graduate diploma program. Thanks in advance Muhammad Tahseen Histology Supervisor SKMCH & RC. From info <@t> ihcworld.com Tue Aug 17 18:46:55 2004 From: info <@t> ihcworld.com (IHC World) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] tris-HCl antigen retrieval In-Reply-To: <081720041955.20554.41226295000E8D740000504A2200735446CECECD02019C9D0A9F02@comcast.net> Message-ID: IHC World website has many different antigen retrieval protocols including Tris-HCl pH10. Here is the link to the protocols: http://www.ihcworld.com/epitope%20retrieval.htm Hope this helps. Richard IHC World Online Information Center for Immunohistochemistry Website: http://ihcworld.com -----Original Message----- From: nperson211@comcast.net [mailto:nperson211@comcast.net] Sent: Tuesday, August 17, 2004 2:55 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] tris-HCl antigen retrieval Histonetters, Would any of you be so kind as to share any and all formulae for buffers utilizing Tris base and HCl for antigen retrieval, with a pH of 10.0? I have one formula but it is giving me uneven results. Thanks in advance, Nancy Lemke Hermelin Brain Tumor Center Henry Ford Hospital Detroit _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Penguindeb <@t> aol.com Tue Aug 17 18:57:36 2004 From: Penguindeb <@t> aol.com (Penguindeb@aol.com) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] CD 2 on Ventana Message-ID: <78.5e7277bd.2e53f570@aol.com> Hello Histonetters, Is anyone running CD 2 on the Ventana? If so, who is your vendor and what is your protocol. Thanks for all your help. Deb McGee HT,HTL(ASCP) HVMA Boston,MA Deborah_McGee@vmed.org From JQB7 <@t> CDC.GOV Tue Aug 17 19:39:31 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] silver question Message-ID: That hasn't changed: same mountant for years. And I need to correct something in my initial email. The stain faded after only 2 weeks. -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Tue 8/17/2004 6:25 PM To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu Cc: Subject: RE: [Histonet] silver question Check the mountant you are using. Some will fade Spirochete silver stains very quickly Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Bartlett, Jeanine [mailto:JQB7@CDC.GOV] Sent: Wednesday, 18 August 2004 2:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] silver question All: Has anyone experienced a problem with their Steiner's "fading" after a period of time? We had a slide that had been stained with the Steiner procedure and it was very positive, however when it was reviewed at a much later date it appeared negative. Another section was stained and it was positive. I know that leaving the sections for a long time in xylene before they are coverslipped can lead to fading but wouldn't this be apparent when the slides were originally viewed? Any suggestions? Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From dmikita <@t> wmcnet.org Wed Aug 18 06:14:34 2004 From: dmikita <@t> wmcnet.org (Daryl Mikita) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Thanks you all. Message-ID: Thanks for all the responses to my questions. Thanks again, Daryl Mikita From cgfields <@t> lexhealth.org Wed Aug 18 08:14:09 2004 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Recycling Systems Message-ID: THX Dianne, Which machine, 2-10 gal., do you have? Carole -----Original Message----- From: Dndsomi@aol.com [mailto:Dndsomi@aol.com] Sent: Tuesday, August 17, 2004 5:22 PM To: Carole Fields Subject: Re: [Histonet] Recycling Systems Carole, We have had the CBG system for more than a year and haven't had any problems other than sometimes we have a fishy odor when we recycle xylene. It has really saved a tremendous amount of $$ since we don't have to pay for xylene disposal. Very simple to operate ! Dianne Dietz _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From cgfields <@t> lexhealth.org Wed Aug 18 08:16:28 2004 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Recycling Systems Message-ID: Hi Amy, We would like to recycle xylene and alcohol. At the present time we are having it hauled and we are about to go to the next level...it is getting expensive. THX Carole -----Original Message----- From: Amy Self [mailto:ASelf@gmhsc.com] Sent: Tuesday, August 17, 2004 5:07 PM To: Carole Fields Subject: RE: [Histonet] Recycling Systems Carole, we have a CBG Biotech recycler. Right now we only recycle xylene since that is a very pricy chemical. I think that we were paying something like 200.00(+) for a case of 4 (1-gal. jugs). It has been a very good machine I can't say that we have any problems with it - hold on let me knock on wood real quick - I think that you woule like the CBG - and they have good tech. support. What are you planning on recycling? This one that we have will recycle alcohol also, but right now we only recycle xylene. Good Luck..... Amy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Carole Fields Sent: Tuesday, August 17, 2004 4:39 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Recycling Systems Hello Everyone, We are looking at reagent recycling systems at our hospital and would like to know the different companies techs are using. CBG Biotech came to talk with us today and I know of R&B but would like to know some of the pros and cons to some of these systems. I would really appreciate any information you can give us, good or bad. THX in advance Carole Fields Lexington Medical Center W.Columbia, SC _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From thallada <@t> noch.org Wed Aug 18 08:37:26 2004 From: thallada <@t> noch.org (Hallada, Teri) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Recycling Systems Message-ID: We are using CBG and have had wonderful service. We recycle formalin, xylene, and alcohol wth the smaller unit. Teri Hallada BS MT CT (ASCP) thallada@noch.org > -----Original Message----- > From: Carole Fields [SMTP:cgfields@lexhealth.org] > Sent: Wednesday, August 18, 2004 09:16 > To: 'Amy Self' > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Recycling Systems > > Hi Amy, > > We would like to recycle xylene and alcohol. At the present time we are > having it hauled and we are about to go to the next level...it is getting > expensive. > THX > Carole > > -----Original Message----- > From: Amy Self [mailto:ASelf@gmhsc.com] > Sent: Tuesday, August 17, 2004 5:07 PM > To: Carole Fields > Subject: RE: [Histonet] Recycling Systems > > > > Carole, we have a CBG Biotech recycler. Right now we only recycle > xylene since that is a very pricy chemical. I think that we were paying > something like 200.00(+) for a case of 4 (1-gal. jugs). It has been a very > good machine I can't say that we have any problems with it - hold on let me > knock on wood real quick - I think that you woule like the CBG - and they > have good tech. support. What are you planning on recycling? This one that > we have will recycle alcohol also, but right now we only recycle xylene. > Good Luck..... Amy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Carole > Fields > Sent: Tuesday, August 17, 2004 4:39 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Recycling Systems > > > Hello Everyone, > > We are looking at reagent recycling systems at our hospital and would like > to know the different companies techs are using. CBG Biotech came to talk > with us today and I know of R&B but would like to know some of the pros and > cons to some of these systems. I would really appreciate any information > you can give us, good or bad. > > THX in advance > > Carole Fields > Lexington Medical Center > W.Columbia, SC > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of its > attachments, please be advised that you have received this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you > have received this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed above if one is provided. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > NOTE: > The information contained in this message may be privileged, confidential > and protected from disclosure. If the reader of this message is not the > intended recipient, or an employee or agent responsible for delivering this > message to the intended recipient, you are hereby notified that any > dissemination, distribution or copying of this communication is strictly > prohibited. If you have received this communication in error, please notify > us immediately by replying to this message and deleting it from your > computer. Thank you. > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of its > attachments, please be advised that you have received this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you> > have received this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed above if one is provided. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Statement: This message is intended only for the individual or entity to which it is addressed. It may contain privileged, confidential information which is exempt from disclosure under applicable laws. If you are not the intended recipient, please note that you are strictly prohibited from disseminating or distributing this information (other than to the intended recipient) or copying this information. If you have received this communication in error, please notify us immediately by e-mail or by telephone at (616) 842-3600. Thank you. From JPCOLEMA <@t> sentara.com Wed Aug 18 09:08:11 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Spectrin staining in megs Message-ID: Does anyone use spectrin for red cell precursors and does anyone get megakaryocyte staining? We do and we would like to not have megs light up. From LINDA.MARGRAF <@t> childrens.com Wed Aug 18 12:02:09 2004 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Regulations for TB exposure in histology Message-ID: Dear Histonetters: We are reviewing our safety procedures and policies for tuberculosis exposure in unfixed tissue in the histology lab. We have arranged for annual fit testing for N95 respirators but our hospital infection control people are also asking if we need negative air pressure in the room and additional safety precautions such as special hoods. Does anyone know of specific reulations regarding these issues. Thanks so much, Linda M Histonet administrator From weneng2004 <@t> yahoo.com Wed Aug 18 12:16:43 2004 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] ImageJ Message-ID: <20040818171643.39187.qmail@web53410.mail.yahoo.com> Dear histonetters, Please forgive me the duplicate email. Due to my address problem I couldn't see the responses to my question regarding trichrome analysis using ImageJ. I downloaded ImageJ but couldn't figure out how to use it on measuring collagen stained blue. Could anybody please help me with this? For those kind people who replied my previous email send me again, please! Many thanks, Wendy __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From gtesdall <@t> yahoo.com Wed Aug 18 12:51:44 2004 From: gtesdall <@t> yahoo.com (greg tesdall) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Sentinel lymph node and CAP Message-ID: <20040818175144.95463.qmail@web14205.mail.yahoo.com> Hi. Would anyone like to share a policy/procedure that addresses CAP ANP.11275 regarding the safe handling of radioactive sentinel lymph nodes. Thank you. I didn't find anything in the archives. __________________________________ Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. http://mobile.yahoo.com/maildemo From dlbroo01 <@t> gwise.louisville.edu Wed Aug 18 13:01:50 2004 From: dlbroo01 <@t> gwise.louisville.edu (Donna L Brooks) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Re: Mouse monoclonal antibody for p-c-Jun (the rest of the message) Message-ID: Hello again Dr. WL Huang, I am not sure why all of my message did not appear. So, here's the rest. I suggested you do a titration method to check for the best concentrations of your antibodies to see which combination of concentrations yielded the best signal by settting up an experiment changing only the primary concentration, establish that perimeter and then further experimentation changing only the secondary concentration to establish that perimeter. I suggest starting with say 1:250, then 1:500, 1:1000 and so on with each and see what happens. Also, look at www.vectorlabs.com. They have the type of "rat adsorbed" products I described. Jackson Immunologicals does as well. Hope this helps and I was clear. Donna Brooks HT(ASCP) Histotechnologist Specialist University of Louisville Department of Anatomical Sciences and Neurobiology >>> histonet-request@lists.utsouthwestern.edu 08/17/04 1:11 PM >>> Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: pHing of eosin (Gary Gill) 2. Muscle biopsies (Vivian.King@CLS.ab.ca) 3. Re: Histonet Digest, Vol 9, Issue 27 (Donna L Brooks) 4. RE: Muscle biopsies (Mitchell (Jean)) 5. Re: Muscle biopsies (Mildred Fail) 6. silver question (Bartlett, Jeanine) 7. Re: Muscle biopsies (Mary North) 8. Cutting fresh "squishy" rat brain (Kim Merriam) 9. T blue for mucin veronal acetate buffer (Kathleen Cormier) 10. Re: Cutting fresh "squishy" rat brain (Gayle Callis) ---------------------------------------------------------------------- Message: 1 Date: Tue, 17 Aug 2004 09:49:43 -0500 From: Gary Gill Subject: RE: [Histonet] pHing of eosin To: histonet@lists.utsouthwestern.edu Message-ID: <0B11620AEE5EFB4FA7A7558339972A8307049A@HALIBUT.dcla.com> Content-Type: text/plain; charset="iso-8859-1" Re dye content variation in biological dyes certified by the Biological Stain Commission, eosin must contain at least 90% dye to be certified. The threshold used to be 80% until about 1996 or so. Dye content is printed on the labels of all certified dyes -- a notable exception being hematoxylin, as it is not a dye per se. The amount of dye that should be weighed out to produce the same total dye content (TDC) solution will vary, therefore. For example, to prepare 1 liter of 0.5% TDC eosin solution (raw dye content = 90% [0.9]), dissolve 5.55 gm eosin in 1 liter solvent (i.e., 5 gm ? 0.9 = 5.55 gm). If the raw dye content were 95%, one would weigh out 5.26 gm (i.e., 5 gm ? 0.95 = 5.26 gm). A difference of 0.29 gm will not make any visually appreciable difference. If one were using dyes with lower certification dye content thresholds, the differences between a corrected and uncorrected amount of dye could be visibly appreciable (e.g., light green SF yellowish = 65%, orange G = 80%). Gary Gill -----Original Message----- From: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: Monday, August 16, 2004 6:37 AM To: histonet@lists.utsouthwestern.edu Subject: FW: [Histonet] pHing of eosin I believe that care should be used in universally adding specific amounts of acetic acid to acidify eosin solutions that differ. I asume that different batches of dye have different dye contents so that the 5% stain that one user prepares is a little different from that used by others. Each lab should prepare their eosin with this is mind. I usually add a low percentage of acetic acid to the eosin until a faint opalescence occurs. I have been told that this is the point at which the dye acid just starts to precipitate. For our eosin solutions this involves 5 drops or so of 2% acetic acid per 100 ml of 5% eosin. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tony Henwood Sent: Sun 8/15/2004 6:27 PM To: 'Daryl Mikita'; histonet@lists.utsouthwestern.edu Cc: Subject: RE: [Histonet] pHing of eosin We use 3% acetic acid Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Daryl Mikita [mailto:dmikita@wmcnet.org] Sent: Saturday, 14 August 2004 5:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pHing of eosin Hello, We are looking at pHing our eosin to the correct pH, instead of just using it straight from the bottle. What should we use to adjust the pH? Should it be 1N sodium hydroxide or 1N hydrochloric acid? Thanks Daryl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Tue, 17 Aug 2004 09:45:19 -0600 From: Vivian.King@CLS.ab.ca Subject: [Histonet] Muscle biopsies To: histonet@lists.utsouthwestern.edu Message-ID: <30C050525B881C4AAFF41E6D16543E681927CF@mail3.cls.ab.ca> Content-Type: text/plain; charset="iso-8859-1" Does anyone out there get muscle biopsies sent to them from far away? (ie: a few hours transport time) How are they sent to you? (Fresh? or Frozen in proper orientation?) I am wondering if anyone has any idea how long a muscle bx can be held on ice before the diagnostic value of the tissue is compromised. Any information would be greatly appreciated. Vivian King Tech II - Neuropathology FMC Calgary, Alberta 403-944-1576 vivian.king@cls.ab.ca ------------------------------ Message: 3 Date: Tue, 17 Aug 2004 11:50:59 -0400 From: "Donna L Brooks" Subject: [Histonet] Re: Histonet Digest, Vol 9, Issue 27 To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hello Dr. WL Huang, In regards to the mouse monoclonal antibody for p-c-Jun: Be sure to check if your secondary antibody is "rat adsorbed". I suspect that you may be experiencing a cross reactivity due to the species (rat/mouse) being so closely related. I have also done quite a bit of immunoflourescent and immunoperoxidase staining of rodent neural tissue from spinal cord injured rodents, and had this very same problem. Also, I would suggest checking the Ig groups. For example, IgG versus IgA. Did you use a protein blocking solution? If not you may want to try that as well. Be sure to watch the concentration of your antibodies and blocking serums/protein solutions. If the concentrations are too high (say 1:250 [high] vs 1:1000[lower]) you will get a high background. I suggest doing a titration method to see which concentrations give th ------------------------------ Message: 4 Date: Tue, 17 Aug 2004 11:22:39 -0500 From: "Mitchell \(Jean\)" Subject: RE: [Histonet] Muscle biopsies To: , Message-ID: Content-Type: text/plain; charset="us-ascii" There is an article in December 1998 Journal of Histotechnology "Delayed Processing of Muscle Biopsy Specimens: Does it Really Compromise Enzyme Histochemistry?" The paper addresses extended delay (up to 48 hours) in processing muscle biopsies. I can fax a copy if you would like. When muscle biopsies are transported to us, ideally we like to receive the tissue on regular ice (not dry ice) within 3 hours. At times it has taken up to 6 hours to receive samples. The staining & diagnostic value of the muscle tissue has remained intact in all cases. With extended transport time usually PAS staining is the most compromised due to glycogan leakage. But one can always note the difference in the section quality of a muscle biopsy that was frozen & sectioned on site vs a biopsy that was transported to us for processing. We tend to see more problems with muscle samples when they are frozen on site then transported to us on dry ice. Much more artifact with that method - but they are still diagnostic. Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vivian.King@CLS.ab.ca Sent: Tuesday, August 17, 2004 10:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Muscle biopsies Does anyone out there get muscle biopsies sent to them from far away? (ie: a few hours transport time) How are they sent to you? (Fresh? or Frozen in proper orientation?) I am wondering if anyone has any idea how long a muscle bx can be held on ice before the diagnostic value of the tissue is compromised. Any information would be greatly appreciated. Vivian King Tech II - Neuropathology FMC Calgary, Alberta 403-944-1576 vivian.king@cls.ab.ca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Tue, 17 Aug 2004 12:19:44 -0400 From: "Mildred Fail" Subject: Re: [Histonet] Muscle biopsies To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Vivian, We have outside clients wrap the fresh muscle in saline dampened gauze put it in a biohazard bag then place that bag in another containing ice. Most of the time the biopsies are sent by cab or by one of their security personnel. But they are sometimes shipped overnight on ice. We did run some tests on Autopsy muscle and saw only minimal loss of enzyme histochemistry after 72 hours of refrigerator storage. But we prefer than none go longer than 48 before snap freezing, and specify that the muscles must be received within 24 hours. Rena Fail >>> 08/17/04 11:45AM >>> Does anyone out there get muscle biopsies sent to them from far away? (ie: a few hours transport time) How are they sent to you? (Fresh? or Frozen in proper orientation?) I am wondering if anyone has any idea how long a muscle bx can be held on ice before the diagnostic value of the tissue is compromised. Any information would be greatly appreciated. Vivian King Tech II - Neuropathology FMC Calgary, Alberta 403-944-1576 vivian.king@cls.ab.ca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Tue, 17 Aug 2004 12:34:41 -0400 From: "Bartlett, Jeanine" Subject: [Histonet] silver question To: Message-ID: Content-Type: text/plain; charset="us-ascii" All: Has anyone experienced a problem with their Steiner's "fading" after a period of time? We had a slide that had been stained with the Steiner procedure and it was very positive, however when it was reviewed at a much later date it appeared negative. Another section was stained and it was positive. I know that leaving the sections for a long time in xylene before they are coverslipped can lead to fading but wouldn't this be apparent when the slides were originally viewed? Any suggestions? Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 ------------------------------ Message: 7 Date: Tue, 17 Aug 2004 09:35:36 -0700 From: "Mary North" Subject: Re: [Histonet] Muscle biopsies To: Vivian.King@CLS.ab.ca, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii In our experience, if the specimen has been kept moist (not soaked in saline) and cold, the enzyme histochemisty stains still yield diagnostic results even after many hours following surgery. Some of our fresh specimens are not received until the day after surgery due to a faraway location. These have been refrigerated at the source hospital and then sent on ice, FedEx overnight. Mary North, HT(ASCP), HTL Neuromuscular Laboratory Oregon Health & Science University Portland, OR >>> 8/17/2004 8:45:19 AM >>> Does anyone out there get muscle biopsies sent to them from far away? (ie: a few hours transport time) How are they sent to you? (Fresh? or Frozen in proper orientation?) I am wondering if anyone has any idea how long a muscle bx can be held on ice before the diagnostic value of the tissue is compromised. Any information would be greatly appreciated. Vivian King Tech II - Neuropathology FMC Calgary, Alberta 403-944-1576 vivian.king@cls.ab.ca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Tue, 17 Aug 2004 09:38:45 -0700 (PDT) From: Kim Merriam Subject: [Histonet] Cutting fresh "squishy" rat brain To: Histonet Message-ID: <20040817163845.22190.qmail@web52505.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hello everyone, I need to cut sections of fresh rat brain, in certain stereotactic coordinates and then embed them in OCT. Does anyone have any tips as to how to do this without squashing the brain? We are trying to cut coronal sections of the brain to precisely expose a particular region, and we are using a fresh disposable microtome blade to do the cuts, but the brain is still very squishy when fresh and is very difficult to cut. We are doing LCM on these cryosections, so any type of fixation is out of the question. Kim Merriam Novartis Cambridge, MA Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Mail is new and improved - Check it out! ------------------------------ Message: 9 Date: Tue, 17 Aug 2004 12:47:58 -0400 From: Kathleen Cormier Subject: [Histonet] T blue for mucin veronal acetate buffer To: histonet@pathology.swmed.edu Message-ID: <5.2.1.1.2.20040817123802.00af5b40@hesiod> Content-Type: text/plain; charset="us-ascii"; format=flowed Hello All! I am looking for a supplier for the veronal acetate buffer needed in the T blue for mucin (Theory and Practice ver 1980 pg 169-70) Since the buffer uses sodium barbiturate (alas! a controlled substance!) it would be much easier to get this buffer pre made. Any one know of anyone who sells this buffer or kit? Is there a similar stain that does not have sodium barbiturate? Is this a lost cause? This researcher is looking at gel membranes with glycosaminoglycans chains, and would like the T blue to stain the sulfated and nonsulfated mucopolysaccharides... Thanks! Kathy ------------------------------ Message: 10 Date: Tue, 17 Aug 2004 10:53:23 -0600 From: Gayle Callis Subject: Re: [Histonet] Cutting fresh "squishy" rat brain To: Kim Merriam , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20040817104743.01b1d888@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed What about holding brain in one of the rat brain matrices holders. It allows holding the brain for precise slices, coronal orientation. www.Myneurolab.com has these, for coronal, sagittal, and different age and size of rats. You can cut as thin as 1 mm At 10:38 AM 8/17/2004, you wrote: >Hello everyone, > >I need to cut sections of fresh rat brain, in certain stereotactic >coordinates and then embed them in OCT. Does anyone have any tips as to >how to do this without squashing the brain? We are trying to cut coronal >sections of the brain to precisely expose a particular region, and we are >using a fresh disposable microtome blade to do the cuts, but the brain is >still very squishy when fresh and is very difficult to cut. We are doing >LCM on these cryosections, so any type of fixation is out of the question. > >Kim Merriam >Novartis >Cambridge, MA > > >Kim Merriam >Novartis >Cambridge, MA > >--------------------------------- >Do you Yahoo!? >Yahoo! Mail is new and improved - Check it out! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 9, Issue 28 *************************************** From Dorothy.L.Webb <@t> HealthPartners.Com Wed Aug 18 13:41:00 2004 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] recyclers Message-ID: We have the CBG Biotech that recycles formalin, xylene, and alcohol. We use it only for xylene and it does a GREAT job. We have tried formalin, as we will have to do this in the near future, due to EPA standrads in our counties. It worked well, although, it is always a little cumbersome to get the PH to the exact as it should be, mostly doing the appropriate math! Their customer service is great and we have really appreciated their support and help throughout our learning curves!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From Dorothy.L.Webb <@t> HealthPartners.Com Wed Aug 18 13:53:31 2004 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] paraffin Message-ID: We recently switched to Surgipath's "Blue Ribbon" paraffin and the techs loved it! Within the past two weeks, they are noticing that the ribbon is more compressed and harder to stretch out on the waterbath. Any ideas as to why this would be happening when the product was used for about two months with no problem whatsoever?? We are using it in both the processor and the embedder and it is a multfunction paraffin. I like the "blueness" of the paraffin, as all of the small biopsies stand out in the paraffin when embedded for easier sectioning visualness! Thanks ahead of time..LOVE the histonet!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From froyer <@t> bitstream.net Wed Aug 18 14:22:44 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] paraffin In-Reply-To: References: Message-ID: <4123AC84.2080104@bitstream.net> Dorothy, You have probably already checked this and eliminated it as the cause, but has the temperature of your water bath(s) changed? Do you continuously monitor your water temp? i.e. does you water bath(s) have a digital temp. display? ~ Ford Ford M. Royer, MT(ASCP) Midwest Science & Biocenter 6551 Jansen Ave. NE Alberville, MN 55301 email Dorothy.L.Webb@HealthPartners.Com wrote: >We recently switched to Surgipath's "Blue Ribbon" paraffin and the techs >loved it! Within the past two weeks, they are noticing that the ribbon is >more compressed and harder to stretch out on the waterbath. Any ideas as to >why this would be happening when the product was used for about two months >with no problem whatsoever?? We are using it in both the processor and the >embedder and it is a multfunction paraffin. I like the "blueness" of the >paraffin, as all of the small biopsies stand out in the paraffin when >embedded for easier sectioning visualness! Thanks ahead of time..LOVE the >histonet!! > >________________________________________ > >This e-mail and any files transmitted with it are confidential and are >intended solely for the use of the individual or entity to whom they >are addressed. If you are not the intended recipient or the individual >responsible for delivering the e-mail to the intended recipient, please >be advised that you have received this e-mail in error and that any >use, dissemination, forwarding, printing, or copying of this e-mail >is strictly prohibited. > >If you have received this e-mail in error, please immediately notify >the HealthPartners Support Center by telephone at (952) 967-6600. >You will be reimbursed for reasonable costs incurred in notifying us. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From JNocito <@t> Pathreflab.com Wed Aug 18 14:28:36 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] paraffin In-Reply-To: Message-ID: Dorothy, Have you tried changing the angle of the blade? Has the humidity level in your lab changed? I know this time of year can play havoc in the histology lab with all the heat and humidity, even in a HVAC environment. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dorothy.L.Webb@HealthPartners.Com Sent: Wednesday, August 18, 2004 1:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] paraffin We recently switched to Surgipath's "Blue Ribbon" paraffin and the techs loved it! Within the past two weeks, they are noticing that the ribbon is more compressed and harder to stretch out on the waterbath. Any ideas as to why this would be happening when the product was used for about two months with no problem whatsoever?? We are using it in both the processor and the embedder and it is a multfunction paraffin. I like the "blueness" of the paraffin, as all of the small biopsies stand out in the paraffin when embedded for easier sectioning visualness! Thanks ahead of time..LOVE the histonet!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LINDA.MARGRAF <@t> childrens.com Wed Aug 18 15:28:48 2004 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] fibrin for cell blocks Message-ID: Dear Histonetters: Does anyone have experience using fibrin or another substance as a matrix to add a cell suspension to when making cell blocks. My colleague wants to make cell blocks to section and stain for a research study but there will only be a small amount of cellular material. She remembers seeing this done in the past but needs to know where to obtain the reagnets and what procedures to follow. Thanks so much Linda M Histonet administrator From gcallis <@t> montana.edu Wed Aug 18 15:33:09 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Re: to paraffin problems In-Reply-To: References: Message-ID: <6.0.0.22.1.20040818140057.01b2aa90@gemini.msu.montana.edu> BE sure you stir your paraffin BEFORE embedding. Polymers and other additives can settle to bottom of melted paraffin and this changes nature of paraffin, you need to redistribute additives before embedding to have your paraffin at its mixture capacity. If you melt paraffin in a separate oven and use this pre-melted paraffin in processor, the same thing needs to be done, stir before filling your paraffin processing pots/stations. A grocery store has plastic spoons that look like wooden spoons or a good wide spatula works nicely. Check you knife angles, it may NOT be the paraffin, but knife settings. A different lot of knives can mean tiny readjustment is needed. Keep track of your lot numbers on paraffin, it helps if you need to contact Surgipaths tech services. At 12:53 PM 8/18/2004, you wrote: >We recently switched to Surgipath's "Blue Ribbon" paraffin and the techs >loved it! Within the past two weeks, they are noticing that the ribbon is >more compressed and harder to stretch out on the waterbath. Any ideas as to >why this would be happening when the product was used for about two months >with no problem whatsoever?? We are using it in both the processor and the >embedder and it is a multfunction paraffin. I like the "blueness" of the >paraffin, as all of the small biopsies stand out in the paraffin when >embedded for easier sectioning visualness! Thanks ahead of time..LOVE the >histonet!! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From PWebster <@t> hei.org Wed Aug 18 16:40:08 2004 From: PWebster <@t> hei.org (Webster, Paul) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] fibrin for cell blocks Message-ID: <4E8C1F1E4E8FA748B5487C50C91695F0862FC5@heimail.hei.org> This article describs the fibrin embedding method in detail. Raska I, Pliss A, Mandys V, Risueno MC, Lojda Z. 1998. Processing of free cells for electron microscopy using a fibrin clot. Acta Histochem. 1998 Jul;100(3):309-13. Regards, Paul Webster. Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 phone (213) 273 8026 fax (213) 413 6739 email: pwebster@hei.org > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of LINDA MARGRAF > Sent: Wednesday, August 18, 2004 1:28 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] fibrin for cell blocks > > Dear Histonetters: > Does anyone have experience using fibrin or another substance as a > matrix to add a cell suspension to when making cell blocks. My > colleague wants to make cell blocks to section and stain for a research > study but there will only be a small amount of cellular material. She > remembers seeing this done in the past but needs to know where to obtain > the reagnets and what procedures to follow. Thanks so much > Linda M > Histonet administrator > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From rkrug <@t> sial.com Wed Aug 18 16:35:58 2004 From: rkrug <@t> sial.com (Robert Krug) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] fibrin for cell blocks Message-ID: Linda: See Histonet archive http://histosearch.com/histonet/Jan03A/RE.clotpreparationsofperi.html. If you include your cell suspension into the mix, the same process should work. You should be able to get the reagents from any company selling coagulation reagents. Best Regards, Bob Krug St Louis, Missouri "LINDA MARGRAF" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/18/2004 03:28 PM To: cc: Subject: [Histonet] fibrin for cell blocks Dear Histonetters: Does anyone have experience using fibrin or another substance as a matrix to add a cell suspension to when making cell blocks. My colleague wants to make cell blocks to section and stain for a research study but there will only be a small amount of cellular material. She remembers seeing this done in the past but needs to know where to obtain the reagnets and what procedures to follow. Thanks so much Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From immunogal3 <@t> yahoo.com Wed Aug 18 16:16:47 2004 From: immunogal3 <@t> yahoo.com (MICHELE M) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] PAS with diastase Message-ID: <20040818211647.75597.qmail@web51807.mail.yahoo.com> Hi All, We're looking for some help with our PAS with diastase stain. For some reason, we can not get it to work. The PAS works fine but the diastase does not seem to be working. We are using diastase of malt from Fisher and a 0.1M phosphate buffer at pH 6.0 from Polyscientific. Our tech also tried using distilled water with the diastase instead of the buffer. The diastase solution is heated in a waterbath at 37 degrees for one hour. We have used liver, colon and esophagus tissue controls. If anyone has any ideas what might be the problem, please let us know. Thanks, Michele __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From lizchlipala <@t> premierhistology.com Wed Aug 18 16:43:45 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] fibrin for cell blocks In-Reply-To: Message-ID: <000001c4856c$73caa8c0$74d48a80@LIZ> Linda I use a 2% agarose solution. Basically, you spin the cells down, remove the supernatant and then re-suspend in the agarose and spin again. You can remove the agarose pellet, process and embed in paraffin. I'll send you the pdf of the reference I have used. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LINDA MARGRAF Sent: Wednesday, August 18, 2004 1:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fibrin for cell blocks Dear Histonetters: Does anyone have experience using fibrin or another substance as a matrix to add a cell suspension to when making cell blocks. My colleague wants to make cell blocks to section and stain for a research study but there will only be a small amount of cellular material. She remembers seeing this done in the past but needs to know where to obtain the reagnets and what procedures to follow. Thanks so much Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dmccaig <@t> ckha.on.ca Wed Aug 18 16:56:11 2004 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] special stains on slides that have been to water overnight Message-ID: <3E5A3F039F0BD8118B4700C00D00202404325A@CKHA9> I was hoping I could get some advice regarding an issue. Would a special stain or immuno stain on a section that has been brought to water and left to sit overnight in water yield reliable results. It gets cut, dried and taken to water the previous day and the staining procedure is not carried out until the next day. Your opinions would be appreciated. Diana McCaig, From bryand <@t> netbistro.com Wed Aug 18 17:19:06 2004 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] PAS with diastase In-Reply-To: <20040818211647.75597.qmail@web51807.mail.yahoo.com> References: <20040818211647.75597.qmail@web51807.mail.yahoo.com> Message-ID: <4123D5DA.3030708@netbistro.com> I had the same problem some years ago. I traced it to the malt diastase. I switched to hog pancreatic alpha amylase from Sigma (product # A3176)and the problem immediately disappeared. I used enough powder to cover a 1 cm circle about 1 mm deep dissolved in 15 mL (a test tube full) of distilled water and applied at room temperature for 30 minutes. That's not histochemically precise, but plenty good enough to remove all glycogen so the underlying PAS positive material can be demonstrated. Buffers and increased temperature should increase the efficiency. Bryan Llewellyn MICHELE M wrote: > Hi All, > > We're looking for some help with our PAS with diastase stain. For some reason, we can not get it to work. The PAS works fine but the diastase does not seem to be working. We are using diastase of malt from Fisher and a 0.1M phosphate buffer at pH 6.0 from Polyscientific. Our tech also tried using distilled water with the diastase instead of the buffer. The diastase solution is heated in a waterbath at 37 degrees for one hour. We have used liver, colon and esophagus tissue controls. If anyone has any ideas what might be the problem, please let us know. > > Thanks, > > Michele > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From scoop <@t> mail.nih.gov Wed Aug 18 17:42:00 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] paraffin block and slide storage Message-ID: Dear Histonetters, My lab does IHC on FFPE mouse brain and other tissues. We would like to store our tissue blocks and unstained slides so we can use them in the future. Is there any limit on the length of time that tissue blocks and unstained slides can be stored and is it ok to just store them in air at room temperature or is there some other way we should store them? Thanks in advance for any advice. Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From AnthonyH <@t> chw.edu.au Wed Aug 18 18:14:25 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] PAS with diastase Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E08F@simba.kids> Try the method described in V-M. Mangan, V. Farago, M. Kelly, and A. F. Henwood (2002) " An Amylase Reagent with a Long Shelf Life for the Removal of Glycogen from Tissue Sections" J Histotechnol. 25(3): 153-154. We have used it for over 5 years and have had no problems. Method as follows: Document Procedure: Diastase Removal of Glycogen Pathnet Code: S DPAS Principle: The enzyme solution is applied to one of two sections of the tissue (preferably consecutive sections) and then both are stained by the PAS method. The presence and relative amount of glycogen in the sections can be determined by examining the extent of loss of staining in the enzyme treated section as compared with the untreated section. This amylase reagent has a long shelf life and uses an incubation time of 10 minutes at room temperature. It is suitable for formalin and Brazil's fixed paraffin sections as well as air-dried and ethanol fixed frozen sections. Controls: Liver containing glycogen Reagents: 1. Amylase Reagent Warning: Harmful, contains azide - see MSDS Alpha Amylase from Bacillus Subtilis (Fluka Cat No 10070,) 1g Oxoid PBS Tablets (Cat No BR14a) 1 tablet Distilled water 100ml Sodium Azide 0.1g This solution, once prepared is stored at 4oC when not in use. A recycled antibody dropper bottle (often used in commercial immunoperoxidase kits) is useful for storage and application. 2. PAS Reagents (see PAS Stain) Warning: Suspected Carcinogen - see MSDS Procedure: 1. Dewax and hydrate paraffin sections, hydrate frozen sections. 2. For amylase digestion, place slides on a rack, cover sections with amylase solution and allow to incubate for 10 minutes at room temperature. 3. Wash slides well in water. 4. Place slides in 1% periodic acid 10 minutes. 5. Wash slides well in water. 6. Rinse slides in distilled water. 7. Place in Schiff's reagent 10 minutes. 8. Rinse slides in distilled water and then wash slides in tap water 3 minutes. 9. Counterstain slides with haematoxylin, differentiate and blue. 10. Dehydrate, clear and mount. Results: * Glycogen is extracted and so loss of PAS positive staining will occur in the enzyme treated section. * Mucopolysaccharides are not extracted and so staining will be the same in both sections. Reference: V-M. Mangan, V. Farago, M. Kelly, and A. F. Henwood (2002) " An Amylase Reagent with a Long Shelf Life for the Removal of Glycogen from Tissue Sections" J Histotechnol. 25(3): 153-4. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: MICHELE M [mailto:immunogal3@yahoo.com] Sent: Thursday, 19 August 2004 7:17 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PAS with diastase Hi All, We're looking for some help with our PAS with diastase stain. For some reason, we can not get it to work. The PAS works fine but the diastase does not seem to be working. We are using diastase of malt from Fisher and a 0.1M phosphate buffer at pH 6.0 from Polyscientific. Our tech also tried using distilled water with the diastase instead of the buffer. The diastase solution is heated in a waterbath at 37 degrees for one hour. We have used liver, colon and esophagus tissue controls. If anyone has any ideas what might be the problem, please let us know. Thanks, Michele __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From ynwang <@t> u.washington.edu Wed Aug 18 18:49:14 2004 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] fibrin for cell blocks In-Reply-To: <000001c4856c$73caa8c0$74d48a80@LIZ> References: <000001c4856c$73caa8c0$74d48a80@LIZ> Message-ID: Linda, We have also used a 2% agarose solution and have been sucessful embedding the agarose-cell mixture in OCT for frozen sections. Yak-Nam Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 On Wed, 18 Aug 2004, Elizabeth Chlipala wrote: > Linda > > I use a 2% agarose solution. Basically, you spin the cells down, remove > the supernatant and then re-suspend in the agarose and spin again. You > can remove the agarose pellet, process and embed in paraffin. I'll send > you the pdf of the reference I have used. > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > Premier Histology Laboratory, LLC > P.O. Box 18592 > Boulder, Colorado 80308 > Office: (303) 735-5001 > Fax: (303) 735-3540 > lizchlipala@premierhistology.com > www.premierhistology.com > > Ship to Address: > Premier Histology Laboratory > University of Colorado > MCBD, Room A3B40 > Boulder, Colorado 80309 > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of > its attachments, please be advised that you have received this email in > error and that any use, dissemination, distribution, forwarding, > printing, or copying of this email or any attached files is strictly > prohibited. If you have received this email in error, please immediately > purge it and all attachments and notify the sender by reply email or > contact the sender at the number listed above if one is provided. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LINDA > MARGRAF > Sent: Wednesday, August 18, 2004 1:29 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] fibrin for cell blocks > > Dear Histonetters: > Does anyone have experience using fibrin or another substance as a > matrix to add a cell suspension to when making cell blocks. My > colleague wants to make cell blocks to section and stain for a research > study but there will only be a small amount of cellular material. She > remembers seeing this done in the past but needs to know where to obtain > the reagnets and what procedures to follow. Thanks so much > Linda M > Histonet administrator > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jstaruk <@t> masshistology.com Wed Aug 18 19:05:42 2004 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] special stains on slides that have been to waterovernight In-Reply-To: <3E5A3F039F0BD8118B4700C00D00202404325A@CKHA9> Message-ID: <000301c48580$456d36f0$6401a8c0@yourw04gtxld67> I tried that once and the immuno was a total failure the next day. I never investigated it as to why, assuming everything washed out overnight. Jim ________________________ James E. Staruk, HT(ASCP) www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: Wednesday, August 18, 2004 4:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] special stains on slides that have been to waterovernight I was hoping I could get some advice regarding an issue. Would a special stain or immuno stain on a section that has been brought to water and left to sit overnight in water yield reliable results. It gets cut, dried and taken to water the previous day and the staining procedure is not carried out until the next day. Your opinions would be appreciated. Diana McCaig, _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From franzef <@t> gmx.de Wed Aug 18 19:42:53 2004 From: franzef <@t> gmx.de (=?ISO-8859-1?Q?Franz-Josef_M=FCller?=) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] FluorojadeB combined with Immunhistochemistry Message-ID: Dear Histonetters, does anybody have experience with combining FluorojadeB with Immunhistochemistry? We tried it according to the methodpaper but saw so'n'so-results at best. Would antigenretrival be possible? Any suggestions? Thanks a lot in advance! Cheers Franzef Mueller From bhewlett <@t> cogeco.ca Wed Aug 18 19:46:54 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] special stains on slides that have been to waterovernight References: <3E5A3F039F0BD8118B4700C00D00202404325A@CKHA9> Message-ID: <005c01c48586$06c12690$6400a8c0@mainbox> Hi Diana, Provided that the tissues were NBF fixed for a minimum of 24 hours, you should have NO problem with the common special stains and with the immuno stains. In fact, back at the beginning of IHC techniques this was a fairly standard procedure with us and provided a measure of antigen retrieval for many difficult antigens( basically it's AR at RT!). Been there, done that! Regards, Bryan ----- Original Message ----- From: "Diana McCaig" To: Sent: Wednesday, August 18, 2004 5:56 PM Subject: [Histonet] special stains on slides that have been to waterovernight > I was hoping I could get some advice regarding an issue. Would a special > stain or immuno stain on a section that has been brought to water and left > to sit overnight in water yield reliable results. It gets cut, dried and > taken to water the previous day and the staining procedure is not carried > out until the next day. Your opinions would be appreciated. > Diana McCaig, > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From barbara.bublava <@t> meduniwien.ac.at Thu Aug 19 06:58:12 2004 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] urgent problem - cryoprotection Message-ID: <009401c485e3$cdbde0a0$1201a8c0@GERICHTS9XOZZ8> Dear Histonetters I have formalin fixed specimens which are in 20% succrose since tuesday. I wanted to section them today but my cryostat stopped working. Now I do not know how long the specimens can be stored in the succrose or if it would be better to put them back to formalin and cryoprotect again when the cryostat is repaired? Or do you have other deas how to handle this situation??? Thanks in advance for any advice Barbara Bublava, Vienna From JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk Thu Aug 19 08:33:19 2004 From: JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk (JOHN PHILLIPS) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Spirit fixation of tissue Message-ID: <166A1E642B5B644DA694C08FD29D0ADC3E1247@ztroy.new-tr.wales.nhs.uk> Hello Histonet users, I would really appreciate knowing where to find an article, author unknown, published, probably somewhere in the time period 1972 to 1986, about the use of spirit for the fixation of skin biopsies and the like. I think it may have been published in the Institute of Medical Laboratory Technology journal (UK) or may be the Journal of Clinical Pathology (UK). The article investigated the effects of rum, whisky, gin and vodka etc. on tissue. I think rum came out on top. I am presenting a talk to GP's and would like to mention this as part of my presentation. I shall of course advise them not to use a Scottish malt under any circumstances that would be sacrilage. American bourbon may be! Thanks, John Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r rheolwr systemau wybod drwy ddefnyddio'r manylion isod. Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain Cymru. Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau’r Ddeddf Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru ar www.newalesnhstrust.org.uk This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager using the details below. The contents of this email represent the views of the individual(s) named above and do not necessarily represent the views of the North East Wales NHS Trust. Please be aware that, under the terms of the Freedom of Information Act 2000, the North East Wales NHS Trust may be required to make public the content of any emails or correspondence received. For futher information on Freedom of Information, please refer to the North East Wales NHS Trust website at www.newalesnhstrust.org.uk For further assistance, please contact system.administrator@new-tr.wales.nhs.uk. From ryaskovich <@t> dir.nidcr.nih.gov Thu Aug 19 08:41:34 2004 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] special stains on slides that have been to waterov ernight Message-ID: <8F3AB322628548428A992EFB0E80F5D3A2F0FB@nihexchange8.nih.gov> Jim, That slide will work fine for special stains. Ruth Yaskovich National Institutes of Health National Institute of Dental and Crainiofacial Research Neuronal Gene Expression Section > ---------- > From: Jim Staruk > Sent: Wednesday, August 18, 2004 7:05 PM > To: 'Diana McCaig'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] special stains on slides that have been to > waterovernight > > I tried that once and the immuno was a total failure the next day. I > never investigated it as to why, assuming everything washed out > overnight. > > Jim > > ________________________ > James E. Staruk, HT(ASCP) > www.masshistology.com > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana > McCaig > Sent: Wednesday, August 18, 2004 4:56 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] special stains on slides that have been to > waterovernight > > I was hoping I could get some advice regarding an issue. Would a > special > stain or immuno stain on a section that has been brought to water and > left > to sit overnight in water yield reliable results. It gets cut, dried > and > taken to water the previous day and the staining procedure is not > carried > out until the next day. Your opinions would be appreciated. > Diana McCaig, > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From mcauliff <@t> umdnj.edu Thu Aug 19 12:06:51 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Spirit fixation of tissue In-Reply-To: <166A1E642B5B644DA694C08FD29D0ADC3E1247@ztroy.new-tr.wales.nhs.uk> References: <166A1E642B5B644DA694C08FD29D0ADC3E1247@ztroy.new-tr.wales.nhs.uk> Message-ID: <4124DE2B.7010103@umdnj.edu> I think it is high time that study was repeated and the results verified. Contact all of the distillers and have them donate a case of their best spirits for the investigation. I for one, would be very interested in the effects of the various single malts (highlands, lowlands, Speyside, etc) on tissues. With a little luck the cheap, rot-gut would give the best results, leaving the good stuff for other investigations. Geoff JOHN PHILLIPS wrote: >Hello Histonet users, > >I would really appreciate knowing where to find an article, author unknown, published, probably somewhere in the time period 1972 to 1986, about the use of spirit for the fixation of skin biopsies and the like. I think it may have been published in the Institute of Medical Laboratory Technology journal (UK) or may be the Journal of Clinical Pathology (UK). The article investigated the effects of rum, whisky, gin and vodka etc. on tissue. I think rum came out on top. > >I am presenting a talk to GP's and would like to mention this as part of my presentation. I shall of course advise them not to use a Scottish malt under any circumstances that would be sacrilage. American bourbon may be! > >Thanks, > >John > >Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn >gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu >atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r >rheolwr systemau wybod drwy ddefnyddio'r manylion isod. > >Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, >felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain >Cymru. > >Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i >Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys >unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau???r Ddeddf >Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, >cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru >ar www.newalesnhstrust.org.uk > > >This email and any files transmitted with it are confidential and >intended solely for the use of the individual or entity to whom they >are addressed. If you have received this email in error please notify >the system manager using the details below. > >The contents of this email represent the views of the individual(s) >named above and do not necessarily represent the views of the >North East Wales NHS Trust. > >Please be aware that, under the terms of the Freedom of Information Act >2000, the North East Wales NHS Trust may be required to make public the >content of any emails or correspondence received. For futher >information on Freedom of Information, please refer to the North East >Wales NHS Trust website at www.newalesnhstrust.org.uk > >For further assistance, please contact >system.administrator@new-tr.wales.nhs.uk. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From galinadeyneko <@t> yahoo.com Thu Aug 19 09:52:09 2004 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] (no subject) Message-ID: <20040819145209.54931.qmail@web14526.mail.yahoo.com> Dear colleagues, Could you explain why you prefer to use not commercial 1% alcoholic eosin. Please , send me recipe of 5% eosin solution. What do you use as a solvent : D water or 70 %ethanol.Now I set up histology lab in cardiovascular department at Novartis, my previous experience is Laser Microdissection using Arcturus system (first model) and PALM from Zeiss. I will appreciate any advises from experienced in cardiovascular research people. Galina Deyneko Histology Core Novartis,Cambridge phone:617-871-7613 --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - Send 10MB messages! From galinadeyneko <@t> yahoo.com Thu Aug 19 09:52:54 2004 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] (no subject) Message-ID: <20040819145254.55084.qmail@web14526.mail.yahoo.com> Dear colleagues, Could you explain why you prefer to use not commercial 1% alcoholic eosin. Please , send me recipe of 5% eosin solution. What do you use as a solvent : D water or 70 %ethanol.Now I set up histology lab in cardiovascular department at Novartis, my previous experience is Laser Microdissection using Arcturus system (first model) and PALM from Zeiss. I will appreciate any advises from experienced in cardiovascular research people. Galina Deyneko Histology Core Novartis,Cambridge phone:617-871-7613 --------------------------------- Do you Yahoo!? Yahoo! Mail is new and improved - Check it out! From Jackie.O'Connor <@t> abbott.com Thu Aug 19 10:13:12 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Caspase 3 on mouse spleen Message-ID: Good Morning Histonetters - Does anyone out there have experience with caspase-3 on mouse spleen? I routinely perform Caspase-3 staining on xenografts, some gut, and a liver now and then - but just started looking at some spleens. They're a mess. I see what appears to be a lot of nonspecific staining of histiocytes in the red pulp. I'm staining immunocompetent mice, and repeated the stain with a naive SCID spleen - same staining pattern. Can anyone give me any insight? I'm using Pharmingen's 55965, Rabbit anti active Caspase-3. I've tried DAKO Envision, as well as an SABC method and get exactly the same staining. I'm a bit baffled. Any tidbits? Thanks! Jackie O' Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics 847.938.4919 Fax 847.938.3266 From lizchlipala <@t> premierhistology.com Thu Aug 19 11:05:33 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Caspase 3 on mouse spleen In-Reply-To: Message-ID: <000e01c48606$5ebb5e80$74d48a80@LIZ> Jackie I have done quite a bit of work with cleaved caspase 3 on rat, mouse and human tissue. The antibody that I use is from cell signaling technologies. I have not performed the stain on mouse spleen, but I have stained thymus, and mesenteric lymph node and I never have had a problem with background staining. In my hands this particular antibody requires high pH retrieval and I do use dako's envision in my protocol. I can provide you with a more detailed protocol and some images of thymus and lymph node if you like. But this is the basic protocol minus rinses. High pH retrieval H2O2 Block CC3 antibody 1:200 30 minutes RT Envision + Rabbit 30 minutes RT DAB + Counterstain with Hematoxylin. Hope this helps Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie.O'Connor@abbott.com Sent: Thursday, August 19, 2004 8:13 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Caspase 3 on mouse spleen Good Morning Histonetters - Does anyone out there have experience with caspase-3 on mouse spleen? I routinely perform Caspase-3 staining on xenografts, some gut, and a liver now and then - but just started looking at some spleens. They're a mess. I see what appears to be a lot of nonspecific staining of histiocytes in the red pulp. I'm staining immunocompetent mice, and repeated the stain with a naive SCID spleen - same staining pattern. Can anyone give me any insight? I'm using Pharmingen's 55965, Rabbit anti active Caspase-3. I've tried DAKO Envision, as well as an SABC method and get exactly the same staining. I'm a bit baffled. Any tidbits? Thanks! Jackie O' Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics 847.938.4919 Fax 847.938.3266 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JEllin <@t> yumaregional.org Thu Aug 19 11:20:59 2004 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] C-kit Message-ID: Hello everyone I thought that I would reach out to the world and ask a question. Currently we are trying to get CD-117\C-kit on line with our immunostainer, and are having some difficulties. We are trying to run this on our Ventanna stainer and are coming up with little to no results. Wondering if anyone can help. Jesus Ellin From Barbara_Lentz <@t> dahlchase.com Thu Aug 19 11:58:46 2004 From: Barbara_Lentz <@t> dahlchase.com (Barbara Lentz) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] C-kit Message-ID: Which Ventana stainer are you using? We are currently using the Benchmark and the XT, and until about a year ago we were using the ES and Nexes. So we have protocols for any of them. Barb From sa.drew <@t> hosp.wisc.edu Thu Aug 19 12:15:07 2004 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] C-kit Message-ID: We have been successfully using Dako's polyclonal CD117 on the Ventana instruments with no pretreatment and a 1:100 dilution, 32" titration. We have also sampled Ventana's monoclonal CD117 and found it worked well after a heat treatment with an EDTA buffer. We do keep our GI stromal tumor control slides in the freezer... -----Original Message----- From: Jesus Ellin [mailto:JEllin@yumaregional.org] Sent: Thursday, August 19, 2004 11:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] C-kit Hello everyone I thought that I would reach out to the world and ask a question. Currently we are trying to get CD-117\C-kit on line with our immunostainer, and are having some difficulties. We are trying to run this on our Ventanna stainer and are coming up with little to no results. Wondering if anyone can help. Jesus Ellin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Thu Aug 19 12:04:44 2004 From: mprice26 <@t> juno.com (Marsha R Price) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Re: Blade Holder Message-ID: <20040819.120444.3168.0.mprice26@juno.com> Histonetters, Does anyone have a used blade holder that will hold high profile disposable blades, that will fit in a Leitz 1512. One that they could sell. Thank you. Marsha Price On Thu, 19 Aug 2004 07:52:54 -0700 (PDT) Galina Deyneko writes: > Dear colleagues, > Could you explain why you prefer to use not commercial 1% alcoholic > eosin. Please , send me recipe of 5% eosin solution. What do you > use as a solvent : D water or 70 %ethanol.Now I set up histology lab > in cardiovascular department at Novartis, my previous experience is > Laser Microdissection using Arcturus system (first model) and PALM > from Zeiss. I will appreciate any advises from experienced in > cardiovascular research people. > > Galina Deyneko > Histology Core > Novartis,Cambridge > phone:617-871-7613 > > > > --------------------------------- > Do you Yahoo!? > Yahoo! Mail is new and improved - Check it out! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From dardofer <@t> hotmail.com Thu Aug 19 12:39:18 2004 From: dardofer <@t> hotmail.com (Dardo Ferrara) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] MOM Message-ID: Hi histoneters, I am doing immunohisto with mouse tissue and using mouse antihuman as a first AB. My secondary is a goat anti mouse fab fragment. With this combination, do I have to use a MOM blocking kit to prevent nonspecific binding??? Thanks Dardo Ferrara Cardiology Department Emory University. Atlanta _________________________________________________________________ De todo para la Mujer Latina http://latino.msn.com/mujer/ From Jackie.O'Connor <@t> abbott.com Thu Aug 19 12:55:58 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] MOM Message-ID: If your mouse tissue is from a SCID, no - (you can get some rare leaking, tho) If your mouse tissue is from a nude, or immunocompetent - yes. Does your anti-human antibody cross react with mouse? Jackie O' "Dardo Ferrara" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/19/2004 12:39 PM To: Histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] MOM Hi histoneters, I am doing immunohisto with mouse tissue and using mouse antihuman as a first AB. My secondary is a goat anti mouse fab fragment. With this combination, do I have to use a MOM blocking kit to prevent nonspecific binding??? Thanks Dardo Ferrara Cardiology Department Emory University. Atlanta _________________________________________________________________ De todo para la Mujer Latina http://latino.msn.com/mujer/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Pathologyarts <@t> aol.com Thu Aug 19 12:59:05 2004 From: Pathologyarts <@t> aol.com (Pathologyarts@aol.com) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] doing immunos by hand Message-ID: <145.315e1b8f.2e564469@aol.com> does anyone still do immunos by hand or are they all done with these $40 billion automated machines? i would like to see what the options are as far as doing these things manually. primarily for derm specimens, HMB 45, S 100, KI 67. any help is appreciated, curt From ynwang <@t> u.washington.edu Thu Aug 19 13:24:52 2004 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] fibrin for cell blocks (fwd) Message-ID: Dear Stan, Here is our protocol for the cell-agarose gel: 1. Make 6% agarose gel (we use type VII low gelling temperature agarose) and maintain at 37% until use 2. Spin down cells from culture and remove supernatant 3. resuspend pelet in media or PBS (you can do a cell count at this point to see how many cells you have) 4. carefully (make sure you do not introduce air bubbles) mix equal amounts of the cell suspension and the aragose (end up with an agarose concentraiton of 3%). 5. put cell-agarose suspension in small mould and place at 4 deg C for 1/2 hour to let gel. 6. remove gel and snap freeze in OCT. We have also use 2% agarose and gelatin for this. For smaller cell numbers we have also resuspended in small amounts of OCT and snap frozen in very small moulds and then embedded the small cell-OCT block in OCT. They all worked pretty well for what we wanted. I hope this is of help Best regards Yak-Nam > Dear Yak-Nam > I have just seen your post stating you have used 2% agarose cell blocks in OCT for frozen sections. > Do you have a specific protocol for this ? > Regards > Stan > > Stan Stylli > Department of Surgery / Neurosurgery > Royal Melbourne Hospital > Parkville Australia 3052 > Tel : 61-3-93427616 > Fax : 61-3-93477695 > > > > -----Original Message----- > From: Y. Wang [mailto:ynwang@u.washington.edu] > Sent: Thursday, 19 August 2004 9:49 AM > To: Elizabeth Chlipala > Cc: histonet@lists.utsouthwestern.edu; 'LINDA MARGRAF' > Subject: RE: [Histonet] fibrin for cell blocks > > > Linda, > > We have also used a 2% agarose solution and have been sucessful embedding > the agarose-cell mixture in OCT for frozen sections. > > Yak-Nam > > Senior Fellow > Department of Bioengineering > University of Washington > Box 357962 > Seattle, WA 98195 > > Tel.: (206)-221-5873 > Fax.: (206)-221-5874 > > On Wed, 18 Aug 2004, Elizabeth Chlipala wrote: > > > Linda > > > > I use a 2% agarose solution. Basically, you spin the cells down, remove > > the supernatant and then re-suspend in the agarose and spin again. You > > can remove the agarose pellet, process and embed in paraffin. I'll send > > you the pdf of the reference I have used. > > > > Liz > > > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > > Premier Histology Laboratory, LLC > > P.O. Box 18592 > > Boulder, Colorado 80308 > > Office: (303) 735-5001 > > Fax: (303) 735-3540 > > lizchlipala@premierhistology.com > > www.premierhistology.com > > > > Ship to Address: > > Premier Histology Laboratory > > University of Colorado > > MCBD, Room A3B40 > > Boulder, Colorado 80309 > > > > _________________________________ > > This email and any files transmitted with it may contain PRIVILEGED or > > CONFIDENTIAL information and may be read or used only by the intended > > recipient. If you are not the intended recipient of the email or any of > > its attachments, please be advised that you have received this email in > > error and that any use, dissemination, distribution, forwarding, > > printing, or copying of this email or any attached files is strictly > > prohibited. If you have received this email in error, please immediately > > purge it and all attachments and notify the sender by reply email or > > contact the sender at the number listed above if one is provided. > > > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LINDA > > MARGRAF > > Sent: Wednesday, August 18, 2004 1:29 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] fibrin for cell blocks > > > > Dear Histonetters: > > Does anyone have experience using fibrin or another substance as a > > matrix to add a cell suspension to when making cell blocks. My > > colleague wants to make cell blocks to section and stain for a research > > study but there will only be a small amount of cellular material. She > > remembers seeing this done in the past but needs to know where to obtain > > the reagnets and what procedures to follow. Thanks so much > > Linda M > > Histonet administrator > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From STEGTM <@t> samcstl.org Thu Aug 19 13:34:06 2004 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] You may want to hit the delete button with this,then again, maybe not Message-ID: I agree with the main banter re: the freedom of speech as it relates to experience, particularly with products, services, et al......after all, aren't we using this media to try to communicate/educate with our fellows? I also know one good lawyer, but I know one great lawyer joke; seems the number of them will soon have them replacing lab rats, you don't get so attached to lawyers as cute little rats, and, there's some things a rat won't do. Peace, Terre >>> "GUTIERREZ, JUAN" 8/13/2004 3:12:37 PM >>> Hey at least they tried to talk to you(a year later)but they tried YOU! Like I told you on my last e-mail my behind is about two pounds lighter after my boss bit some of it off because of my comments on the histonet about a certain German company who should remain nameless. Apparently my speaking the truth, the whole truth, and nothing but the truth on this forum made them a little mad. If they made good products to begin with, we wouldn't be having this conversation. Have anybody noticed how many pages of the phonebook are dedicated to lawyers? There is about twice the number of lawyers compared to doctors. Aint that scary? I wonder when lawyer season is going to open, heck there's more of them than there is deer. Just kidding. I know one good lawyer, one. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My views are my own and do not reflect those of my employer. Long live free speech! "We have nothing to fear but fear itself". -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Thursday, August 12, 2004 3:35 PM To: Histonet Subject: [Histonet] You may want to hit the delete button with this,then again, maybe not Afternoon Netter's, Remember about a week or two the topic was getting "flamed"? Well, here's a new one. Sometime last year, I made several postings about a certain company and their product that I used and it did not work for me. I wrote them a letter and returned all the merchandise back to them saying I was not happy with their product. I waited several months and never got a response, so I named the company on the Histonet. Since then, I have been throwing away their flyers, catalogs and any other correspondence I receive. Obviously, they are doing fine without my business and I am doing fine without their products. Today, I receive a letter from the company with a copy of my Histonet postings from over a year ago, telling me that this has been in customer service for about a year (hmmmmmmmm, I'm trying to find out where the customer service is in all of this. (One year, okay, I'll get back to that), that their customer service has tried several times to contact me but to no avail. (Hmmmmm, one way would have been to respond to my email address, but I'll come back to this also). So after all this time, they want to do business with me. (So close to the NSH meeting, hmmmmmmmmm, another point to ponder). My big beef is that these companies read this forum and try to use strong arm tactics on people who post negative comments about them or their products. I have many friends that are sales reps in this field (I'm talking like come over my house and eat type of friends) and I love these people. I must reiterate that this forum was set up to exchange ideas, theories and experiences, both positive and negative. If this was such a "hot item", point #1, why did it take a whole year to get a response when they had my company and address on file? Is this how customer service works? I don't think so!! Point #2 If you are a vendor and don't like a particular posting, why don't you reply back to that person via email. Point #3 don't wait until it's almost time for the NSH annual S/C to fix a problem, it's probably too late by then. Point #4 If you haven't had a purchase order or have not corresponded with a customer for over a year, chances are that customer does not want to do business with you. Let lying dogs lie and don't open old scars. All you did was get me going on a tangent. I know, there goes Nocito again, but I refuse to be treated like this from any one. Like they say in the movies, "Don't call me, I'll call you". Okay, I've said my piece now let the FLAMING BEGIN!!!!!!!!! As always, the opinions stated by this author in no way reflects the opinions of his employer, subordinates, friends or relatives. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hborgeri <@t> wfubmc.edu Thu Aug 19 13:42:10 2004 From: hborgeri <@t> wfubmc.edu (Hermina Borgerink) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] doing immunos by hand Message-ID: <9AEEF1FB6254224AA355ED285F8491650999C88D@EXCHVS2.medctr.ad.wfubmc.edu> Curt, I do all my immunos by hand (up to 80 slides at a time) using the MicroProbe Staining system from Fisher. The system operates by means of special slides that have a painted surface and a special slide holder. When paired together (facing each other) a capillary gap is created that allows fluids to rise between the two slides using capillary action. You don't need to purchase the complete set up either, just the special slide holders and isolons (rubber pads with wells to dispense reagents). I think each slide holder (to stain 20 slides at once) runs around $250.00. I'm not sure about the isolons, but they are not that expense and are re-usable. I routinely use a large panel of antibodies, including the ones you mentioned. Check the Fisher catalog for more details. Hermina Hermina M. Borgerink, BA, HTL(ASCP)QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax (336) 716-1515 e-mail hborgeri@wfubmc.edu "Treat a man as he appears to be, and you make him worse. But treat a man as if he already were what he potentially could be, and you make him what he should be." (Goethe) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pathologyarts@aol.com Sent: Thursday, August 19, 2004 1:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] doing immunos by hand does anyone still do immunos by hand or are they all done with these $40 billion automated machines? i would like to see what the options are as far as doing these things manually. primarily for derm specimens, HMB 45, S 100, KI 67. any help is appreciated, curt _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rpw4 <@t> psu.edu Thu Aug 19 14:15:42 2004 From: rpw4 <@t> psu.edu (Ronald P. Wilson) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Caspace-3 on rat tissue Message-ID: In response to a question about Caspace-3 for mouse spleen, Liz Chlipala mentions using this antibody on rat, mouse and human tissue. My question to Liz and others, anyone have experience with this antibody and rat skin. We tried it a few weeks ago using rat intestine and skin and pH 6.0 citrate buffer HIER. We did get positive staining on the intestine tissue but not on the skin. The skin is our tissue of interest. It was suggested to us by the manufacturer that perhaps Caspace-3 is not expressed or has very low expression in the skin. Anyone know the answer. I would love to hear if others have any experience with rat skin. Ronald P. Wilson, V.M.D., M.S. Associate Professor Department of Comparative Medicine, HO54 Penn State University College of Medicine M. S. Hershey Medical Center 500 University Drive Hershey, PA 17033 phone: 717-531-8460 fax: 717-531-5001 e-mail: rpw4@psu.edu From rpw4 <@t> psu.edu Thu Aug 19 14:15:43 2004 From: rpw4 <@t> psu.edu (Ronald P. Wilson) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] IHC polymer detection system for rat tissues Message-ID: Has anyone found a successful polymer detection system (i.e., Envision or others) directed against a mouse monoclonal on rat tissue. We tried one but got a lot of background staining, I suspect from cross reactivity with closely related antigens. Our standard IHC approach uses an anti-mouse, rat-absorbed secondary antibody, but I would like to get the added sensitivity of a polymer-based system. Ronald P. Wilson, V.M.D., M.S. Associate Professor Department of Comparative Medicine, HO54 Penn State University College of Medicine M. S. Hershey Medical Center 500 University Drive Hershey, PA 17033 phone: 717-531-8460 fax: 717-531-5001 e-mail: rpw4@psu.edu From lizchlipala <@t> premierhistology.com Thu Aug 19 14:51:27 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] IHC polymer detection system for rat tissues In-Reply-To: Message-ID: <000001c48625$eda91690$74d48a80@LIZ> Ronald We have been able to amplify rat anti mouse antibodies with a polymer based system such as Envision. In this case we use a rabbit anti-rat (mouse adsorbed) secondary and then we amplify by using the rabbit envision instead of strep ABC or ABC. I'm assuming that this can be done with mouse antibodies if there is a secondary antibody of rabbit anti-mouse that can be used to link the rabbit envision to your primary. I hope this helps Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ronald P. Wilson Sent: Thursday, August 19, 2004 12:16 PM To: Histonet Subject: [Histonet] IHC polymer detection system for rat tissues Has anyone found a successful polymer detection system (i.e., Envision or others) directed against a mouse monoclonal on rat tissue. We tried one but got a lot of background staining, I suspect from cross reactivity with closely related antigens. Our standard IHC approach uses an anti-mouse, rat-absorbed secondary antibody, but I would like to get the added sensitivity of a polymer-based system. Ronald P. Wilson, V.M.D., M.S. Associate Professor Department of Comparative Medicine, HO54 Penn State University College of Medicine M. S. Hershey Medical Center 500 University Drive Hershey, PA 17033 phone: 717-531-8460 fax: 717-531-5001 e-mail: rpw4@psu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Aug 19 14:51:28 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] MOM In-Reply-To: References: Message-ID: <6.0.0.22.1.20040819133522.01b1fa70@gemini.msu.montana.edu> What mouse on mouse kit are you using? Vector M.O.M. ? Normally you use the biotinylated antimouse antibody provided with these special kits or at least find out from kit manufacturers what their secondary antibody concentration is in terms of ug/ml. Once you know what they recommend in terms of Their tech services often are helpful with this. Then it may be possible to substitute your biotinylated antimouse secondary for theirs. Yes, you would still have to do a mouse on mouse block since you have a secondary that is anti-mouse? Is this a new thing? A new, separate mouse on mouse blocking kit? Or are the ms on ms blocking reagents part of a mouse on mouse staining kit? M.O.M. indicates the abbreviation for mouse on mouse kit from VECTOR. At 11:39 AM 8/19/2004, you wrote: >Hi histoneters, > >I am doing immunohisto with mouse tissue and using mouse antihuman as a >first AB. My secondary is a goat anti mouse fab fragment. With this >combination, do I have to use a MOM blocking kit to prevent nonspecific >binding??? > >Thanks > >Dardo Ferrara >Cardiology Department >Emory University. >Atlanta > >_________________________________________________________________ >De todo para la Mujer Latina http://latino.msn.com/mujer/ > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From ploykasek <@t> phenopath.com Thu Aug 19 16:12:30 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] IHC polymer detection system for rat tissues In-Reply-To: Message-ID: I had an email today from my Signet rep that they have a new mouse on mouse polymer kit. I have not tried it, but you may want to contact them. Patti Loykasek PhenoPath Laboratories Seattle, WA> Has anyone found a successful polymer detection system (i.e., Envision or > others) directed against a mouse monoclonal on rat tissue. We tried one but > got a lot of background staining, I suspect from cross reactivity with > closely related antigens. Our standard IHC approach uses an anti-mouse, > rat-absorbed secondary antibody, but I would like to get the added > sensitivity of a polymer-based system. > > Ronald P. Wilson, V.M.D., M.S. > Associate Professor > Department of Comparative Medicine, HO54 > Penn State University College of Medicine > M. S. Hershey Medical Center > 500 University Drive > Hershey, PA 17033 > phone: 717-531-8460 > fax: 717-531-5001 > e-mail: rpw4@psu.edu > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wickedmiss <@t> earthlink.net Thu Aug 19 17:36:37 2004 From: wickedmiss <@t> earthlink.net (Jacqueline Cruz) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] Required retention of frozen tissues Message-ID: <000a01c4863c$fed44360$2633e504@jac1> Can anyone tell me the length of time I am required to keep frozen tissues from IMF procedures? We currently keep them for 2 yrs. I contacted U of M and they keep theirs for 1 yr. Is there a CAP requirement? From Jacqueline.Miller <@t> UTSouthwestern.edu Thu Aug 19 18:34:15 2004 From: Jacqueline.Miller <@t> UTSouthwestern.edu (Jacqueline Miller) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] transferring paraffin ribbon Message-ID: Hi everyone, I just wanted to say thanks to everybody who gave me suggestions/advice for transferring paraffin ribbons to the water bath. Some hints that proved particularly helpful were 1) Keeping the blocks cold--virtually eliminates wrinkles 2) Simply making sure that the plate of the knife holder and the knife itself are very, very clean. This helps in getting smooth, unwrinkled sections off of the knife 3) Keeping the pair of forceps used to remove the sections ice cold and keeping them very, very clean. This helps keep the paraffin from sticking to the forceps, thus allowing that first section to be kept if necessary. My sectioning is going much, much better, so thank you all very much. Now, if someone could solve my chatter problem on the cryostat..... It has all the service techs, including a specialist who flew in from California to look at it today, baffled. They're saying it might be the bearings, but they just don't know. Basically, the chatter occurs when cutting at a fast speed, and hardly at all when cutting slowly. We've tightened just about every nut, bolt, and lever on the cryostat, tried different blades, adjusted the clearance angle, and we're running out of ideas. It's a Leica CM1850. Any ideas are greatly appreciated. Thanks, Jackie >>> "Sarah Jones" 08/01/04 2:11 PM >>> Hi Jackie, I use a wooden applicator stick that I angle cut, at the tip, with a single edge razor blade, making a fine point. I dip the tip of the stick into my waterbath before touching the ribbon and the water makes the ribbon adhere to the stick. I use them for both the beginning and the end of the ribbon. When doing serial sections, it's difficult to save every section if you need more than one ribbon. At the best of times, I'll usually lose one section at the beginning and one at the end. If I have trouble getting the ribbon started, I may lose even more than that. If researchers are doing 3D reconstruction, I just keep a record of where sections were lost and how many sections were lost. I'm doing that right now with some laser induced retinal lesions in the eye of the rat. If you need thin sections (3-5 microns), the blocks need to be cold. If the block isn't cold, the ribbon will be compressed. I always cut thin sections off of an ice tray. I use the dissecting pans (without wax) available from Fisher for my ice trays. If you are doing thicker sections, you can cut at room temperature. I once had a disposable knife holder where the sections would stick to the metal. I ended up putting a piece of wide, clear, plastic packaging tape over it and that kept the ribbon from sticking to it. You might try Paraguard too, a spray used for keeping paraffin from sticking to surfaces. It has a nasty smell that makes me nauseous so I don't use it myself. Also, make sure there isn't any water on the face of the block or on the knife before you start a ribbon. Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Jacqueline Miller" 7/30/2004 2:16:00 PM >>> Hi everyone, I would like to know about the techniques that histotechs use to cut paraffin sections--especially how the ribbon is transferred from microtome to water bath while preserving the first and last sections. I've been sectioning paraffin blocks for almost 4 years now. However, I've never needed to cut serial sections and the samples were always large. So, I never worried about losing the first section of each ribbon (and sometimes the last). I would like to know what techniques the histologists out there use to transfer the ribbon from the microtome to the water bath. I've used metal forceps and my fingers (gloved and not gloved) to grab the first section (but the section sticks to the forceps), and I'm using the wooden part of a cotton swab to lift off the last section, which is working fairly well. I'm also using a new microtome, Leica RM2235, and having some trouble with the first section coming off the knife just wrinkling up. And, the surface below the knife is such that even if I grab the edge of the section with a paintbrush, it won't budge. Is there anything I can do to correct these problems? Also, which is preferred, to cut the blocks chilled or to leave them RT. I used to cut blocks chilled all the time, but here it's preferred that I cut them RT unless I have a problem. Thanks, Jackie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Thu Aug 19 21:36:49 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:23:55 2005 Subject: [Histonet] doing immunos by hand References: <145.315e1b8f.2e564469@aol.com> Message-ID: <000f01c4865e$8b906380$5d6bce44@yourxhtr8hvc4p> Curt, Biocare Medical has some equipment that you might be able to use. They have these slide shakers (Kinetic Stainers) that can hold either 24 or 36 slides. They have heaters that heat up to 95 degrees C. I used these until my pathologists decided to go to the $40 billion machines. I still have them as a back up, just in case. They also have a wide variety of primary antibodies and detection kits. Their number is 1-800-799-9499. They also have great technical support, in my opinion, which in no way reflects the opinions of my employers. I know I'm gonna get "flamed" on this one, but I like excitement. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: To: Sent: Thursday, August 19, 2004 12:59 PM Subject: [Histonet] doing immunos by hand > does anyone still do immunos by hand or are they all done with these $40 > billion automated machines? > > i would like to see what the options are as far as doing these things > manually. > > primarily for derm specimens, HMB 45, S 100, KI 67. > > > any help is appreciated, > curt > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From chesarato <@t> hotmail.com Thu Aug 19 21:51:45 2004 From: chesarato <@t> hotmail.com (Cesar Francisco Romero) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] High pH AR Message-ID: Dear Histonetters I have had terrible results with Tris-Clorhidric. I prefer de following buffer: Tris/EDTA pH 9 (10/1 mM) Buffer may be produced as follows: 1,21 g Tris, Merck 1.08387 0,37 g EDTA, Fluka Chemika 03680 1000 ml destilled water Buffer pH is stable at 9.0 - 9.1, usually without adjustment. The buffer is stable for approximately 4 weeks at room temperature. I keep it in the icebox for several weeks. I add two drops per 1000 ml of tween 20. _________________________________________________________________ MSN Amor: busca tu ? naranja http://latam.msn.com/amor/ From rocan <@t> mac.com Thu Aug 19 23:24:48 2004 From: rocan <@t> mac.com (Rocan) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Cre recombinase Message-ID: Can anyone recommend a good anti-Cre recombinase antibody for IHC? Your protocol and antibody dilutions would also be appreciated. Thanks Dr.Rocio Sierra-Honigmann Engineered Wound Repair Laboratory Cedars Sinai Research Institute Honigmannr@cshs.org From John.Auld <@t> whnt.nhs.uk Fri Aug 20 02:51:03 2004 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Re: Spirit fixation Message-ID: John This is probably of no help what so ever, but your message rang a bell. The paper that I have vague recollections of I think was written by people from London Zoo and published in the early 90s. Around that time I was probably too interested in the fixative effect of commercially available alcohols on liver and brain tissue in situ, Ah fun times. Scottish malts have no fixative effect for histological use and it would not be worth using anymore in this type of experiment. It does however have beneficial effects on the human body and is particularly beneficial psycologically. Cheers ( sl?inte mhath ) John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral External Tel 0151 604 7025 Message: 1 Date: Thu, 19 Aug 2004 14:33:19 +0100 From: "JOHN PHILLIPS" Subject: [Histonet] Spirit fixation of tissue To: Message-ID: <166A1E642B5B644DA694C08FD29D0ADC3E1247@ztroy.new-tr.wales.nhs.uk> Content-Type: text/plain; charset="US-ASCII" Hello Histonet users, I would really appreciate knowing where to find an article, author unknown, published, probably somewhere in the time period 1972 to 1986, about the use of spirit for the fixation of skin biopsies and the like. I think it may have been published in the Institute of Medical Laboratory Technology journal (UK) or may be the Journal of Clinical Pathology (UK). The article investigated the effects of rum, whisky, gin and vodka etc. on tissue. I think rum came out on top. I am presenting a talk to GP's and would like to mention this as part of my presentation. I shall of course advise them not to use a Scottish malt under any circumstances that would be sacrilage. American bourbon may be! Thanks, John From juan.gutierrez <@t> christushealth.org Fri Aug 20 08:41:50 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] transferring paraffin ribbon Message-ID: Hi Jackie, I have the same problem periodically with my cryostat. The problem seems to be the little metal piece on the back of the blade holder plate. When you take the lever off and release the plate, turn it over and you will see a small, shiny metal plate bent at an angle. With time the angle gets too wide and the pressure decreases, so all you have to do is gently bend it back close to a 90 degree angle and re-install. Hope this works. Call me if you have any questions. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My views are my own and do not reflect those of my employer. Long live free speech! "We have nothing to fear but fear itself". -----Original Message----- From: Jacqueline Miller [mailto:Jacqueline.Miller@UTSouthwestern.edu] Sent: Thursday, August 19, 2004 6:34 PM To: SJones@cvm.tamu.edu; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] transferring paraffin ribbon Hi everyone, I just wanted to say thanks to everybody who gave me suggestions/advice for transferring paraffin ribbons to the water bath. Some hints that proved particularly helpful were 1) Keeping the blocks cold--virtually eliminates wrinkles 2) Simply making sure that the plate of the knife holder and the knife itself are very, very clean. This helps in getting smooth, unwrinkled sections off of the knife 3) Keeping the pair of forceps used to remove the sections ice cold and keeping them very, very clean. This helps keep the paraffin from sticking to the forceps, thus allowing that first section to be kept if necessary. My sectioning is going much, much better, so thank you all very much. Now, if someone could solve my chatter problem on the cryostat..... It has all the service techs, including a specialist who flew in from California to look at it today, baffled. They're saying it might be the bearings, but they just don't know. Basically, the chatter occurs when cutting at a fast speed, and hardly at all when cutting slowly. We've tightened just about every nut, bolt, and lever on the cryostat, tried different blades, adjusted the clearance angle, and we're running out of ideas. It's a Leica CM1850. Any ideas are greatly appreciated. Thanks, Jackie >>> "Sarah Jones" 08/01/04 2:11 PM >>> Hi Jackie, I use a wooden applicator stick that I angle cut, at the tip, with a single edge razor blade, making a fine point. I dip the tip of the stick into my waterbath before touching the ribbon and the water makes the ribbon adhere to the stick. I use them for both the beginning and the end of the ribbon. When doing serial sections, it's difficult to save every section if you need more than one ribbon. At the best of times, I'll usually lose one section at the beginning and one at the end. If I have trouble getting the ribbon started, I may lose even more than that. If researchers are doing 3D reconstruction, I just keep a record of where sections were lost and how many sections were lost. I'm doing that right now with some laser induced retinal lesions in the eye of the rat. If you need thin sections (3-5 microns), the blocks need to be cold. If the block isn't cold, the ribbon will be compressed. I always cut thin sections off of an ice tray. I use the dissecting pans (without wax) available from Fisher for my ice trays. If you are doing thicker sections, you can cut at room temperature. I once had a disposable knife holder where the sections would stick to the metal. I ended up putting a piece of wide, clear, plastic packaging tape over it and that kept the ribbon from sticking to it. You might try Paraguard too, a spray used for keeping paraffin from sticking to surfaces. It has a nasty smell that makes me nauseous so I don't use it myself. Also, make sure there isn't any water on the face of the block or on the knife before you start a ribbon. Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Jacqueline Miller" 7/30/2004 2:16:00 PM >>> Hi everyone, I would like to know about the techniques that histotechs use to cut paraffin sections--especially how the ribbon is transferred from microtome to water bath while preserving the first and last sections. I've been sectioning paraffin blocks for almost 4 years now. However, I've never needed to cut serial sections and the samples were always large. So, I never worried about losing the first section of each ribbon (and sometimes the last). I would like to know what techniques the histologists out there use to transfer the ribbon from the microtome to the water bath. I've used metal forceps and my fingers (gloved and not gloved) to grab the first section (but the section sticks to the forceps), and I'm using the wooden part of a cotton swab to lift off the last section, which is working fairly well. I'm also using a new microtome, Leica RM2235, and having some trouble with the first section coming off the knife just wrinkling up. And, the surface below the knife is such that even if I grab the edge of the section with a paintbrush, it won't budge. Is there anything I can do to correct these problems? Also, which is preferred, to cut the blocks chilled or to leave them RT. I used to cut blocks chilled all the time, but here it's preferred that I cut them RT unless I have a problem. Thanks, Jackie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jerry.lyons <@t> ucd.ie Fri Aug 20 09:28:41 2004 From: jerry.lyons <@t> ucd.ie (Jeremiah Lyons) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Endothelial cell staining Message-ID: <003001c486c1$fdae5f90$6e892b89@DFFRCL0J> I just wanted to thank all histonet users for replying to my e-mail on endothelial cell staining. To summarise my findings, von Willebrand Factor would work on Carnoy's fixed normal mouse tissue but I could not get it to work on my tumour sections. The tumour sections for some reason would not tolerate any antigen retrieval even proteinase K for only 30 seconds would destroy the tumour section. Maybe I didn't fix the tissue in the Carnoy's for long enough. I got no staining with no antigen retrieval. I also tried the CD31 rat anti-mouse which is available from PharMingen. I got this to work on frozen sections but with alot of background staining which proved difficult to eliminate. Maybe my protocol was wrong or there is alot of endogenous peroxidases in my tumours. I am especially grateful to John McGinley and Liz Chlipala who recommended the CD31 from Santa Cruz which worked exceptionally well first time of asking on formalin-fixed paraffin embedded tissue. Thanks, Jerry From jerry.lyons <@t> ucd.ie Fri Aug 20 09:31:19 2004 From: jerry.lyons <@t> ucd.ie (Jeremiah Lyons) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Flk-1 staining Message-ID: <003f01c486c2$5c22d2e0$6e892b89@DFFRCL0J> Is anyone familiar with staining for Flk-1 in mice? I am especially interested if there is an antibody that works in formalin-fixed paraffin embedded tissue. Thanks, Jerry From gcallis <@t> montana.edu Fri Aug 20 10:08:20 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Cryostat chatter problems (long!) In-Reply-To: References: Message-ID: <6.0.0.22.1.20040820082209.01aed4a8@gemini.msu.montana.edu> One additional area to clean on a knife holder (paraffin microtomy) is the BACK of the holder. If there is any paraffin buildup, you can have defective ribbons plus icky sticky problems. Protect your all edges on knife holder, and do not use solvents to clean off paraffin - this removes lubricants put there by manufacturer, use a rough towel, and correct cleaning materials to remove paraffin. Ask your manufacturer what they recommend. We have three 1850's, and dearly love them. Cutting fast is a huge No No in our lab - we teach a slow, steady motion as knife passes through tissue. If you hesitate going through a tissue, you get BIG chatter or uneven section thickness. Remember, the tissue is NOT embedded in hard paraffin, and has only itself as support with OCT, a softer embedding media than the paraffin ONLY surrounding the tissue. Cutting fast on the cryostat is a No No in our lab! How you hold the flywheel is important, if you grab it like driving a "Harley" down the road,( too tightly!) - you can impart chatter through flywheel mechanism, use a light hold to get rid of the VAROOM grip! Relax, and cut slowly. If the temperature setting is too cold or block is not firmly attached to metal block holder with a generous amount of OCT, you can get chatter. Bubbles in or underneath a frozen block is the enemy! -20C is a general cutting temperature, but it will depend on the tissue. For fatty tissue, colder, for liver, brain, and spleen we warm to -16C -to -17C. Very dense tissue (connective tissue) can cause chatter - if really cold, harder to cut. You must be willing to change temperatures for any given tissue. Thick prefixed sucrose cryoprotected brain (50um) is cut at -19C or so, can be warmer, slowly and with antiroll device - no problems. Metal chucks for 1850's are little mushrooms with shallow circle depressions. These are fine for smaller, less dense tissues. You can buy better designed metal chuck in several sizes from Thermo Electron aka Shandon, but you must cut the stem to have the same stem length as the original holders in order to fit in microtome block holder. Thermo chucks have deeper, waffle weave depressions, basically an old fashioned style used for paraffin blocks years ago. The waffle weave holds blocks far better and more firmly than the Leica holders. Achieving stability in block and knife is important for cryostat work. If you take the block directly from the Peltier or cooling bars in the 1850, the block will be 10 degrees colder than the desired cutting temperature at the block and knife. Mount the block, and set it NEXT to the cryostat (use a microcentrifuge rack, bright colors and plastic for many blocks) and allow the block to equilibrate to you chosen cutting temperature - it takes about 20 minutes. If you set the cryostat cutting temperature to -20C, it will be approx that at knife and block, but not on those bars. Know your temperature differentials inside the cryostat - test with thermometer to locate warm and cold areas, record the temperatures for any given readout setting, then use these areas to your advantage. How you orient block in holder can make a difference. If you orient a tissue so the broadest, widest part of sample is cut by the knife FIRST, you get a lot of resistance and the knife will "spring" no matter if everything else is tight. Create a path of least resistance. Example, if block is a perfect square, place narrowest part of tissue pointed towards blade edge so it is cut first. In other words, the face of the block looks like a diamond and we cut the narrowest point first to get that less resistant pathway. One can trim rectangular blocks so you achieve a similar look, and proper orientation of tissue in block at embedding helps too. Change the knife frequently and try OTHER brands of disposable blades and try high profile. Low profile are thinner and narrower, and not as sturdy overall as high profiles. We use high profile disposable AccuEdge blades (Sakura Finetek, VWR has them) and do not get chatter - when we change to low profile AccuEdge and cutting denser tissue, chatter tends to return - so we stay with high, since they are just as sharp but stable. We use low profile occasionally but only for small, less dense tissues. Make sure the levers one pulls out when cleaning any part of holders and even with paraffin microtomes are reoiled so a dry metal surface is not tightened down on another dry metal surface. My Leica rep indicated that this is a source of poor tightening, and lubricated surfaces allows for properly tight fits and not the "dry metal on dry metal" syndrome. Whew, what a lecture!! Good luck At 05:34 PM 8/19/2004, you wrote: >Hi everyone, > >I just wanted to say thanks to everybody who gave me suggestions/advice >for transferring paraffin ribbons to the water bath. Some hints that >proved particularly helpful were 1) Keeping the blocks cold--virtually >eliminates wrinkles 2) Simply making sure that the plate of the knife >holder and the knife itself are very, very clean. This helps in getting >smooth, unwrinkled sections off of the knife 3) Keeping the pair of >forceps used to remove the sections ice cold and keeping them very, very >clean. This helps keep the paraffin from sticking to the forceps, thus >allowing that first section to be kept if necessary. > >My sectioning is going much, much better, so thank you all very much. > >Now, if someone could solve my chatter problem on the cryostat..... >It has all the service techs, including a specialist who flew in from >California to look at it today, baffled. They're saying it might be the >bearings, but they just don't know. Basically, the chatter occurs when >cutting at a fast speed, and hardly at all when cutting slowly. We've >tightened just about every nut, bolt, and lever on the cryostat, tried >different blades, adjusted the clearance angle, and we're running out of >ideas. It's a Leica CM1850. Any ideas are greatly appreciated. > >Thanks, >Jackie > > >>> "Sarah Jones" 08/01/04 2:11 PM >>> >Hi Jackie, > I use a wooden applicator stick that I angle cut, at the tip, with a >single edge razor blade, making a fine point. I dip the tip of the >stick into my waterbath before touching the ribbon and the water makes >the ribbon adhere to the stick. I use them for both the beginning and >the end of the ribbon. When doing serial sections, it's difficult to >save every section if you need more than one ribbon. At the best of >times, I'll usually lose one section at the beginning and one at the >end. If I have trouble getting the ribbon started, I may lose even more >than that. If researchers are doing 3D reconstruction, I just keep a >record of where sections were lost and how many sections were lost. I'm >doing that right now with some laser induced retinal lesions in the eye >of the rat. > If you need thin sections (3-5 microns), the blocks need to be cold. >If the block isn't cold, the ribbon will be compressed. I always cut >thin sections off of an ice tray. I use the dissecting pans (without >wax) available from Fisher for my ice trays. If you are doing thicker >sections, you can cut at room temperature. > I once had a disposable knife holder where the sections would stick >to the metal. I ended up putting a piece of wide, clear, plastic >packaging tape over it and that kept the ribbon from sticking to it. >You might try Paraguard too, a spray used for keeping paraffin from >sticking to surfaces. It has a nasty smell that makes me nauseous so I >don't use it myself. Also, make sure there isn't any water on the face >of the block or on the knife before you start a ribbon. > > > >Sarah Jones HT(ASCP) >Dept. of Vet. Anatomy & Public Health >Histology Lab >Texas A&M University >College Station, TX 77843-4458 >phone: 979-845-3177 >fax: 979-458-3499 > > > >>> "Jacqueline Miller" >7/30/2004 2:16:00 PM >>> >Hi everyone, > >I would like to know about the techniques that histotechs use to cut >paraffin sections--especially how the ribbon is transferred from >microtome to water bath while preserving the first and last sections. > >I've been sectioning paraffin blocks for almost 4 years now. However, >I've never needed to cut serial sections and the samples were always >large. So, I never worried about losing the first section of each >ribbon (and sometimes the last). I would like to know what techniques >the histologists out there use to transfer the ribbon from the >microtome >to the water bath. I've used metal forceps and my fingers (gloved and >not gloved) to grab the first section (but the section sticks to the >forceps), and I'm using the wooden part of a cotton swab to lift off >the >last section, which is working fairly well. > >I'm also using a new microtome, Leica RM2235, and having some trouble >with the first section coming off the knife just wrinkling up. And, >the surface below the knife is such that even if I grab the edge of >the >section with a paintbrush, it won't budge. Is there anything I can do >to correct these problems? Also, which is preferred, to cut the >blocks >chilled or to leave them RT. I used to cut blocks chilled all the >time, >but here it's preferred that I cut them RT unless I have a problem. > > >Thanks, >Jackie > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Linke_Noelle <@t> Allergan.com Fri Aug 20 10:10:16 2004 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] doing immunos by hand Message-ID: I do all my immunos by hand and haven't spent a penny on 'stainers', be they large or small. Question is, how many will you be doing at one time? If you're only doing a small amount, an old slide box works wonderfully, put some moist paper towels in the bottom, shut the lid and you've got yourself an incubation chamber. I would only recommend kits if you are a beginner. They are a HUGE waste of money. Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, August 19, 2004 7:37 PM To: Pathologyarts@aol.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] doing immunos by hand Curt, Biocare Medical has some equipment that you might be able to use. They have these slide shakers (Kinetic Stainers) that can hold either 24 or 36 slides. They have heaters that heat up to 95 degrees C. I used these until my pathologists decided to go to the $40 billion machines. I still have them as a back up, just in case. They also have a wide variety of primary antibodies and detection kits. Their number is 1-800-799-9499. They also have great technical support, in my opinion, which in no way reflects the opinions of my employers. I know I'm gonna get "flamed" on this one, but I like excitement. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: To: Sent: Thursday, August 19, 2004 12:59 PM Subject: [Histonet] doing immunos by hand > does anyone still do immunos by hand or are they all done with these $40 > billion automated machines? > > i would like to see what the options are as far as doing these things > manually. > > primarily for derm specimens, HMB 45, S 100, KI 67. > > > any help is appreciated, > curt > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wasielewski.reinhard.von <@t> mh-hannover.de Fri Aug 20 10:42:45 2004 From: wasielewski.reinhard.von <@t> mh-hannover.de (wasielewski.reinhard.von@mh-hannover.de) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] CK7 and CK19 on mouse tissue Message-ID: <41263815.9358.F6C0D4D@localhost> Dear Histonetters, I want to do Cytokeratin staining 7 and 19 (CK 7, CK19) on FFPE mouse tissue. Can anyone give me some advice which antibodies work best. Pretreatment ? Our regular anti CK7 and CK19 for human tissue didn't work. Thanks in advance Reinhard von Wasielewski PD Dr. med. Reinhard von Wasielewski From pedro.louro <@t> spcorp.com Fri Aug 20 10:53:11 2004 From: pedro.louro <@t> spcorp.com (Louro, Pedro) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] decal solution on Immuno. stains Message-ID: <4508920F80C0D411B90200508BF9A9F4062B4700@LAFMSG30.us.schp.com> Any suggestions on what type of Decal. solution is good for Immuno. stains? I will have mouse bones to stain Thanks Pedro ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From gcallis <@t> montana.edu Fri Aug 20 10:59:21 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Endothelial cell staining In-Reply-To: <003001c486c1$fdae5f90$6e892b89@DFFRCL0J> References: <003001c486c1$fdae5f90$6e892b89@DFFRCL0J> Message-ID: <6.0.0.22.1.20040820093452.01b4b728@gemini.msu.montana.edu> First of all, you do NOT NEED ANY retrieval or digestion on Carnoys fixed tissue - there is NO formalin involved with this fixative - we had total success with DAKO Factor VIII, at 1:200 - 1:250. Tissue was fCarnoys fixed overnight (we took out the chloroform since it is not a fixative, just there to remove fat/lipids). Your retrieval may have ruined the antigen. As for CD31 rat antimouse, we get no background staining with rat antimouse antibodies and we use mainly BD Pharmingen antibodies. It can be done two ways with either a biotinylated primary or CD31 (dilution panel needed) come back with goat antiRat F(ab')2 biotinylated, ADSORBED TO MOUSE, and Strepavidin HRP. For frozen section, 5 um air dry overnight, fix in 75ml acetone/25ml absolute ethanol mixture for 5 minutes at room temperature, DO NOT AIR DRY again, go directly to buffer 3 changes. Use appropriate rinses throughout, rinse buffer contains 0.05 - 0.005% Tween 20 and 0.2% goat serum. Room temperature staining, no added temperature. This fixative will give you good morphology and excellent staining. Do DAKO peroxidase block (S2001) for frozen sections (for excessive endogenous Px, glucose oxidase method) Normal serum block 10% goat/2.5% mouse serum mixture, 30 min Strepavidin/biotin block (Vector), rather than avidin/biotin blocking Primary antibody diluted in 5% goat- 30 min , negative control is either Rat IgG or isotype matched IgG Secondary antibody diluted in NORMAL SERUM BLOCK with the mouse serum - we use Biosource/TAGO secondary, diluted 1:250 (0.5mg/ml stock) for 30 min Strepavidin-HRP - Biosource, diluted 1:500 in buffer - 20 min AEC+ from DAKO and control color development with a microscope on the positive control. If you use biotinylated primary (negative control is Rat IgG-biotinylated from Jackson at exact concentration in ug/ml as primary) just dilute this in the NORMAL SERUM BLOCK with mouse serum, 30 min - Rinse and use Strepavidin-HRP or AP. If you use DAB, you will have to adjust all antibody dilutions accordingly. If you have too much endogenous peroxidase, there is a superior method to block peroxidase/pseudoperoxidase, called Glucose oxidase method. I will be happy to attach via private email OR use an alkaline phos method instead. DAKO has a new red chromogen for alk phos that is very sensitive, try that instead. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman Bozeman MT 59717-3610 At 08:28 AM 8/20/2004, you wrote: >I just wanted to thank all histonet users for replying to my e-mail on >endothelial cell staining. > >To summarise my findings, von Willebrand Factor would work on Carnoy's >fixed normal mouse tissue but I could not get it to work on my tumour >sections. The tumour sections for some reason would not tolerate any >antigen retrieval even proteinase K for only 30 seconds would destroy the >tumour section. Maybe I didn't fix the tissue in the Carnoy's for long >enough. I got no staining with no antigen retrieval. > >I also tried the CD31 rat anti-mouse which is available from PharMingen. I >got this to work on frozen sections but with alot of background staining >which proved difficult to eliminate. Maybe my protocol was wrong or there >is alot of endogenous peroxidases in my tumours. > >I am especially grateful to John McGinley and Liz Chlipala who recommended >the CD31 from Santa Cruz which worked exceptionally well first time of >asking on formalin-fixed paraffin embedded tissue. > >Thanks, > >Jerry >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bjhighto <@t> utmb.edu Fri Aug 20 11:15:05 2004 From: bjhighto <@t> utmb.edu (Hightower, Barbara J.) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] UNSCRIBE Message-ID: <977F35BB611E644F9C099E1B660D19F62030D7@EXCH2K5.utmb.edu> From David.Edmondson <@t> christie-tr.nwest.nhs.uk Fri Aug 20 11:09:25 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Spirit fixation of tissue Message-ID: John, I have all the back issues of the IMLT/IMLS Journal so will have look, but slowly, over a glass. ( I inherited them, honest ) Dave -----Original Message----- From: JOHN PHILLIPS [mailto:JOHN.PHILLIPS@new-tr.wales.nhs.uk] Sent: 19 August 2004 14:33 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Spirit fixation of tissue Hello Histonet users, I would really appreciate knowing where to find an article, author unknown, published, probably somewhere in the time period 1972 to 1986, about the use of spirit for the fixation of skin biopsies and the like. I think it may have been published in the Institute of Medical Laboratory Technology journal (UK) or may be the Journal of Clinical Pathology (UK). The article investigated the effects of rum, whisky, gin and vodka etc. on tissue. I think rum came out on top. I am presenting a talk to GP's and would like to mention this as part of my presentation. I shall of course advise them not to use a Scottish malt under any circumstances that would be sacrilage. American bourbon may be! Thanks, John Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r rheolwr systemau wybod drwy ddefnyddio'r manylion isod. Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain Cymru. Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau?EUR(tm)r Ddeddf Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru ar www.newalesnhstrust.org.uk This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager using the details below. The contents of this email represent the views of the individual(s) named above and do not necessarily represent the views of the North East Wales NHS Trust. Please be aware that, under the terms of the Freedom of Information Act 2000, the North East Wales NHS Trust may be required to make public the content of any emails or correspondence received. For futher information on Freedom of Information, please refer to the North East Wales NHS Trust website at www.newalesnhstrust.org.uk For further assistance, please contact system.administrator@new-tr.wales.nhs.uk. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Aug 20 11:21:17 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] decal solution on Immuno. stains In-Reply-To: <4508920F80C0D411B90200508BF9A9F4062B4700@LAFMSG30.us.schp. com> References: <4508920F80C0D411B90200508BF9A9F4062B4700@LAFMSG30.us.schp.com> Message-ID: <6.0.0.22.1.20040820101738.01b6f8f8@gemini.msu.montana.edu> What are you planning to stain for with IHC? In research, you have some luxury of time, then EDTA is good 14% tetrasodium EDTA, pH 7.6 adjusted with acetic acid. We make this up in Dulbeccos PBS (Sigma). Decalcify in refrigerator, although room temperature is fine. or buffered formic acid, buy the slower acid decalcifiers that contain formic acid with either sodium formate or sodium citrate. Do endpoint checks to know when decalcification is done. At 09:53 AM 8/20/2004, you wrote: >Any suggestions on what type of Decal. solution is good for Immuno. stains? > >I will have mouse bones to stain > >Thanks > >Pedro > > >********************************************************************* >This message and any attachments are solely for the intended recipient. If >you are not the intended recipient, disclosure, copying, use or >distribution of the information included in this message is prohibited -- >Please immediately and permanently delete. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From David.Edmondson <@t> christie-tr.nwest.nhs.uk Fri Aug 20 11:28:44 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] doing immunos by hand Message-ID: Hi, We use Shando's "Sequenza" to do only a small number of slides these days since a DAKO Techmate came along. The Sequenza has the option of an organiser for several sets of slides but this is not necessary for small numbers of slides The slides are arrayed each against plastic plates, in groups of ten. The plates forms a well at one end and solutions slowly drain past, accross the face of the slides, buffers antibodies, chromogens , whatever. In terms of a manual system it is sweeter than using trays, boxes. Once set in the clip there is no handling until the end. The waste is held in the container, staining is even , no ringing of the sections with a pen, sweet. Having done batches of a hundred with trays and then the sequenza, my money is on the latter. David Edmondson Christie Hospital Manchester UK -----Original Message----- From: Pathologyarts@aol.com [mailto:Pathologyarts@aol.com] Sent: 19 August 2004 18:59 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] doing immunos by hand does anyone still do immunos by hand or are they all done with these $40 billion automated machines? i would like to see what the options are as far as doing these things manually. primarily for derm specimens, HMB 45, S 100, KI 67. any help is appreciated, curt _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janice.Fightmaster <@t> HCAhealthcare.com Fri Aug 20 11:44:55 2004 From: Janice.Fightmaster <@t> HCAhealthcare.com (Fightmaster Janice) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] pancreas tissue Message-ID: <1734C20AE655CD449599A15EDC589D89017CA599@orlex05.hca.corpad.net> I am in need of a block of normal pancreas tissue for IHC controls. Can anyone help me? Janice Fightmaster Pathology Supervisor Oak Hill Hospital 11375 Cortez Blvd. Brooksville, FL 34613 (352)597-6363 Ext. 3636 From scoop <@t> mail.nih.gov Fri Aug 20 11:47:39 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] decal solution on Immuno. stains In-Reply-To: <4508920F80C0D411B90200508BF9A9F4062B4700@LAFMSG30.us.schp.com> References: <4508920F80C0D411B90200508BF9A9F4062B4700@LAFMSG30.us.schp.com> Message-ID: I (a rank amateur compared to most of the people on the Histonet) have used BBC Rapid-Cal Immuno on mouse bones for Immuno and was very pleased with the results. It has formic acid in it. Their phone number is (360)629-4477 and they are very helpful with suggestions. They told me to use around 90 minutes for my mouse bones (at room temp) after fixation and that was perfect. Although I'm not as knowledgable as many others on the Histonet and there are probably better suggestions (which I would love to hear about) the fact that a person at my level of experience could get things to work well the first time with this reagent is probably a good recommendation for it. Good Luck! Sharon >Any suggestions on what type of Decal. solution is good for Immuno. stains? > >I will have mouse bones to stain > >Thanks > >Pedro > > >********************************************************************* >This message and any attachments are solely for the intended >recipient. If you are not the intended recipient, disclosure, >copying, use or distribution of the information included in this >message is prohibited -- Please immediately and permanently delete. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From Janet.Bonner <@t> FLHOSP.ORG Fri Aug 20 11:47:40 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Decal for immunos Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB401A@fh2k093.fhmis.net> -----Original Message----- From: Bonner, Janet Sent: Tuesday, August 17, 2004 8:34 AM To: 'lpwenk@sbcglobal.net '; 'WWmn916@aol.com '; 'histonet@pathology.swmed.edu ' Subject: RE: [Histonet] Iron stain question Just a note on the side about the core being negative - this no longer has to be! BCC rapidcal immuno decals Bone Marrow cores without compromising the positivity of the iron (no- I don't work for them). We just discovered it and the immuno and iron results are beautiful. -----Original Message----- From: lpwenk@sbcglobal.net To: WWmn916@aol.com; histonet@pathology.swmed.edu Sent: 8/15/2004 7:39 AM Subject: Re: [Histonet] Iron stain question Usually, it's the bone core that is negative due to the decalcification, while the clot and the smear are positive. Couple off the wall thoughts. Any chance that the smear slide was in the same coplin jar slot as the control, so that the smear was stuck against the back of the previous slide? That would prevent the ferrocyanide solution from "touching" the smear. If this is possible, remove the coverslip, remove the mounting media with xylene (or whatever you use), run the slide back down through alcohols to water, and restain. Any chance that the smear was fixed in something containing an acid, like acetic acid? That might remove the iron. No way to correct this problem. Did they do a bone core and/or clot? Were they negative? If so, then the person IS iron deficient, regardless of whether the person has leukemia or not. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Saturday, August 14, 2004 11:49 PM Subject: [Histonet] Iron stain question > Hello again histonetters, > > Can anyone give me any insight as to why some smear slides stained for iron > would not stains positive as it was expected they would? Doctor say's the > slides should have stained positive for iron because the patient has leukemia. > The control stained positive as it should have. Staining tech is sure all her > slides were exposed to solution. Thanks for any help. > > Deb King, HT(ASCP) > Sacramento, CA > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From ploykasek <@t> phenopath.com Fri Aug 20 11:58:45 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:56 2005 Subject: FW: [Histonet] doing immunos by hand In-Reply-To: Message-ID: ------ Forwarded Message From: Patti Loykasek Date: Fri, 20 Aug 2004 09:35:03 -0700 To: Hermina Borgerink Subject: Re: [Histonet] doing immunos by hand Curt, there are a couple of ways to go about it. Several involve little expense. The way we do it most frequently is to use either micro culture trays that have a lid or cafeteria trays. Either way, you will need to use some bench top paper that is a foam type material on one side & absorbent type material on the other. Cut the paper to fit your tray. Wet down the paper side with a water squirt bottle & smooth out the paper to eliminate bubbles by rolling a small tube or pipette over the surface. If using the micro trays, lay the paper inside the tray, lay your slides in the tray, cover with lid for incubation. These micro trays hold about 20-22 slides. For more slides, lay the bench paper on the counter top, wet & smooth. Lay your slides on top, cover with the cafeteria tray to create a "lid" for your incubation chamber. If you need more info, please contact me. Hope this helps & gives you some ideas. For ease of handling, you may want to investigate the MicroProbe from Fisher. It operates on the capillary gap principle. You must use cap. Gap slides. It holds 10 pairs of slides. Not very expensive. Has heat, can do in-situs, too. Patti Loykasek PhenoPath Laboratories Seattle, WA Curt, > > I do all my immunos by hand (up to 80 slides at a time) using the > MicroProbe Staining system from Fisher. The system operates by means of > special slides that have a painted surface and a special slide holder. > When paired together (facing each other) a capillary gap is created that > allows fluids to rise between the two slides using capillary action. > You don't need to purchase the complete set up either, just the special > slide holders and isolons (rubber pads with wells to dispense reagents). > I think each slide holder (to stain 20 slides at once) runs around > $250.00. I'm not sure about the isolons, but they are not that expense > and are re-usable. I routinely use a large panel of antibodies, > including the ones you mentioned. Check the Fisher catalog for more > details. > Hermina > > > Hermina M. Borgerink, BA, HTL(ASCP)QIHC > > Wake Forest University Health Sciences > Department of Pathology > Medical Center Blvd. > Winston-Salem, NC 27157 > Tel. (336) 716-1538 > Fax (336) 716-1515 > e-mail hborgeri@wfubmc.edu > > "Treat a man as he appears to be, and you make him worse. But treat a > man as if he already were what he potentially could be, and you make him > what he should be." (Goethe) > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Pathologyarts@aol.com > Sent: Thursday, August 19, 2004 1:59 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] doing immunos by hand > > does anyone still do immunos by hand or are they all done with these $40 > > billion automated machines? > > i would like to see what the options are as far as doing these things > manually. > > primarily for derm specimens, HMB 45, S 100, KI 67. > > > any help is appreciated, > curt > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------ End of Forwarded Message From pmarcum <@t> polysciences.com Fri Aug 20 12:26:30 2004 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Spirit fixation of tissue In-Reply-To: Message-ID: <000e01c486da$d4bdfd70$4f00a8c0@PMARCUM2K> The Journals, the glass or the spirits, which of theses was the inheritance here? By the way a long time ago, in the 80's I remember a specimen being sent (or at least it is what I was told) to the AFIP in tequila as fixative and dehydrant. We all mourned the lose of a good tequila as that is what was used. I think a cheap one would have been just as good. Pam Marcum > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Edmondson > David (RBV) NHS Christie Tr > Sent: Friday, August 20, 2004 12:09 PM > To: 'JOHN PHILLIPS' > Cc: Histonet (E-mail 2) > Subject: RE: [Histonet] Spirit fixation of tissue > > > John, > I have all the back issues of the IMLT/IMLS Journal so will have look, but > slowly, over a glass. ( I inherited them, honest ) > Dave > > -----Original Message----- > From: JOHN PHILLIPS [mailto:JOHN.PHILLIPS@new-tr.wales.nhs.uk] > Sent: 19 August 2004 14:33 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Spirit fixation of tissue > > > Hello Histonet users, > > I would really appreciate knowing where to find an article, > author unknown, > published, probably somewhere in the time period 1972 to 1986, > about the use > of spirit for the fixation of skin biopsies and the like. I think it may > have been published in the Institute of Medical Laboratory Technology > journal (UK) or may be the Journal of Clinical Pathology (UK). The article > investigated the effects of rum, whisky, gin and vodka etc. on tissue. I > think rum came out on top. > > I am presenting a talk to GP's and would like to mention this as > part of my > presentation. I shall of course advise them not to use a Scottish > malt under > any circumstances that would be sacrilage. American bourbon may be! > > Thanks, > > John > > Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn > gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu > atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r > rheolwr systemau wybod drwy ddefnyddio'r manylion isod. > > Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, > felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain > Cymru. > > Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i > Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys > unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag > amodau?EUR(tm)r Ddeddf > Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, > cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru > ar www.newalesnhstrust.org.uk > > > This email and any files transmitted with it are confidential and > intended solely for the use of the individual or entity to whom they > are addressed. If you have received this email in error please notify > the system manager using the details below. > > The contents of this email represent the views of the individual(s) > named above and do not necessarily represent the views of the > North East Wales NHS Trust. > > Please be aware that, under the terms of the Freedom of Information Act > 2000, the North East Wales NHS Trust may be required to make public the > content of any emails or correspondence received. For futher > information on Freedom of Information, please refer to the North East > Wales NHS Trust website at www.newalesnhstrust.org.uk > > For further assistance, please contact > system.administrator@new-tr.wales.nhs.uk. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Rcartun <@t> harthosp.org Fri Aug 20 12:30:51 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] pancreas tissue Message-ID: I can. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> Fightmaster Janice 08/20/04 12:44PM >>> I am in need of a block of normal pancreas tissue for IHC controls. Can anyone help me? Janice Fightmaster Pathology Supervisor Oak Hill Hospital 11375 Cortez Blvd. Brooksville, FL 34613 (352)597-6363 Ext. 3636 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janice.Fightmaster <@t> HCAhealthcare.com Fri Aug 20 12:31:55 2004 From: Janice.Fightmaster <@t> HCAhealthcare.com (Fightmaster Janice) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] pancreas tissue Message-ID: <1734C20AE655CD449599A15EDC589D89017CA59D@orlex05.hca.corpad.net> Thanks for the responses; I have plenty on the way now!! Janice Fightmaster Pathology Supervisor Oak Hill Hospital 11375 Cortez Blvd. Brooksville, FL 34613 (352)597-6363 Ext. 3636 From hymclab <@t> hyhc.com Fri Aug 20 13:30:45 2004 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] doing immunos by hand Message-ID: I have also used the Shandon sequenza for my manual immunos. We have used it for 10 years and love it!!! Dawn Schneider, HT(ASCP) Howard Young Medical Center Woodruff, WI -----Original Message----- From: Edmondson David (RBV) NHS Christie Tr [mailto:David.Edmondson@christie-tr.nwest.nhs.uk] Sent: Friday, August 20, 2004 11:29 AM To: 'Pathologyarts@aol.com' Cc: Histonet (E-mail 2) Subject: RE: [Histonet] doing immunos by hand Hi, We use Shando's "Sequenza" to do only a small number of slides these days since a DAKO Techmate came along. The Sequenza has the option of an organiser for several sets of slides but this is not necessary for small numbers of slides The slides are arrayed each against plastic plates, in groups of ten. The plates forms a well at one end and solutions slowly drain past, accross the face of the slides, buffers antibodies, chromogens , whatever. In terms of a manual system it is sweeter than using trays, boxes. Once set in the clip there is no handling until the end. The waste is held in the container, staining is even , no ringing of the sections with a pen, sweet. Having done batches of a hundred with trays and then the sequenza, my money is on the latter. David Edmondson Christie Hospital Manchester UK -----Original Message----- From: Pathologyarts@aol.com [mailto:Pathologyarts@aol.com] Sent: 19 August 2004 18:59 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] doing immunos by hand does anyone still do immunos by hand or are they all done with these $40 billion automated machines? i would like to see what the options are as far as doing these things manually. primarily for derm specimens, HMB 45, S 100, KI 67. any help is appreciated, curt _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Fri Aug 20 13:28:42 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] RE: IHC polymer detection system for rat tissues Message-ID: Dear Ronald, Just use the anti-mouse EnVision for use on rat tissue specimens. However, before using it you have to mix the EnVision with 10-15% (final concentration) normal rat serum. Leave the mixture for 60 min at room temp (or overnight 4C) then use it. All the cross-reactions are abolished now. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Ronald P. Wilson" Date Thu, 19 Aug 2004 15:15:43 -0400 To Histonet Subject [Histonet] IHC polymer detection system for rat tissues Has anyone found a successful polymer detection system (i.e., Envision or others) directed against a mouse monoclonal on rat tissue. We tried one but got a lot of background staining, I suspect from cross reactivity with closely related antigens. Our standard IHC approach uses an anti-mouse, rat-absorbed secondary antibody, but I would like to get the added sensitivity of a polymer-based system. Ronald P. Wilson, V.M.D., M.S. Associate Professor Department of Comparative Medicine, HO54 Penn State University College of Medicine M. S. Hershey Medical Center 500 University Drive Hershey, PA 17033 phone: 717-531-8460 fax: 717-531-5001 e-mail: rpw4@psu.edu From madaryhtl <@t> juno.com Fri Aug 20 13:33:22 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Hand IHC Message-ID: <20040820.113415.3885.451884@webmail23.lax.untd.com> 40 Billion is alot to spend on a machine, I would shop around. Sequenza chambers serve us quite well as well as the automation. All the stains you mentioned are easily accomplished using hand techniqes. Nick(Rocky) Madary, HT,HTL(ASCP)QIHC Histology Manager, Medimmune Inc Cell-3012334950 Work-3013984745 Fax-3013989745 From lsb <@t> jax.org Fri Aug 20 13:36:48 2004 From: lsb <@t> jax.org (Lesley S. Bechtold) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Desperately seeking..information Message-ID: <6.0.3.0.0.20040820142237.0261f600@aretha.jax.org> Hi Everyone, Up here in Bar Harbor, Maine, we are trying to get an idea of what other Histology Labs are doing - how many people they have working in them, what they consider reasonable capacity, how much work they do in a year, how diversified they are, etc. We are not interested in salary information. We have created a bench-marking survey on our home website at http://www.jax.org/sci_survey/ . Click on the "Histology" link and it will take you right to the survey. You'll notice there are other surveys on there as well so if your institution/hospital/business carries out any other activities (such as Electron Microscopy), please feel free to complete those surveys as well. It only takes a few minutes to complete and will remain completely confidential. Anyone who participates in the survey will receive a copy of the results (with names, etc. deleted) within a few weeks. We're aiming for the beginning of October to send the results to all the participants. We will need your email address in order to send the results to you so make sure you include that. So we'd really like to hear from you. I know how active the Histonet is so I'm hoping for a good response as I'm sure everyone will be interested in the results. For anyone who is not interested in participating, I apologize for using up your time. Please delete this email. Thank you! Lesley Bechtold Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191 From Charlotte.Kopczynski <@t> baycare.org Fri Aug 20 13:42:32 2004 From: Charlotte.Kopczynski <@t> baycare.org (Kopczynski, Charlotte) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Drying slides before Filing Message-ID: <7C3A384EF3ABD611B3270008023E3E030481CC1C@mphexch01.mpm.baycare.internal> Hello fellow Histonetters, We are having a big problem with air bubbles under our coverslips before filing. We use Richard Allan mounting media and the slides in the trays are placed in an oven set at 58 degrees C overnight. I am thinking the oven may be part of the problem but was wondering what the process is out there in Histo-land. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 ************************************************************************************************** The contents of this email and any attachments are confidential. They are intended for the named recipient(s) only. If you have received this email in error please notify the system manager or the sender immediately and do not disclose the contents to anyone or make copies. ** eSafe scanned this email for viruses, vandals and malicious content. ** ************************************************************************************************** From garygill <@t> dcla.com Fri Aug 20 13:54:08 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Drying slides before Filing Message-ID: <0B11620AEE5EFB4FA7A7558339972A830704F3@HALIBUT.dcla.com> Air bubbles form because there is not enough solid material remaining under the cover glass after the solvent has evaporate. Among possible causes: * RAS put out a bad batch of mounting medium (i.e., more solvent than usual) * Not enough mounting medium being applied * Accelerated drying (e.g., oven too hot [58 degrees C is not too hot, in my experience]) -- how soon after coverslipping are slides put in oven? Right away could make a negative difference. Simplest first solution is to add more mounting medium, while appreciating that way too much degrades image quality under 40x objectives -- which would be the subject of another discussion. Little useful information has ever been published about mounting media. The best information was published as a 2-part article in Stain Technology in the 1950s. Authors were a committee chaired by the late and great Dr. Ralph D. Lillie. Gary Gill -----Original Message----- From: Kopczynski, Charlotte [mailto:Charlotte.Kopczynski@baycare.org] Sent: Friday, August 20, 2004 1:43 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Drying slides before Filing Hello fellow Histonetters, We are having a big problem with air bubbles under our coverslips before filing. We use Richard Allan mounting media and the slides in the trays are placed in an oven set at 58 degrees C overnight. I am thinking the oven may be part of the problem but was wondering what the process is out there in Histo-land. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 **************************************************************************** ********************** The contents of this email and any attachments are confidential. They are intended for the named recipient(s) only. If you have received this email in error please notify the system manager or the sender immediately and do not disclose the contents to anyone or make copies. ** eSafe scanned this email for viruses, vandals and malicious content. ** **************************************************************************** ********************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BriggsK <@t> drmc.drhsi.org Fri Aug 20 14:21:37 2004 From: BriggsK <@t> drmc.drhsi.org (Kevin Briggs) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Drying slides before Filing Message-ID: <8A37FE7D529ED411983E00D0B75D01FC0663A79C@sirius.drmc.drhsi.org> Charlotte, I think Gary's got it. 58 degrees C should only be too hot if the slides were placed in the oven too soon after mounting as we have found--although, decreasing the temperature will indeed slow the solvent's rate of evaporation. Try delaying your oven cure time and see if that helps. Good luck! Kevin D. Briggs, MS,CT(ASCP) Team Leader- Cytopathology/Histopathology Services Danville Regional Medical Center 142 South Main Street Danville, VA 24541 Telephone: (434) 799-4470 ext.5451 Fax: (434) 799-2118 E-mail: briggsk@drhsi.org -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Friday, August 20, 2004 2:54 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Drying slides before Filing Air bubbles form because there is not enough solid material remaining under the cover glass after the solvent has evaporate. Among possible causes: * RAS put out a bad batch of mounting medium (i.e., more solvent than usual) * Not enough mounting medium being applied * Accelerated drying (e.g., oven too hot [58 degrees C is not too hot, in my experience]) -- how soon after coverslipping are slides put in oven? Right away could make a negative difference. Simplest first solution is to add more mounting medium, while appreciating that way too much degrades image quality under 40x objectives -- which would be the subject of another discussion. Little useful information has ever been published about mounting media. The best information was published as a 2-part article in Stain Technology in the 1950s. Authors were a committee chaired by the late and great Dr. Ralph D. Lillie. Gary Gill -----Original Message----- From: Kopczynski, Charlotte [mailto:Charlotte.Kopczynski@baycare.org] Sent: Friday, August 20, 2004 1:43 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Drying slides before Filing Hello fellow Histonetters, We are having a big problem with air bubbles under our coverslips before filing. We use Richard Allan mounting media and the slides in the trays are placed in an oven set at 58 degrees C overnight. I am thinking the oven may be part of the problem but was wondering what the process is out there in Histo-land. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 **************************************************************************** ********************** The contents of this email and any attachments are confidential. They are intended for the named recipient(s) only. If you have received this email in error please notify the system manager or the sender immediately and do not disclose the contents to anyone or make copies. ** eSafe scanned this email for viruses, vandals and malicious content. ** **************************************************************************** ********************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histojock <@t> hotmail.com Fri Aug 20 21:26:42 2004 From: histojock <@t> hotmail.com (Histo Jock) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Image analysis and slide scanning machine Message-ID: Can you describe more about how this works? How does it create the image if it doesn't tile? What kind of analysis can it do? You're exhibiting with Beecher - can it run Beecher's new software? HistoJock. >Message: 9 >Date: Mon, 16 Aug 2004 16:11:45 -0400 >From: "Glenn Smith" >Subject: [Histonet] Image analysis and slide scanning machine (Wester, > Martha) >To: >Message-ID: <00c101c483cd$45edc040$1f05a8c0@confocal.com> >Content-Type: text/plain; charset="US-ASCII" > >Hi Kim, > >We supply such an instrument, our TISSUEscope - combining confocal scanning >laser microscopy and a wide field of view. Depending on your resolution >requirements, the TISSUEscope can acquire a single image at up to 1 um >resolution with no mosaic and no tiling of images. A standard slide (1" x >3") is loaded into the machine and the rest of operations is via software. >We have another model which offers submicron resolution as well. Both >brightfield and fluorescence detection is provided within the same >instrument. Please feel free to contact me offline for more details. > >Also, we will have one of these instruments at the NSH meeting in Toronto - >we'll be exhibiting in conjunction with Beecher Instruments. > >Regards >Glenn Smith, P.Eng. >519.886.9013 x38 > >Biomedical Photometrics Inc/GeneFocus www.GeneFocus.com> >Widefield Confocal Scanning Instruments and Software _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar – get it now! http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/ From David.Edmondson <@t> christie-tr.nwest.nhs.uk Sat Aug 21 05:14:12 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] RE: Old IMLT journals Message-ID: OK Bryan, It will take a little while, but I shall start. I wonder that someone has already scanned some journals, like "path and bact", stain tech and histochem journal. Or at least found the annual indexes to scan. There are Stain tech 69-83 and Histochen J 79-86 beside me now, and I wonder at the quantity of published material. Maybe I shall ask the IBMS, I remember there is/was a historical section that may have moved from preservation to digitising. Not the same as "Brass and Glass", but. Dave -----Original Message----- From: Bryan Llewellyn [mailto:bryand@netbistro.com] Sent: 20 August 2004 20:41 To: Edmondson David (RBV) NHS Christie Tr Subject: Old IMLT journals Hi, I have a big favour to ask. I am the author of the StainsFile web site (http://stainsfile.info). One of my aims for this site is to include all published staining methods. I know that's quite futile, but I would like to get as many as possible. I have the IMLT journals between 1962 and about 1975, when I let my membership finally lapse after emigrating to Canada. Is there any possibility that I could get you to scan or photocopy any histological methods involving techniques or theory about dyes or metallic impregnations (almost everything except immuno- or enzyme histochemistry as these are outside my limits for the site) from any journals you have apart from those years? I would like to be able to include the material on StainsFile, and I have been wondering how I might be able to get details from Canada. Unfortunately, none of the British Techs I know over here has kept their copies, or have let their membership lapse. Here's hoping. Bryan Llewellyn Edmondson David (RBV) NHS Christie Tr wrote: > John, > I have all the back issues of the IMLT/IMLS Journal so will have look, but > slowly, over a glass. ( I inherited them, honest ) > Dave > > -----Original Message----- > From: JOHN PHILLIPS [mailto:JOHN.PHILLIPS@new-tr.wales.nhs.uk] > Sent: 19 August 2004 14:33 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Spirit fixation of tissue > From terribraud <@t> msn.com Sun Aug 22 09:36:17 2004 From: terribraud <@t> msn.com (TERRI BRAUD) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Hand IHC Message-ID: When I worked in a smaller volume lab, we easily and consistently stained IHC with the assist of the Shandon Sequenza. Anyone's reagent's worked, but those buffers with surfactants seemed a wee bit better. And for any Shandon folks out there reading this message, whose totally unbrilliant idea was it to lose the Shandon-Lipshaw names? You will NEVER be Thermo to me, and from the looks of this list, to a lot of other people, too. Terri Braud Lab Rat From barbara.bublava <@t> meduniwien.ac.at Mon Aug 23 01:09:43 2004 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] urgent problem - cryoprotection References: <009401c485e3$cdbde0a0$1201a8c0@GERICHTS9XOZZ8> <4124B17B.6050402@bms.com> Message-ID: <001401c488d7$c8422220$1201a8c0@GERICHTS9XOZZ8> Thank you very much for your help. My cryostat will soon be repaired and I decided to leave the specimes in succrose. But now there appears another problem. I will do oil red O with these specimens and my boss says now, that the succrose and/or the O.C.T. is washing out the fat. He also says that oil red O is not state of the art for diagnosing thrombosis of fat in lungtissue. I do not think so, but just to be shure I would be interested in your opinion. I know methods using Sudan Black or other dyes but they use the same principle as oil red O. The only "real" alternative I read about here in Histonet is using osmiumtetroxide and doing paraffinsections. But as I remember Osmiumtetroxide is not very healthy and has to be collected for disposal. Thank you all for your help!!! Barbara Bublava, Vienna From barbara.bublava <@t> meduniwien.ac.at Mon Aug 23 02:13:32 2004 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] urgent problem - cryoprotection References: <009401c485e3$cdbde0a0$1201a8c0@GERICHTS9XOZZ8><4124B17B.6050402@bms.com> <001401c488d7$c8422220$1201a8c0@GERICHTS9XOZZ8> Message-ID: <003401c488e0$b276d2c0$1201a8c0@GERICHTS9XOZZ8> sorry - I do mean embolism of fat not thrombosis :o) ----- Original Message ----- From: "Barbara Bublava" To: Sent: Monday, August 23, 2004 8:09 AM Subject: Re: [Histonet] urgent problem - cryoprotection > Thank you very much for your help. My cryostat will soon be repaired and I > decided to leave the specimes in succrose. But now there appears another > problem. I will do oil red O with these specimens and my boss says now, that > the succrose and/or the O.C.T. is washing out the fat. He also says that oil > red O is not state of the art for diagnosing thrombosis of fat in > lungtissue. > > I do not think so, but just to be shure I would be interested in your > opinion. > > I know methods using Sudan Black or other dyes but they use the same > principle as oil red O. The only "real" alternative I read about here in > Histonet is using osmiumtetroxide and doing paraffinsections. But as I > remember Osmiumtetroxide is not very healthy and has to be collected for > disposal. > > Thank you all for your help!!! > > Barbara Bublava, Vienna > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rentonlf <@t> bru.wits.ac.za Mon Aug 23 03:08:15 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Spirit fixation of tissue Message-ID: <1093248495.921a1660rentonlf@bru.wits.ac.za> Not much use this but... I can confirm that it would have been a UK lab journal, as the info was reported to me by my senior tech (straight off the boat from Cardiff) in about 1982/3. The essence was that cheap gin was the best fixative....make you think! -- ---Original Message----- From: "JOHN PHILLIPS" To: Date: Thu, 19 Aug 2004 14:33:19 +0100 Subject: [Histonet] Spirit fixation of tissue Hello Histonet users, I would really appreciate knowing where to find an article, author unknown, published, probably somewhere in the time period 1972 to 1986, about the use of spirit for the fixation of skin biopsies and the like. I think it may have been published in the Institute of Medical Laboratory Technology journal (UK) or may be the Journal of Clinical Pathology (UK). The article investigated the effects of rum, whisky, gin and vodka etc. on tissue. I think rum came out on top. I am presenting a talk to GP's and would like to mention this as part of my presentation. I shall of course advise them not to use a Scottish malt under any circumstances that would be sacrilage. American bourbon may be! Thanks, John Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r rheolwr systemau wybod drwy ddefnyddio'r manylion isod. Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain Cymru. Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau?r Ddeddf Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru ar www.newalesnhstrust.org.uk This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager using the details below. The contents of this email represent the views of the individual(s) named above and do not necessarily represent the views of the North East Wales NHS Trust. Please be aware that, under the terms of the Freedom of Information Act 2000, the North East Wales NHS Trust may be required to make public the content of any emails or correspondence received. For futher information on Freedom of Information, please refer to the North East Wales NHS Trust website at www.newalesnhstrust.org.uk For further assistance, please contact system.administrator@new-tr.wales.nhs.uk. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From rentonlf <@t> bru.wits.ac.za Mon Aug 23 03:13:47 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] doing immunos by hand Message-ID: <1093248827.921a1660rentonlf@bru.wits.ac.za> Doing immuno manually is entirely dependent on the volume. In a lab where I worked previously, we would do up to 400 slides by hand (we had a panel of over 100 diffent immunos to choose from). This was 8hrs HARD work. We had specially constructed perspex boxes to use as a moisture chamber, but in emergencies, large oblong Tupperware containers with moistened filter paper or roller towel were also used. The slides would be raised from the paper by means of blank slides placed at right angles to the immuno slide so as to avoid breaking nails when trying to overcome the surface tensiion that would suck the slide down onto the surface. best regards -----Original Message----- From: Pathologyarts@aol.com To: histonet@lists.utsouthwestern.edu Date: Thu, 19 Aug 2004 13:59:05 EDT Subject: [Histonet] doing immunos by hand does anyone still do immunos by hand or are they all done with these $40 billion automated machines? i would like to see what the options are as far as doing these things manually. primarily for derm specimens, HMB 45, S 100, KI 67. any help is appreciated, curt _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From rosemarie_g_behan <@t> groton.pfizer.com Mon Aug 23 09:03:48 2004 From: rosemarie_g_behan <@t> groton.pfizer.com (Behan, Rosemarie G) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] RE: Histonet Digest, Vol 9, Issue 34 Message-ID: <9DF8DDFE3736D3119D6D0008C79148D60A5BFED3@groexmbcr17.pfizer.com> also seeking information--does anyone use "dry acetone"? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Saturday, August 21, 2004 1:11 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 9, Issue 34 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: Spirit fixation of tissue (Pamela Marcum) 2. Re: pancreas tissue (Richard Cartun) 3. pancreas tissue (Fightmaster Janice) 4. RE: doing immunos by hand (hymclab) 5. RE: IHC polymer detection system for rat tissues (C.M. van der Loos) 6. Hand IHC (madaryhtl@juno.com) 7. Desperately seeking..information (Lesley S. Bechtold) 8. Drying slides before Filing (Kopczynski, Charlotte) 9. RE: Drying slides before Filing (Gary Gill) 10. RE: Drying slides before Filing (Kevin Briggs) 11. RE: Image analysis and slide scanning machine (Histo Jock) 12. RE: Old IMLT journals (Edmondson David (RBV) NHS Christie Tr) ---------------------------------------------------------------------- Message: 1 Date: Fri, 20 Aug 2004 13:26:30 -0400 From: "Pamela Marcum" Subject: RE: [Histonet] Spirit fixation of tissue To: "Edmondson David \(RBV\) NHS Christie Tr" , "'JOHN PHILLIPS'" Cc: "Histonet \(E-mail 2\)" Message-ID: <000e01c486da$d4bdfd70$4f00a8c0@PMARCUM2K> Content-Type: text/plain; charset="iso-8859-1" The Journals, the glass or the spirits, which of theses was the inheritance here? By the way a long time ago, in the 80's I remember a specimen being sent (or at least it is what I was told) to the AFIP in tequila as fixative and dehydrant. We all mourned the lose of a good tequila as that is what was used. I think a cheap one would have been just as good. Pam Marcum > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Edmondson > David (RBV) NHS Christie Tr > Sent: Friday, August 20, 2004 12:09 PM > To: 'JOHN PHILLIPS' > Cc: Histonet (E-mail 2) > Subject: RE: [Histonet] Spirit fixation of tissue > > > John, > I have all the back issues of the IMLT/IMLS Journal so will have look, but > slowly, over a glass. ( I inherited them, honest ) > Dave > > -----Original Message----- > From: JOHN PHILLIPS [mailto:JOHN.PHILLIPS@new-tr.wales.nhs.uk] > Sent: 19 August 2004 14:33 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Spirit fixation of tissue > > > Hello Histonet users, > > I would really appreciate knowing where to find an article, > author unknown, > published, probably somewhere in the time period 1972 to 1986, > about the use > of spirit for the fixation of skin biopsies and the like. I think it may > have been published in the Institute of Medical Laboratory Technology > journal (UK) or may be the Journal of Clinical Pathology (UK). The article > investigated the effects of rum, whisky, gin and vodka etc. on tissue. I > think rum came out on top. > > I am presenting a talk to GP's and would like to mention this as > part of my > presentation. I shall of course advise them not to use a Scottish > malt under > any circumstances that would be sacrilage. American bourbon may be! > > Thanks, > > John > > Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn > gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu > atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r > rheolwr systemau wybod drwy ddefnyddio'r manylion isod. > > Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, > felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain > Cymru. > > Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i > Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys > unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag > amodau?EUR(tm)r Ddeddf > Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, > cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru > ar www.newalesnhstrust.org.uk > > > This email and any files transmitted with it are confidential and > intended solely for the use of the individual or entity to whom they > are addressed. If you have received this email in error please notify > the system manager using the details below. > > The contents of this email represent the views of the individual(s) > named above and do not necessarily represent the views of the > North East Wales NHS Trust. > > Please be aware that, under the terms of the Freedom of Information Act > 2000, the North East Wales NHS Trust may be required to make public the > content of any emails or correspondence received. For futher > information on Freedom of Information, please refer to the North East > Wales NHS Trust website at www.newalesnhstrust.org.uk > > For further assistance, please contact > system.administrator@new-tr.wales.nhs.uk. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 2 Date: Fri, 20 Aug 2004 13:30:51 -0400 From: "Richard Cartun" Subject: Re: [Histonet] pancreas tissue To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I can. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> Fightmaster Janice 08/20/04 12:44PM >>> I am in need of a block of normal pancreas tissue for IHC controls. Can anyone help me? Janice Fightmaster Pathology Supervisor Oak Hill Hospital 11375 Cortez Blvd. Brooksville, FL 34613 (352)597-6363 Ext. 3636 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Fri, 20 Aug 2004 12:31:55 -0500 From: Fightmaster Janice Subject: [Histonet] pancreas tissue To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <1734C20AE655CD449599A15EDC589D89017CA59D@orlex05.hca.corpad.net> Content-Type: text/plain; charset="iso-8859-1" Thanks for the responses; I have plenty on the way now!! Janice Fightmaster Pathology Supervisor Oak Hill Hospital 11375 Cortez Blvd. Brooksville, FL 34613 (352)597-6363 Ext. 3636 ------------------------------ Message: 4 Date: Fri, 20 Aug 2004 13:30:45 -0500 From: hymclab Subject: RE: [Histonet] doing immunos by hand To: "'Edmondson David (RBV) NHS Christie Tr'" , "'Pathologyarts@aol.com'" Cc: "Histonet \(E-mail 2\)" Message-ID: Content-Type: text/plain I have also used the Shandon sequenza for my manual immunos. We have used it for 10 years and love it!!! Dawn Schneider, HT(ASCP) Howard Young Medical Center Woodruff, WI -----Original Message----- From: Edmondson David (RBV) NHS Christie Tr [mailto:David.Edmondson@christie-tr.nwest.nhs.uk] Sent: Friday, August 20, 2004 11:29 AM To: 'Pathologyarts@aol.com' Cc: Histonet (E-mail 2) Subject: RE: [Histonet] doing immunos by hand Hi, We use Shando's "Sequenza" to do only a small number of slides these days since a DAKO Techmate came along. The Sequenza has the option of an organiser for several sets of slides but this is not necessary for small numbers of slides The slides are arrayed each against plastic plates, in groups of ten. The plates forms a well at one end and solutions slowly drain past, accross the face of the slides, buffers antibodies, chromogens , whatever. In terms of a manual system it is sweeter than using trays, boxes. Once set in the clip there is no handling until the end. The waste is held in the container, staining is even , no ringing of the sections with a pen, sweet. Having done batches of a hundred with trays and then the sequenza, my money is on the latter. David Edmondson Christie Hospital Manchester UK -----Original Message----- From: Pathologyarts@aol.com [mailto:Pathologyarts@aol.com] Sent: 19 August 2004 18:59 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] doing immunos by hand does anyone still do immunos by hand or are they all done with these $40 billion automated machines? i would like to see what the options are as far as doing these things manually. primarily for derm specimens, HMB 45, S 100, KI 67. any help is appreciated, curt _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Fri, 20 Aug 2004 20:28:42 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: IHC polymer detection system for rat tissues To: histonet@lists.utsouthwestern.edu Cc: rpw4@psu.edu Message-ID: Content-Type: text/plain; charset=us-ascii Dear Ronald, Just use the anti-mouse EnVision for use on rat tissue specimens. However, before using it you have to mix the EnVision with 10-15% (final concentration) normal rat serum. Leave the mixture for 60 min at room temp (or overnight 4C) then use it. All the cross-reactions are abolished now. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Ronald P. Wilson" Date Thu, 19 Aug 2004 15:15:43 -0400 To Histonet Subject [Histonet] IHC polymer detection system for rat tissues Has anyone found a successful polymer detection system (i.e., Envision or others) directed against a mouse monoclonal on rat tissue. We tried one but got a lot of background staining, I suspect from cross reactivity with closely related antigens. Our standard IHC approach uses an anti-mouse, rat-absorbed secondary antibody, but I would like to get the added sensitivity of a polymer-based system. Ronald P. Wilson, V.M.D., M.S. Associate Professor Department of Comparative Medicine, HO54 Penn State University College of Medicine M. S. Hershey Medical Center 500 University Drive Hershey, PA 17033 phone: 717-531-8460 fax: 717-531-5001 e-mail: rpw4@psu.edu ------------------------------ Message: 6 Date: Fri, 20 Aug 2004 18:33:22 GMT From: "madaryhtl@juno.com" Subject: [Histonet] Hand IHC To: histonet@lists.utsouthwestern.edu Message-ID: <20040820.113415.3885.451884@webmail23.lax.untd.com> Content-Type: text/plain 40 Billion is alot to spend on a machine, I would shop around. Sequenza chambers serve us quite well as well as the automation. All the stains you mentioned are easily accomplished using hand techniqes. Nick(Rocky) Madary, HT,HTL(ASCP)QIHC Histology Manager, Medimmune Inc Cell-3012334950 Work-3013984745 Fax-3013989745 ------------------------------ Message: 7 Date: Fri, 20 Aug 2004 14:36:48 -0400 From: "Lesley S. Bechtold" Subject: [Histonet] Desperately seeking..information To: histonet@Pathology.swmed.edu Message-ID: <6.0.3.0.0.20040820142237.0261f600@aretha.jax.org> Content-Type: text/plain; charset="us-ascii"; format=flowed Hi Everyone, Up here in Bar Harbor, Maine, we are trying to get an idea of whato ther Histology Labs are doing - how many people they have working in them,w hat they consider reasonable capacity, how much work they do in a year, how diversified they are, etc. We are not interested in salary information. We have created a bench-marking survey on our home website ath ttp://www.jax.org/sci_survey/ . Click on the "Histology" link and it willt ake you right to the survey. You'll notice there are other surveys on there as well so if your institution/hospital/business carries out any other activities (such as Electron Microscopy), please feel free to complete those surveys as well. It only takes a few minutes to complete and will remain completely confidential. Anyone who participates in the survey will receive a copy of the results (with names, etc. deleted) withina few weeks. We're aiming for the beginning of October to send the resultst o all the participants. We will need your email address in order to sendt he results to you so make sure you include that. So we'd really like to hear from you. I know how active the Histonet is so I'm hoping for a good response as I'm sure everyone will bei nterested in the results. For anyone who is not interested in participating, I apologize for using up your time. Please delete this email. Thank you! Lesley Bechtold Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191 ------------------------------ Message: 8 Date: Fri, 20 Aug 2004 14:42:32 -0400 From: "Kopczynski, Charlotte" Subject: [Histonet] Drying slides before Filing To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <7C3A384EF3ABD611B3270008023E3E030481CC1C@mphexch01.mpm.baycare.internal> Hello fellow Histonetters, We are having a big problem with air bubbles under our coverslips before filing. We use Richard Allan mounting media and the slides in the trays are placed in an oven set at 58 degrees C overnight. I am thinking the oven may be part of the problem but was wondering what the process is out there in Histo-land. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 **************************************************************************** ********************** The contents of this email and any attachments are confidential. They are intended for the named recipient(s) only. If you have received this email in error please notify the system manager or the sender immediately and do not disclose the contents to anyone or make copies. ** eSafe scanned this email for viruses, vandals and malicious content. ** **************************************************************************** ********************** ------------------------------ Message: 9 Date: Fri, 20 Aug 2004 13:54:08 -0500 From: Gary Gill Subject: RE: [Histonet] Drying slides before Filing To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <0B11620AEE5EFB4FA7A7558339972A830704F3@HALIBUT.dcla.com> Content-Type: text/plain Air bubbles form because there is not enough solid material remaining under the cover glass after the solvent has evaporate. Among possible causes: * RAS put out a bad batch of mounting medium (i.e., more solvent than usual) * Not enough mounting medium being applied * Accelerated drying (e.g., oven too hot [58 degrees C is not too hot, in my experience]) -- how soon after coverslipping are slides put in oven? Right away could make a negative difference. Simplest first solution is to add more mounting medium, while appreciating that way too much degrades image quality under 40x objectives -- which would be the subject of another discussion. Little useful information has ever been published about mounting media. The best information was published as a 2-part article in Stain Technology in the 1950s. Authors were a committee chaired by the late and great Dr. Ralph D. Lillie. Gary Gill -----Original Message----- From: Kopczynski, Charlotte [mailto:Charlotte.Kopczynski@baycare.org] Sent: Friday, August 20, 2004 1:43 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Drying slides before Filing Hello fellow Histonetters, We are having a big problem with air bubbles under our coverslips before filing. We use Richard Allan mounting media and the slides in the trays are placed in an oven set at 58 degrees C overnight. I am thinking the oven may be part of the problem but was wondering what the process is out there in Histo-land. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 **************************************************************************** ********************** The contents of this email and any attachments are confidential. They are intended for the named recipient(s) only. If you have received this email in error please notify the system manager or the sender immediately and do not disclose the contents to anyone or make copies. ** eSafe scanned this email for viruses, vandals and malicious content. ** **************************************************************************** ********************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Fri, 20 Aug 2004 15:21:37 -0400 From: Kevin Briggs Subject: RE: [Histonet] Drying slides before Filing To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <8A37FE7D529ED411983E00D0B75D01FC0663A79C@sirius.drmc.drhsi.org> Content-Type: text/plain; charset="iso-8859-1" Charlotte, I think Gary's got it. 58 degrees C should only be too hot if the slides were placed in the oven too soon after mounting as we have found--although, decreasing the temperature will indeed slow the solvent's rate of evaporation. Try delaying your oven cure time and see if that helps. Good luck! Kevin D. Briggs, MS,CT(ASCP) Team Leader- Cytopathology/Histopathology Services Danville Regional Medical Center 142 South Main Street Danville, VA 24541 Telephone: (434) 799-4470 ext.5451 Fax: (434) 799-2118 E-mail: briggsk@drhsi.org -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Friday, August 20, 2004 2:54 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Drying slides before Filing Air bubbles form because there is not enough solid material remaining under the cover glass after the solvent has evaporate. Among possible causes: * RAS put out a bad batch of mounting medium (i.e., more solvent than usual) * Not enough mounting medium being applied * Accelerated drying (e.g., oven too hot [58 degrees C is not too hot, in my experience]) -- how soon after coverslipping are slides put in oven? Right away could make a negative difference. Simplest first solution is to add more mounting medium, while appreciating that way too much degrades image quality under 40x objectives -- which would be the subject of another discussion. Little useful information has ever been published about mounting media. The best information was published as a 2-part article in Stain Technology in the 1950s. Authors were a committee chaired by the late and great Dr. Ralph D. Lillie. Gary Gill -----Original Message----- From: Kopczynski, Charlotte [mailto:Charlotte.Kopczynski@baycare.org] Sent: Friday, August 20, 2004 1:43 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Drying slides before Filing Hello fellow Histonetters, We are having a big problem with air bubbles under our coverslips before filing. We use Richard Allan mounting media and the slides in the trays are placed in an oven set at 58 degrees C overnight. I am thinking the oven may be part of the problem but was wondering what the process is out there in Histo-land. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 **************************************************************************** ********************** The contents of this email and any attachments are confidential. They are intended for the named recipient(s) only. If you have received this email in error please notify the system manager or the sender immediately and do not disclose the contents to anyone or make copies. ** eSafe scanned this email for viruses, vandals and malicious content. ** **************************************************************************** ********************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Fri, 20 Aug 2004 22:26:42 -0400 From: "Histo Jock" Subject: RE: [Histonet] Image analysis and slide scanning machine To: histonet@lists.utsouthwestern.edu Cc: gsmith@confocal.com Message-ID: Content-Type: text/plain; format=flowed Can you describe more about how this works? How does it create the image ifi t doesn't tile? What kind of analysis can it do? You're exhibiting with Beecher - can it run Beecher's new software? HistoJock. >Message: 9 >Date: Mon, 16 Aug 2004 16:11:45 -0400 >From: "Glenn Smith" >Subject: [Histonet] Image analysis and slide scanning machine (Wester, > Martha) >To: >Message-ID: <00c101c483cd$45edc040$1f05a8c0@confocal.com> >Content-Type: text/plain; charset="US-ASCII" > >Hi Kim, > >We supply such an instrument, our TISSUEscope - combining confocal scanning >laser microscopy and a wide field of view. Depending on your resolution >requirements, the TISSUEscope can acquire a single image at up to 1 um >resolution with no mosaic and no tiling of images. A standard slide (1" x >3") is loaded into the machine and the rest of operations is via software. >We have another model which offers submicron resolution as well. Both >brightfield and fluorescence detection is provided within the same >instrument. Please feel free to contact me offline for more details. > >Also, we will have one of these instruments at the NSH meeting in Toronto - >we'll be exhibiting in conjunction with Beecher Instruments. > >Regards >Glenn Smith, P.Eng. >519.886.9013 x38 > >Biomedical Photometrics Inc/GeneFocus www.GeneFocus.com> >Widefield Confocal Scanning Instruments and Software _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar - get it now! http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/ ------------------------------ Message: 12 Date: Sat, 21 Aug 2004 11:14:12 +0100 From: "Edmondson David (RBV) NHS Christie Tr" Subject: [Histonet] RE: Old IMLT journals To: 'Bryan Llewellyn' Cc: "Histonet \(E-mail 2\)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" OK Bryan, It will take a little while, but I shall start. I wonder that someone has already scanned some journals, like "path and bact", stain tech and histochem journal. Or at least found the annual indexes to scan. There are Stain tech 69-83 and Histochen J 79-86 beside me now, and I wonder at the quantity of published material. Maybe I shall ask the IBMS, I remember there is/was a historical section that may have moved from preservation to digitising. Not the same as "Brass and Glass", but. Dave -----Original Message----- From: Bryan Llewellyn [mailto:bryand@netbistro.com] Sent: 20 August 2004 20:41 To: Edmondson David (RBV) NHS Christie Tr Subject: Old IMLT journals Hi, I have a big favour to ask. I am the author of the StainsFile web site (http://stainsfile.info). One of my aims for this site is to include all published staining methods. I know that's quite futile, but I would like to get as many as possible. I have the IMLT journals between 1962 and about 1975, when I let my membership finally lapse after emigrating to Canada. Is there any possibility that I could get you to scan or photocopy any histological methods involving techniques or theory about dyes or metallic impregnations (almost everything except immuno- or enzyme histochemistry as these are outside my limits for the site) from any journals you have apart from those years? I would like to be able to include the material on StainsFile, and I have been wondering how I might be able to get details from Canada. Unfortunately, none of the British Techs I know over here has kept their copies, or have let their membership lapse. Here's hoping. Bryan Llewellyn Edmondson David (RBV) NHS Christie Tr wrote: > John, > I have all the back issues of the IMLT/IMLS Journal so will have look, but > slowly, over a glass. ( I inherited them, honest ) > Dave > > -----Original Message----- > From: JOHN PHILLIPS [mailto:JOHN.PHILLIPS@new-tr.wales.nhs.uk] > Sent: 19 August 2004 14:33 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Spirit fixation of tissue > ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 9, Issue 34 *************************************** LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From Ian.Bernard <@t> LACKLAND.AF.MIL Mon Aug 23 09:15:52 2004 From: Ian.Bernard <@t> LACKLAND.AF.MIL (Bernard Ian R SSgt 59 CRES/MSROP) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Please unsubscribe. Message-ID: <0F2AB4D3260BB1428CE3928748F7DA1060F08F@fsmpls17.whmc.af.mil> Please unsubscribe. I have this access on my homew email address, for those who wish to keep in contact. From dlcowie <@t> prodigy.net Mon Aug 23 09:56:35 2004 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] histology position open Message-ID: <20040823145635.34362.qmail@web81003.mail.yahoo.com> hello netters, I have a histotechnician position open in Pensacola Florida. Full time, Monday thru Friday with rotating Saturdays (1 in every 5). Day time hours. Excellent pay and benefits. Florida license as HTL (preferred) or HT. ASCP certification preferred. We are located in the panhandle area of Florida with beautiful beaches and lots of sunshine. We are a full service lab with 6 pathologists, 3 PA's, 6 histotechs, 5 cytotechs and a variety of lab aides. Experience with embedding, routine H&E, special stains and IHC required. If interested or for more information, please contact me as follows. Dawn Cowie, HT (ASCP) Histology Supv Pensacola Pathologists, PA 850-416-7251 phone 850-416-4471 fax or email me at dlcowie@prodigy.net From gsmith <@t> confocal.com Mon Aug 23 10:32:05 2004 From: gsmith <@t> confocal.com (Glenn Smith) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Image analysis and slide scanning machine In-Reply-To: Message-ID: <002801c48926$5d07eea0$1f05a8c0@confocal.com> Sure Histo! First the slide is inserted much the same way you would introduce your bank card to an ATM. From there, everything is software driven including defining the area you want to scan - entire slide or some subsection - , the resolution settings, which light sources (various fluorescence or brightfield) and other relevant parameters. Then simply hit scan and the image is acquired (time for acquisition is seconds or minutes depending on the resolution and area desired). I would say that image analysis offered with the software is limited - we often bundle other software as desired. Hence our desire to work with partners in various segments like Beecher. The aquired image is a TIFF format file importable to other preferred software. Re how the TISSUEscope creates images without tiling, our proprietary MACROscope lens technology gives us the ability to aquire a single image over 10-20X scan area of that of high-end microscopes at the same resolution. A scanning mirror is placed at the entrance pupil position and the beam is scanned from left to right (x-direction) across the slide. While the beam moves from left to right, we take data at a constant rate. At the same time, the slide is slowly moved under the scanning beam, resulting in a raster scan of the slide contents. I can provide more details via a ppt or white paper that I can send offline (~ 0.6 Mb) if you're hotmail account can accommodate? Re Beecher, they ran a first test of their software on some of our Tissue Array images generated with our 2 um resolution lens earlier this summer. On a first test on their automated analysis software with our images, they could detect cells and get measurements like nuclei count and identify general tissue regions. Based on the quality of the images we sent them, this speaks well to their software. (there was no IHC staining to quantitate on the sample we sent). We have been grateful for their feedback to improve the instrument characteristics and continue to work with them. Glenn Smith, P.Eng. 519.886.9013 x38 Biomedical Photometrics Inc/GeneFocus Widefield Confocal Scanning Instruments and Software ------------------------------ Message: 11 Date: Fri, 20 Aug 2004 22:26:42 -0400 From: "Histo Jock" Subject: RE: [Histonet] Image analysis and slide scanning machine To: histonet@lists.utsouthwestern.edu Cc: gsmith@confocal.com Message-ID: Content-Type: text/plain; format=flowed Can you describe more about how this works? How does it create the image if it doesn't tile? What kind of analysis can it do? You're exhibiting with Beecher - can it run Beecher's new software? HistoJock. >Message: 9 >Date: Mon, 16 Aug 2004 16:11:45 -0400 >From: "Glenn Smith" >Subject: [Histonet] Image analysis and slide scanning machine (Wester, > Martha) >To: >Message-ID: <00c101c483cd$45edc040$1f05a8c0@confocal.com> >Content-Type: text/plain; charset="US-ASCII" > >Hi Kim, > >We supply such an instrument, our TISSUEscope - combining confocal >scanning laser microscopy and a wide field of view. Depending on your >resolution requirements, the TISSUEscope can acquire a single image at >up to 1 um resolution with no mosaic and no tiling of images. A >standard slide (1" x >3") is loaded into the machine and the rest of operations is via software. >We have another model which offers submicron resolution as well. Both >brightfield and fluorescence detection is provided within the same >instrument. Please feel free to contact me offline for more details. > >Also, we will have one of these instruments at the NSH meeting in >Toronto - we'll be exhibiting in conjunction with Beecher Instruments. > >Regards >Glenn Smith, P.Eng. >519.886.9013 x38 > >Biomedical Photometrics Inc/GeneFocus www.GeneFocus.com> Widefield Confocal Scanning Instruments and Software _________________________________________________________________ From settembr <@t> umdnj.edu Mon Aug 23 11:31:50 2004 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] doing immunos by hand Message-ID: Go with Shandon. Although I believe their name is not ThermoShandon or even ThermoElectric. Buy their racks and coverplates. Get a squirt bottle. It is sweet. I use it when not using my $40 billion machine Dana Settembre University Hospital - UMDNJ Newark, NJ >>> 8/19/2004 1:59:05 PM >>> does anyone still do immunos by hand or are they all done with these $40 billion automated machines? i would like to see what the options are as far as doing these things manually. primarily for derm specimens, HMB 45, S 100, KI 67. any help is appreciated, curt _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dorothy.L.Webb <@t> HealthPartners.Com Mon Aug 23 11:47:51 2004 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Leica 1850 Message-ID: We had nothing but "chatter" trouble and other cutting concerns with a new Leica 1850 at the facility we used to work at. They even had the rep from Germany in and ended up giving us a new cryostat, same model. We still had periodic problems with it and have been told that the new knife holder design for this cryostat has some design flaws! So, we went back to an older model knife holder and that kept our cutting concerns to a minimum..still not perfect! At the facility I work at now, they purchased the same model and we have the same problems, mostly with skin specimens. The tech who works on this cryostat is very good from a local company and he has given us an older knife holder for this one, too, and recently found another internal problem that he is ordering parts for. I can let you know exactly what that is when he comes back to fix it. It is a nice cryostat, but it bothers me why there are so many cutting concerns with this model??? ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From LBlack <@t> carilion.com Mon Aug 23 12:44:31 2004 From: LBlack <@t> carilion.com (Lisa Black) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Leica 1850 Message-ID: There are 2 Leica cryostats in my facility. They are used frequently and are very reliable. Lisa Black Histology Supervisor Carilion Consolidated Laboratory Roanoke, VA LBLACK@CARILION.COM From mark.lewis <@t> thermo.com Mon Aug 23 13:43:44 2004 From: mark.lewis <@t> thermo.com (mark.lewis@thermo.com) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] doing immunos by hand Message-ID: We are Thermo Electron ...formerly Thermo Shandon, Shandon Lipshaw, Shandon..... The racks are called Sequenza racks. Any questions, give me a call. Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com Dana Settembre To: Pathologyarts@aol.com, histonet@lists.utsouthwestern.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: Re: [Histonet] doing immunos by hand western.edu 08/23/2004 12:31 PM Go with Shandon. Although I believe their name is not ThermoShandon or even ThermoElectric. Buy their racks and coverplates. Get a squirt bottle. It is sweet. I use it when not using my $40 billion machine Dana Settembre University Hospital - UMDNJ Newark, NJ >>> 8/19/2004 1:59:05 PM >>> does anyone still do immunos by hand or are they all done with these $40 billion automated machines? i would like to see what the options are as far as doing these things manually. primarily for derm specimens, HMB 45, S 100, KI 67. any help is appreciated, curt _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From HornHV <@t> archildrens.org Mon Aug 23 14:16:06 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Leica 1850 Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B92D@EMAIL.archildrens.org> I have an 1850 and have had none of these concerns. Ours has worked well for us. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dorothy.L.Webb@HealthPartners.Com Sent: Monday, August 23, 2004 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Leica 1850 We had nothing but "chatter" trouble and other cutting concerns with a new Leica 1850 at the facility we used to work at. They even had the rep from Germany in and ended up giving us a new cryostat, same model. We still had periodic problems with it and have been told that the new knife holder design for this cryostat has some design flaws! So, we went back to an older model knife holder and that kept our cutting concerns to a minimum..still not perfect! At the facility I work at now, they purchased the same model and we have the same problems, mostly with skin specimens. The tech who works on this cryostat is very good from a local company and he has given us an older knife holder for this one, too, and recently found another internal problem that he is ordering parts for. I can let you know exactly what that is when he comes back to fix it. It is a nice cryostat, but it bothers me why there are so many cutting concerns with this model??? ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From waltersk <@t> mail.medicine.uiowa.edu Mon Aug 23 14:18:24 2004 From: waltersk <@t> mail.medicine.uiowa.edu (Walters, Katherine S) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] looking to purchase sliding microtome and slide dryer Message-ID: Our lab is in the market to purchase a sliding microtome with cold knife capabilities. I am also looking for a slide dryer for drying slides (flat) in about 1/2 an hour. The more slides the better. I remember seeing a tri-level contraption (for lack of a better name) at an NSH meeting that could heat slides at 3 different temperatures, but I don't remember the company, does anyone else remember that? Thanks for any info, Kathy From t-sherman <@t> comcast.net Mon Aug 23 14:46:29 2004 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Re: doing immunos by hand (renton louise mrs) Message-ID: <412A4995.2090104@comcast.net> -----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Well I must say, I'm impressed! How many people did you have performing this many immunos? And how many different proteins were you trying to detect on a "typical" run? With all of the slide handling and epitope retrieval steps, not to mention blocking and counterstaining, I don't see how this was done in an 8hr day. Did you all wear capes? Regards, Todd Todd Sherman President HistoSoft Corporation "Biology in a new form..." Home: www.histosoft.com Member Services: www.myhistosoft.com histonet-request@lists.utsouthwestern.edu wrote: | | Today's Topics: | 4. Re: doing immunos by hand (renton louise mrs) | | ---------------------------------------------------------------------- | | Message: 4 | Date: Mon, 23 Aug 2004 10:13:47 +0200 | From: "renton louise mrs" | Subject: Re: [Histonet] doing immunos by hand | To: histonet@lists.utsouthwestern.edu | Cc: histonet@lists.utsouthwestern.edu | Message-ID: <1093248827.921a1660rentonlf@bru.wits.ac.za> | Content-Type: text/plain; charset="UTF-8" | | Doing immuno manually is entirely dependent on the volume. In a lab where | I worked previously, we would do up to 400 slides by hand (we had a panel | of over 100 diffent immunos to choose from). This was 8hrs HARD work. We | had specially constructed perspex boxes to use as a moisture chamber, but | in emergencies, large oblong Tupperware containers with moistened filter | paper or roller towel were also used. The slides would be raised from the | paper by means of blank slides placed at right angles to the immuno slide | so as to avoid breaking nails when trying to overcome the surface | tensiion that would suck the slide down onto the surface. | | best regards | | | -----Original Message----- | From: Pathologyarts@aol.com | To: histonet@lists.utsouthwestern.edu | Date: Thu, 19 Aug 2004 13:59:05 EDT | Subject: [Histonet] doing immunos by hand | | does anyone still do immunos by hand or are they all done with these $40 | billion automated machines? | | i would like to see what the options are as far as doing these things | manually. | | primarily for derm specimens, HMB 45, S 100, KI 67. | | | any help is appreciated, | curt | _______________________________________________ | Histonet mailing list | Histonet@lists.utsouthwestern.edu | http://lists.utsouthwestern.edu/mailman/listinfo/histonet | | Louise Renton | Bone Research Unit | University of the Witwatersrand | Johannesburg | South Africa | .......so what IS the speed of dark? | | ------------------------------ -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.2.3 (MingW32) Comment: Using GnuPG with MultiZilla - http://enigmail.mozdev.org iD8DBQFBKkmTEmHXdrslGRcRAtKvAKDMYgJGqsWquGWfgLBgpQnIkJA4sQCfSj43 VlxVX16eKZYr7TkWvARNjQ0= =Wh6X -----END PGP SIGNATURE----- From POWELL_SA <@t> Mercer.edu Mon Aug 23 15:09:00 2004 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Re: doing immunos by hand (renton louise mrs) In-Reply-To: <412A4995.2090104@comcast.net> Message-ID: As a matter of fact, yes. :) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Todd Sherman Sent: Monday, August 23, 2004 2:46 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: doing immunos by hand (renton louise mrs) -----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Well I must say, I'm impressed! How many people did you have performing this many immunos? And how many different proteins were you trying to detect on a "typical" run? With all of the slide handling and epitope retrieval steps, not to mention blocking and counterstaining, I don't see how this was done in an 8hr day. Did you all wear capes? Regards, Todd Todd Sherman President HistoSoft Corporation "Biology in a new form..." Home: www.histosoft.com Member Services: www.myhistosoft.com histonet-request@lists.utsouthwestern.edu wrote: | | Today's Topics: | 4. Re: doing immunos by hand (renton louise mrs) | | ---------------------------------------------------------------------- | | Message: 4 | Date: Mon, 23 Aug 2004 10:13:47 +0200 | From: "renton louise mrs" | Subject: Re: [Histonet] doing immunos by hand | To: histonet@lists.utsouthwestern.edu | Cc: histonet@lists.utsouthwestern.edu | Message-ID: <1093248827.921a1660rentonlf@bru.wits.ac.za> | Content-Type: text/plain; charset="UTF-8" | | Doing immuno manually is entirely dependent on the volume. In a lab where | I worked previously, we would do up to 400 slides by hand (we had a panel | of over 100 diffent immunos to choose from). This was 8hrs HARD work. We | had specially constructed perspex boxes to use as a moisture chamber, but | in emergencies, large oblong Tupperware containers with moistened filter | paper or roller towel were also used. The slides would be raised from the | paper by means of blank slides placed at right angles to the immuno slide | so as to avoid breaking nails when trying to overcome the surface | tensiion that would suck the slide down onto the surface. | | best regards | | | -----Original Message----- | From: Pathologyarts@aol.com | To: histonet@lists.utsouthwestern.edu | Date: Thu, 19 Aug 2004 13:59:05 EDT | Subject: [Histonet] doing immunos by hand | | does anyone still do immunos by hand or are they all done with these $40 | billion automated machines? | | i would like to see what the options are as far as doing these things | manually. | | primarily for derm specimens, HMB 45, S 100, KI 67. | | | any help is appreciated, | curt | _______________________________________________ | Histonet mailing list | Histonet@lists.utsouthwestern.edu | http://lists.utsouthwestern.edu/mailman/listinfo/histonet | | Louise Renton | Bone Research Unit | University of the Witwatersrand | Johannesburg | South Africa | .......so what IS the speed of dark? | | ------------------------------ -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.2.3 (MingW32) Comment: Using GnuPG with MultiZilla - http://enigmail.mozdev.org iD8DBQFBKkmTEmHXdrslGRcRAtKvAKDMYgJGqsWquGWfgLBgpQnIkJA4sQCfSj43 VlxVX16eKZYr7TkWvARNjQ0= =Wh6X -----END PGP SIGNATURE----- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Aug 23 15:54:30 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Problems with Leica 1850 In-Reply-To: References: Message-ID: <6.0.0.22.1.20040823144156.01b2c9d0@gemini.msu.montana.edu> Interesting, we have three 1850's and never have cutting concerns with them. We cut snap frozen bovine skin (tough leathery type) innumerable tissues from mouse, rat, hamster, decalcified and undecalcified bone, thick 50 um brain and have sectioned human tissues without problems and some exotic other things like biofilms. Maybe it is more than just instrument problems and your cryosectioning technics (blades, temperatures, etc) that needs reevaluation. If you use low profile blades exclusively and have not tried high profile blades, you should TRY the high profile, and try other brands of knife blades. Some blades perform much better than others. Accuedge high profiles are just as sharp as their low profile blades but much sturdier for cryomicrotomy. At 10:47 AM 8/23/2004, you wrote: >We had nothing but "chatter" trouble and other cutting concerns with a new >Leica 1850 at the facility we used to work at. They even had the rep from >Germany in and ended up giving us a new cryostat, same model. We still had >periodic problems with it and have been told that the new knife holder >design for this cryostat has some design flaws! So, we went back to an >older model knife holder and that kept our cutting concerns to a >minimum..still not perfect! At the facility I work at now, they purchased >the same model and we have the same problems, mostly with skin specimens. >The tech who works on this cryostat is very good from a local company and he >has given us an older knife holder for this one, too, and recently found >another internal problem that he is ordering parts for. I can let you know >exactly what that is when he comes back to fix it. > >It is a nice cryostat, but it bothers me why there are so many cutting >concerns with this model??? > >________________________________________ > >This e-mail and any files transmitted with it are confidential and are >intended solely for the use of the individual or entity to whom they >are addressed. If you are not the intended recipient or the individual >responsible for delivering the e-mail to the intended recipient, please >be advised that you have received this e-mail in error and that any >use, dissemination, forwarding, printing, or copying of this e-mail >is strictly prohibited. > >If you have received this e-mail in error, please immediately notify >the HealthPartners Support Center by telephone at (952) 967-6600. >You will be reimbursed for reasonable costs incurred in notifying us. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From rockbeki <@t> ufl.edu Mon Aug 23 16:07:12 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] MMP-1 immuno labelling of placenta Message-ID: <970777571.1093295232555.JavaMail.osg@osgjas02.cns.ufl.edu> I've been doing eNOS staining on placenta and I found it helpful to block in 1:10 30%peroxide to methanol for 1hr (to get rid of endogenous peroxidase-I know most people probably don't do it for that long though, but shorter times didn't help) and NaOH for 15 min to deal with the blood cells staining.Good luck! I feel your pain:-) Rebekah Smith On Mon Aug 16 11:30:51 EDT 2004, "Y. Wang" wrote: > Hello Louise, > > Thank you for your suggestions. > 1) Seeing as we could not go to the farm to collect the tissue > the tissue > was kept on ice (~5 hours) while it was shipped to us and > immediately > embedded. Do you think this is a problem? > 2)We have not done any biotin blocking for these samples. Thank > you for > suggesting that. We will try this next. > > Thanks again > Yak-Nam Wang > > > On Mon, 16 Aug 2004, renton louise mrs wrote: > >> 1. Was the tissue fresh when sampled? autolysis can alter IHC >> dramatically >> 2.Placenta contains a lot of biotin...are you blocking >> adequately? >> >> Regards >> >> >> >> -----Original Message----- >> From: "Y. Wang" >> To: Histonet@lists.utsouthwestern.edu >> Date: Fri, 13 Aug 2004 14:35:07 -0700 (PDT) >> Subject: [Histonet] MMP-1 immuno labelling of placenta >> >> Hello, >> >> Has anyone had any experience immuno labelling placenta tissue >> with >> anti-MMP1? We have tried this (cryosections with ABC kit with >> DAB) but are >> getting alot of (what we assume to be) background and nothing >> that looks >> like images we have seen on-line. Seeing as we have not had any >> experience >> with placenta tissue we are questioning if we are took the >> tissue from the >> correct place from the placenta (we were given a whole one which >> we took >> random samples from). >> >> Thank you for your help >> Yak-Nam Wang >> >> Senior Fellow >> Department of Bioengineering >> University of Washington >> Box 357962 >> Seattle, WA 98195 >> >> Tel.: (206)-221-5873 >> Fax.: (206)-221-5874 >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> Louise Renton >> Bone Research Unit >> University of the Witwatersrand >> Johannesburg >> South Africa >> .......so what IS the speed of dark? >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From NSEARCY <@t> swmail.sw.org Tue Aug 24 06:59:26 2004 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Screened Cassettes Message-ID: How many manufacturers supply screened cassettes? I am now using Stat Lab and have requested samples from Richard Allan (Cardinal) and Surgipath. Any others out there? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD From Julie.Sanders <@t> med.va.gov Tue Aug 24 07:17:22 2004 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] decal of bone marrows Message-ID: <457381D92B01BD44B21CF37CC02EBDFD28E912@vhacinexc2.v10.med.va.gov> I am wondering what everyone uses to decal bone marrow cores. We have been having trouble with immunos and losing cell architecture and I think it's because of how we decal. What is everyone else using? Also, we use B-5 at the pathologist's request, but is anyone using anything comparable (same cell clarity, etc.)? Thanks, Julie Julie Sanders Supervisor, Anatomic Pathology VAMC, Cincinnati, Ohio From failm <@t> musc.edu Tue Aug 24 08:12:50 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] decal of bone marrows Message-ID: Julie, We are using Formic acid, with good results on IHC. Rena >>> 08/24/04 08:17AM >>> I am wondering what everyone uses to decal bone marrow cores. We have been having trouble with immunos and losing cell architecture and I think it's because of how we decal. What is everyone else using? Also, we use B-5 at the pathologist's request, but is anyone using anything comparable (same cell clarity, etc.)? Thanks, Julie Julie Sanders Supervisor, Anatomic Pathology VAMC, Cincinnati, Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Myri37 <@t> aol.com Tue Aug 24 08:36:41 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] BrdU Message-ID: <74B4ECC8.53625E49.0005167B@aol.com> Hi everyone Does someone have a protocol of detection of BrdU incorporation in DNA synthesizing cells ? I would like to try it on fresh fixed cells. Thank you very much in advance. Myriam Baali Natural implant From mprice26 <@t> juno.com Tue Aug 24 09:39:58 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] RE: Microwaveable slide racks Message-ID: <20040824.074044.6762.398869@webmail27.nyc.untd.com> Histonetters, Does anyone know where I can purchase some microwaveable slide racks to dry slides in for IHC? Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From gcallis <@t> montana.edu Tue Aug 24 09:57:48 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] decal of bone marrows In-Reply-To: References: Message-ID: <6.0.0.22.1.20040824085645.01b35300@gemini.msu.montana.edu> Question: Buffered formic acid, commercial preparations? or do you make up an inhouse preparation? At 07:12 AM 8/24/2004, you wrote: >Julie, > We are using Formic acid, with good results on IHC. >Rena > > >>> 08/24/04 08:17AM >>> Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From failm <@t> musc.edu Tue Aug 24 10:10:01 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] decal of bone marrows Message-ID: Gayle, 10% aqueous solution made from 90% purchased Formic acid Rena Fail >>> Gayle Callis 08/24/04 10:57AM >>> Question: Buffered formic acid, commercial preparations? or do you make up an inhouse preparation? At 07:12 AM 8/24/2004, you wrote: >Julie, > We are using Formic acid, with good results on IHC. >Rena > > >>> 08/24/04 08:17AM >>> Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From NSEARCY <@t> swmail.sw.org Tue Aug 24 10:13:33 2004 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Powerpath Users Message-ID: Could I hear from Powerpath users? I know that here are many users out there. We are wanting to customize our histology worksheet (again) and want to see how others are being set-up. I have no name contacts presently, except for maybe Kim Simmons @ U of Washington?? Thanks From wecare <@t> qualityhistology.com Tue Aug 24 10:20:15 2004 From: wecare <@t> qualityhistology.com (wecare@qualityhistology.com) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] RE: Microwaveable slide racks References: <20040824.074044.6762.398869@webmail27.nyc.untd.com> Message-ID: <007601c489ee$5bbf4520$0efea8c0@internetconnect.net> Dear Ms. Price, We have some prototype slide racks right now and within next 2-3 weeks we will be in full production with microwavable slide carriers for our slide stainer. The rack will hold 30 slides and is fully microwavable. it will also fit in Hacker Instruments' as well as TBS' automatic coverslipper. For more information please call Preyas Shah @ 215-491-0081 1#. Preyas ----- Original Message ----- From: To: Sent: Tuesday, August 24, 2004 10:39 AM Subject: [Histonet] RE: Microwaveable slide racks > > Histonetters, > Does anyone know where I can purchase some microwaveable slide racks to dry slides in for IHC? > > Thank you. > > Marsha Price > > ________________________________________________________________ > The best thing to hit the Internet in years - Juno SpeedBand! > Surf the Web up to FIVE TIMES FASTER! > Only $14.95/ month - visit www.juno.com to sign up today! > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From RStaskiewicz <@t> agr.state.il.us Tue Aug 24 10:43:43 2004 From: RStaskiewicz <@t> agr.state.il.us (RStaskiewicz@agr.state.il.us) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Any teleconference schools out there? Message-ID: Are there any internet or teleconference schools for histology out there now? Please respond if there are any to jbordyn@aol.com. Thanks, Rae Ann Staskiewicz From Marjorie.Lehman <@t> unilever.com Tue Aug 24 10:53:03 2004 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Any teleconference schools out there? Message-ID: Please respond to Histonet also. Thanks -----Original Message----- From: RStaskiewicz@agr.state.il.us [SMTP:RStaskiewicz@agr.state.il.us] Sent: Tuesday, August 24, 2004 11:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Any teleconference schools out there? Are there any internet or teleconference schools for histology out there now? Please respond if there are any to jbordyn@aol.com. Thanks, Rae Ann Staskiewicz _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pcfanti <@t> buffalo.edu Tue Aug 24 11:05:50 2004 From: pcfanti <@t> buffalo.edu (pcfanti@buffalo.edu) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] IHC for Vit e Message-ID: <1093363550.412b675e47b49@mail3.buffalo.edu> I am doing IHC looking for Vitamin E in the corpus luteum of the rat ovary. The tissue is already paraffin embedded. We are attempting to locate a protocol to salvage the vitamin e during the deparifinization process. I would appreciate any advice. Thank you. Peter Fanti From gu.lang <@t> gmx.at Tue Aug 24 11:13:23 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] decal of bone marrows References: <457381D92B01BD44B21CF37CC02EBDFD28E912@vhacinexc2.v10.med.va.gov> Message-ID: <005a01c489f5$4d0a9af0$eeeea8c0@server> We use Tris-buffered EDTA 7,2 (Titriplex), 3-4 days. Gudrun Akh, Linz, Austria ----- Original Message ----- From: To: Sent: Tuesday, August 24, 2004 2:17 PM Subject: [Histonet] decal of bone marrows > > > I am wondering what everyone uses to decal bone marrow cores. We have been > having trouble with immunos and losing cell architecture and I think it's > because of how we decal. What is everyone else using? Also, we use B-5 at > the pathologist's request, but is anyone using anything comparable (same > cell clarity, etc.)? > Thanks, > Julie > > Julie Sanders > Supervisor, Anatomic Pathology > VAMC, Cincinnati, Ohio > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Ben.Shelkowsky <@t> chomp.org Tue Aug 24 12:03:10 2004 From: Ben.Shelkowsky <@t> chomp.org (Shelkowsky, Ben) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Screened Cassettes Message-ID: <384DA2BD670CE34DA8D3B60F4ED7157F4D6139@exchsrvr.chomp.org> Also... CMS (Fisher) has them in their inventory. I know I see about a dozen distributors carrying them at the National meeting. After the meeting in September I can give you the long list. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Nita Searcy Sent: Tue 8/24/2004 4:59 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Screened Cassettes How many manufacturers supply screened cassettes? I am now using Stat Lab and have requested samples from Richard Allan (Cardinal) and Surgipath. Any others out there? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 Confidentiality Notice: This is a transmission from Community Hospital of the Monterey Peninsula. This message and any attached documents may be confidential and contain information protected by state and federal medical privacy statutes. They are intended only for the use of the addressee. If you are not the intended recipient, any disclosure, copying, or distribution of this information is strictly prohibited. If you received this transmission in error, please accept our apologies and notify the sender. Thank you. From JonesLy <@t> mir.wustl.edu Tue Aug 24 12:53:17 2004 From: JonesLy <@t> mir.wustl.edu (JonesLy@mir.wustl.edu) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Questions on cryotome for large samples and autoradiography Message-ID: Hello - I'm new to the list and have little experience with thin slicing of tissues. (Our lab does some autoradiography but usually with 1 mm or larger slices.) We may be getting a new scanner and to go with it plan to purchase a cryotome. We definitely need to slice whole rat brains (fresh frozen) and would like the option of slicing whole mice. (Rats would be nice, but space and money are limited, which rules out the Leica Macrotomes.) Because we work with short-lived radioisotopes, speed is a very important factor, so I think we need to be able to do both thin slices and thicker ones (100-300 micron). We are considering the Hacker-Bright OTF cryotome. I don't know if there are other comparable instruments. I would appreciate any advice on options for freezing and sample handling as well as cryotome options. Suggestions for reference reading material would also be very helpful. The Macro Tape Transfer system looks nice, and seems to be a larger version of the CryoJane system. Any feedback from folks who use either system would be handy. How useful are they? I also have some very basic questions: How long does it take process a rat brain if you are doing 30 micron slices? Can you readily switch from 30 micron slices to 100-300 micron slices and back again? Can/how should/how long are brain slices stored for later use with receptor binding assays or autoradiography? (Since one rat brain should give lots of good 30 micron slices.) About how long does it take to slice a whole mouse, and what slice thickness do people use? Is there a fast and eays way to trim off the areas that aren't interesting? Thank you very much for your time, Lynne Jones Senior Research Technician Dept. of Radiological Sciences Washington University Medical School St. Louis, MO From caleri <@t> buffalo.edu Tue Aug 24 14:03:15 2004 From: caleri <@t> buffalo.edu (Katie Caleri) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Villanueva Osteochrome Bone Stain Message-ID: <412B90F3.5060807@buffalo.edu> Has anyone done the Villanueva Bone Stain from Polysciences, Inc. on decaled paraffin embedded sections? From Dndsomi <@t> aol.com Tue Aug 24 14:17:33 2004 From: Dndsomi <@t> aol.com (Dndsomi@aol.com) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] IHC Controls Message-ID: <1d8.29d3121c.2e5cee4d@aol.com> According to CAP guidelines, should you run a negative control with each case or can you use one negative control for each batch. Thanks for your help. Dianne D From RossS <@t> BaylorHealth.edu Tue Aug 24 14:22:46 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Blocks and slides for Research use Message-ID: What are other folks doing about requests for blocks or slides for research use? I am not talking about the patient getting approved for a clinical trial of a drug or a second opinion. We are getting swamped with requests for straight research from various groups. Some are internal to our organization, but most are from outside research groups. They all seem to want 30 unstained slides or the blocks. The latest one wants this from over 90 cases. We won't release the blocks at this time although our lawyers are looking into that possibility. These all have signed consents or I wouldn't even consider it. We have started trying to bill these research groups (not the patient) for our time. It is not for the patient's benefit, so if we can bill Lawyers for legal cases we can bill these places that want to create tissue banks on our slides. I have not one problem doing this, especially with the outrageous demands we have received lately. The other issue is that the patient's tissue that we are required to save for CAP regulations is being exhausted for these research institutions. Of course the reason for the regulations is so that if new drugs and tests are developed that the tissue is available to be tested, just like we all did when Her2Neu became available. Even with a consent, isn't it irresponsible to allow all of the tissue to be used by these research companies? Of course the other argument is without the research there will be no new tests to do on the tissue. Is anyone else experiencing this volume of research requests? What are you doing about it? Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From WWmn916 <@t> aol.com Tue Aug 24 15:18:05 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Re: Shurmark labeler Message-ID: <2843F00A.010C0D63.001A2AA1@aol.com> Hi Karen, I'm sorry to report (and hopefully do so without complaining) that I have not had a very good initial impression of Shurmark. The sales rep (when I can get a hold of him and trust what he says he will follow through on) has been difficult to see the machine. Too, bad, because the equipment itself looks good on paper. But sales reps do, in my opinion, sell themselves too. He tells me one thing, I update my boss, and then next time I talk with the sales rep, he tells me something different. After this much effort, I've decided to look at others. I will be able to tell you more about some of the competing labeling systems then. If you decide to check out Shurmark yourself, I'd be interested in what you experience with them is. Deb King, HT(ASCP) Sacramento, CA From gcallis <@t> montana.edu Tue Aug 24 15:41:45 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Villanueva Osteochrome Bone Stain In-Reply-To: <412B90F3.5060807@buffalo.edu> References: <412B90F3.5060807@buffalo.edu> Message-ID: <6.0.0.22.1.20040824143640.01b8bdf8@gemini.msu.montana.edu> No, it was designed for plastic sections (methylmethacrylate embedded). What do you want to demonstrate in bone? There are other stains that work well with decalcified bone instead of Villanueva stain. Movat's pentachrome is one. At 01:03 PM 8/24/2004, you wrote: >Has anyone done the Villanueva Bone Stain from Polysciences, Inc. on >decaled paraffin embedded sections? > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From WWmn916 <@t> aol.com Tue Aug 24 16:36:08 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] bar coding in histology Message-ID: <24118602.4D7B44C4.001A2AA1@aol.com> Greetings everyone, I've turned to the histonet a lot these days as I set up a new lab. I so appreciate the voluminous information that is shared on this site! On to my next question: are there many places using barcoding for cassettes/slides in histology? Do you like it? In what ways has barcoding made a difference? If not too involved to answer, what kinds of things do I need to consider for our software and lab info system? (We have an in-house developed software) I'm realizing what a major task this is to think about barcoding.....any help appreciated!!!! Deb King, HT(ASCP) Sacramento, CA From garygill <@t> dcla.com Tue Aug 24 16:45:02 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] bar coding in histology Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070522@HALIBUT.dcla.com> Bar Code Primer (40 pages, general, not histology per se): http://www.barcodehq.com/primer.pdf Gary Gill -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Tuesday, August 24, 2004 4:36 PM To: histonet@pathology.swmed.edu Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] bar coding in histology Greetings everyone, I've turned to the histonet a lot these days as I set up a new lab. I so appreciate the voluminous information that is shared on this site! On to my next question: are there many places using barcoding for cassettes/slides in histology? Do you like it? In what ways has barcoding made a difference? If not too involved to answer, what kinds of things do I need to consider for our software and lab info system? (We have an in-house developed software) I'm realizing what a major task this is to think about barcoding.....any help appreciated!!!! Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Tue Aug 24 16:45:02 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] bar coding in histology Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070522@HALIBUT.dcla.com> Bar Code Primer (40 pages, general, not histology per se): http://www.barcodehq.com/primer.pdf Gary Gill -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Tuesday, August 24, 2004 4:36 PM To: histonet@pathology.swmed.edu Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] bar coding in histology Greetings everyone, I've turned to the histonet a lot these days as I set up a new lab. I so appreciate the voluminous information that is shared on this site! On to my next question: are there many places using barcoding for cassettes/slides in histology? Do you like it? In what ways has barcoding made a difference? If not too involved to answer, what kinds of things do I need to consider for our software and lab info system? (We have an in-house developed software) I'm realizing what a major task this is to think about barcoding.....any help appreciated!!!! Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Tue Aug 24 17:08:13 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] decal of bone marrows Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E098@simba.kids> Gayle, What are the advantages of buffering an acid. We use a formic acid/formalin/NaCl mix to decal after fixation in 10% formalin/ethanol. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Wednesday, 25 August 2004 12:58 AM To: Mildred Fail; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] decal of bone marrows Question: Buffered formic acid, commercial preparations? or do you make up an inhouse preparation? At 07:12 AM 8/24/2004, you wrote: >Julie, > We are using Formic acid, with good results on IHC. >Rena > > >>> 08/24/04 08:17AM >>> Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From lpwenk <@t> sbcglobal.net Tue Aug 24 19:05:55 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Any teleconference schools out there? References: Message-ID: <003f01c48a37$4c351c80$80f4ff44@domainnotset.invalid> Indiana University is the only NAACLS-accredited HT program that runs a teleconference program. Students listen to a teleconference once a week (1 1/2 - 2 hours), do homework via email and snail mail, mail in slides to IU, take exams under the watchful eye of an ASCP registered histotech at the lab. Have to pay tuition to IU, so earn college credit at IU. It's a one year program, that starts in August, I believe. So I think it is too late to sign up this year, but contact Debra anyway for more information. Indiana University School of Medicine Coleman Hall, Room 322 1140 West Michigan Street Indianapolis, IN 46202-5113 Ms. Debra Wood, BS, HT(ASCP) demwood@iupui.edu (317) 278-1599 There are several NAACLS-accredited HT programs that have a lot of their lessons on the internet. Students have a password to get to the lessons, read them on line, follow the links, answer questions, etc. However, the students still have to go on campus of these programs, to do the lab work, and then have to do rotations at hospital or private histology labs. To date, that I know of, none of them are NAACLS-accredited for training histotechs in other states, via only the internet. There is one NAACLS-accredited program does make their lectures/lessons that are on line available to other people out of state for a fee. You can also take the exams and have them graded on line, for that fee. So if you need help studying for the written part of the HT/HTL exam, this would be helpful. However, you are NOT a student in the NAACLS-accredited program, because you are not attending the college labs nor are you doing a rotation. If interested, contact: Harford Community College Allied Health Department 401 Thomas Run Road Bel Air, MD 21014 Mr. Floyd Grimm, III, M.Ed. fgrimm@harford.edu (410) 836-4372 Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Tuesday, August 24, 2004 11:43 AM Subject: [Histonet] Any teleconference schools out there? > Are there any internet or teleconference schools for histology out there > now? Please respond if there are any to jbordyn@aol.com. > > > Thanks, > > Rae Ann Staskiewicz > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dpeppera <@t> mail.newcastle.edu.au Tue Aug 24 22:44:10 2004 From: dpeppera <@t> mail.newcastle.edu.au (Debbie Pepperal) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] insurance coverage Message-ID: <3.0.6.32.20040825134410.00b406d0@mail.newcastle.edu.au> Hi Guy's, Just wondering does anyone know of an insurance company within Australia who covers histo technicians for injury whilst doing private work. Thanking you Cheer's Debbie From arvind <@t> nbrc.res.in Wed Aug 25 04:29:03 2004 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] RE: mol wt of mu, kappa,delta opiod receptors Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A6C2@mail.nbrc.res.in> can any one tell mol. wt of mu,kappa,delta opiod receptors arvind@nbrc.res.in From mikael.niku <@t> helsinki.fi Wed Aug 25 08:27:20 2004 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] doing immunos by hand In-Reply-To: Message-ID: <003801c48aa7$41f4cbb0$8c0fd680@ekk1116> Yes, Shandon racks & coverplates are great. We're using them with squirt bottles & an automatic electric pipette, for both immunohistochemistry and in situ hybridization (most steps, not everything). I'd say up to 100 slides at a time goes nicely. No need for the Sequenza (?) workstation itself. And you can use the disposable coverplates several times, although they say "single use only". At least for academic research like mine :) //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf > Of Dana Settembre > Sent: 23. elokuuta 2004 19:32 > To: Pathologyarts@aol.com; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] doing immunos by hand > > > Go with Shandon. Although I believe their name is not > ThermoShandon or even ThermoElectric. Buy their racks and > coverplates. Get a squirt bottle. It is sweet. I use it > when not using my > $40 billion machine > Dana Settembre > University Hospital - UMDNJ > Newark, NJ From Dorothy.L.Webb <@t> HealthPartners.Com Wed Aug 25 10:30:58 2004 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] slide dryer and decal Message-ID: We are currently using the "Decal Stat" product from the Decal Corporation out of Texas. It is a gentler decalifier and we love it on the bone marrow cores. We stain IHC slides off of these cores and have great results! We have recently purchased the "Roto" dryer from Mopec. We LOVE it!! It holds multiple racks of slides and has consistent drying, as it has a turntabale inside the unit!! The temperature is controlled by a touchpad on the outside. It is great for IHC's, as it is so uniform and gentle!! If anyone wants the order numbers for these products, EMail me and I will be happy to furnish them!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From dbpiontek <@t> hsc.wvu.edu Wed Aug 25 10:53:41 2004 From: dbpiontek <@t> hsc.wvu.edu (Denise Bland-Piontek) Date: Fri Sep 16 15:23:56 2005 Subject: [Histonet] Research AP Manager related position Message-ID: West Virginia University is seeking a candidate with the following skills: Job Title(s): > Administrator ; Chemistry ; Cytology ; Director ; Histology ; > Immunology ; Pathology ; Technologist > > Job Summary: > Tissue Bank Administrator > Robert C. Byrd Health Sciences Center > West Virginia University University Health Associates invites applications for the full-time position of Tissue Bank Administrator. This position manages, coordinates and evaluates all elements of laboratory services in assigned specialties of the Department of pathology Tissue Bank and Outreach Laboratory located in the Robert C. Byrd Health Sciences Center of West Virginia University. The position is available immediately. Salary will be commensurate with professional experience. The person in this position will be expected to: Operate as the technical advisor for the Pathology Laboratory for Translational Research; immunohistochemistry, in situ hybridization,TUNEL, immunofluorescence, tissue microarray, laser capture microdissection and routine histology. Be knowledgeable of biohazard shipping regulations and tissue grossing techniques. Oversees autopsy requirements for Tissue Bank collections. Deal one-on-one with surgeons, nurses and Cancer Center Administration to harmonize Tissue Banking at WVU. Coordinate proper documentation for relevant case study and obtains patient consent to retain tissue specimen(s) for future research. Coordinates specimen handling, preparation and transfer from surgical and/ or pathologic sites; maintaining chain-of-custody documentation, including requisite data and computer/ software logs. Qualifications: The successful candidate must have a Bachelors Degree in Sciences. Five years supervisory and Anatomic Pathology and/ or human anatomy related fields, i.e., HT/ HTL, CT, Pathologist Assistant, surgical RN experience. Two years experience or working knowledge of Tissue Banking/ Tissue Archiving preferred, but not required. Applicants must be a Certified Histotechnologist (HT/ HTL) currently. If a candidate is not currently certified as a Histotechnologist, successful certification is required within one year. Other certifications may substitute for the Histotechnologist as follows: Pathologist Assistant (PA), Cytotechnologist (CT), Medical Technologist (MT) with research experience, or Certified Operating RN. To be considered for this exciting and innovative opportunity, send a resume to: Tissue Bank Administrator, c/o University Health Associates, PO Box 785, Morgantown, WV 26507-0785. For immediate consideration, fax or email your resume to: (304) 293-6678 or thornp@rcbhsc.wvu.edu. EOE From cannaria <@t> usc.edu Wed Aug 25 11:16:06 2004 From: cannaria <@t> usc.edu (Kevin Cannariato) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] disposable blade holder and blades for A.O. Spencer 820 Microtome Message-ID: <11BEF20E-F6B2-11D8-9DFB-000393B64516@usc.edu> First off, forgive me for asking what is probably a novice question. I am a geologist that would like to sample tree rings at sub-annual resolution. This requires taking ~20 um thick slices of tree cores. People who have done this before use a microtome. I have obtained an American Optical Spencer 820 microtome that appears to be in working order. We have been told that it would probably be easier to use disposable blades rather than the old one that came with it. I would like to know where we could purchase a disposable blade holder and disposable blades for this American Optical Spencer 820 microtome. Thanks, Kevin _________________________________________ Kevin G. Cannariato, Ph.D. Post-Doctoral Research Associate Department of Earth Sciences University of Southern California 3651 Trousdale Parkway Los Angeles, CA 90089-0740 tel: (213) 740-6733 fax: (213) 740-8801 email: cannaria@usc.edu web: http://earth.usc.edu/~cannaria From SJones <@t> cvm.tamu.edu Wed Aug 25 11:37:05 2004 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] disposable blade holder and blades for A.O. Spencer 820Microtome Message-ID: Hi Kevin, I would suggest using high profile disposable blades, they are sturdier for cutting hard tissues. Both VWR and Fisher carry the TBS Shur/Sharp blade holder for high profile blades. If the microtome sectioning doesn't work, I would try the Buehler Isomet low speed saw. Just FYI, in histo circles your microtome is called the "grey AO", a little histo history for you. Good luck, Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> Kevin Cannariato 8/25/2004 11:16:06 AM >>> First off, forgive me for asking what is probably a novice question. I am a geologist that would like to sample tree rings at sub-annual resolution. This requires taking ~20 um thick slices of tree cores. People who have done this before use a microtome. I have obtained an American Optical Spencer 820 microtome that appears to be in working order. We have been told that it would probably be easier to use disposable blades rather than the old one that came with it. I would like to know where we could purchase a disposable blade holder and disposable blades for this American Optical Spencer 820 microtome. Thanks, Kevin _________________________________________ Kevin G. Cannariato, Ph.D. Post-Doctoral Research Associate Department of Earth Sciences University of Southern California 3651 Trousdale Parkway Los Angeles, CA 90089-0740 tel: (213) 740-6733 fax: (213) 740-8801 email: cannaria@usc.edu web: http://earth.usc.edu/~cannaria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Wed Aug 25 12:24:27 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:23:57 2005 Subject: [BULK] - Re: [Histonet] disposable blade holder and blades for A.O. Spencer 820Microtome Message-ID: Ahhh, Old Grey. Kevin, it's lucky you didn't end up with a "black AO". Finding a blade holder that accepts Pterodactyl talons is difficult. >>> "Sarah Jones" 08/25/04 12:37PM >>> Hi Kevin, I would suggest using high profile disposable blades, they are sturdier for cutting hard tissues. Both VWR and Fisher carry the TBS Shur/Sharp blade holder for high profile blades. If the microtome sectioning doesn't work, I would try the Buehler Isomet low speed saw. Just FYI, in histo circles your microtome is called the "grey AO", a little histo history for you. Good luck, Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> Kevin Cannariato 8/25/2004 11:16:06 AM >>> First off, forgive me for asking what is probably a novice question. I am a geologist that would like to sample tree rings at sub-annual resolution. This requires taking ~20 um thick slices of tree cores. People who have done this before use a microtome. I have obtained an American Optical Spencer 820 microtome that appears to be in working order. We have been told that it would probably be easier to use disposable blades rather than the old one that came with it. I would like to know where we could purchase a disposable blade holder and disposable blades for this American Optical Spencer 820 microtome. Thanks, Kevin _________________________________________ Kevin G. Cannariato, Ph.D. Post-Doctoral Research Associate Department of Earth Sciences University of Southern California 3651 Trousdale Parkway Los Angeles, CA 90089-0740 tel: (213) 740-6733 fax: (213) 740-8801 email: cannaria@usc.edu web: http://earth.usc.edu/~cannaria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histophilhuff <@t> yahoo.com Wed Aug 25 12:46:00 2004 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Stable DAB - Usage Conditions - Phoenix BioTechnologies - Message-ID: <20040825174601.70449.qmail@web50309.mail.yahoo.com> We have decided to try the Stable DAB from Phoenix and we would like to know what staining conditions others on histonet have used. The protocol calls for 5 + 5 minutes at 55C. Is this the condition that everyone is using or are people incubating at room temperature? Please let me know as we are attempting to optimize a new protocol for an antibody we hope to use regularly. Thanks, Phil Huff http://www.phxbio.com/ --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! From tbourm <@t> olympicmedical.org Wed Aug 25 12:46:47 2004 From: tbourm <@t> olympicmedical.org (Tasha Bourm) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] BOUINS AND B-5 PROCESSING Message-ID: CAN ANYONE HELP ME WITH PROCESSING SOULUTIONS AND TIMES FOR BOUINS AND B-5 FIXATIVE? I'M A STUDENT AND CAN NOT FIND THIS INFORMATION IN ANY PRINTED MATERIAL. THANKS FOR YOUR HELP! TASHA BOURM HISTOLOGY OLYMPIC MEDICAL CENTER PORT ANGELES, WA 98362 From SJones <@t> cvm.tamu.edu Wed Aug 25 13:44:56 2004 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:23:57 2005 Subject: [BULK] - Re: [Histonet] disposable blade holder and blades forA.O. Spencer 820Microtome Message-ID: Hey Fred, Those black AO's were indestructible and you could hook them up to a Singer sewing machine treadle and make them foot-a-mated because they had that groove in the wheel. At least Kevin isn't using a pen knife and a piece of cork. Of course, I'm jealous of the UK guys that could go to the pub at noon and then come back to the lab and stain and smoke pipes........ are those jobs still available? lol Sarah >>> "Fred Underwood" 8/25/2004 12:24:27 PM >>> Ahhh, Old Grey. Kevin, it's lucky you didn't end up with a "black AO". Finding a blade holder that accepts Pterodactyl talons is difficult. >>> "Sarah Jones" 08/25/04 12:37PM >>> Hi Kevin, I would suggest using high profile disposable blades, they are sturdier for cutting hard tissues. Both VWR and Fisher carry the TBS Shur/Sharp blade holder for high profile blades. If the microtome sectioning doesn't work, I would try the Buehler Isomet low speed saw. Just FYI, in histo circles your microtome is called the "grey AO", a little histo history for you. Good luck, Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> Kevin Cannariato 8/25/2004 11:16:06 AM >>> First off, forgive me for asking what is probably a novice question. I am a geologist that would like to sample tree rings at sub-annual resolution. This requires taking ~20 um thick slices of tree cores. People who have done this before use a microtome. I have obtained an American Optical Spencer 820 microtome that appears to be in working order. We have been told that it would probably be easier to use disposable blades rather than the old one that came with it. I would like to know where we could purchase a disposable blade holder and disposable blades for this American Optical Spencer 820 microtome. Thanks, Kevin _________________________________________ Kevin G. Cannariato, Ph.D. Post-Doctoral Research Associate Department of Earth Sciences University of Southern California 3651 Trousdale Parkway Los Angeles, CA 90089-0740 tel: (213) 740-6733 fax: (213) 740-8801 email: cannaria@usc.edu web: http://earth.usc.edu/~cannaria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From waltersk <@t> mail.medicine.uiowa.edu Wed Aug 25 14:31:01 2004 From: waltersk <@t> mail.medicine.uiowa.edu (Walters, Katherine S) Date: Fri Sep 16 15:23:57 2005 Subject: [BULK] - Re: [Histonet] disposable blade holder and blades forA.O. Spencer 820Microtome Message-ID: Hey now, I LOVE my "Black Beauty"-never needs repair! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Wednesday, August 25, 2004 12:24 PM To: histonet@lists.utsouthwestern.edu Subject: [BULK] - Re: [Histonet] disposable blade holder and blades forA.O. Spencer 820Microtome Ahhh, Old Grey. Kevin, it's lucky you didn't end up with a "black AO". Finding a blade holder that accepts Pterodactyl talons is difficult. >>> "Sarah Jones" 08/25/04 12:37PM >>> Hi Kevin, I would suggest using high profile disposable blades, they are sturdier for cutting hard tissues. Both VWR and Fisher carry the TBS Shur/Sharp blade holder for high profile blades. If the microtome sectioning doesn't work, I would try the Buehler Isomet low speed saw. Just FYI, in histo circles your microtome is called the "grey AO", a little histo history for you. Good luck, Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> Kevin Cannariato 8/25/2004 11:16:06 AM >>> First off, forgive me for asking what is probably a novice question. I am a geologist that would like to sample tree rings at sub-annual resolution. This requires taking ~20 um thick slices of tree cores. People who have done this before use a microtome. I have obtained an American Optical Spencer 820 microtome that appears to be in working order. We have been told that it would probably be easier to use disposable blades rather than the old one that came with it. I would like to know where we could purchase a disposable blade holder and disposable blades for this American Optical Spencer 820 microtome. Thanks, Kevin _________________________________________ Kevin G. Cannariato, Ph.D. Post-Doctoral Research Associate Department of Earth Sciences University of Southern California 3651 Trousdale Parkway Los Angeles, CA 90089-0740 tel: (213) 740-6733 fax: (213) 740-8801 email: cannaria@usc.edu web: http://earth.usc.edu/~cannaria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From syedab <@t> totalise.co.uk Wed Aug 25 14:38:23 2004 From: syedab <@t> totalise.co.uk (Anila Syed) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Atherosclerosis and freshly cut tissue References: Message-ID: <002601c48adb$34fec800$f5744ed5@pbncomputer> Dear All, We are currently collecting carotid endarterectomies in order to stain them for the presence of inflammatory markers and various endothelial receptors. We collect the tissue into 10% formalin and it has been left like that for about two months while we gather sufficient numbers. My question is, we have been told that once they are wax embedded, they will need to be freshly cut each time in order for the IHC to work. Is this true? Could I cut a batch of, say 50 sections and keep them until they need to be stained? Or will I need to cut fresh sections each time. I want to stain for MMP's, TIMP1 etc. Also, in general, is it true that receptor proteins start to become oxidised in wax-embedded tissue and therefore staining starts to diminish? I hope someone has some experience of this. Thanks in advance for your help. Best wishes Anila Syed --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.744 / Virus Database: 496 - Release Date: 8/24/04 From pcfanti <@t> buffalo.edu Wed Aug 25 14:40:56 2004 From: pcfanti <@t> buffalo.edu (pcfanti@buffalo.edu) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Immunohistochemistry for Vitamen E? Message-ID: <1093462856.412ceb4809d11@mail3.buffalo.edu> I am trying to do immunohistochemistry for vitamen e in parrafin embeded specimens. I'm worried that the vitamin e will be leached out of the tissue during the deparrafinization step. I would appreciate any help on this topic. Thank you. Peter Fanti University at Buffalo OB/GYN department pcfanti@buffalo.edu From mcauliff <@t> umdnj.edu Wed Aug 25 18:01:59 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Immunohistochemistry for Vitamen E? In-Reply-To: <1093462856.412ceb4809d11@mail3.buffalo.edu> References: <1093462856.412ceb4809d11@mail3.buffalo.edu> Message-ID: <412D1A67.4030505@umdnj.edu> If vitamin E is fat-soluble, it may be removed by the alcohols and xylenes during dehydration and clearing. If it survives embedding, I would not worry so much about it coming out during dewaxing. Why not try frozen sections of fixed material? Geoff pcfanti@buffalo.edu wrote: >I am trying to do immunohistochemistry for vitamen e in parrafin >embeded specimens. I'm worried that the vitamin e will be leached out >of the tissue during the deparrafinization step. I would appreciate >any help on this topic. Thank you. >Peter Fanti >University at Buffalo >OB/GYN department >pcfanti@buffalo.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From cleger <@t> seattlecca.org Wed Aug 25 15:05:22 2004 From: cleger <@t> seattlecca.org (Leger, Carolyn A) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] cassette printer Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B94841BB38@wala01.seattlecca.org> I need some help in determining what cassette printer is worth taking the time to demo. Does anyone out there in histo-land have a preference or really like the one they are using. I need simple and easy. Thanks Carolyn A. Leger Pathology Department 206/288-2232 206/288-1355 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please notify me by telephone at (206) 288-2232 or by electronic reply, and delete this message. Thank you This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From RossS <@t> BaylorHealth.edu Wed Aug 25 15:05:30 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Atherosclerosis and freshly cut tissue Message-ID: 1. More than 48 to 72 hours in formalin can cause big problems for immunos in my experience. 2. My former lab had major problems with certain antibodies when slides were cut in advance and we cut those antibodies fresh, or had to use certain others within 2 weeks. Slides to be stained with other antibodies were good for months. It was a case of experience. When we had a problem we found that the same tissue block stained fresh was much better than when it had been cut a month or sometimes a month before. For other antibodies there was no difference. Here we are able to cut everything ahead, but we rarely cut more than 10 slides per antibody ahead. Some of those are refrigerated to slow down the loss of antigenicity. My advice is that if someone told you to cut them fresh and if they have worked with the antibody you are dealing with or at least do immuno's reguarly in your institution, then trust their word. The need to cut fresh seems to vary some by climate and fixation/processing protocal as well as antibody. Refrigeration or freezing of the slides may help you to store them longer. I would be much more worried about your fixation times. 2 months is a long time in formalin. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anila Syed Sent: Wednesday, August 25, 2004 2:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Atherosclerosis and freshly cut tissue Dear All, We are currently collecting carotid endarterectomies in order to stain them for the presence of inflammatory markers and various endothelial receptors. We collect the tissue into 10% formalin and it has been left like that for about two months while we gather sufficient numbers. My question is, we have been told that once they are wax embedded, they will need to be freshly cut each time in order for the IHC to work. Is this true? Could I cut a batch of, say 50 sections and keep them until they need to be stained? Or will I need to cut fresh sections each time. I want to stain for MMP's, TIMP1 etc. Also, in general, is it true that receptor proteins start to become oxidised in wax-embedded tissue and therefore staining starts to diminish? I hope someone has some experience of this. Thanks in advance for your help. Best wishes Anila Syed --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.744 / Virus Database: 496 - Release Date: 8/24/04 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From madaryhtl <@t> juno.com Wed Aug 25 15:10:06 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] brdu on fresh cells Message-ID: <20040825.131038.23867.522891@webmail19.lax.untd.com> the animal has to be injected with Brdu and taken down at a very specific time frame like 1-2 hours after injection. contact me at my work email and I can scrounge one up. Nick(Rocky) Madary, HT,HTL(ASCP)QIHC Histology Manager, Medimmune Inc Cell-3012334950 Work-3013984745 Fax-3013989745\ company email madaryj@medimmune.com From mari.ann.mailhiot <@t> leica-microsystems.com Wed Aug 25 15:23:31 2004 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] BOUINS AND B-5 PROCESSING Message-ID: Tasha We usually fixed our bone bx 's in B5 for about 2 hrs to 4hrs depending on the size, and then went into decal. We washed the bone bx after decal and placed it in formalin on the tissue processor. Four lymph nodes we fixed no more than 4 hrs. The nodes were cut in half and placed in B5. As far as Bouin's goes, the maximum time for tissue in Bouin's should be less than 24 hrs. You must remove the picric acid out with 50% to 70% alcohol, or I beleive you can use 70% alcohol saturated with lithium carbonate to removed the picric acid. I beleive Freida Carson has all of this information in her book. Hope this helps. Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com Tasha Bourm To: "'histonet@lists.utsouthwestern.edu'" Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] BOUINS AND B-5 PROCESSING western.edu 08/25/2004 12:46 PM CAN ANYONE HELP ME WITH PROCESSING SOULUTIONS AND TIMES FOR BOUINS AND B-5 FIXATIVE? I'M A STUDENT AND CAN NOT FIND THIS INFORMATION IN ANY PRINTED MATERIAL. THANKS FOR YOUR HELP! TASHA BOURM HISTOLOGY OLYMPIC MEDICAL CENTER PORT ANGELES, WA 98362 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From Patty.Lott <@t> ORTHO.UAB.EDU Wed Aug 25 16:02:38 2004 From: Patty.Lott <@t> ORTHO.UAB.EDU (Patty Lott) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] CD11c Message-ID: <85F6C7A1330E794DB8540AFD001CC77E0532898E@rosco.ortho.uab.edu> Does anyone have a source for this primary antibody? I am staining mouse tissue. Thanks, Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 From gcallis <@t> montana.edu Wed Aug 25 16:19:01 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] CD11c In-Reply-To: <85F6C7A1330E794DB8540AFD001CC77E0532898E@rosco.ortho.uab.e du> References: <85F6C7A1330E794DB8540AFD001CC77E0532898E@rosco.ortho.uab.edu> Message-ID: <6.0.0.22.1.20040825151157.01b22a10@gemini.msu.montana.edu> CD11c N418 clone from SEROTEC. BD PharminGen has another clone. Be careful, this is an Armenian hamster clone and your negative control must be Armenian hamster IgG1 from BD Pharmingen. Mouse spleen is a good positive control. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From syedab <@t> totalise.co.uk Wed Aug 25 16:55:28 2004 From: syedab <@t> totalise.co.uk (Anila Syed) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Atherosclerosis and freshly cut tissue References: Message-ID: <006701c48aee$5b9111e0$f5744ed5@pbncomputer> Hi Ross, Thank you very much for your reply. The trouble is that I need to collect a sufficient quantity to give to my pathology department to put into their machine. How do people usually get around this problem of tiem in formalin? The machine is on a three day cycle and they wont run through a small number of tissue samples for me, they will only do it if there are enough to fill one bucket. Would you say manual processing is a viable option? ----- Original Message ----- From: "Stapf, Ross" To: "Anila Syed" ; Sent: Wednesday, August 25, 2004 9:05 PM Subject: RE: [Histonet] Atherosclerosis and freshly cut tissue 1. More than 48 to 72 hours in formalin can cause big problems for immunos in my experience. 2. My former lab had major problems with certain antibodies when slides were cut in advance and we cut those antibodies fresh, or had to use certain others within 2 weeks. Slides to be stained with other antibodies were good for months. It was a case of experience. When we had a problem we found that the same tissue block stained fresh was much better than when it had been cut a month or sometimes a month before. For other antibodies there was no difference. Here we are able to cut everything ahead, but we rarely cut more than 10 slides per antibody ahead. Some of those are refrigerated to slow down the loss of antigenicity. My advice is that if someone told you to cut them fresh and if they have worked with the antibody you are dealing with or at least do immuno's reguarly in your institution, then trust their word. The need to cut fresh seems to vary some by climate and fixation/processing protocal as well as antibody. Refrigeration or freezing of the slides may help you to store them longer. I would be much more worried about your fixation times. 2 months is a long time in formalin. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anila Syed Sent: Wednesday, August 25, 2004 2:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Atherosclerosis and freshly cut tissue Dear All, We are currently collecting carotid endarterectomies in order to stain them for the presence of inflammatory markers and various endothelial receptors. We collect the tissue into 10% formalin and it has been left like that for about two months while we gather sufficient numbers. My question is, we have been told that once they are wax embedded, they will need to be freshly cut each time in order for the IHC to work. Is this true? Could I cut a batch of, say 50 sections and keep them until they need to be stained? Or will I need to cut fresh sections each time. I want to stain for MMP's, TIMP1 etc. Also, in general, is it true that receptor proteins start to become oxidised in wax-embedded tissue and therefore staining starts to diminish? I hope someone has some experience of this. Thanks in advance for your help. Best wishes Anila Syed --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.744 / Virus Database: 496 - Release Date: 8/24/04 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sharon.osborn <@t> dnax.org Wed Aug 25 16:57:07 2004 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] RE: screened cassettes Message-ID: <29B25753F6B1D51196110002A589D44401A177E6@PALMSG30.us.schp.com> Nita, Leica also carries screened cassettes. Please check with your Leica rep for information/prices/samples. Have a wonderful day! Sharon Osborn ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From bryand <@t> netbistro.com Wed Aug 25 18:16:22 2004 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] medical abbreviations Message-ID: <412D1DC6.2000104@netbistro.com> There have been some requests for where information about medical abbreviations can be found. I have just downloaded a freeware medical dictionary for Windows that seems to be pretty good. The address is http://www.medical-dictionary.ro/ Bryan Llewellyn From pathrm35 <@t> adelphia.net Wed Aug 25 20:52:13 2004 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Pensacola, Florida histotechs Message-ID: <20040826015212.HCWW24693.mta13.adelphia.net@mail.adelphia.net> Could someone from the Pensacola, Florida area please contact me? Thanks Ron From jnocito <@t> satx.rr.com Wed Aug 25 20:53:55 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Here we go again Message-ID: <01cb01c48b0f$8bed1fc0$0863ce44@yourxhtr8hvc4p> Hello netters, I've been contemplating whether or not to post this, but then again, it is what I do best. Once again, a posting I did several weeks ago got the attention of a certain vendor (like this hasn't happened in the past). Today, I received a telephone call from a representative from this company. This person didn't want to "flame" me. Actually, it was an interesting conversation. He said his piece and (of course) I said mine. I just wanted to let y'all know, especially the vendors, because I know you're reading this, I thought this company showed some definite class. Instead of writing letters, (albeit a year after the fact), contacting my CEO, threatening me with tar and feathers or syrup and fire ants, this representative and I had a constructive discussion. See, I'm not a bad guy at all (if you don't believe, just ask me, I'll tell you). My point is there is a right way and a wrong way to handle criticism. This company chose the right way. No flaming, no threatening, no yelling, just a professional discussion about concerns. This company is Erie Scientific. There, I said it. Correct me if I'm wrong (I know some one will) but isn't this called "customer service" and "customer satisfaction"? Let The Flaming Begin If this keeps up, I won't have anyone to talk to in Toronto nor will there be any vendor booths I can safely visit. Oh well, never did things the easy way. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX From AnthonyH <@t> chw.edu.au Wed Aug 25 22:22:25 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] decal of bone marrows Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E09A@simba.kids> Gayle, A few comments below (in blue): -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu ] Sent: Wednesday, 25 August 2004 8:51 AM To: Tony Henwood Subject: RE: [Histonet] decal of bone marrows Well for one thing, you are going backwards with some potential problems. Buffered apprx 4.5% formic acid is very gentle for immunostaining, less destructive to antigen. I have noticed several postings for "buffered Formic acid" but what is the pH? and is there a reference for this method? I assume it would need to be an acidic pH, otherwise Ca2+ would not ionise. Also wouldn't an acetic acid/sodium acetate buffer at pH 4 or there-abouts work just as well? If you fix in alcoholic formalin, you create some problems in that acid cannot ionize calcium in alcoholic conditions. The biopsy needs to rehydrate from alcoholic solution to decalcify effectively. The trephines are fixed in alcoholic formalin (ethanol aids the penetration of formalin into fatty tissues as well as defats the specimen). The biopsies are then decalcified in AQUEOUS Formic acid/formalin/Sodium Chloride solution. You would be better off to fix with NBF totally, then go to buffered formic acid. We tried 10% buffered formalin followed by Formic acid/formalin/Sodium Chloride solution but the results were not as good as fixing with alcoholic formalin (better fixation eg clearer nuclear details, well preserved cytoplasm & granules) Formic acid with formalin continues to cross link antigens strongly, and the formic acid here is approx 10% so here you may compromise antigens further by more exposure unnecessarily to formaldehyde. But isn't the rule of thumb to have complete (at least adequate) fixation before the deleterious effects of decalcification begins. Formalin helps to protect the tissues from subsequent processing. Also HIER is great tool to apply in these cases. HCl is a mineral acid and is never buffered. Formic acid can be buffered since it is an organic acid. 10% formic acid is generally not buffered and made from 90% stock formic, is not bad as it will decalcify faster on the basis it is more concentrated than buffered formic acid solution. If your bone is well fixed, (but you said this would be deleterious to immunohistochemical staining!) you want speed, then 10% formic is not bad, just do not overexpose to higher conc of any acid. some people decalcify in buffered formic overnight (maybe not needle biopsy though). It is a safer bet than HCl at any concentration. If antigen preservation is an issue, then I would probably recommend one of the EDTA decalcification methods. Whew, what a lecture - But was it? Gayle At 04:08 PM 8/24/2004, you wrote: >Gayle, > >What are the advantages of buffering an acid. >We use a formic acid/formalin/NaCl mix to decal after fixation in 10% >formalin/ethanol. > > >Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) >Laboratory Manager >The Children's Hospital at Westmead, >Locked Bag 4001, Westmead, 2145, AUSTRALIA. >Tel: 612 9845 3306 >Fax: 612 9845 3318 > > >-----Original Message----- >From: Gayle Callis [ mailto:gcallis@montana.edu] >Sent: Wednesday, 25 August 2004 12:58 AM >To: Mildred Fail; Histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] decal of bone marrows > > >Question: Buffered formic acid, commercial preparations? or do you >make up an inhouse preparation? > >At 07:12 AM 8/24/2004, you wrote: > >Julie, > > We are using Formic acid, with good results on IHC. Rena > > > > >>> 08/24/04 08:17AM >>> > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From abright <@t> brightinstruments.com Thu Aug 26 06:23:25 2004 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] disposable blade holder and blades for A.O. Spencer 820Microtome Message-ID: Dear Kevin, If I was you I would firstly try to section with the standard knife as you need to see if the microtome sections well at 20?, if it does not section well, and the knife is sharp, you will need to have the microtome checked out to make sure the slideways are adjusted properly to eliminate any play. I am sure disposable blades will not section wood samples at 20?. If you find the microtome sections well then the preferred choice of knife would be a Tungsten Carbide tipped knife if the edge of the standard knife blunted rapidly. Good luck Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Kevin Cannariato [mailto:cannaria@usc.edu] Sent: 25 August 2004 17:16 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] disposable blade holder and blades for A.O. Spencer 820Microtome First off, forgive me for asking what is probably a novice question. I am a geologist that would like to sample tree rings at sub-annual resolution. This requires taking ~20 um thick slices of tree cores. People who have done this before use a microtome. I have obtained an American Optical Spencer 820 microtome that appears to be in working order. We have been told that it would probably be easier to use disposable blades rather than the old one that came with it. I would like to know where we could purchase a disposable blade holder and disposable blades for this American Optical Spencer 820 microtome. Thanks, Kevin _________________________________________ Kevin G. Cannariato, Ph.D. Post-Doctoral Research Associate Department of Earth Sciences University of Southern California 3651 Trousdale Parkway Los Angeles, CA 90089-0740 tel: (213) 740-6733 fax: (213) 740-8801 email: cannaria@usc.edu web: http://earth.usc.edu/~cannaria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From travers.3 <@t> osu.edu Thu Aug 26 10:48:38 2004 From: travers.3 <@t> osu.edu (Susan Travers) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Fos aby NOT IN RABBIT for IHC Message-ID: -- I am looking for an antibody against c-fos that is reactive in rat and NOT MADE in rabbit for immunohistochemistry. We have tried the goat anti-fos from Santa Cruz and it did not work well for us (we have alot of experience with ihc for fos and the rabbot aby's from both santa cruz and oncogene have worked well in the past). Does anyone have any suggestions? We normally use para/PLP, para/acrolein, or para-fixed tissue (in decreasing order of preference) Thanks very much for any suggestions. Susan Travers, Ph.D. Professor, Oral Biology The Ohio State University College of Dentistry 305 W. 12th Avenue Columbus, Ohio 43210-1267 (614)292-6366(V) (614)247-6945(F) From Angel.Enniss <@t> pfizer.com Thu Aug 26 08:22:15 2004 From: Angel.Enniss <@t> pfizer.com (Enniss, Angel) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Bacteria in eye lens Message-ID: <4E2F83D5DEE32C41B4816FE9D63462420B3CC734@anagrdexm01.research.aa.wl.com> Pathologists are noting bacterial contamination in the lens of eyes. We had thought it might be from water baths or stain solutions, but it does not appear in other tissues. Can anyone tell me what this is caused from or ideas?? Anything will be much appreciated. Kindest Regards, Angel C. Enniss World Wide Safety Sciences AA Angel.Enniss@Pfizer.com _____ Upgrade Your Email - Click here! LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From SAllen <@t> exchange.hsc.mb.ca Thu Aug 26 08:35:28 2004 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] FW: Salt-split skit technique for immunofluorescence Message-ID: <304B5A264EC9974E8121B0EA14A9C3936619A2@hsc01mx1.hsc.mb.ca> > -----Original Message----- > From: Sharon Allen > Sent: August 23, 2004 9:37 AM > To: Histonet (E-mail) > Subject: Salt-split skit technique for immunofluorescence > > Hi, > Does anyone have a method for the "Salt-split skin technique for > immunofluorescence"? > I would appreciate any help with this method. > Thanks > Sharon Allen > sallen@hsc.mb.ca > Winnipeg, Manitoba, Canada This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. From browning <@t> HHSC.CA Thu Aug 26 08:59:34 2004 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] cytology question Message-ID: <3AADFB88753AD31189C100902786B91C0E2783E6@hch_nt_exchange.hhsc.ca> I would like to know how many institutions make an effort to produce cell blocks from FNA specimens, and whether the 50% alcohol that is used to collect the FNA affects any IHC results. Thanx Debra Browning, ART Technical Specialist, Immunohistochemistry Anatomic Pathology Hamilton Health Sciences phone: (905) 527-4322 ext 46131 e-mail: browning@hhsc.ca fax: (905) 524-2681 This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From tpschaer <@t> vet.upenn.edu Thu Aug 26 09:20:18 2004 From: tpschaer <@t> vet.upenn.edu (Tom Schaer) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] alcian blue pH Message-ID: <07f201c48b77$d0ac2c10$38595b82@vet.upenn.edu> Greetings List: I would like to stain some intervertebral disc x-sections (rat, decalcified) with alcian blue. Interest in cellular morphology of nucleus pulposus, transition and annulus cells. What pH of the stain should I use. Many thanks, tom -------------------------------------------------- Thomas P. Sch?r, VMD Asst Prof School of Biomedical Engineering, Drexel University & Comparative Orthopaedic Research & Tissue Engineering Department of Clinical Studies University of Pennsylvania New Bolton Center 382 West Street Road Kennett Square, PA 19348 tel. 610-444-5800 (x6261 office) tel. 610-444-5800 (x6131 lab) fax. 610-925-8100 tpschaer@mail.vet.upenn.edu http://www2.vet.upenn.edu/labs/corl/drtps.html From kerry.l.crabb <@t> gsk.com Thu Aug 26 09:45:28 2004 From: kerry.l.crabb <@t> gsk.com (kerry.l.crabb@gsk.com) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Kerry L Crabb/PharmRD/GSK is out of the office. Message-ID: I will be out of the office starting 26-Aug-2004 and will not return until 30-Aug-2004. Contact Eve about necropsy issue and contact Teresa about histology issues. I will respond to your message when I return. From pcfanti <@t> buffalo.edu Thu Aug 26 10:02:23 2004 From: pcfanti <@t> buffalo.edu (pcfanti@buffalo.edu) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Using Cy2 immunofluorescence? Message-ID: <1093532543.412dfb7f6c865@mail3.buffalo.edu> We are using a cy2 immunofluorescent secondary antibody for IHC, and we would like to know if there are any complications using it on a biotin conjugated primary. We are not sure if there will be interferance with the signal or if there will be no sites on the primary ab for the secondary to bind. I appreciate your help. Peter Fanti University at Buffalo OB/GYN department pcfanti@buffalo.edu From jlinda <@t> ces.clemson.edu Thu Aug 26 10:14:56 2004 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Re: Bacteria in eye lens Message-ID: <5.2.1.1.2.20040826110740.00a951f0@mailhost.ces.clemson.edu> Dear Angel, Is this an artificial lens implant that is inserted during cataract surgery? There have been numerous problems with bacterial growth with these implants. It was theorized that the design of the lens implant was partly responsible for the bacterial growth and when a textured surface was added to the design, the bacteria was eliminated. Again, it was theorized that the textured surface promoted quicker cellular attachment. Hope you find this useful, Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo/histo.htm From Barry.R.Rittman <@t> uth.tmc.edu Thu Aug 26 10:13:58 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] FW: Salt-split skit technique for immunofluorescence Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0E2E974@UTHEVS3.mail.uthouston.edu> Are you referring to the separation of epithelium from the underlying connective tissue by splitting at the basement membrane? If so I can send you the EDTA separation technique. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Allen Sent: Thursday, August 26, 2004 8:35 AM To: Histonet (E-mail) Subject: [Histonet] FW: Salt-split skit technique for immunofluorescence > -----Original Message----- > From: Sharon Allen > Sent: August 23, 2004 9:37 AM > To: Histonet (E-mail) > Subject: Salt-split skit technique for immunofluorescence > > Hi, > Does anyone have a method for the "Salt-split skin technique for > immunofluorescence"? > I would appreciate any help with this method. > Thanks > Sharon Allen > sallen@hsc.mb.ca > Winnipeg, Manitoba, Canada This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Thu Aug 26 13:18:27 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] alcian blue pH In-Reply-To: <07f201c48b77$d0ac2c10$38595b82@vet.upenn.edu> References: <07f201c48b77$d0ac2c10$38595b82@vet.upenn.edu> Message-ID: <412E2973.9080706@umdnj.edu> Hi Tom: Alcian Blue will stain the glycosaminoglycans in the extracellular matrix, it won't show much cellular morphology. Still, the traditional method is 1% Alcian Blue in 3% acetic acid for a pH of 2.5. At 2.5 both sulfated and carboxylated GAGs are stained. A red nuclear stain, Scarba red or nuclear fast red, gives a nice contrasty counterstain. You might try 0.05% Toluidine blue in dist water with a pH of about 4.0 for 10-15 minutes, dist water rinse and dehydration in 3 changes of actone followed by clearing in fresh xylene. Nice metachromasia of extracellular matrix and you will see some cellular morphology as well. Geoff Tom Schaer wrote: >Greetings List: > >I would like to stain some intervertebral disc x-sections (rat, decalcified) with alcian blue. Interest in cellular morphology of nucleus pulposus, transition and annulus cells. > >What pH of the stain should I use. > >Many thanks, > >tom >-------------------------------------------------- >Thomas P. Sch?r, VMD >Asst Prof School of Biomedical Engineering, Drexel University >& Comparative Orthopaedic Research & Tissue Engineering >Department of Clinical Studies >University of Pennsylvania New Bolton Center >382 West Street Road >Kennett Square, PA 19348 >tel. 610-444-5800 (x6261 office) >tel. 610-444-5800 (x6131 lab) >fax. 610-925-8100 >tpschaer@mail.vet.upenn.edu >http://www2.vet.upenn.edu/labs/corl/drtps.html >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From d.gregg <@t> juno.com Thu Aug 26 10:34:00 2004 From: d.gregg <@t> juno.com (Douglas A Gregg) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Atherosclerosis and freshly cut tissue Message-ID: <20040826.113401.3676.0.d.gregg@juno.com> I would suggest that in future studies to try PLP fixative (paraformaldehyde-lysine-periodate). I have had good experience with a number of antigens that I have been able to preserve in wet tissue for years. I work mainly with viruses. About half of the viral antigens tried could be preserved for years. The other could only stand a few days to a couple of weeks in fixative before losing their antigenicity. Retrieval is needed in almost every case but staining can be as good as fresh tissue even years later. Formalin fixed tissue usually failed completely after a year even with the most robust antigens. PLP will turn yellow and stain the tissues yellow to some extent after a few weeks or months, but don't worry about it. This does not seem to affect the immunostaining. I might add that with some antigens it works better to leave the tissue in the wet PLP until needed and then embed what you need to cut shortly before immunostaining. I have lost staining in paraffin blocks in as little as one month when the wet tissue re-embedded was fine. My record is 12 years. Best of Luck, Douglas Gregg Veterinary Pathologist Plum Island Animal Disease Center Greenport,k NY 11944 From aviancourt <@t> propathlab.com Thu Aug 26 10:41:43 2004 From: aviancourt <@t> propathlab.com (aviancourt@propathlab.com) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Open Position - IP Supervisor Message-ID: <795594536E92D4118C800050046449F3020D24CF@mailhost.propathlab.com> ProPath, a high volume Anatomic Pathology practice in Dallas, Texas has an opening for a Supervisor in the Immunohistochemistry Laboratory. The position reports directly to Dr. Rodney Miller. Candidates with HT/HTL and QIHC registry and supervisory experience are encouraged to send a resume to: ProPath 8267 Elmbrook Dr, Suite 100 Dallas, TX 75247 Fax: 214/237-1820 Email: jobs@propath.com ______________________________________________________________________________ This e-mail may contain confidential or privileged information. If you think you have received this e-mail in error, please advise the sender by reply e-mail and then delete this e-mail immediately. From Barry.R.Rittman <@t> uth.tmc.edu Thu Aug 26 11:31:26 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] EDTA separation MO Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0E2E988@UTHEVS3.mail.uthouston.edu> General procedure The use of EDTA chelates calcium in the basement membrane allowing the basement membrane to split and thus separates the epidermis from the dermis. 1. Carefully shave hair from the skin using a stiff backed commercial blade. The blade should first be rinsed in acetone to remove any grease that may be present. It is best to use commercial blades and to try not to cut the epidermis but to just remove the hair. If super sharp blades are used then the epidermis generally will become cut in areas and this affects subsequent treatment. If skin is from animal ears then it is best to first separate both sides at the cartilage plate. Remove excess fat and connective tissue as this will decrease penetration of EDTA. 2. Dip for few seconds in 70% ethanol followed by 2- 3 changes of phosphate buffered saline (PBS). This to remove debris or loose hair that may be on the skin and does not have any fixing action. 3. Place in a small closed jar containing 3mM EDTA in PBS (see below) in a shaking water bath for 2 hrs at 37 degrees C. For a piece of abdominal skin 1 by 1 cm probably use around 40 - 50 ml. of solution. In addition, the jar should be removed every half hour and briefly shaken to ensure all pieces of skin remain separate from each other. 4. Pour contents of jar into a shallow Petri dish with a small amount of fluid. 5. Arrange pieces so that epithelium side is uppermost. 6. Using a dissecting microscope at low power and Dumont style #7 forceps, hold the edge of the connective tissue onto the Petri dish with one pair of forceps. With the second pair gentle ease an edge of epithelium. Once an edge is free then the epithelium can be peeled taking care to go in one direction. This takes a little practice. Use pieces of skin from an understanding relative to start with until you become proficient. 7. The separated epithelium should float on the solution. It can be lifted using a glass hockey stick and treated as a free floating section, with the remnants of the basement membrane down. If there are tears in the epithelium it will tend to allow fluid to gain access to the surface and affect staining in that area. 8. The epidermal sheet can be floated onto PBS and used for cell separation, histochemistry, immunohistochemistry etc. It is important for any staining that sheets are not submerged in reagents but that they float on the reagents with the surface of epidermis uppermost. This provide more uniform staining. 3 mM EDTA 3 mM disodium EDTA (0.558 gms disodium EDTA in phosphate buffered saline with pH adjusted to 7.3 to 7.4. Separation of some mucosae such as dog palate and thick skin samples may require 10-20 mM EDTA instead of 3mM or longer times. These conditions do however decrease the uniformity of staining. Barry From MGomez <@t> ameripath.com Thu Aug 26 12:11:32 2004 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] digestive system biopsy embedding protocol Message-ID: Dear Histonetters: What are your recommendations for embedding this type of biopsies? Do you have a protocol you would like to share? My embedding team is having difficulty embedding the smaller biopsies on edge. I've heard that there's a company out there that sells tissue cassettes to help with orientation of specimens. Thank you, Milton From gcallis <@t> montana.edu Thu Aug 26 14:26:53 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Using Cy2 immunofluorescence? In-Reply-To: <1093532543.412dfb7f6c865@mail3.buffalo.edu> References: <1093532543.412dfb7f6c865@mail3.buffalo.edu> Message-ID: <6.0.0.22.1.20040826132057.01b10110@gemini.msu.montana.edu> For a biotinylated primary, you should come back with Strepavidin conjugated to the CY2 fluorophore. I don't think the secondary conjugated to Cy2 will like all that biotin in the way when you try to connect primary to secondary. Jackson Immunoresearch sells SA-Cy2, not terribly expensive and they have excellent products. If you use a biotinylated primary, be sure to use an avidin/biotin block or better yet, Strepavidin/biotin blocking kit (both available from Vector). At 09:02 AM 8/26/2004, you wrote: >We are using a cy2 immunofluorescent secondary antibody for IHC, and >we would like to know if there are any complications using it on a >biotin conjugated primary. We are not sure if there will be >interferance with the signal or if there will be no sites on the >primary ab for the secondary to bind. I appreciate your help. >Peter Fanti >University at Buffalo >OB/GYN department >pcfanti@buffalo.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From JWEEMS <@t> sjha.org Thu Aug 26 14:46:34 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] "Predictation" Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A63DD@sjhaexc02.sjha.org> This may sound like a stupid question, but do your pathologists ever predictate results of immuno and special stains before they receive them? In other words, dictate what they anticipate the results of the stain to be and then dictate changes later if necessary? I have been asked to pose this question. You guys are the first I turn to! Thanks much, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From Charlotte.Kopczynski <@t> baycare.org Thu Aug 26 15:01:59 2004 From: Charlotte.Kopczynski <@t> baycare.org (Kopczynski, Charlotte) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Drying Slides before Filing Message-ID: <7C3A384EF3ABD611B3270008023E3E030481CC41@mphexch01.mpm.baycare.internal> Just wanted to thank those who responded to my question about drying slides before filing. Was wondering if there are more out there who can share what they are doing. Method of drying and length of time.... Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: None To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 9, Issue 40 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Kerry L Crabb/PharmRD/GSK is out of the office. (kerry.l.crabb@gsk.com) 2. Using Cy2 immunofluorescence? (pcfanti@buffalo.edu) 3. Re: Bacteria in eye lens (Linda Jenkins) 4. RE: FW: Salt-split skit technique for immunofluorescence (Barry R Rittman) 5. Re: alcian blue pH (Geoff McAuliffe) 6. Atherosclerosis and freshly cut tissue (Douglas A Gregg) 7. Open Position - IP Supervisor (aviancourt@propathlab.com) 8. EDTA separation MO (Barry R Rittman) ---------------------------------------------------------------------- Message: 1 Date: Thu, 26 Aug 2004 10:45:28 -0400 From: kerry.l.crabb@gsk.com Subject: [Histonet] Kerry L Crabb/PharmRD/GSK is out of the office. To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii I will be out of the office starting 26-Aug-2004 and will not return until 30-Aug-2004. Contact Eve about necropsy issue and contact Teresa about histology issues. I will respond to your message when I return. ------------------------------ Message: 2 Date: Thu, 26 Aug 2004 11:02:23 -0400 From: pcfanti@buffalo.edu Subject: [Histonet] Using Cy2 immunofluorescence? To: histonet@lists.utsouthwestern.edu Message-ID: <1093532543.412dfb7f6c865@mail3.buffalo.edu> Content-Type: text/plain We are using a cy2 immunofluorescent secondary antibody for IHC, and we would like to know if there are any complications using it on a biotin conjugated primary. We are not sure if there will be interferance with the signal or if there will be no sites on the primary ab for the secondary to bind. I appreciate your help. Peter Fanti University at Buffalo OB/GYN department pcfanti@buffalo.edu ------------------------------ Message: 3 Date: Thu, 26 Aug 2004 11:14:56 -0400 From: Linda Jenkins Subject: [Histonet] Re: Bacteria in eye lens To: histonet@lists.utsouthwestern.edu Message-ID: <5.2.1.1.2.20040826110740.00a951f0@mailhost.ces.clemson.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Dear Angel, Is this an artificial lens implant that is inserted during cataract surgery? There have been numerous problems with bacterial growth with these implants. It was theorized that the design of the lens implant was partly responsible for the bacterial growth and when a textured surface was added to the design, the bacteria was eliminated. Again, it was theorized that the textured surface promoted quicker cellular attachment. Hope you find this useful, Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo/histo.htm ------------------------------ Message: 4 Date: Thu, 26 Aug 2004 10:13:58 -0500 From: "Barry R Rittman" Subject: RE: [Histonet] FW: Salt-split skit technique for immunofluorescence To: "histonet" Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0E2E974@UTHEVS3.mail.uthouston.edu> Content-Type: text/plain; charset="us-ascii" Are you referring to the separation of epithelium from the underlying connective tissue by splitting at the basement membrane? If so I can send you the EDTA separation technique. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Allen Sent: Thursday, August 26, 2004 8:35 AM To: Histonet (E-mail) Subject: [Histonet] FW: Salt-split skit technique for immunofluorescence > -----Original Message----- > From: Sharon Allen > Sent: August 23, 2004 9:37 AM > To: Histonet (E-mail) > Subject: Salt-split skit technique for immunofluorescence > > Hi, > Does anyone have a method for the "Salt-split skin technique for > immunofluorescence"? > I would appreciate any help with this method. > Thanks > Sharon Allen > sallen@hsc.mb.ca > Winnipeg, Manitoba, Canada This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Thu, 26 Aug 2004 11:18:27 -0700 From: Geoff McAuliffe Subject: Re: [Histonet] alcian blue pH To: Tom Schaer Cc: Histonet@lists.utsouthwestern.edu Message-ID: <412E2973.9080706@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi Tom: Alcian Blue will stain the glycosaminoglycans in the extracellular matrix, it won't show much cellular morphology. Still, the traditional method is 1% Alcian Blue in 3% acetic acid for a pH of 2.5. At 2.5 both sulfated and carboxylated GAGs are stained. A red nuclear stain, Scarba red or nuclear fast red, gives a nice contrasty counterstain. You might try 0.05% Toluidine blue in dist water with a pH of about 4.0 for 10-15 minutes, dist water rinse and dehydration in 3 changes of actone followed by clearing in fresh xylene. Nice metachromasia of extracellular matrix and you will see some cellular morphology as well. Geoff Tom Schaer wrote: >Greetings List: > >I would like to stain some intervertebral disc x-sections (rat, decalcified) with alcian blue. Interest in cellular morphology of nucleus pulposus, transition and annulus cells. > >What pH of the stain should I use. > >Many thanks, > >tom >-------------------------------------------------- >Thomas P. Schär, VMD >Asst Prof School of Biomedical Engineering, Drexel University >& Comparative Orthopaedic Research & Tissue Engineering >Department of Clinical Studies >University of Pennsylvania New Bolton Center >382 West Street Road >Kennett Square, PA 19348 >tel. 610-444-5800 (x6261 office) >tel. 610-444-5800 (x6131 lab) >fax. 610-925-8100 >tpschaer@mail.vet.upenn.edu >http://www2.vet.upenn.edu/labs/corl/drtps.html >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** ------------------------------ Message: 6 Date: Thu, 26 Aug 2004 11:34:00 -0400 From: Douglas A Gregg Subject: [Histonet] Atherosclerosis and freshly cut tissue To: histonet@lists.utsouthwestern.edu Cc: syedab@totalise.co.uk Message-ID: <20040826.113401.3676.0.d.gregg@juno.com> Content-Type: text/plain; charset=us-ascii I would suggest that in future studies to try PLP fixative (paraformaldehyde-lysine-periodate). I have had good experience with a number of antigens that I have been able to preserve in wet tissue for years. I work mainly with viruses. About half of the viral antigens tried could be preserved for years. The other could only stand a few days to a couple of weeks in fixative before losing their antigenicity. Retrieval is needed in almost every case but staining can be as good as fresh tissue even years later. Formalin fixed tissue usually failed completely after a year even with the most robust antigens. PLP will turn yellow and stain the tissues yellow to some extent after a few weeks or months, but don't worry about it. This does not seem to affect the immunostaining. I might add that with some antigens it works better to leave the tissue in the wet PLP until needed and then embed what you need to cut shortly before immunostaining. I have lost staining in paraffin blocks in as little as one month when the wet tissue re-embedded was fine. My record is 12 years. Best of Luck, Douglas Gregg Veterinary Pathologist Plum Island Animal Disease Center Greenport,k NY 11944 ------------------------------ Message: 7 Date: Thu, 26 Aug 2004 10:41:43 -0500 From: aviancourt@propathlab.com Subject: [Histonet] Open Position - IP Supervisor To: Histonet@lists.utsouthwestern.edu Message-ID: <795594536E92D4118C800050046449F3020D24CF@mailhost.propathlab.com> Content-Type: text/plain; charset="iso-8859-1" ProPath, a high volume Anatomic Pathology practice in Dallas, Texas has an opening for a Supervisor in the Immunohistochemistry Laboratory. The position reports directly to Dr. Rodney Miller. Candidates with HT/HTL and QIHC registry and supervisory experience are encouraged to send a resume to: ProPath 8267 Elmbrook Dr, Suite 100 Dallas, TX 75247 Fax: 214/237-1820 Email: jobs@propath.com ____________________________________________________________________________ __ This e-mail may contain confidential or privileged information. If you think you have received this e-mail in error, please advise the sender by reply e-mail and then delete this e-mail immediately. ------------------------------ Message: 8 Date: Thu, 26 Aug 2004 11:31:26 -0500 From: "Barry R Rittman" Subject: [Histonet] EDTA separation MO To: "histonet" Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0E2E988@UTHEVS3.mail.uthouston.edu> Content-Type: text/plain; charset="us-ascii" General procedure The use of EDTA chelates calcium in the basement membrane allowing the basement membrane to split and thus separates the epidermis from the dermis. 1. Carefully shave hair from the skin using a stiff backed commercial blade. The blade should first be rinsed in acetone to remove any grease that may be present. It is best to use commercial blades and to try not to cut the epidermis but to just remove the hair. If super sharp blades are used then the epidermis generally will become cut in areas and this affects subsequent treatment. If skin is from animal ears then it is best to first separate both sides at the cartilage plate. Remove excess fat and connective tissue as this will decrease penetration of EDTA. 2. Dip for few seconds in 70% ethanol followed by 2- 3 changes of phosphate buffered saline (PBS). This to remove debris or loose hair that may be on the skin and does not have any fixing action. 3. Place in a small closed jar containing 3mM EDTA in PBS (see below) in a shaking water bath for 2 hrs at 37 degrees C. For a piece of abdominal skin 1 by 1 cm probably use around 40 - 50 ml. of solution. In addition, the jar should be removed every half hour and briefly shaken to ensure all pieces of skin remain separate from each other. 4. Pour contents of jar into a shallow Petri dish with a small amount of fluid. 5. Arrange pieces so that epithelium side is uppermost. 6. Using a dissecting microscope at low power and Dumont style #7 forceps, hold the edge of the connective tissue onto the Petri dish with one pair of forceps. With the second pair gentle ease an edge of epithelium. Once an edge is free then the epithelium can be peeled taking care to go in one direction. This takes a little practice. Use pieces of skin from an understanding relative to start with until you become proficient. 7. The separated epithelium should float on the solution. It can be lifted using a glass hockey stick and treated as a free floating section, with the remnants of the basement membrane down. If there are tears in the epithelium it will tend to allow fluid to gain access to the surface and affect staining in that area. 8. The epidermal sheet can be floated onto PBS and used for cell separation, histochemistry, immunohistochemistry etc. It is important for any staining that sheets are not submerged in reagents but that they float on the reagents with the surface of epidermis uppermost. This provide more uniform staining. 3 mM EDTA 3 mM disodium EDTA (0.558 gms disodium EDTA in phosphate buffered saline with pH adjusted to 7.3 to 7.4. Separation of some mucosae such as dog palate and thick skin samples may require 10-20 mM EDTA instead of 3mM or longer times. These conditions do however decrease the uniformity of staining. Barry ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 9, Issue 40 *************************************** From la.sebree <@t> hosp.wisc.edu Thu Aug 26 15:04:44 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] "Predictation" Message-ID: Hi Joyce, Our pathologists don't usually go that far but one in particular (that doesn't dictate) will write up his results and diagnosis on the report but does at least wait until he sees the immunos before he gives the report to the transcriptionists to finish. He's rarely wrong but on those few instances that he is, we do give him a hard time about it! Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Thursday, August 26, 2004 2:47 PM To: Histonet Subject: [Histonet] "Predictation" This may sound like a stupid question, but do your pathologists ever predictate results of immuno and special stains before they receive them? In other words, dictate what they anticipate the results of the stain to be and then dictate changes later if necessary? I have been asked to pose this question. You guys are the first I turn to! Thanks much, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From japoteete <@t> saintfrancis.com Thu Aug 26 15:07:03 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] "Predictation" Message-ID: Our nine pathologists have been known to "predict" results, but several were burned doing it, so the practice has stopped. If they predict anything now, it's on a notepad or in their head. Our various hospital committees look closely at supplemental reports, especially the corrected variety. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Weems, Joyce [SMTP:JWEEMS@sjha.org] > Sent: Thursday, August 26, 2004 2:47 PM > To: Histonet > Subject: [Histonet] "Predictation" > > This may sound like a stupid question, but do your pathologists ever > predictate results of immuno and special stains before they receive them? > In other words, dictate what they anticipate the results of the stain to > be and then dictate changes later if necessary? > I have been asked to pose this question. You guys are the first I turn to! > > > Thanks much, j > > Joyce Weems > Pathology Manager > Saint Joseph's Hospital > 5665 Peachtree Dunwoody Rd NE > Atlanta, GA 30342 > 404-851-7376 - Phone > 404-851-7831 - Fax > > > > Confidentiality Notice ** The information contained in this message may be > privileged and is confidential information intended for the use of the > addressee listed above. If you are neither the intended recipient nor the > employee or agent responsible for delivering this message to the intended > recipient, you are hereby notified that any disclosure, copying, > distribution or the taking of any action in reliance on the contents of > this information is strictly prohibited. If you have received this > communication in error, please notify us immediately by replying to the > message and deleting it from your computer. > Thank you. Saint Josephs Health System, Inc. > << File: disclaimer.txt >> << File: ATT1072420.txt >> > > > > ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From tpmorken <@t> labvision.com Thu Aug 26 16:21:29 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Barcoding for histology lab Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2D94BD7@usca0082k08.labvision.apogent.com> Deb, Dr Steve McClain has done extensive work in barcoding for the path lab and give talks on it at many meetings. You can contact Dr McClaine at: Steve A. McClain, MD Director of Dermatopathology Director of Pathology Informatics Montefiore Medical Center-Central 207 111 E. 210th Street, Bronx, NY 10467 718 920-2874 FAX 718 324-2948 Good luck! Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm Tue Aug 24 16:36:08 CDT 2004 ---------------------------------------------------------------------------- ---- Greetings everyone, I've turned to the histonet a lot these days as I set up a new lab. I so appreciate the voluminous information that is shared on this site! On to my next question: are there many places using barcoding for cassettes/slides in histology? Do you like it? In what ways has barcoding made a difference? If not too involved to answer, what kinds of things do I need to consider for our software and lab info system? (We have an in-house developed software) I'm realizing what a major task this is to think about barcoding.....any help appreciated!!!! Deb King, HT(ASCP) Sacramento, CA From KYAMAMOTO <@t> chromavision.com Thu Aug 26 16:54:30 2004 From: KYAMAMOTO <@t> chromavision.com (Yamamoto, Karen) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] unsubscribe Message-ID: > -----Original Message----- > From: HistoNet Server [SMTP:histonet@pathology.swmed.edu] > Sent: Tuesday, March 12, 2002 7:40 AM > To: Yamamoto, Karen > Subject: re: subscribe > > Your address has been added to the addresses that comprise this Listserv > List. > Welcome to HISTONET. This is an electronic mailing list for the exchange > of > information pertaining to histotechnology and related fields. > > PLEASE SAVE THIS MESSAGE. > It contains useful information about how to use the list and what to do if > you > experience problems. It also includes some basic rules for email etiquette > (Netiquette) which will be helpful to those who are new to this form of > communication. > > WHAT IS A LISTSERVER? > A list server is a computer that runs software which will receive incoming > electronic mail (email) messages and reroute them automatically to > everyone on > the subscriber list. Email uses the vast expanse of the Internet to allow > almost instantaneous communication between networked computers around the > world. Our system uses the LISTSTAR software from Quarterdeck Corporation > (California) and can currently send about 30 messages a minute. With the > present number of subscribers, we are processing about 10,000 outbound > messages a day. > > WHO SHOULD SUBSCRIBE? > Anyone interested in research or clinical applications of histology, > immunohistochemistry, in-situ hybridization pathology, and electron > microscopy > may find Histonet informative and useful. Currently, there are more than > 850 > subscribers from all over the world. Subscribers include hospital > employees > from major urban centers and obscure remote locales, university > researchers, > botanists and the employees of commercial laboratories, government > agencies, > veterinary facilities and a wide variety of commercial industrial > ventures. > > WHO RUNS HISTONET? > The list is run by Linda R. Margraf, M.D. and Herb K. Hagler, Ph.D. using > hardware and software owned by the University of Texas Southwestern > Medical > School, Department of Pathology in Dallas, Texas. If you have any > questions or > problems with Histonet please contact Linda Margraf at > LMargraf@childmed.dallas.tx.us. > > HOW DOES THE LIST WORK? > This server, unlike many systems, uses ONLY ONE ADDRESS to send commands > to > the computer and to post messages. The server will recognize commands sent > in > the SUBJECT line of the message and only when they are spelled exactly as > listed below. Anything not identified as a command will be circulated to > EVERYONE on the list. > > The following is a list of commands the server recognizes: > > subscribe > Your address will be added to the list of subscribers. You will then be > able > to send messages to this list that will be forwarded to all other list > subscribers. You will begin to receive all messages sent to the list by > other > subscribers. > > subscribe digest > Your address will be added to the list of subscribers who receive a > digest > instead of each forwarded message. A digest is a compilation of all the > messages received in a 24 hour period. It is sent to the digest > subscribers > every night after midnight. Digest subscribers can post and respond to > messages the same as "real-time" subscribers. > > digests > A list of available digests will be returned to you. Histonet stores old > messages as daily digests for approximately three months. To read > previous > messages, copy the list of available digests, mark the dates of interest > and > return it to the server. > > unsubscribe > Your address will be removed from the list of subscribers. > You will no longer be able to send messages to the members > of the list. > > help > A list of the commands recognized by the server will be returned to > you. > > WHAT ARE THE RULES? > You may post any questions you wish pertaining to histology, pathology, > in-situ hybridization, immunohistochemistry etc. Equipment and reagent > evaluations, laboratory management issues, government regulations, and job > opportunities are all appropriate topics. The University asks that we > restrict > the use of its hardware and software to business purposes only (occasional > jokes do slip through but PLEASE use restraint). Vendors and those with > commercial interests in histology products are welcome contributors > however, > we ask that blatant advertisements be avoided at all times. It is fine to > refer to product that your company produces if it is pertinent to a topic > being discussed on the list. Unsolicited advertisements are poorly > tolerated > by the members and you will likely receive a number of negative comments > if > you overstep the boundaries. Please contact Linda Margraf at > LMargraf@childmed.dallas.tx.us if you are not sure about the > appropriateness > if a message you wish to post. > > > BASIC HISTONET "NETIQUETTE" > It is most helpful to the list members if you post your responses to > queries > to everyone on the list and not just as a personal reply to the person > asking > the question. That way duplicate messages are minimized and we all learn > from > each other's comments. > > Likewise, if you post a question and get a number of responses back > directly > to you, it is helpful to everyone if you could send out a summary of the > replies you got to Histonet. > > Please avoid abbreviations unless they are explained in your message. For > example: immunohistochemistry (IHC). This list circulates to a wide > variety of > individuals and what seems obvious to you may have no meaning on the other > side of the world. > > Please sign your letter and include your institution or affiliation and > location. Not all email systems have headers which identify the sender. > > Do use the subject line to indicate the topic of your message. > > DON'T USE ONLY CAPITAL LETTERS -it is considered shouting. > > Please send questions and problems about the list directly to Linda > Margraf at > LMargraf@childmed.dallas.tx.us and don't circulate them to the >850 > subscribers on the list. Be careful when sending commands to the server to > put > the command in the SUBJECT LINE and spell it correctly. > > Please do not send images as attachments with your message. We can now > post > images at our web site (http://pathcuri1.swmed.edu). To have an image > posted > send it to Herb Hagler at herb.hagler@email.swmed.edu. > > > > > es From Jan.Minshew <@t> leica-microsystems.com Thu Aug 26 19:07:56 2004 From: Jan.Minshew <@t> leica-microsystems.com (Jan.Minshew@leica-microsystems.com) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Here we go again Message-ID: Hi Joe, Since we've known each other for a long while, I feel like it's ok to respond to your email. I've been following this thread with great interest, and I suspect most of the other vendors have as well. As you know, I started my career in the lab so I'm well aware of the frustrations that can be caused when an instrument or product does not perform the way you envisioned. I also understand that the frustration level is escalated when you feel that you aren't getting an appropriate response from the company from which you purchased. Now that I am one of the "company people", I also see the other side. I know full well that there are instances when problems are not fully communicated to an appropriate person within the company and that, on rare occasion, the problem is associated with the way the product is being used. To your other point, there are indeed vendors who monitor the Histonet religiously. We are extremely interested in what goes on in the field because it helps us determine your likes and dislikes. It also gives us a glimpse of where technology is going so we can anticipate and prepare for your future needs. Sometimes we get to revel in the fact that someone appreciates something they have purchased from us. Other times we find out that there is a customer who is really unhappy. On those occasions, most of us reply off-line, directly to the individual, to successfully resolve the problem. The unfortunate aspect of this situation is that when the problems are resolved and the person is happy again, none of the people who read the original posting are aware that there was a happy ending. As far as they know, the company or representative (sometimes mentioned by name) has been given a bad review forever. Remember, everything is stored in the archives. I'm all for free speech, but I think we should all give careful consideration to potentially damaging remarks, especially if they are directed toward specific individuals. We all know that things aren't always going to be perfect. Occasionally, there are products or instruments that should not have made it through the QC process and sometimes end users make mistakes when learning to use a new product. This happens in our professional and home life. Problems don't always get fixed right away, even though we all strive for that, but they will almost always get resolved if both parties keep communicating with each other. We all need to remember that communication is two-way and, if everything goes just right, it should result in an action from one or both parties. If you have a problem, the best way to resolve it is to contact the appropriate person, begin a meaningful dialog in a professional manner and remember that complaints and recommendations for improvements are almost always better received when they are done with tact and an open mind. Joe, let me thank you for letting your fellow Histonetters know that you were satisfied with your communications with the great people at Erie. And, I have no doubt that you will be welcomed into any booth at NSH, especially if you continue to show support for the effectiveness of the "no flaming, no threatening, no yelling" form of professional discussions. While you're in the exhibit area, take a look at all of the new and innovative products and show pride in the fact that effective communication has brought about the wonderful advances in our technology. I'll look forward to seeing you there. Best wishes, Jan Minshew Marketing Manager Leica Microsystems, Inc. 2345 Waukegan Road Bannockburn, IL 60015 800.248.0123 x7051 "Joe Nocito" To: "histonet" Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Here we go again western.edu 08/25/2004 08:53 PM Hello netters, I've been contemplating whether or not to post this, but then again, it is what I do best. Once again, a posting I did several weeks ago got the attention of a certain vendor (like this hasn't happened in the past). Today, I received a telephone call from a representative from this company. This person didn't want to "flame" me. Actually, it was an interesting conversation. He said his piece and (of course) I said mine. I just wanted to let y'all know, especially the vendors, because I know you're reading this, I thought this company showed some definite class. Instead of writing letters, (albeit a year after the fact), contacting my CEO, threatening me with tar and feathers or syrup and fire ants, this representative and I had a constructive discussion. See, I'm not a bad guy at all (if you don't believe, just ask me, I'll tell you). My point is there is a right way and a wrong way to handle criticism. This company chose the right way. No flaming, no threatening, no yelling, just a professional discussion about concerns. This company is Erie Scientific. There, I said it. Correct me if I'm wrong (I know some one will) but isn't this called "customer service" and "customer satisfaction"? Let The Flaming Begin If this keeps up, I won't have anyone to talk to in Toronto nor will there be any vendor booths I can safely visit. Oh well, never did things the easy way. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From n_hedgecock <@t> hotmail.com Thu Aug 26 19:50:49 2004 From: n_hedgecock <@t> hotmail.com (Nicole Hedgecock) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] fixation and decal of cortical bone Message-ID: Hi, All I would like to fix, decalcify, and embed rabbit cortical bone sections in order to do TUNEL staining for osteocyte apoptosis, but I've come across conflicts between the literature and an actual, working protocol a colleague has shared with me. That being said, can you store the sample in a fixative for long periods of time? i thought that if left in formaldehye or formalin for too long then there would be too much cross-linking and antibodies (for IHC) could not penetrate. Is over-cross-linking not an issue for TUNEL? Also, don't formalin and paraformaldehyde break down into formic acid after a period of time? I read that formic acid is no good for TUNEL, EDTA is the decalcifier of choice. finally, if you can't store your sample in the fixative for extended periods of time (weeks to months), then how can I store it until I am ready to decalcify them? Thanks, Nicky Nicole Hedgecock Orthopedic Research Lab UCD Medical Center University of California, Davis _________________________________________________________________ Visita MSN Latino Noticias: Todo lo que pasa en el mundo y en tu pa?n, ?en tu idioma! http://latino.msn.com/noticias/ From jnocito <@t> satx.rr.com Thu Aug 26 21:51:25 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] "Predictation" References: <83AACDB0810528418AA106F9AE9B7F7E0A63DD@sjhaexc02.sjha.org> Message-ID: <021101c48be0$be5d3090$0863ce44@yourxhtr8hvc4p> Joyce, in a word YES. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Weems, Joyce" To: "Histonet" Sent: Thursday, August 26, 2004 2:46 PM Subject: [Histonet] "Predictation" > This may sound like a stupid question, but do your pathologists ever predictate results of immuno and special stains before they receive them? In other words, dictate what they anticipate the results of the stain to be and then dictate changes later if necessary? > I have been asked to pose this question. You guys are the first I turn to! > > Thanks much, j > > Joyce Weems > Pathology Manager > Saint Joseph's Hospital > 5665 Peachtree Dunwoody Rd NE > Atlanta, GA 30342 > 404-851-7376 - Phone > 404-851-7831 - Fax > > > > Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. > Thank you. Saint Josephs Health System, Inc. > > ---------------------------------------------------------------------------- ---- > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From eileen_dusek <@t> yahoo.com Fri Aug 27 05:13:21 2004 From: eileen_dusek <@t> yahoo.com (eileen dusek) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] CD3 on BM Message-ID: <20040827101321.627.qmail@web11901.mail.yahoo.com> Good Morning, Is anyone having trouble with CD3 on BM? Our stain works great on regular tissue but weak on BM. We pressure cook the slides and yesterday I tried using an amp, still no staining. I was thinking of trying no pretreatment or protease instead of PC. Are there any other suggestions? Sorry about the font change, I'm not sure how it happened. I appreciate all your help Eileen Edward Hospital --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! From Terry.Marshall <@t> rothgen.nhs.uk Fri Aug 27 07:20:54 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] "Predictation" Message-ID: I have done it in one circumstance only, and that is when I "know" a lesion is a melanoma, but there is no pigment + some other confusing feature. I have then said something like "will confirm with immunochemistry - further report if negative". So, even then, I'm "coming clean". In general, if you don't need the stain, why ask? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: 26 August 2004 20:47 To: Histonet Subject: [Histonet] "Predictation" This may sound like a stupid question, but do your pathologists ever predictate results of immuno and special stains before they receive them? In other words, dictate what they anticipate the results of the stain to be and then dictate changes later if necessary? I have been asked to pose this question. You guys are the first I turn to! Thanks much, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From la.sebree <@t> hosp.wisc.edu Fri Aug 27 08:35:29 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] CD3 on BM Message-ID: Eileen, Can you be more specific? Are you staining manually or automated? Is your CD3 polyclonal or monoclonal? Predilute or at what concentration? Which HIER buffer are you using? Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: eileen dusek [mailto:eileen_dusek@yahoo.com] Sent: Friday, August 27, 2004 5:13 AM To: histonet Subject: [Histonet] CD3 on BM Good Morning, Is anyone having trouble with CD3 on BM? Our stain works great on regular tissue but weak on BM. We pressure cook the slides and yesterday I tried using an amp, still no staining. I was thinking of trying no pretreatment or protease instead of PC. Are there any other suggestions? Sorry about the font change, I'm not sure how it happened. I appreciate all your help Eileen Edward Hospital --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histo <@t> ambree.com Fri Aug 27 08:55:14 2004 From: histo <@t> ambree.com (histo@ambree.com) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Golgi Cox staining Message-ID: I try to establish a Golgi Cox staining for mouse brain tissue. After the first trials, I have two major problems: 1) The slices (between 20 and 80?m) cut from paraffin blocks using a sliding microtome are rather fragile. 2) There are so many neurones stained, that I do not know, how to distinguish between them and how to quantify the data. Does anybody have an idea to solve these problems? Thank you, and kind regards Oliver Ambr?e --- Oliver Ambr?e Institute of Neuropathology University Hospital M?nster Domagkstr. 19 D - 48149 M?nster phone: +49 251/ 83-52359 fax: +49 251/ 83-56971 ambree@uni-muenster.de --- From Rcartun <@t> harthosp.org Fri Aug 27 08:55:10 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] cytology question Message-ID: It is my experience that initial fixation in alcohol can affect immunoreactivity for those proteins that are highly soluble (e.g. S-100 and calretinin). We also have a difficult time identifying synaptophysin and chromogranin in alcohol-fixed cytology specimens. We have seen several specimens where the cytology preparations have been negative, but tissue submitted to histology has been positive. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> Browning Deb 08/26/04 09:59AM >>> I would like to know how many institutions make an effort to produce cell blocks from FNA specimens, and whether the 50% alcohol that is used to collect the FNA affects any IHC results. Thanx Debra Browning, ART Technical Specialist, Immunohistochemistry Anatomic Pathology Hamilton Health Sciences phone: (905) 527-4322 ext 46131 e-mail: browning@hhsc.ca fax: (905) 524-2681 This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Fri Aug 27 08:59:14 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] "Predictation" Message-ID: "Some do; most do not". Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Weems, Joyce" 08/26/04 03:46PM >>> This may sound like a stupid question, but do your pathologists ever predictate results of immuno and special stains before they receive them? In other words, dictate what they anticipate the results of the stain to be and then dictate changes later if necessary? I have been asked to pose this question. You guys are the first I turn to! Thanks much, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Rcartun <@t> harthosp.org Fri Aug 27 09:02:10 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] CD3 on BM Message-ID: Are you using a mAb or pAb? We were using a mAb for awhile, but we noticed that it did not do well on bone marrow specimens so we went back to the pAb. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> eileen dusek 08/27/04 06:13AM >>> Good Morning, Is anyone having trouble with CD3 on BM? Our stain works great on regular tissue but weak on BM. We pressure cook the slides and yesterday I tried using an amp, still no staining. I was thinking of trying no pretreatment or protease instead of PC. Are there any other suggestions? Sorry about the font change, I'm not sure how it happened. I appreciate all your help Eileen Edward Hospital --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histomjans <@t> yahoo.com Fri Aug 27 10:46:05 2004 From: histomjans <@t> yahoo.com (Melissa Jans) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] roommate needed for NSH Message-ID: <20040827154605.31930.qmail@web50310.mail.yahoo.com> Hello all!! I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. Melissa Jans University of Iowa Healthcare 1-319-356-2140 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Nancy.Lowen <@t> med.va.gov Fri Aug 27 11:02:54 2004 From: Nancy.Lowen <@t> med.va.gov (Lowen, Nancy) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] frozen sections on bone Message-ID: I am doing frozen sections on fresh, undecalcified mouse femurs. My sections seem to be losing all of the marrow and many are torn . I have tried disposable cryostat knives as well as the tungsten carbide, but the sections are still not good. If anyone else is doing this procedure, could you share any tips or suggestions on your technique, cutting temperatures, or overall procedure. I have tried different temperatures, but none of them seem to give me the sections I want. Any help would be greatly appreciated. Nancy.Lowen@med.va.gov From paw555 <@t> yahoo.com Fri Aug 27 11:04:56 2004 From: paw555 <@t> yahoo.com (pam plumlee) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Coverslipping tape Message-ID: <20040827160456.1827.qmail@web11607.mail.yahoo.com> Hi all: I know this has been asked before, is there a quick fix for removing tissue from coverslipping tape, reattaching to the slide, covering with a new glass coverslip so the tissue can be photographed? Also, does anyone know a good alertnate fixative for osmium tetroxide? We are trying to keep it out of the lab. We have been asked to use it for a post-fix for fatty tissue and on an EM processor. Thanks, Pam __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail From Terry.Marshall <@t> rothgen.nhs.uk Fri Aug 27 11:14:13 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] roommate needed for NSH Message-ID: Come on guys - you don't get an offer like this everyday:-) Dr Terry L (too far to travel) Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Melissa Jans [mailto:histomjans@yahoo.com] Sent: 27 August 2004 16:46 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] roommate needed for NSH Hello all!! I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. Melissa Jans University of Iowa Healthcare 1-319-356-2140 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Aug 27 11:30:02 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] roommate needed for NSH Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0A63E8@sjhaexc02.sjha.org> I knew that was coming! Have a good weekend everybody! j:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Friday, August 27, 2004 12:14 PM To: Melissa Jans; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] roommate needed for NSH Come on guys - you don't get an offer like this everyday:-) Dr Terry L (too far to travel) Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Melissa Jans [mailto:histomjans@yahoo.com] Sent: 27 August 2004 16:46 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] roommate needed for NSH Hello all!! I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. Melissa Jans University of Iowa Healthcare 1-319-356-2140 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From gcallis <@t> montana.edu Fri Aug 27 11:32:49 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Successful frozen sections on bone In-Reply-To: References: Message-ID: <6.0.0.22.1.20040827102138.01b1a5a0@gemini.msu.montana.edu> The ONLY way we have successful cryosections of any kind of bone is using the Instrumedics CryoJane tape transfer system. Check out their website for information, how it works. Bone frozen sections are destroyed by disposable or c profile steel blade, tungsten carbide blades are ideal as you are finding out. However, I have never had a bone frozen section cut with a TC knife which remains intact unless the tape transfer method is used to maintains bone integrity. Cutting temperature must be -28C up to -32C or so. Your tungsten carbide knife edge must be in perfectly, sharp condition even with tape transfer method. For us, cryosectioning bone without the Cryojane has been a waste of time. Although a pricey bit of equipment, it has paid for itself many times over with good bone frozen sections. I am going to attach a publication to you privately so you can see advantages. At 10:02 AM 8/27/2004, you wrote: >I am doing frozen sections on fresh, undecalcified mouse femurs. My >sections seem to be losing all of the marrow and many are torn . I have >tried disposable cryostat knives as well as the tungsten carbide, but the >sections are still not good. If anyone else is doing this procedure, could >you share any tips or suggestions on your technique, cutting temperatures, >or overall procedure. I have tried different temperatures, but none of them >seem to give me the sections I want. Any help would > >be greatly appreciated. > >Nancy.Lowen@med.va.gov > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From juan.gutierrez <@t> christushealth.org Fri Aug 27 11:50:39 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] FW: Salt-split skit technique for immunofluorescence Message-ID: Where I use to work, we thawed the frozen block in cold PBS and let the sections sit in it over the weekend( or at least two days) until the epithelium separated from the basement membrane. Then pad dry carefully and refreeze. By the way, this was all done in the refrigerator. Good luck. Juan -----Original Message----- From: Sharon Allen [mailto:SAllen@exchange.hsc.mb.ca] Sent: Thursday, August 26, 2004 8:35 AM To: Histonet (E-mail) Subject: [Histonet] FW: Salt-split skit technique for immunofluorescence > -----Original Message----- > From: Sharon Allen > Sent: August 23, 2004 9:37 AM > To: Histonet (E-mail) > Subject: Salt-split skit technique for immunofluorescence > > Hi, > Does anyone have a method for the "Salt-split skin technique for > immunofluorescence"? > I would appreciate any help with this method. > Thanks > Sharon Allen > sallen@hsc.mb.ca > Winnipeg, Manitoba, Canada This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri Aug 27 11:54:36 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] "Predictation" Message-ID: Yes, repeatedly, but you didn't hear it from me. ;) -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Thursday, August 26, 2004 2:47 PM To: Histonet Subject: [Histonet] "Predictation" This may sound like a stupid question, but do your pathologists ever predictate results of immuno and special stains before they receive them? In other words, dictate what they anticipate the results of the stain to be and then dictate changes later if necessary? I have been asked to pose this question. You guys are the first I turn to! Thanks much, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From froyer <@t> bitstream.net Fri Aug 27 12:04:36 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] roommate needed for NSH In-Reply-To: <20040827154605.31930.qmail@web50310.mail.yahoo.com> References: <20040827154605.31930.qmail@web50310.mail.yahoo.com> Message-ID: <412F69A4.1060003@bitstream.net> Could you tell me a little bit more about this person? What's her sign? Can she cook? ...etc. ~ Ford (SWM) Royer ;-) M.S.B. Minneapolis, MN 800-745-4869 Melissa Jans wrote: >Hello all!! > >I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... >Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. > >Melissa Jans >University of Iowa Healthcare >1-319-356-2140 > >__________________________________________________ >Do You Yahoo!? >Tired of spam? Yahoo! Mail has the best spam protection around >http://mail.yahoo.com >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From JQB7 <@t> CDC.GOV Fri Aug 27 12:11:32 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] roommate needed for NSH Message-ID: She has a real good personality and makes her own clothes! :) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ford Royer Sent: Friday, August 27, 2004 1:05 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] roommate needed for NSH Could you tell me a little bit more about this person? What's her sign? Can she cook? ...etc. ~ Ford (SWM) Royer ;-) M.S.B. Minneapolis, MN 800-745-4869 Melissa Jans wrote: >Hello all!! > >I am looking for someone who wants to save a little money at NSH this >year!!! I have a female co-worker who is looking for someone to share >a room with in Toronto. She already has a room reserved..... Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. > >Melissa Jans >University of Iowa Healthcare >1-319-356-2140 > >__________________________________________________ >Do You Yahoo!? >Tired of spam? Yahoo! Mail has the best spam protection around >http://mail.yahoo.com >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eca9 <@t> georgetown.edu Fri Aug 27 12:19:48 2004 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Coverslipping? Message-ID: <412F6D34.6010907@georgetown.edu> Hi, I have a question about coverslipping slides. Is there any harm in leaving slides without coverslipping them overnight? With this I mean after dehydration and xylenes...just removing them from the xylenes and letting them dry....redipping them in xylenes before coverslipping the following morning. Thanks in advance for you help, Eva From lucyb <@t> biocare.net Fri Aug 27 12:29:05 2004 From: lucyb <@t> biocare.net (Lucy Brooks) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] roommate needed for NSH References: <20040827154605.31930.qmail@web50310.mail.yahoo.com> <412F69A4.1060003@bitstream.net> Message-ID: <001501c48c5b$5ab461a0$0a01a8c0@LUCYSALES> Does NSH stand for Nice Single Histologists?Histotechs? :) ----- Original Message ----- From: "Ford Royer" To: Sent: Friday, August 27, 2004 10:04 AM Subject: Re: [Histonet] roommate needed for NSH > Could you tell me a little bit more about this person? What's her > sign? Can she cook? ...etc. > > ~ Ford (SWM) Royer ;-) > M.S.B. > Minneapolis, MN > 800-745-4869 > > Melissa Jans wrote: > > >Hello all!! > > > >I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... > >Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. > > > >Melissa Jans > >University of Iowa Healthcare > >1-319-356-2140 > > > >__________________________________________________ > >Do You Yahoo!? > >Tired of spam? Yahoo! Mail has the best spam protection around > >http://mail.yahoo.com > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cforster <@t> umn.edu Fri Aug 27 12:35:09 2004 From: cforster <@t> umn.edu (Colleen Forster) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] roomate for NSH Message-ID: <412F70CD.7060808@umn.edu> Just to give a little more information on the roomsharing, you can also contact the NSH office. They have a list of people needing room sharing and can hook you up with someones information. I used this when we were in Louisville and it worked very well. www.nsh.org. Colleen Forster U of Mn From Jason.Wiese <@t> med.va.gov Fri Aug 27 12:33:58 2004 From: Jason.Wiese <@t> med.va.gov (Jason.Wiese@med.va.gov) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Coverslipping? Message-ID: It would be better to leave them in Xylene overnight then to let them dry out... Jason Wiese, HT(ASCP) VARHS Histology/Pathology 913 Garden Valley Blvd Roseburg, OR 97470 (541)-440-1000 x44751 -----Original Message----- From: Eva C Andersson [mailto:eca9@georgetown.edu] Sent: Friday, August 27, 2004 10:20 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Coverslipping? Hi, I have a question about coverslipping slides. Is there any harm in leaving slides without coverslipping them overnight? With this I mean after dehydration and xylenes...just removing them from the xylenes and letting them dry....redipping them in xylenes before coverslipping the following morning. Thanks in advance for you help, Eva _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Fri Aug 27 12:42:12 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] roommate needed for NSH Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070563@HALIBUT.dcla.com> How's about N for Naughty...? Sorry, couldn't resist. TGIF. Gary Gill -----Original Message----- From: Lucy Brooks [mailto:lucyb@biocare.net] Sent: Friday, August 27, 2004 12:29 PM To: Ford Royer; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] roommate needed for NSH Does NSH stand for Nice Single Histologists?Histotechs? :) ----- Original Message ----- From: "Ford Royer" To: Sent: Friday, August 27, 2004 10:04 AM Subject: Re: [Histonet] roommate needed for NSH > Could you tell me a little bit more about this person? What's her > sign? Can she cook? ...etc. > > ~ Ford (SWM) Royer ;-) > M.S.B. > Minneapolis, MN > 800-745-4869 > > Melissa Jans wrote: > > >Hello all!! > > > >I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... > >Please contact me privately if you or someone you know is interested. > >I can be contacted via email (please send your phone # so I may call you) or the phone # below. > > > >Melissa Jans > >University of Iowa Healthcare > >1-319-356-2140 > > > >__________________________________________________ > >Do You Yahoo!? > >Tired of spam? Yahoo! Mail has the best spam protection around > >http://mail.yahoo.com _______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Fri Aug 27 12:46:50 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Coverslipping? Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070564@HALIBUT.dcla.com> Why remove them from xylene? Don't let cells and tissues dry -- ever -- unless standard protocol (e.g., air-dried blood films) calls for otherwise. Immersing slides overnight in xylene is no different, and no more "harmful", than coverslipping slides with xylene-based mounting medium. The xylene solvent remains in contact with the cells or tissue for many days until it is able to evaporate completely but ever so slowly through the edge seal of mounting medium that borders the cover glass. Gary Gill -----Original Message----- From: Eva C Andersson [mailto:eca9@georgetown.edu] Sent: Friday, August 27, 2004 12:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Coverslipping? Hi, I have a question about coverslipping slides. Is there any harm in leaving slides without coverslipping them overnight? With this I mean after dehydration and xylenes...just removing them from the xylenes and letting them dry....redipping them in xylenes before coverslipping the following morning. Thanks in advance for you help, Eva _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Fri Aug 27 12:59:43 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] roommate needed for NSH In-Reply-To: <001501c48c5b$5ab461a0$0a01a8c0@LUCYSALES> Message-ID: it could be Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Lucy Brooks Sent: Friday, August 27, 2004 12:29 PM To: Ford Royer; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] roommate needed for NSH Does NSH stand for Nice Single Histologists?Histotechs? :) ----- Original Message ----- From: "Ford Royer" To: Sent: Friday, August 27, 2004 10:04 AM Subject: Re: [Histonet] roommate needed for NSH > Could you tell me a little bit more about this person? What's her > sign? Can she cook? ...etc. > > ~ Ford (SWM) Royer ;-) > M.S.B. > Minneapolis, MN > 800-745-4869 > > Melissa Jans wrote: > > >Hello all!! > > > >I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... > >Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. > > > >Melissa Jans > >University of Iowa Healthcare > >1-319-356-2140 > > > >__________________________________________________ > >Do You Yahoo!? > >Tired of spam? Yahoo! Mail has the best spam protection around > >http://mail.yahoo.com > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From heiroe <@t> hotmail.com Fri Aug 27 14:07:42 2004 From: heiroe <@t> hotmail.com (heidi roehrich) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] rat eyes Message-ID: I am having huge problems with unperfused rat eyes. Shortly after the eyes are enucleated I place them in oct and snap freeze the tissue in 2-methylbutane placed in liquid nitrogen. After sectioning the tissue, I fix it for 10 min. in 4% paraformaldyhde and follow the HNE antibody protocol. The results are that the photoreceptors are almost gone and the rest of the tissue looks completely degenerated. What went wrong? Heidi Röhrich Histology Core Lab Department of Ophthalmology University of Minnesota 370 Lions research Building _________________________________________________________________ [1]Is your PC infected? Get a FREE online computer virus scan from McAfeeŽ Security. References 1. http://g.msn.com/8HMBENUS/2755??PS=47575 From MTitford <@t> aol.com Fri Aug 27 14:34:16 2004 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] More on medical abbreviations Message-ID: <7783CE9A.3E7CBD7C.00762DB1@aol.com> Brian Llewellyn gives some pointers as to where medical abbreviations can be found. Here in hospitals in the USA, the JCAHO has a campaign going to abolish some abbreviations in the interests of patient safety. They mainly concern written notes in patients charts but also have applications in the histology lab. Some of the banned abbreviations and their corrections are: 1) instead of 1.0 mg write 1 mg (no trailing zero's) 2) Instead of MgSO4 write magnesium sulphate 3) Instead of .1 mg write 0.1 mg (always use a zero before a decimal point) Etc Another thing they recommend is reading back of telephone orders which has applications in histology. For example if a pathologist phones up to request a special stain on a certain block, read back the request to confirm you have recorded the right block, etc. Mike Titford USA Pathology Mobile AL USA From funderwood <@t> mcohio.org Fri Aug 27 14:35:38 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:23:57 2005 Subject: [BULK] - [Histonet] Coverslipping tape Message-ID: Hi Pam, Immerse the slide in acetone for about 5 minutes. The film will swell up and come right off the slide. Swish the slide in alcohol then back into xylene(a couple of changes) and coverslip. Fred >>> pam plumlee 08/27/04 12:04PM >>> Hi all: I know this has been asked before, is there a quick fix for removing tissue from coverslipping tape, reattaching to the slide, covering with a new glass coverslip so the tissue can be photographed? Also, does anyone know a good alertnate fixative for osmium tetroxide? We are trying to keep it out of the lab. We have been asked to use it for a post-fix for fatty tissue and on an EM processor. Thanks, Pam __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Fri Aug 27 14:41:19 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:23:57 2005 Subject: [BULK] - Re: [Histonet] roommate needed for NSH Message-ID: 'round these parts it stands for Nymphomaniacal Sadistic Histosleezes. >>> "Lucy Brooks" 08/27/04 01:29PM >>> Does NSH stand for Nice Single Histologists?Histotechs? :) ----- Original Message ----- From: "Ford Royer" To: Sent: Friday, August 27, 2004 10:04 AM Subject: Re: [Histonet] roommate needed for NSH > Could you tell me a little bit more about this person? What's her > sign? Can she cook? ...etc. > > ~ Ford (SWM) Royer ;-) > M.S.B. > Minneapolis, MN > 800-745-4869 > > Melissa Jans wrote: > > >Hello all!! > > > >I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... > >Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. > > > >Melissa Jans > >University of Iowa Healthcare > >1-319-356-2140 > > > >__________________________________________________ > >Do You Yahoo!? > >Tired of spam? Yahoo! Mail has the best spam protection around > >http://mail.yahoo.com > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lchung <@t> ppmh.org Fri Aug 27 14:57:37 2004 From: lchung <@t> ppmh.org (Chung, Luong) Date: Fri Sep 16 15:23:57 2005 Subject: [BULK] - Re: [Histonet] roommate needed for NSH Message-ID: Enough already... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Fred Underwood Sent: Friday, August 27, 2004 3:41 PM To: histonet@lists.utsouthwestern.edu Subject: [BULK] - Re: [Histonet] roommate needed for NSH 'round these parts it stands for Nymphomaniacal Sadistic Histosleezes. >>> "Lucy Brooks" 08/27/04 01:29PM >>> Does NSH stand for Nice Single Histologists?Histotechs? :) ----- Original Message ----- From: "Ford Royer" To: Sent: Friday, August 27, 2004 10:04 AM Subject: Re: [Histonet] roommate needed for NSH > Could you tell me a little bit more about this person? What's her > sign? Can she cook? ...etc. > > ~ Ford (SWM) Royer ;-) > M.S.B. > Minneapolis, MN > 800-745-4869 > > Melissa Jans wrote: > > >Hello all!! > > > >I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... > >Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. > > > >Melissa Jans > >University of Iowa Healthcare > >1-319-356-2140 > > > >__________________________________________________ > >Do You Yahoo!? > >Tired of spam? Yahoo! Mail has the best spam protection around > >http://mail.yahoo.com > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lucyb <@t> biocare.net Fri Aug 27 15:04:41 2004 From: lucyb <@t> biocare.net (Lucy Brooks) Date: Fri Sep 16 15:23:57 2005 Subject: [BULK] - Re: [Histonet] roommate needed for NSH References: Message-ID: <002901c48c71$176e8a90$0a01a8c0@LUCYSALES> Well Fred, "These Parts" sounds a little too wild for me! ----- Original Message ----- From: "Fred Underwood" To: Sent: Friday, August 27, 2004 12:41 PM Subject: [BULK] - Re: [Histonet] roommate needed for NSH > 'round these parts it stands for Nymphomaniacal Sadistic Histosleezes. > > >>> "Lucy Brooks" 08/27/04 01:29PM >>> > Does NSH stand for Nice Single Histologists?Histotechs? > > :) > ----- Original Message ----- > From: "Ford Royer" > To: > Sent: Friday, August 27, 2004 10:04 AM > Subject: Re: [Histonet] roommate needed for NSH > > > > Could you tell me a little bit more about this person? What's her > > sign? Can she cook? ...etc. > > > > ~ Ford (SWM) Royer ;-) > > M.S.B. > > Minneapolis, MN > > 800-745-4869 > > > > Melissa Jans wrote: > > > > >Hello all!! > > > > > >I am looking for someone who wants to save a little money at NSH > this > year!!! I have a female co-worker who is looking for someone to share > a > room with in Toronto. She already has a room reserved..... > > >Please contact me privately if you or someone you know is > interested. I > can be contacted via email (please send your phone # so I may call you) > or > the phone # below. > > > > > >Melissa Jans > > >University of Iowa Healthcare > > >1-319-356-2140 > > > > > >__________________________________________________ > > >Do You Yahoo!? > > >Tired of spam? Yahoo! Mail has the best spam protection around > > >http://mail.yahoo.com > > >_______________________________________________ > > >Histonet mailing list > > >Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri Aug 27 15:11:36 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:57 2005 Subject: [BULK] - Re: [Histonet] roommate needed for NSH Message-ID: People frequently ask me what HTASCP stands for - Hot Tempered Anti Social Castrating Piranna. Jacqueline M. O'Connor HT(ASCP) and proud of it. "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/27/2004 02:41 PM To: cc: Subject: [BULK] - Re: [Histonet] roommate needed for NSH 'round these parts it stands for Nymphomaniacal Sadistic Histosleezes. >>> "Lucy Brooks" 08/27/04 01:29PM >>> Does NSH stand for Nice Single Histologists?Histotechs? :) ----- Original Message ----- From: "Ford Royer" To: Sent: Friday, August 27, 2004 10:04 AM Subject: Re: [Histonet] roommate needed for NSH > Could you tell me a little bit more about this person? What's her > sign? Can she cook? ...etc. > > ~ Ford (SWM) Royer ;-) > M.S.B. > Minneapolis, MN > 800-745-4869 > > Melissa Jans wrote: > > >Hello all!! > > > >I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... > >Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. > > > >Melissa Jans > >University of Iowa Healthcare > >1-319-356-2140 > > > >__________________________________________________ > >Do You Yahoo!? > >Tired of spam? Yahoo! Mail has the best spam protection around > >http://mail.yahoo.com > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bwhitaker <@t> brownpathology.com Fri Aug 27 15:58:03 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:23:57 2005 Subject: [BULK] - Re: [Histonet] roommate needed for NSH In-Reply-To: Message-ID: <000001c48c78$8bfe00a0$3601a8c0@brownpathology.net> It must be Friday...... Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie.O'Connor@abbott.com Sent: Friday, August 27, 2004 3:12 PM To: Fred Underwood Cc: histonet@lists.utsouthwestern.edu Subject: Re: [BULK] - Re: [Histonet] roommate needed for NSH People frequently ask me what HTASCP stands for - Hot Tempered Anti Social Castrating Piranna. Jacqueline M. O'Connor HT(ASCP) and proud of it. "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/27/2004 02:41 PM To: cc: Subject: [BULK] - Re: [Histonet] roommate needed for NSH 'round these parts it stands for Nymphomaniacal Sadistic Histosleezes. >>> "Lucy Brooks" 08/27/04 01:29PM >>> Does NSH stand for Nice Single Histologists?Histotechs? :) ----- Original Message ----- From: "Ford Royer" To: Sent: Friday, August 27, 2004 10:04 AM Subject: Re: [Histonet] roommate needed for NSH > Could you tell me a little bit more about this person? What's her > sign? Can she cook? ...etc. > > ~ Ford (SWM) Royer ;-) > M.S.B. > Minneapolis, MN > 800-745-4869 > > Melissa Jans wrote: > > >Hello all!! > > > >I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... > >Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. > > > >Melissa Jans > >University of Iowa Healthcare > >1-319-356-2140 > > > >__________________________________________________ > >Do You Yahoo!? > >Tired of spam? Yahoo! Mail has the best spam protection around > >http://mail.yahoo.com > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From michael_keysock <@t> merck.com Sat Aug 28 10:32:20 2004 From: michael_keysock <@t> merck.com (Keysock, Michael A) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Coverslipping? Message-ID: Do not let slides air dry. Slides will be fine if they are just simply left in xylene overnight. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C Andersson Sent: Friday, August 27, 2004 1:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Coverslipping? Hi, I have a question about coverslipping slides. Is there any harm in leaving slides without coverslipping them overnight? With this I mean after dehydration and xylenes...just removing them from the xylenes and letting them dry....redipping them in xylenes before coverslipping the following morning. Thanks in advance for you help, Eva _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From asmith <@t> mail.barry.edu Sat Aug 28 10:49:34 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] More on medical abbreviations Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3BC9@exchsrv01.barrynet.barry.edu> Reading or reciting an instruction back is a good practice in face-to-face communication as well as in telephone conversation. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of MTitford@aol.com Sent: Friday, August 27, 2004 3:34 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] More on medical abbreviations Brian Llewellyn gives some pointers as to where medical abbreviations can be found. Here in hospitals in the USA, the JCAHO has a campaign going to abolish some abbreviations in the interests of patient safety. They mainly concern written notes in patients charts but also have applications in the histology lab. Some of the banned abbreviations and their corrections are: 1) instead of 1.0 mg write 1 mg (no trailing zero's) 2) Instead of MgSO4 write magnesium sulphate 3) Instead of .1 mg write 0.1 mg (always use a zero before a decimal point) Etc Another thing they recommend is reading back of telephone orders which has applications in histology. For example if a pathologist phones up to request a special stain on a certain block, read back the request to confirm you have recorded the right block, etc. Mike Titford USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From Krat18 <@t> aol.com Sat Aug 28 11:24:54 2004 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Coverslipping? Message-ID: <84.3203b3ea.2e620bd6@aol.com> I've heard for many years that slides shouldn't be allowed to dry out, but we have one tech that always lets hers dry before coverslipping, and I've never heard any complaints from the docs about her stains. So what is the problem with letting the slides air dry? One other question for everyone: what's the best (and possibly quickest) way to decontaminate a cryostat? Karen_Raterman@ssmhc.com St. Mary's Health Center From medjnw <@t> emory.edu Sat Aug 28 13:42:32 2004 From: medjnw <@t> emory.edu (Josiah N. Wilcox, Ph.D.) Date: Fri Sep 16 15:23:57 2005 Subject: [Histonet] Histopathology positions at Medtronic Message-ID: <057B8DF3-F922-11D8-BEB2-000393DBA4AA@emory.edu> I would like to post the following job opportunities at Medtronic There are currently two openings at Medtronic Vascular in Santa Rosa, California for a histopathologist (senior manager, PhD, DVM or equivalent) and histology technician/histotechnologist (BS or MS with experience) to help establish a new histopathology laboratory to work on studies related to drug eluting stents and other vascular projects. Individuals with experience in general histology, paraffin or plastic embedding and sectioning or previous experience in cardiovascular histopathology and morphometric analysis are encouraged to apply. Josiah N. Wilcox, Ph.D. Senior Director and Resident Scholar Science and Technology Medtronic Vascular 3576 Unocal Place SS-48, FBMC Santa Rosa, CA 95403 Office 707-591-2256 Cy.Wilcox@medtronic.com From asmith <@t> mail.barry.edu Sat Aug 28 15:25:23 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] fixatives Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3BCD@exchsrv01.barrynet.barry.edu> If you mean a fixative that is also an electron stain, potassium permanganate (Luft,1956, J. Biophys. Biochem. Cytol. 2:799) and sodium permanganate (Wetzel, 1961, J. Biophys. Biochem. Cytol. 9:711) have been uses with some success. While both are safer than osmium tetroxide, neither gives good results as consistently. If you just want a fixative for electron microscopy, Karnovsky's (1965) paraformaldehyde-glutaraldehyde mixture is one of the best (J. Cell Biol. 27:137A or any EM handbook). Plain glutaraldehyde also works: any EM handbook will give a formula for 2.5% glutaraldehyde in pH 7.2-7.4 phosphate or cacodylate buffer. (Note that cacodylate is very poisonous.) Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pam plumlee Sent: Friday, August 27, 2004 12:05 PM To: HistoNet Server Subject: [Histonet] Coverslipping tape Hi all: I know this has been asked before, is there a quick fix for removing tissue from coverslipping tape, reattaching to the slide, covering with a new glass coverslip so the tissue can be photographed? Also, does anyone know a good alertnate fixative for osmium tetroxide? We are trying to keep it out of the lab. We have been asked to use it for a post-fix for fatty tissue and on an EM processor. Thanks, Pam __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From peoshel <@t> wisc.edu Sat Aug 28 17:56:01 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] fixatives In-Reply-To: <4C051EAE581BB646BF53A749A73FBA2D1F3BCD@exchsrv01.barrynet.barry.edu> References: <4C051EAE581BB646BF53A749A73FBA2D1F3BCD@exchsrv01.barrynet.barry.edu> Message-ID: Pam, There isn't a very good substitute for osmium tetroxide, but Maunsback and Aufzelius, "Biomedical Electron Microscopy" shows various fixitive alternatives, and is a good place to look. It is for TEM, though, and not light microscopy. DON'T use OsO4 in a tissue processor unless it is in a **very** well vented fume hood and will continue so. I wouldn't use OsO4 in a processor period, but there are processors for EM work that do use it. It can be used safely, though, so I wouldn't worry about having in the lab, just use it properly. It does do a reasonable job preserving fat, but it won't prevent extract in the dehydration steps, especially of the unsaturated fats -- OsO4 likes to bind to C=C double bonds, and isn't that great a fixative for unsaturated lipids. Cryostat sections and no dehydration would be better. K or NaMnO4 can work, and in the TEM it does a nice job of preserving membranes -- the cells can look like line drawings of the membrane systems. But this is because the cytoplasm is heavily extracted. Not what you want, I think. Sorry I can't help with the sections-stuck-to-coverslip-tape problem, I've only used glass. Phil >Hi all: I know this has been asked before, is there a >quick fix for removing tissue from coverslipping tape, >reattaching to the slide, covering with a new glass >coverslip so the tissue can be photographed? Also, >does anyone know a good alertnate fixative for osmium >tetroxide? We are trying to keep it out of the lab. >We have been asked to use it for a post-fix for fatty >tissue and on an EM processor. Thanks, Pam -- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From heytallguy2000 <@t> hotmail.com Sat Aug 28 23:19:28 2004 From: heytallguy2000 <@t> hotmail.com (john clark) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] (no subject) Message-ID: One of the CAP questions asks how a lab documents that its glassware is detergent free. We hand wash our glassware. Is it adequate to randomly select something and pH it to show that the detergent is absent? Any suggestions? John Clark From lpwenk <@t> sbcglobal.net Sun Aug 29 06:11:11 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] (no subject) References: Message-ID: <000c01c48db8$e5293b40$fb35d445@domainnotset.invalid> After washing and rinsing, rinse again with d. water. Use a pH strip to test the pH of the d. water out of the tap. Record that. Then randomly pull a glassware from the batch you just washed, while it is still wet. Touch a pH strip to a wet area. It should be the same pH as the d. water. Depending on the soap or bleach you are using in the wash water, the pH of the wash water will be higher or low than the pH of the d. water. So if your random glassware is the same pH as the d. water, then all the wash water must have been removed from the glassware Record the pH of the d. water AND the pH of the glassware. For EACH time you wash dishes. So batching is better in this case. Of course, since it is for CAP, write up this procedure in the proper format, and have it dated and since (and then reviewed after that) by your pathologist. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "john clark" To: Sent: Sunday, August 29, 2004 12:19 AM Subject: [Histonet] (no subject) One of the CAP questions asks how a lab documents that its glassware is detergent free. We hand wash our glassware. Is it adequate to randomly select something and pH it to show that the detergent is absent? Any suggestions? John Clark _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Sun Aug 29 06:22:36 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Coverslipping? References: <84.3203b3ea.2e620bd6@aol.com> Message-ID: <001601c48dba$7cf23ca0$fb35d445@domainnotset.invalid> STAINS: Well, for H&E, if allowed to dry out first before coverslipping, there appear these little blackish refractile particles on the tissue, randomly placed. For other special stains, I've seen the tissue shrink and buckle/warp, when allowed to dry out first. Not all of them, just some, but I can't remember which ones, as I don't usually allow this to happen. DECONTAMINATE CRYOSTAT: A quick way, after each use (which is what we do with the FS of muscles from enzyme histochemistry or kidneys for immunofluorescence) or at the end of the day (which is what we do in the FS rooms, since it is our pathologists and residents who cut FS from surgery) - Clean out as much of the FS debris with a gauze, and throw in the biohazard bin. Pour some 95-100% reagent alcohol on another gauze, and wipe down the inside of the cryostat and blade (careful!). Throw the gauze in the biohazard bin (in all likelihood would be OK in regular trash, but we like people being in the habit of using the biohazard bin). The alcohol will decontaminate everything except spores, will evaporate rather quickly, will not get too much water into the cryostat so moving parts won't "freeze up", and won't corrode the inside the of cryostat. This is from a Journal of Histotechology article, many 15 years ago. Don't have the information on hand at home. Still have to occasionally do a complete defrost. Again, write it up as a procedure, and have it signed by pathologist. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: ; ; Sent: Saturday, August 28, 2004 12:24 PM Subject: Re: [Histonet] Coverslipping? > I've heard for many years that slides shouldn't be allowed to dry > out, but we have one tech that always lets hers dry before coverslipping, and > I've never heard any complaints from the docs about her stains. So what is the > problem with letting the slides air dry? > > One other question for everyone: what's the best (and possibly > quickest) way to decontaminate a cryostat? > > Karen_Raterman@ssmhc.com > St. Mary's Health Center > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Krat18 <@t> aol.com Sun Aug 29 07:23:20 2004 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Coverslipping? Message-ID: Thanks for the response. We always use the absolute alcohol too, but I'd heard in the past that we should put an open dish of 37% formaldehyde in the closed cryostat overnight (after defrosting) to kill all organisms. And I remember long ago we used to use phenol. So I wasn't sure if the alcohol alone was sufficient. Karen_Raterman@ssmhc.com St. Mary's Health Center From pruegg <@t> ihctech.net Sun Aug 29 10:07:09 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] TB in Lung Specimens Message-ID: This is alarming. I have not been on histonet for a while, was there responses to this? I have been working with formalin fixed paraffin embedded tb infected tissue for years without using these kind of precautions. Am I at risk for TB infection? Patsy Ruegg -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Saturday, August 28, 2004 9:51 AM To: pruegg@ihctech.net Subject: FW: [Histonet] TB in Lung Specimens -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrew Kennedy Sent: Monday, August 09, 2004 2:19 PM To: Histonet Subject: [Histonet] TB in Lung Specimens Hi Histonetters, A recent article in Human Pathology suggests that Mycobacteria in formalin fixed tissue can remain viable and therefore there is a risk of contracting TB from these specimens. It suggests that formalin fixed tissue from suspected TB cases should be handled with gloves, gown and mask. I wonder if we should be using these precautions for every lung specimen at every step in the histological process! If the organisms are still viable, trimmings from blocks should probably be bagged and disposed of in infectious waste. It could quite possibly end up being the same as with CJD brain specimens. What do you all think about this? Reference: Gerston, K.F. Blumberg, L. Tshabalala, V.A. Murray, J. Viability of Mycobacteria in Formalin Fixed Lungs. Human Pathology, Vol 35, No 5. May 2004. pp 571-575 Andrew Kennedy Senior Science Officer Anatomical Pathology Concord Repatriation General Hospital Hospital Road Concord NSW 2139 ph: +612 9767 6115 Fax +612 9767 8427 "corpora non agunt nisi fixata" _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lesley <@t> vancouverbc.net Sun Aug 29 10:14:15 2004 From: lesley <@t> vancouverbc.net (Lesley Weston) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] fixatives In-Reply-To: Message-ID: If you're not interested in membrane, you can dehydrate small samples in a water-miscible resin such as Aquembed from Ladd, rather than alcohols, before embedding in epoxy. This preserves fat. If you do need to see the membrane, OsO4 is your best bet (possibly as well as Aquembed) but be sure to use it in a fume hood and with gloves. It's nasty stuff, and I would agree that you shouldn't use it in a processor but that you can use it safely if you're careful. Lesley Weston. on 28/08/2004 3:56 PM, Philip Oshel at peoshel@wisc.edu wrote: > Pam, > > There isn't a very good substitute for osmium tetroxide, but > Maunsback and Aufzelius, "Biomedical Electron Microscopy" shows > various fixitive alternatives, and is a good place to look. It is for > TEM, though, and not light microscopy. > DON'T use OsO4 in a tissue processor unless it is in a **very** well > vented fume hood and will continue so. I wouldn't use OsO4 in a > processor period, but there are processors for EM work that do use > it. It can be used safely, though, so I wouldn't worry about having > in the lab, just use it properly. It does do a reasonable job > preserving fat, but it won't prevent extract in the dehydration > steps, especially of the unsaturated fats -- OsO4 likes to bind to > C=C double bonds, and isn't that great a fixative for unsaturated > lipids. Cryostat sections and no dehydration would be better. > K or NaMnO4 can work, and in the TEM it does a nice job of preserving > membranes -- the cells can look like line drawings of the membrane > systems. But this is because the cytoplasm is heavily extracted. Not > what you want, I think. > Sorry I can't help with the sections-stuck-to-coverslip-tape problem, > I've only used glass. > > Phil > >> Hi all: I know this has been asked before, is there a >> quick fix for removing tissue from coverslipping tape, >> reattaching to the slide, covering with a new glass >> coverslip so the tissue can be photographed? Also, >> does anyone know a good alertnate fixative for osmium >> tetroxide? We are trying to keep it out of the lab. >> We have been asked to use it for a post-fix for fatty >> tissue and on an EM processor. Thanks, Pam From browning <@t> HHSC.CA Fri Aug 27 13:06:38 2004 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] roommate needed for NSH Message-ID: <3AADFB88753AD31189C100902786B91C0E2783FE@hch_nt_exchange.hhsc.ca> Sad to say, but this is my first time at a conference for more than half a day in my 26 years of working!!! Now I am really wondering what I've missed over the years, and is it possible to catch up? -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Friday, August 27, 2004 12:30 PM To: Marshall Terry Dr, Consultant Histopathologist; Melissa Jans; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] roommate needed for NSH I knew that was coming! Have a good weekend everybody! j:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Friday, August 27, 2004 12:14 PM To: Melissa Jans; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] roommate needed for NSH Come on guys - you don't get an offer like this everyday:-) Dr Terry L (too far to travel) Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Melissa Jans [mailto:histomjans@yahoo.com] Sent: 27 August 2004 16:46 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] roommate needed for NSH Hello all!! I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. Melissa Jans University of Iowa Healthcare 1-319-356-2140 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. 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From bryand <@t> netbistro.com Sun Aug 29 13:10:45 2004 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] TB in Lung Specimens In-Reply-To: References: Message-ID: <41321C25.5070002@netbistro.com> I haven't read the article, so read this with that fact in mind. When I trained in the 1960s TB was fairly common still. We were taught to handle TB infected tissues relatively safely. However, looking back on it we were quite lax compared to today's attitudes towards safety in lab work. There were some things we did that were more likely to protect us than are done today, though. One of the most important is the duration of fixation with formalin mixtures. We used to fix a minimum of overnight, sometimes up to a couple of weeks, mostly because there was no fixation (pun intended) on a 24 hour diagnosis. Even then it was known that TB organisms resisted formalin fixation, so known cases were routinely left for a few days before grossing. Often they would be refixed after that. The usual approach was to expose the organisms to a minimum of 48 hours in formalin, with a week being better. After that they were presumed to be safe. It should be kept in mind, though, that in the past we used longer processing schedules with quite a bit longer in ethanol and xylene. It was presumed (no proof) that the fat solvent nature of those affected any remaining viability of TB organisms by dissolving out the mycolic acid coat. At any rate, with older long fixation and slow processing we considered the organisms to be dead. Today, short fixation and fast processing is the norm so I would suggest that should no longer be considered true, and that you are indeed at risk. On the plus side, modern processors do use heat during processing, and cassettes limit the thickness of the tissue, so they are less likely to come out unprocessed than they were in the past. Since TB is on the rise again I think the whole subject should be reappraised and standard safe working protocols publicised and enforced. I stress the "enforced" since we all know that protocols are established and may then be completely ignored because they are inconvenient. This comes through very clearly with some of the comments about the (US) CAP inspections on Histonet. We have similar inspections in BC, Canada with similar attitudes, I might add, and I presume other countries are the same. Before I retired I had a long term disagreement with our safety committee and lab administration regarding this issue. The lab wanted a 24 hour turnaround time, while the safety committee wanted all lab departments to work with universal precautions, that is, treat all specimens as if they are infected with the worst possible organism. I repeatedly pointed out that these were mutually exclusive for TB, HIV, hepatitis and other things. For universal precautions we would have had to fix all biopsies for 2-3 days before processing just in case a formalin resistant organism was involved. That would preclude a 24 hour diagnosis. I was never able to get an answer to my objections on this issue, and I consider it to be an area of administrative hypocrisy. I did, however, make sure my comments were clearly recorded just in case of a WCB claim by one of the technologists. The reality is that we are expected to section and stain tissues that are infected and which may well contain viable organisms. In practice, the issue then becomes how to do this and work safely. I suggest that the following should form the basis. 1. Get vaccinated for everything you can. This should be at employer's expense. If they refuse to pay, get it done, pay yourself and then kick up one hell of a fuss. 2. Wash your hands frequently. Use soap and hot water and maybe a brush. Be thorough, not cursory. This helps with squame contamination on sections, too. Don't forget the face, especially around the mouth and nose. We frequently touch those areas without being aware of it. 3. Wear gloves. Don't presume this means you need not wash your hands. 4. Wear masks. 5. NEVER cut frozen sections on unfixed TB, HIV or hepatitis cases, ever. 6. Fix tissues as long as possible as a routine. 7. Use as long a processing schedule as you can get away with. 8. NEVER process known TB, HIV or hepatitis tissues with less than 48 hours fixation. 9. Sacrilege. Put your own health before that of the patients. You won't do anybody any good lying in a hospital bed. Do NOT fall into the trap of making an exception because "the patient needs a diagnosis". Usually that means someone else can't be bothered to wait. 10. Make a nuisance of yourself with any committees or groups that have influence in these areas, and don't stop. This is not exhaustive by any means, and like Patsy, I would like to see this issue discussed far more thoroughly. Bryan Llewellyn Patsy Ruegg wrote: > This is alarming. I have not been on histonet for a while, was there > responses to this? I have been working with formalin fixed paraffin > embedded tb infected tissue for years without using these kind of > precautions. Am I at risk for TB infection? > Patsy Ruegg > > -----Original Message----- > From: Patsy Ruegg [mailto:pruegg@ihctech.net] > Sent: Saturday, August 28, 2004 9:51 AM > To: pruegg@ihctech.net > Subject: FW: [Histonet] TB in Lung Specimens > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrew > Kennedy > Sent: Monday, August 09, 2004 2:19 PM > To: Histonet > Subject: [Histonet] TB in Lung Specimens > > > Hi Histonetters, > > > > A recent article in Human Pathology suggests that Mycobacteria in formalin > fixed tissue can remain viable and therefore there is a risk of contracting > TB from these specimens. It suggests that formalin fixed tissue from > suspected TB cases should be handled with gloves, gown and mask. > > > I wonder if we should be using these precautions for every lung specimen at > every step in the histological process! If the organisms are still viable, > trimmings from blocks should probably be bagged and disposed of in > infectious waste. It could quite possibly end up being the same as with CJD > brain specimens. What do you all think about this? > > > > Reference: Gerston, K.F. Blumberg, L. Tshabalala, V.A. Murray, J. Viability > of Mycobacteria in Formalin Fixed Lungs. Human Pathology, Vol 35, No 5. May > 2004. pp 571-575 > > > > Andrew Kennedy > > Senior Science Officer > > Anatomical Pathology > > Concord Repatriation General Hospital > > Hospital Road > Concord NSW 2139 > > > > ph: +612 9767 6115 > > Fax +612 9767 8427 > > > > "corpora non agunt nisi fixata" > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gu.lang <@t> gmx.at Sun Aug 29 15:45:36 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] TB in Lung Specimens References: Message-ID: <000e01c48e09$2332d400$eeeea8c0@server> It may be a silly question. If we put our positiv-control blocks of Tb on an adequate agar, will the bacteria grow? And if not, are they really killed? Gudrun ----- Original Message ----- From: "Patsy Ruegg" To: Sent: Sunday, August 29, 2004 5:07 PM Subject: [Histonet] TB in Lung Specimens > This is alarming. I have not been on histonet for a while, was there > responses to this? I have been working with formalin fixed paraffin > embedded tb infected tissue for years without using these kind of > precautions. Am I at risk for TB infection? > Patsy Ruegg > > -----Original Message----- > From: Patsy Ruegg [mailto:pruegg@ihctech.net] > Sent: Saturday, August 28, 2004 9:51 AM > To: pruegg@ihctech.net > Subject: FW: [Histonet] TB in Lung Specimens > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrew > Kennedy > Sent: Monday, August 09, 2004 2:19 PM > To: Histonet > Subject: [Histonet] TB in Lung Specimens > > > Hi Histonetters, > > > > A recent article in Human Pathology suggests that Mycobacteria in formalin > fixed tissue can remain viable and therefore there is a risk of contracting > TB from these specimens. It suggests that formalin fixed tissue from > suspected TB cases should be handled with gloves, gown and mask. > > > I wonder if we should be using these precautions for every lung specimen at > every step in the histological process! If the organisms are still viable, > trimmings from blocks should probably be bagged and disposed of in > infectious waste. It could quite possibly end up being the same as with CJD > brain specimens. What do you all think about this? > > > > Reference: Gerston, K.F. Blumberg, L. Tshabalala, V.A. Murray, J. Viability > of Mycobacteria in Formalin Fixed Lungs. Human Pathology, Vol 35, No 5. May > 2004. pp 571-575 > > > > Andrew Kennedy > > Senior Science Officer > > Anatomical Pathology > > Concord Repatriation General Hospital > > Hospital Road > Concord NSW 2139 > > > > ph: +612 9767 6115 > > Fax +612 9767 8427 > > > > "corpora non agunt nisi fixata" > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jluis.palazon <@t> icman.csic.es Mon Aug 30 04:01:52 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] H&E/Alcian Blue Message-ID: <20040830090152.15F21285BFB@perceval.uca.es> Dear all I would like to perform an H&E/Alcian blue staining to differentiate bone and cartilage and I was wondering if I have to do the H&E first and then AB or the contrary or maybe there is a protocole to this staining technique. The acid AB solution will bleach the Hematoxylin?. If the AB is used first why not call the technique AB/H&E instead of H&E/AB? thanks in advance jose luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From Jamie.Dukes <@t> se.amedd.army.mil Mon Aug 30 05:35:34 2004 From: Jamie.Dukes <@t> se.amedd.army.mil (Dukes, Jamie Mr EAMC) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] GROSSROOM Message-ID: <4FB3076916FCD311AF7900805FA7A678077B4B0A@dasmthfdz001.amedd.army.mil> Hi all, Can someone give me any information on whether or not a pregnant person can work in the grossroom. From NSEARCY <@t> swmail.sw.org Mon Aug 30 07:15:40 2004 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Powerpath Message-ID: Last week I aked for names of Tamtron/ Powerpath users- I received one reply. There are supposed to be 350 users out there---where are you??? Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD From rschoon <@t> email.unc.edu Mon Aug 30 07:31:25 2004 From: rschoon <@t> email.unc.edu (rschoon) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] GFP antibodies In-Reply-To: <057B8DF3-F922-11D8-BEB2-000393DBA4AA@emory.edu> References: <057B8DF3-F922-11D8-BEB2-000393DBA4AA@emory.edu> Message-ID: <41331E1D.5060607@email.unc.edu> 'Netter's, I would like to order in some anti-GFP antibodies. I realize that there are several out there and I'd like to get one that will work on both FFPE and frozen sections of murine tissue.... Open to suggestions. THX Robert Schoonhoven, UNC-CH CEHS From JNocito <@t> Pathreflab.com Mon Aug 30 07:38:21 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] H&E/Alcian Blue In-Reply-To: <20040830090152.15F21285BFB@perceval.uca.es> Message-ID: Jose, we perform the Alcian Blue first, then use the H & E as a counterstain. We haven't had any problems with the staining. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jose Luis Palazon Fernandez Sent: Monday, August 30, 2004 4:02 AM To: histonet@pathology.swmed.edu Subject: [Histonet] H&E/Alcian Blue Dear all I would like to perform an H&E/Alcian blue staining to differentiate bone and cartilage and I was wondering if I have to do the H&E first and then AB or the contrary or maybe there is a protocole to this staining technique. The acid AB solution will bleach the Hematoxylin?. If the AB is used first why not call the technique AB/H&E instead of H&E/AB? thanks in advance jose luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Mon Aug 30 07:48:56 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:58 2005 Subject: [BULK] - Re: [Histonet] roommate needed for NSH In-Reply-To: Message-ID: Jacqueline, ouch Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie.O'Connor@abbott.com Sent: Friday, August 27, 2004 3:12 PM To: Fred Underwood Cc: histonet@lists.utsouthwestern.edu Subject: Re: [BULK] - Re: [Histonet] roommate needed for NSH People frequently ask me what HTASCP stands for - Hot Tempered Anti Social Castrating Piranna. Jacqueline M. O'Connor HT(ASCP) and proud of it. "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/27/2004 02:41 PM To: cc: Subject: [BULK] - Re: [Histonet] roommate needed for NSH 'round these parts it stands for Nymphomaniacal Sadistic Histosleezes. >>> "Lucy Brooks" 08/27/04 01:29PM >>> Does NSH stand for Nice Single Histologists?Histotechs? :) ----- Original Message ----- From: "Ford Royer" To: Sent: Friday, August 27, 2004 10:04 AM Subject: Re: [Histonet] roommate needed for NSH > Could you tell me a little bit more about this person? What's her > sign? Can she cook? ...etc. > > ~ Ford (SWM) Royer ;-) > M.S.B. > Minneapolis, MN > 800-745-4869 > > Melissa Jans wrote: > > >Hello all!! > > > >I am looking for someone who wants to save a little money at NSH this year!!! I have a female co-worker who is looking for someone to share a room with in Toronto. She already has a room reserved..... > >Please contact me privately if you or someone you know is interested. I can be contacted via email (please send your phone # so I may call you) or the phone # below. > > > >Melissa Jans > >University of Iowa Healthcare > >1-319-356-2140 > > > >__________________________________________________ > >Do You Yahoo!? > >Tired of spam? Yahoo! Mail has the best spam protection around > >http://mail.yahoo.com > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Mon Aug 30 07:50:16 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] H&E/Alcian Blue Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3BCE@exchsrv01.barrynet.barry.edu> Do the alcian blue first. The acid in the alcian blue solution and the rinse will extract hematoxylin. Nothing other than trifluoroacetic acid (in methylene chloride) can extract alcian blue. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jose Luis Palazon Fernandez Sent: Monday, August 30, 2004 5:02 AM To: histonet@pathology.swmed.edu Subject: [Histonet] H&E/Alcian Blue Dear all I would like to perform an H&E/Alcian blue staining to differentiate bone and cartilage and I was wondering if I have to do the H&E first and then AB or the contrary or maybe there is a protocole to this staining technique. The acid AB solution will bleach the Hematoxylin?. If the AB is used first why not call the technique AB/H&E instead of H&E/AB? thanks in advance jose luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From k.s.bosch <@t> amc.uva.nl Mon Aug 30 08:13:09 2004 From: k.s.bosch <@t> amc.uva.nl (Klazien Bosch) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] BrdU Message-ID: Dear Histonetters, Is there anybody who has experience with the BrdU antibody: BU1/75 (ICR1)? I want to use it on fresh frozen sections of mouse liver. What kind of fixation, dilution, blocking steps needed, etc.? Thanks! Klazien Klazien Bosch Academic Medical Center Dept. Cell Biology and Histology Amsterdam the Netherlands From gcallis <@t> montana.edu Mon Aug 30 09:03:06 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Successful frozen sections on bone In-Reply-To: References: <6.0.0.22.1.20040827102138.01b1a5a0@gemini.msu.montana.edu> Message-ID: <6.0.0.22.1.20040830075759.01b13218@gemini.msu.montana.edu> An addition to Patsy's suggestions, another trick is to flash with UV twice, we have even done 3 times. The capacitor in power source needs to build up adequate charge (I hope I said this correctly) , so do not flash rapidly, do it slowly so you get the "flash". When the green light is on, you have the go signal to flash. Also remove the tape VERY slowly, and diagonally from one corner of slide towards the other, and make sure the slide is cold when you do this. We use 1/2X slides, with success, if you have a great deal of problem with polymer slides then try 1 x or even 4X. The latter are really gooey but with double UV exposure, 1/2X worked nicely. neAt 09:19 AM 8/28/2004, you wrote: >I agree with Gayle on this, I have cut whole rat tibia/femurs without decal >using the Instrumedics tape system and the sections are pretty darn good, it >is sometimes difficult to get the tape off the slide leaving all the bone >section behind, I expose the section to the UV light and then put it on dry >ice for 10 min. or so before trying to remove the tape, you are still going >to get some of the cortex left on the tape but this is the best I could do >(my experience). >Patsy Ruegg > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle >Callis >Sent: Friday, August 27, 2004 9:33 AM >To: Lowen, Nancy; Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Successful frozen sections on bone > > >The ONLY way we have successful cryosections of any kind of bone is using >the Instrumedics CryoJane tape transfer system. Check out their website for >information, how it works. >Bone frozen sections are destroyed by disposable or c profile steel blade, >tungsten carbide blades are ideal as you are finding out. However, I have >never had a bone frozen section cut with a TC knife which remains intact >unless the tape transfer method is used to maintains bone >integrity. Cutting temperature must be -28C up to -32C or so. > >Your tungsten carbide knife edge must be in perfectly, sharp condition even >with tape transfer method. > >For us, cryosectioning bone without the Cryojane has been a waste of >time. Although a pricey bit of equipment, it has paid for itself many >times over with good bone frozen sections. > >I am going to attach a publication to you privately so you can see >advantages. > > > > > >At 10:02 AM 8/27/2004, you wrote: > >I am doing frozen sections on fresh, undecalcified mouse femurs. My > >sections seem to be losing all of the marrow and many are torn . I have > >tried disposable cryostat knives as well as the tungsten carbide, but the > >sections are still not good. If anyone else is doing this procedure, could > >you share any tips or suggestions on your technique, cutting temperatures, > >or overall procedure. I have tried different temperatures, but none of them > >seem to give me the sections I want. Any help would > > > >be greatly appreciated. > > > >Nancy.Lowen@med.va.gov > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From arvind <@t> nbrc.res.in Mon Aug 30 09:17:58 2004 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] FW: mol wt of mu, kappa,delta opiod receptors Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A6CA@mail.nbrc.res.in> can any one tell mol. wt of mu,kappa,delta opiod receptors arvind@nbrc.res.in From info <@t> instrumedics.com Mon Aug 30 09:32:10 2004 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Cleaning the cryostat Message-ID: <015a01c48e9e$2232fb80$6401a8c0@INSTRUMEDICS22> The Cryo-Vac-Away vacuum system does not "decontaminate" the cryostat but it does keep the cryostat free of infectious trimming debris. As the block is faced off a nozzle at the blockface suctions away the trimming debris into a cold filter that is inside the cryostat. Pathogens that might escape the debris are stopped by a second viral/bacterial filter. The cryostat remains spotless! See details on the web www. instrumedics.com Bernice From gcallis <@t> montana.edu Mon Aug 30 09:33:24 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] GFP antibodies In-Reply-To: <41331E1D.5060607@email.unc.edu> References: <057B8DF3-F922-11D8-BEB2-000393DBA4AA@emory.edu> <41331E1D.5060607@email.unc.edu> Message-ID: <6.0.0.22.1.20040830082710.01b2e760@gemini.msu.montana.edu> Try Chemicons polyclonal or Molecular Probes polyclonal. Clontech may have them too and they are the GFP gurus. You could contact Clonetech technical services, gmehta@BD.com. She is Clonetech/BD tech services person. A t 06:31 AM 8/30/2004, you wrote: >'Netter's, > >I would like to order in some anti-GFP antibodies. I realize that there >are several out there and I'd like to get one that will work on both FFPE >and frozen sections of murine tissue.... Open to suggestions. > >THX > >Robert Schoonhoven, >UNC-CH >CEHS > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From RBARNHART <@t> summithealth.org Mon Aug 30 09:38:43 2004 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Trimming blocks Message-ID: Just curious what everyone else is using to trim the excess paraffin off the blocks? Thanks for the input. From gu.lang <@t> gmx.at Mon Aug 30 09:54:08 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] IF troubleshooting Message-ID: <003601c48ea1$396fe5d0$eeeea8c0@server> Can anyone give me a website with IF-troubleshooting, especially on FFPE renal biopsies? Or any other information in electronical form. thanks in advance Gudrun Lang From GDawson <@t> Milw.Dynacare.com Mon Aug 30 10:12:23 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] IF on RPMI tissue Message-ID: All, I have a client that wants to do an Immunofluorescent Study on a skin punch bx. that's been in RPMI medium for 5 days. Does anyone have any thoughts as to whether or not this will work? Can anyone give me an idea on what effect the RPMI may have on the results? Thanx In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Coordinator Milwaukee, WI From Jamie.Dukes <@t> se.amedd.army.mil Mon Aug 30 10:25:59 2004 From: Jamie.Dukes <@t> se.amedd.army.mil (Dukes, Jamie Mr EAMC) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] pregant woman in the grossroom Message-ID: <4FB3076916FCD311AF7900805FA7A678077B4B0C@dasmthfdz001.amedd.army.mil> HELLO ALL, I really need something in writing to clarify that a pregnant woman cannot not work in the grossroom. From cfavara <@t> niaid.nih.gov Mon Aug 30 10:36:21 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] GFP antibodies Message-ID: I did not see the original post on this : Molecular Probes A-11122 work well on FFPE murine tissue can send specifics if you like! Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Monday, August 30, 2004 8:33 AM To: rschoon; Histonet@lists.utsouthwestern.edu Subject: [Histonet] GFP antibodies Try Chemicons polyclonal or Molecular Probes polyclonal. Clontech may have them too and they are the GFP gurus. You could contact Clonetech technical services, gmehta@BD.com. She is Clonetech/BD tech services person. A t 06:31 AM 8/30/2004, you wrote: >'Netter's, > >I would like to order in some anti-GFP antibodies. I realize that >there are several out there and I'd like to get one that will work on >both FFPE and frozen sections of murine tissue.... Open to suggestions. > >THX > >Robert Schoonhoven, >UNC-CH >CEHS > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Aug 30 10:36:33 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Trimming blocks In-Reply-To: References: Message-ID: <6.0.0.22.1.20040830093522.01b26a80@gemini.msu.montana.edu> an old solid scalpel blade. For a nice setup, look at Thermo Electron's (Shandon) hot plate, tilted for this purpose- I would love one. At 08:38 AM 8/30/2004, you wrote: >Just curious what everyone else is using to trim the excess paraffin off >the blocks? Thanks for the input. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From BlazekL <@t> childrensdayton.org Mon Aug 30 10:44:30 2004 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Trimming blocks Message-ID: I use one of those little knives that you see for $1.00 at the hardware store checkout. I've used the same one for 20 + years. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org From HornHV <@t> archildrens.org Mon Aug 30 10:45:56 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Trimming blocks Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B938@EMAIL.archildrens.org> An old butter knife from home.....works great........ Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca Barnhart Sent: Monday, August 30, 2004 9:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Trimming blocks Just curious what everyone else is using to trim the excess paraffin off the blocks? Thanks for the input. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From convmcm <@t> cc.usu.edu Mon Aug 30 10:53:46 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] pregant woman in the grossroom In-Reply-To: <4FB3076916FCD311AF7900805FA7A678077B4B0C@dasmthfdz001.amedd.army.mil> Message-ID: <001a01c48ea9$890ec400$4a737b81@Cygnus> Why? What's in your grossing room that would require you to request support for such a strict ban? There are all kinds of hazards in a laboratory environment that are not good for a fetus, but then, they aren't good for the people who work there, either. Also, each lab is different. Some places have good ventilation and provide the very best in PPE. Other's aren't so good. I've known many pregnant women who have worked in grossing rooms, morgues, labs, etc (I'm one of them) and I see a great and urgent need to provide the very best in protective apparel, safety and proper ventilation, but never have I felt a woman should be banned from any area just because she is pregnant. I feel that is a very personal thing that the woman, the employer and her physician should consult and concur on. So, again, I ask you why? I understand there are liability issues that are a more recent problem, but these are conflicting in nature: prevent her from working just because she's pregnant and that's discrimination. If she has a baby with birth defects, you've got a liability issue. Pick you poiso & have fun. Just a thought Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 email: convmcm@cc.usu.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dukes, Jamie Mr EAMC Sent: Monday, August 30, 2004 8:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pregant woman in the grossroom HELLO ALL, I really need something in writing to clarify that a pregnant woman cannot not work in the grossroom. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lleach <@t> uic.edu Mon Aug 30 11:10:32 2004 From: lleach <@t> uic.edu (Lu Leach) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] injecting tracers Message-ID: <6.1.0.6.2.20040830110655.024a0a08@tigger.uic.edu> Dear Histonetters, I need advice on a fluorescent tracer to use for mouse injections that will hopefully survive tissue processing. If anyone has experience and wisdom in this area, please contact me. Thank you, Lu Leach University of Illinois at Chicago lleach@uic.edu From garygill <@t> dcla.com Mon Aug 30 11:10:37 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] IF on RPMI tissue Message-ID: <0B11620AEE5EFB4FA7A7558339972A83070570@HALIBUT.dcla.com> Generally speaking, RPMI is highly physiologic, so skin punch bx may be fine -- especially if refrigerated in the interim. However, RPMI also readily supports microbial growth. If that latter occurred in this instance, bx may be useless. Gary Gill -----Original Message----- From: Dawson, Glen [mailto:GDawson@Milw.Dynacare.com] Sent: Monday, August 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IF on RPMI tissue All, I have a client that wants to do an Immunofluorescent Study on a skin punch bx. that's been in RPMI medium for 5 days. Does anyone have any thoughts as to whether or not this will work? Can anyone give me an idea on what effect the RPMI may have on the results? Thanx In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Coordinator Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From celebrej <@t> HHSC.CA Mon Aug 30 11:17:04 2004 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Trimming blocks Message-ID: <3AADFB88753AD31189C100902786B91C0E41CE61@hch_nt_exchange.hhsc.ca> We use old steak knives... -----Original Message----- From: Linda Blazek [mailto:BlazekL@childrensdayton.org] Sent: Monday, August 30, 2004 11:45 AM To: Histonet@lists.utsouthwestern.edu; gcallis@montana.edu; RBARNHART@summithealth.org Subject: Re: [Histonet] Trimming blocks I use one of those little knives that you see for $1.00 at the hardware store checkout. I've used the same one for 20 + years. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From Tbarnhart <@t> primecare.org Mon Aug 30 11:16:26 2004 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] pregant woman in the grossroom Message-ID: <1779904B5E82D511914C00D0B793339205BFD841@exchangent> Humm...If your gross room is following OSHA standards (formaldehyde exposure limits) and Universal Precautions (blood and body fluid precautions) than ANYONE can work in your gross room. If not, NOBODY can work in your gross room. Pregnancy is not an issue. -----Original Message----- From: Dukes, Jamie Mr EAMC [mailto:Jamie.Dukes@se.amedd.army.mil] Sent: Monday, August 30, 2004 10:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pregant woman in the grossroom HELLO ALL, I really need something in writing to clarify that a pregnant woman cannot not work in the grossroom. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. From JQB7 <@t> CDC.GOV Mon Aug 30 11:17:39 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Trimming blocks Message-ID: I am lazy so I embed VERY carefully so I don't have any excess to speak of. However, if I bump the base mold as I am placing it on the cold plate I may have a little excess. I just pop it off with my fingernail Jeanine Bartlett CDC -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rebecca Barnhart Sent: Mon 8/30/2004 10:38 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Trimming blocks Just curious what everyone else is using to trim the excess paraffin off the blocks? Thanks for the input. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bill501 <@t> mindspring.com Mon Aug 30 11:30:50 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Processor stuck in 100% ETOH Message-ID: Our processor got stuck in a 100% ethanol station and sat for about 6 hours. The tissue is hard, brittle and very difficult to cut- mostly skin and GI biopsies. Any suggestions on 'softening' the tissue and making cutting easier would be greatly appreciated. TIA, Bill -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board From gu.lang <@t> gmx.at Mon Aug 30 11:37:27 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] IF troubleshooting References: Message-ID: <005301c48eaf$a4b9d680$eeeea8c0@server> RE: [Histonet] IF troubleshootingTeresa, the doctors complain about a misty look of the slides. They believe something like too little deparaffination. There is some reaction, but not clear. I have to ask them about further informations. We did the same procedure like everytime. (this time even with very fresh xylol). We had this problem once before a while. We repeated the whole procedure on the same slides incl. dryer and xylol. Then the doctors were satisfied, but I have my worries about the significance of the test. (Besides every antibody was negativ) We don't run controls. our protocol: 30 min 60 degree dryer 3 x 7 min Xylol each 2 min Alcohol 96-80-50 15 min 0,1% Protease in PBS 37 degree cool in cold PBS ... 30 min fitc-labeled antibodies 3 x 5 min rinse with PBS mounting with IF-medium Can you help us? Gudrun ----- Original Message ----- From: Flores, Teresa To: Gudrun Lang Sent: Monday, August 30, 2004 6:14 PM Subject: RE: [Histonet] IF troubleshooting Gudrum, what kind of trouble are you having? Teresa -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Monday, August 30, 2004 9:54 AM To: Histonetliste Subject: [Histonet] IF troubleshooting Can anyone give me a website with IF-troubleshooting, especially on FFPE renal biopsies? Or any other information in electronical form. thanks in advance Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From flemons <@t> bhset.org Mon Aug 30 11:37:48 2004 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Ethel Macrea Message-ID: Would you be so kind as to contact me, either email or by phone....I need to ask you about a block you sent me. Thanks Fran Walker flemons@bhset.org 865-632-5133 From TJJ <@t> Stowers-Institute.org Mon Aug 30 11:40:25 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] GFP antibodies on murine tissue Message-ID: We use anti-GFP from Novus Biologicals (it's actually an Abcam antibody), rabbit polyclonal NB 600-303 on both frozen and paraffin mouse and chick tissues. It works beautifully at a 1:1000 dilution with Dako Rabbit Envision + HRP. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From Jackie.O'Connor <@t> abbott.com Mon Aug 30 11:42:23 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] pregnant woman in the grossroom Message-ID: This has been discussed before - I don't think there are any LEGAL reasons to keep a pregnant woman out of the gross room. Personally, when I was pregnant (#6) I was ordered in writing by my high risk OB to stay away from Formalin and Xylene, and the large company complied by giving me a medical LOA. When I was pregnant (#3) and worked for a Tox Lab at the University of Tennessee, the tox PhDs suggested I stay away from Formalin and Xylene. When pregnant (#4) and working in a California hospital where they did 2nd term abortions, I couldn't bear seeing the 'products of conception' - so the hospital complied and I didn't have to work in the gross room. There hasn't been a full blown study on birth defects/affects and low level exposure to chemicals in the histology lab (as far as I know). I think it would be a great study for the NSH to sponsor, or maybe by one of the big Histology vendors. Retrospectively, I would not have worked in a histology lab during my pregnancies. Since hindsight is 20/20 - I would have sent all four of my kids to boarding school as soon as they became teenagers, too - - - - - oh well. Jackie O' "Barnhart, Tammy" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/30/2004 11:16 AM To: "'Dukes, Jamie Mr EAMC'" , histonet@lists.utsouthwestern.edu cc: Subject: RE: [Histonet] pregant woman in the grossroom Humm...If your gross room is following OSHA standards (formaldehyde exposure limits) and Universal Precautions (blood and body fluid precautions) than ANYONE can work in your gross room. If not, NOBODY can work in your gross room. Pregnancy is not an issue. -----Original Message----- From: Dukes, Jamie Mr EAMC [mailto:Jamie.Dukes@se.amedd.army.mil] Sent: Monday, August 30, 2004 10:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pregant woman in the grossroom HELLO ALL, I really need something in writing to clarify that a pregnant woman cannot not work in the grossroom. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rschoon <@t> email.unc.edu Mon Aug 30 12:14:15 2004 From: rschoon <@t> email.unc.edu (rschoon) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] THANKS to all on my GFP antibodies on murine tissue quetion In-Reply-To: <200408301641.i7UGfMYI013848@email.unc.edu> References: <200408301641.i7UGfMYI013848@email.unc.edu> Message-ID: <41336067.2020800@email.unc.edu> To all who replied, a huge Thank You!! I have what I needed. Tanks, Bob From tliparul <@t> mix.wvu.edu Mon Aug 30 12:48:19 2004 From: tliparul <@t> mix.wvu.edu (Timothy Lewis Liparulo) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Trypan Blue Removal For Protein Analysis Message-ID: <4778566.1093888099058.JavaMail.tliparul@mix.wvu.edu> I am currently using Trypan Blue as a measure for injury in rat skeletal muscle cells. The animals were preinjected with the dye 24 hrs prior to sacrifice. I would like to perform a protein analysis on the tissue but am afraid that the dye in the tissue will be problematic for western blot analysis. Does anyone know of a way to remove the dye pigment without significantly affecting the integrity of the muscle protein? From John.Garlits <@t> STJUDE.ORG Mon Aug 30 13:04:22 2004 From: John.Garlits <@t> STJUDE.ORG (Garlits, John) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] alkaline phosphatase staining Message-ID: <1E0CC447E59C974CA5C7160D2A2854EC03C40E75@SJMEMXMB04.stjude.sjcrh.local> Hello, I am interested in doing some alkaline phosphatase staining in decalcified, paraffin-fixed mouse long bone sections. Does anyone have any experience with alk phos staining, who could recommend a protocol or even a commercial kit? Thank you! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 Phone: 901-495-3961 Fax: 901-495-2176 From eshields <@t> bhset.org Mon Aug 30 14:00:47 2004 From: eshields <@t> bhset.org (E Sharon Shields) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] AMACR(P504S) Message-ID: Histonetters and vendors, I need contact information for Zeta Corp.,Sierra Madre, CA. Our pathologist are very interested in the prostate marker AMACR (P504S), which was written about recently in The American jJournal of Clinical Pathology. Sharon Shields, CT(ASCP)QIHC Baptist Hospital of E TN Knoxville, TN eshields@bhset.org 865.549.4351 From AFeatherstone <@t> KaleidaHealth.Org Mon Aug 30 14:05:29 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] acetylcholoresterase Message-ID: is anyone familiar with a immuno stain for acetylcholoresterase? Annette Featherstone HT/MLT -----Original Message----- From: Garlits, John [mailto:John.Garlits@STJUDE.ORG] Sent: Monday, August 30, 2004 14:04 To: HistoNet Server Subject: [Histonet] alkaline phosphatase staining Hello, I am interested in doing some alkaline phosphatase staining in decalcified, paraffin-fixed mouse long bone sections. Does anyone have any experience with alk phos staining, who could recommend a protocol or even a commercial kit? Thank you! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 Phone: 901-495-3961 Fax: 901-495-2176 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From failm <@t> musc.edu Mon Aug 30 14:14:20 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] AMACR(P504S) Message-ID: Sharon, We use Biocare for this product with good results Rena Fail >>> "E Sharon Shields" 08/30/04 03:00PM >>> Histonetters and vendors, I need contact information for Zeta Corp.,Sierra Madre, CA. Our pathologist are very interested in the prostate marker AMACR (P504S), which was written about recently in The American jJournal of Clinical Pathology. Sharon Shields, CT(ASCP)QIHC Baptist Hospital of E TN Knoxville, TN eshields@bhset.org 865.549.4351 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lucyb <@t> biocare.net Mon Aug 30 15:12:30 2004 From: lucyb <@t> biocare.net (Lucy Brooks) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] NSH Question Message-ID: <000c01c48ecd$adb1ff60$0a01a8c0@LUCYSALES> Does anyone have a list of Vendor Sponsored Evening Events that are going on at NSH after the booths close each night? Please email me back, and let me know what's going on. Thanks so much everyone for your help. Lucy Brooks Technical Sales Rep & Support Biocare Medical (800) 799-9499 From rockbeki <@t> ufl.edu Mon Aug 30 15:32:05 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Trimming blocks Message-ID: <517870335.1093897925495.JavaMail.osg@osgjas02.cns.ufl.edu> I use safety razors for the sides, and then plop it on the microtome for the front trimming. Rebekah Smith On Mon Aug 30 11:45:56 EDT 2004, "Horn, Hazel V" wrote: > An old butter knife from home.....works great........ > > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > > Phone - 501.364.4240 > Fax - 501.364.3912 -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Rebecca > Barnhart > Sent: Monday, August 30, 2004 9:39 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Trimming blocks > > > Just curious what everyone else is using to trim the excess > paraffin off > the blocks? Thanks for the input. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ------------------------------------------------------------------------------ > The information contained in this message may be privileged and > confidential and protected from disclosure. If the reader of this > message is not the intended recipient, or an employee or agent > responsible for delivering this message to the intended > recipient, you are hereby notified that any dissemination, > distribution or copying of this communication is strictly > prohibited. If you have received this communication in error, > please notify us immediately by replying to the message and > deleting it from your computer. Thank you. > ============================================================================== > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From Dndsomi <@t> aol.com Mon Aug 30 15:38:21 2004 From: Dndsomi <@t> aol.com (Dndsomi@aol.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Trimming blocks Message-ID: <141.32578c29.2e64ea3d@aol.com> We use an old pocket knife ! Works great ! From JNocito <@t> Pathreflab.com Mon Aug 30 16:31:52 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] AMACR(P504S) In-Reply-To: Message-ID: Sharon, we purchase our P504s from Biocare. Their number is 1-800-799-9499. They also have a PIN cocktail that has P504s and p63 that the pathologists' here like. You get two antibodies on one section. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of E Sharon Shields Sent: Monday, August 30, 2004 2:01 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] AMACR(P504S) Histonetters and vendors, I need contact information for Zeta Corp.,Sierra Madre, CA. Our pathologist are very interested in the prostate marker AMACR (P504S), which was written about recently in The American jJournal of Clinical Pathology. Sharon Shields, CT(ASCP)QIHC Baptist Hospital of E TN Knoxville, TN eshields@bhset.org 865.549.4351 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bills <@t> icpmr.wsahs.nsw.gov.au Mon Aug 30 16:40:28 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Trimming blocks In-Reply-To: Message-ID: <000f01c48ed9$f791a020$83a7080a@wsahs.nsw.gov.au> Thermo Electron have this wonderful Trimming plate. Expensive but well worthwhile. We could not be without them now. Purchased as a result of an accident with the solid scalpels we used and a workplace assessment. Now all we worry about is people burning themselves!!!!! Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rebecca Barnhart Sent: Tuesday, 31 August 2004 12:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Trimming blocks Just curious what everyone else is using to trim the excess paraffin off the blocks? Thanks for the input. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From JWEEMS <@t> sjha.org Mon Aug 30 16:44:08 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Roommate Message-ID: <83AACDB0810528418AA106F9AE9B7F7E2781D2@sjhaexc02.sjha.org> After all the jabber on Friday, did the person who needed a roommate for NSH find one? Just curious. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From bamur <@t> alaska.net Mon Aug 30 06:36:35 2004 From: bamur <@t> alaska.net (Barbara Murray) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Histology Position Message-ID: <001b01c48e85$d415a780$ab5ae642@d3b1l9> Greetings from Anchorage, Alaska, The Deptartment of Pathology/Histology @ The Alaska Native Tribal Health Consortium has an opening for a Histologist. This is a full time position with no weekends and holidays. Please visit our website @ www.anthc.org for a complete application. Any questions, please call The Office of Human Resources. The phone number is : 907-729-1301 and the facimile is : 907-729-3638. Thanks and have a great day! Barbara A. Murray, HT.(ASCP) Histology/Pathology Dept. The Alaska Native Medical Center Anchorage, Alaska 99508 907-729-1804 From Andrew.Gray <@t> vcp.monash.edu.au Mon Aug 30 23:03:38 2004 From: Andrew.Gray <@t> vcp.monash.edu.au (Andrew Gray) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Re: mol wt of mu, kappa,delta opiod receptors References: <20040830163727.7000A4FB03@moe.its.monash.edu.au> Message-ID: <130.194.217.111.1093924531.94964@my.monash.edu.au> Hi Arvind, Unfortunately this is not a simple question to answer. You can calculate the theoretical MW of the receptor proteins from their amino acid sequences, but then the party is spoilt by post-traslational modifications, not to mention the many isoforms/splice variants of opioid receptors that exist, all of which have different MW's. Of course you could look at a paper of a Western blots done to opioid receptors, being mindful that the receptors tend to dimerise, producing a second band. What are your requirements? Best wishes, Andrew (Still a PhD student in opioid receptors) > ------------------------------ > > Message: 12 > Date: Mon, 30 Aug 2004 19:47:58 +0530 > From: "Arvind Pundir" > Subject: [Histonet] FW: mol wt of mu, kappa,delta opiod receptors > > can any one tell mol. wt of mu,kappa,delta opiod > receptors > > arvind@nbrc.res.in From JCarpenter764 <@t> aol.com Tue Aug 31 04:23:51 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] cutting tonsil Message-ID: <09DC1D7B.29D6EC80.40C6E687@aol.com> does anyone have any good tips or suggestions on cutting a perfect section of tonsil at 2-3 microns? From JCarpenter764 <@t> aol.com Tue Aug 31 04:24:11 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Fwd: cutting tonsil at 2-3 microns Message-ID: <3F1D1DBD.3597549A.40C6E687@aol.com> From JQB7 <@t> CDC.GOV Tue Aug 31 05:18:33 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] NSH Question Message-ID: Go to www.NSH.org and click on NSH Convention. The click on "Events" (in gold) and there you are! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lucy Brooks Sent: Monday, August 30, 2004 4:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] NSH Question Does anyone have a list of Vendor Sponsored Evening Events that are going on at NSH after the booths close each night? Please email me back, and let me know what's going on. Thanks so much everyone for your help. Lucy Brooks Technical Sales Rep & Support Biocare Medical (800) 799-9499 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From stevemachinuk <@t> yahoo.co.uk Tue Aug 31 07:44:04 2004 From: stevemachinuk <@t> yahoo.co.uk (=?iso-8859-1?q?Steve=20Machin=20UK?=) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Re: Working while pregnant Message-ID: <20040831124404.451.qmail@web25104.mail.ukl.yahoo.com> I can think of three causes for concern when working while prognant. 1 Ionising radiations from the Lab Faxitron and incidental radiation while in the X-Ray department. 2 Chemical hazards identified in the data sheets which are described as ?Evidence of reproductive effects?. Lithium carbonate, Trichloroacetic acid, Physostigmine (seserine), Manganese (II) chloride, Propane 1,2 diol (Propylene glycol), Tartrazine, Formaldehyde, OG6 (Pap stain), Copper (II) sulphate, Ethylene glycol, Silver nitrate, Phenol, Mercuric oxide, Light green SF, Glycerol and 2 Ethoxyethanol (cellosolve). Chemicals identified by Health and Safety Executive: Lead (II) acetate. 3 Physical hazard from lifting sitting, standing and posture especially microtomy and prolonged VDU work. 4 Infectious hazards in the post mortem room and when handling fresh specimens especially placentas and products of conception. Steve Machin UK ___________________________________________________________ALL-NEW Yahoo! Messenger - all new features - even more fun! http://uk.messenger.yahoo.com From chiggerson <@t> memhosp.com Tue Aug 31 07:48:31 2004 From: chiggerson <@t> memhosp.com (chiggerson@memhosp.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Powerpath In-Reply-To: Message-ID: We have PowerPath V8.3.4. Cindy Higgerson HTL(ASCP) Surgical Pathology Supervisor Memorial Hospital Belleville, Illinois office: 618-257-5085 fax: 618-257-6868 "Nita Searcy" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/30/2004 07:15 AM To cc Subject [Histonet] Powerpath Last week I aked for names of Tamtron/ Powerpath users- I received one reply. There are supposed to be 350 users out there---where are you??? Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pengbw <@t> sjtu.edu.cn Tue Aug 31 08:55:31 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Muscle regeneration Message-ID: <20040831135531.87C7B1103C42@sjtu.edu.cn> Dear all, We are currently involved in a project in which satellite cell regenerated skeletal muscle fibers and fibroblasts, which generally in appearance are required to be discriminated. Does anyone have a protocol to identify skeletal muscle regeration? Any comments will be appreciated greatly! Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From DCoulter <@t> Lifespan.org Tue Aug 31 09:29:01 2004 From: DCoulter <@t> Lifespan.org (Coulter, Diane) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Coverslipping tape problems Message-ID: We're having serious problems with older (7-8 yrs) slides coverslipped with tape. When these slides are retrieved form the files more often than not the tape has separated from the slide with the tissue attached to the tape. I've repaired some of these using Sakura's suggested method of immersing the tape in xylene for a few minutes then reattaching to the slide. This is messy, time consuming and usually results in air bubbles. Any suggestions, short of recutting all these slides? Is there any way to prevent this from happening in the first place? Right now I'm ready toss the coverslipper out the window. Thanks in advance, Diane Coulter, RIH Histology From BSylinda <@t> aol.com Tue Aug 31 09:37:08 2004 From: BSylinda <@t> aol.com (BSylinda@aol.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Looking for tissue for registry Message-ID: <4625B630.41C51C5A.00698496@aol.com> Hello to all, I am writing trying to find out if anyone may have these pieces of tissue available in their lab: artery liver lung small intestine If so could you mail me parrafin blocks to Pathology Services 1002 Texas Blvd Texarkana, TX 75501 Suite 500 Thanks in advance, Sylinda Battle From pam <@t> ategra.com Tue Aug 31 10:57:04 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Job Opportunities in Histology Latest Update 09/01/04 Message-ID: Hi Histonetters, Here is the latest update on opportunities with some of my best clients throughout the US who are seeking Histology Supervisors, Histo Technologists and Histo Technicians. These are positions as direct employees of our client. There are fulltime 40 hour per week positions.. As a direct employee of one of our clients you will be provided with full benefits including Health Insurance, Vacation, Sick Pay, Relocation money and a lucrative sign-on bonus. I have part time and temporary positions as well. I have supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are some of my HOTTEST Histology Supervisory positions: 1. Alabama - Histology Supervisor 2. Colorado - Histology Manager/PA (I also need a cytology manager/supervisor for this client if you know anyone) 3. Massachusetts - Histology Manager 4. California - Immunohistochemistry Supervisor 5. Texas - Pathology Assistant Here are some of my HOTTEST Histo Tech and Histology Supervisor positions: 1. Massachusetts - Histo Tech 2. Oregon - Histo Tech (nights) 3. Maine - Histo Tech 4. Maine - IHC Tech 5. N. California - Histo Tech 6. NYC - Lead Histo Tech - evenings 2p-11p 7. NYC - IHC Tech - nights 10p-7a 8. SW Florida - Histo Tech If you are interested in these jobs, please CALL ME ASAP at 800 466 9919 x234. To speed things up, please also send me a copy of your resume, (if you haven't already done so). If you are interested in jobs outside the above-mentioned areas, please send me your resume as well. I have clients throughout the US. I will keep your resume confidential and will not release it to anyone without your permission (This is Ategra policy as well as my own). My services are at no charge to you. Of course, you may be happy in your present job, but it never hurts to to keep an eye open. Also, if you have friends/peers who might be interested as well, if you could pass my query & name on to them I'd be very grateful. However, if you are interested in any of the jobs above, please call me. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Pam Barker Senior Lab Recruiter Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd. Winter Park, FL 32792 VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com To Learn More About Ategra: http://www.ategra.com -------------------------------------------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. -------------------------------------------------------------------------------------------------- From Rcartun <@t> harthosp.org Tue Aug 31 12:22:49 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] acetylcholoresterase Message-ID: Do you mean "Acetylcholinesterase"? RWC >>> "Featherstone, Annette" 08/30/04 03:05PM >>> is anyone familiar with a immuno stain for acetylcholoresterase? Annette Featherstone HT/MLT -----Original Message----- From: Garlits, John [mailto:John.Garlits@STJUDE.ORG] Sent: Monday, August 30, 2004 14:04 To: HistoNet Server Subject: [Histonet] alkaline phosphatase staining Hello, I am interested in doing some alkaline phosphatase staining in decalcified, paraffin-fixed mouse long bone sections. Does anyone have any experience with alk phos staining, who could recommend a protocol or even a commercial kit? Thank you! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 Phone: 901-495-3961 Fax: 901-495-2176 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AFeatherstone <@t> KaleidaHealth.Org Tue Aug 31 12:25:26 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] acetylcholinesterase Message-ID: I am sorry...spelling...yes Acetylcholinesterase -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, August 31, 2004 13:23 To: AFeatherstone@kaleidahealth.org; histonet@lists.utsouthwestern.edu; John.Garlits@STJUDE.ORG Subject: Re: [Histonet] acetylcholoresterase Do you mean "Acetylcholinesterase"? RWC >>> "Featherstone, Annette" 08/30/04 03:05PM >>> is anyone familiar with a immuno stain for acetylcholoresterase? Annette Featherstone HT/MLT -----Original Message----- From: Garlits, John [mailto:John.Garlits@STJUDE.ORG] Sent: Monday, August 30, 2004 14:04 To: HistoNet Server Subject: [Histonet] alkaline phosphatase staining Hello, I am interested in doing some alkaline phosphatase staining in decalcified, paraffin-fixed mouse long bone sections. Does anyone have any experience with alk phos staining, who could recommend a protocol or even a commercial kit? Thank you! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 Phone: 901-495-3961 Fax: 901-495-2176 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From Janet.Bonner <@t> FLHOSP.ORG Tue Aug 31 12:42:04 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Coverslipping tape problems Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB402A@fh2k093.fhmis.net> We have the same problem- and we just recut the slides as they are needed for send out or review boards. The important thing is to go back to glass ASAP. We use the automatic glass coverslippers now. Sorry to hear about your predicament, I know it's a rude situation to confront. -Janet -----Original Message----- From: Coulter, Diane To: 'histonet@lists.utsouthwestern.edu' Sent: 8/31/2004 10:29 AM Subject: [Histonet] Coverslipping tape problems We're having serious problems with older (7-8 yrs) slides coverslipped with tape. When these slides are retrieved form the files more often than not the tape has separated from the slide with the tissue attached to the tape. I've repaired some of these using Sakura's suggested method of immersing the tape in xylene for a few minutes then reattaching to the slide. This is messy, time consuming and usually results in air bubbles. Any suggestions, short of recutting all these slides? Is there any way to prevent this from happening in the first place? Right now I'm ready toss the coverslipper out the window. Thanks in advance, Diane Coulter, RIH Histology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From akbitting <@t> geisinger.edu Tue Aug 31 12:55:46 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Factor XIII Message-ID: Calbiotech has discont'd their Factor XIII Ab. Can anyone tell me of another supplier of this Ab? We're using it on FFPE human tissue for IHC. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From portera203 <@t> yahoo.com Tue Aug 31 13:04:21 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Factor XIII In-Reply-To: Message-ID: <20040831180421.35102.qmail@web40908.mail.yahoo.com> We use Dako's Polyclonal - it works very well. Angela Bitting wrote:Calbiotech has discont'd their Factor XIII Ab. Can anyone tell me of another supplier of this Ab? We're using it on FFPE human tissue for IHC. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet W Amy S.Porter, HT(ASCP) Michigan State University Department of Physiology Division of Human Pathology College of Human Medicine portera203@yahoo.com --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - 100MB free storage! From arvind <@t> nbrc.res.in Tue Aug 31 13:36:15 2004 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] opioid receptor mol wt Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A6CD@mail.nbrc.res.in> Message: 14 Date: Tue, 31 Aug 2004 04:03:38 +0000 From: Andrew Gray Subject: [Histonet] Re: mol wt of mu, kappa,delta opiod receptors To: histonet@lists.utsouthwestern.edu Message-ID: <130.194.217.111.1093924531.94964@my.monash.edu.au> Content-Type: text/plain Hi Arvind, Unfortunately this is not a simple question to answer. You can calculate the theoretical MW of the receptor proteins from their amino acid sequences, but then the party is spoilt by post-traslational modifications, not to mention the many isoforms/splice variants of opioid receptors that exist, all of which have different MW's. Of course you could look at a paper of a Western blots done to opioid receptors, being mindful that the receptors tend to dimerise, producing a second band. What are your requirements? Best wishes, Andrew (Still a PhD student in opioid receptors) \ arvind@nbrc.res.in Hi andrew, thanks for ur suggestions on opioid receptors actually we r doing immuno on zebrafinch bird brain sections 40 micron thick we have got opioid receptors from sigma , but after repeat trials we r not able to localise opioid receptors in brain sections we have tried with mu.delta and kappa but non is working till now , we have done western blot and were able to get two bands nearly 45-49 kd can u tell why there r two bands there............ can u do me a favour by suggesting some protocol in detail, i will be thanfull to u for that, if u can pls mail by earliest at .................................................. arvind@nbrc.res.in Arvind Singh Pundir N B R C, Haryana, INDIA From gcallis <@t> montana.edu Tue Aug 31 14:39:20 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Factor XIII In-Reply-To: References: Message-ID: <6.0.0.22.1.20040831133832.01b273d0@gemini.msu.montana.edu> DAKO's is superb and works on mouse also!! At 11:55 AM 8/31/2004, you wrote: >Calbiotech has discont'd their Factor XIII Ab. Can anyone tell me of >another supplier of this Ab? We're using it on FFPE human tissue for IHC. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From madaryhtl <@t> juno.com Tue Aug 31 14:52:56 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] My tech needs uterus for HT.. Got Uterus? Message-ID: <20040831.125325.3306.609124@webmail11.lax.untd.com> Nick(Rocky) Madary, HT,HTL(ASCP)QIHC Histology Manager, Medimmune Inc Cell-3012334950 Work-3013984745 Fax-3013989745 From Rcartun <@t> harthosp.org Tue Aug 31 16:17:31 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Factor XIII Message-ID: Are we talking Factor XIIIa or VIII here? RWC >>> Gayle Callis 08/31/04 03:39PM >>> DAKO's is superb and works on mouse also!! At 11:55 AM 8/31/2004, you wrote: >Calbiotech has discont'd their Factor XIII Ab. Can anyone tell me of >another supplier of this Ab? We're using it on FFPE human tissue for IHC. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From azdudley <@t> hotmail.com Tue Aug 31 16:25:30 2004 From: azdudley <@t> hotmail.com (anita dudley) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] NSH Question Message-ID: Lucy, if you go to the NSH website and go to convention then events there is a list of the things that are going on. anita dudley, providence hosp. mobile ala >From: "Lucy Brooks" >To: >Subject: [Histonet] NSH Question >Date: Mon, 30 Aug 2004 13:12:30 -0700 > >Does anyone have a list of Vendor Sponsored Evening Events that are going >on at NSH after the booths close each night? Please email me back, and let >me know what's going on. Thanks so much everyone for your help. > >Lucy Brooks >Technical Sales Rep & Support >Biocare Medical >(800) 799-9499 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Get ready for school! Find articles, homework help and more in the Back to School Guide! http://special.msn.com/network/04backtoschool.armx From gcallis <@t> montana.edu Tue Aug 31 16:54:54 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Factor X111a or VIII, a whoops In-Reply-To: References: Message-ID: <6.0.0.22.1.20040831155328.01b14f58@gemini.msu.montana.edu> I was referring to Factor VIII von willebrands. Must be blind and not reading Roman numerals!! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From JNocito <@t> Pathreflab.com Tue Aug 31 18:01:20 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] I have a funny Message-ID: Hello all, for once, I have, what I think is funny, is a short story. A couple of months ago, I purchased a piece of equipment from a vendor (who will remain anonymous, at this time at least). I was told it was 6 weeks for delivery. It's been over 8 weeks. I called the rep and left a voice mail asking when I can expect my equipment. S/he called me right back. I asked when I can expect the equipment. Then I said "Don't make me go on the Histonet". My techs started laughing and I couldn't keep a straight face. The rep was all apologetic and promised me I'll have my equipment in 2 weeks. Gee, I didn't think I was getting that bad. Maybe I should settle down a little bit. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX From lucyb <@t> biocare.net Tue Aug 31 18:10:52 2004 From: lucyb <@t> biocare.net (Lucy Brooks) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] I have a funny References: Message-ID: <000b01c48faf$c3768630$0a01a8c0@LUCYSALES> Joe!! Speak your mind!!! The Histonet is a powerful thing!! ----- Original Message ----- From: "Joe Nocito" To: "Histonet" Sent: Tuesday, August 31, 2004 4:01 PM Subject: [Histonet] I have a funny > Hello all, > for once, I have, what I think is funny, is a short story. A couple of > months ago, I purchased a piece of equipment from a vendor (who will remain > anonymous, at this time at least). I was told it was 6 weeks for delivery. > It's been over 8 weeks. I called the rep and left a voice mail asking when I > can expect my equipment. S/he called me right back. I asked when I can > expect the equipment. Then I said "Don't make me go on the Histonet". > My techs started laughing and I couldn't keep a straight face. The rep was > all apologetic and promised me I'll have my equipment in 2 weeks. Gee, I > didn't think I was getting that bad. Maybe I should settle down a little > bit. > > Joe Nocito, BS, HT(ASCP) QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Tue Aug 31 20:15:43 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] I have a funny In-Reply-To: <000b01c48faf$c3768630$0a01a8c0@LUCYSALES> Message-ID: so 6 weeks promise turned into ? at least 10 wks by now, that's not much of a promise. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Lucy Brooks Sent: Tuesday, August 31, 2004 4:11 PM To: Joe Nocito; Histonet Subject: Re: [Histonet] I have a funny Joe!! Speak your mind!!! The Histonet is a powerful thing!! ----- Original Message ----- From: "Joe Nocito" To: "Histonet" Sent: Tuesday, August 31, 2004 4:01 PM Subject: [Histonet] I have a funny > Hello all, > for once, I have, what I think is funny, is a short story. A couple of > months ago, I purchased a piece of equipment from a vendor (who will remain > anonymous, at this time at least). I was told it was 6 weeks for delivery. > It's been over 8 weeks. I called the rep and left a voice mail asking when I > can expect my equipment. S/he called me right back. I asked when I can > expect the equipment. Then I said "Don't make me go on the Histonet". > My techs started laughing and I couldn't keep a straight face. The rep was > all apologetic and promised me I'll have my equipment in 2 weeks. Gee, I > didn't think I was getting that bad. Maybe I should settle down a little > bit. > > Joe Nocito, BS, HT(ASCP) QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Tue Aug 31 20:23:13 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] CD3 on BM In-Reply-To: <20040827101321.627.qmail@web11901.mail.yahoo.com> Message-ID: i have really good results with the new rabbit monoclonal cd3/ the one i use is from Lab Vision/Neomarkers, works great on mouse tissue as well. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of eileen dusek Sent: Friday, August 27, 2004 3:13 AM To: histonet Subject: [Histonet] CD3 on BM Good Morning, Is anyone having trouble with CD3 on BM? Our stain works great on regular tissue but weak on BM. We pressure cook the slides and yesterday I tried using an amp, still no staining. I was thinking of trying no pretreatment or protease instead of PC. Are there any other suggestions? Sorry about the font change, I'm not sure how it happened. I appreciate all your help Eileen Edward Hospital --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SJones <@t> cvm.tamu.edu Tue Aug 31 20:56:34 2004 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] I have a funny Message-ID: Ah, this begs an ethical question.... should the histonet be used as a threat to intimidate companies? Is this an Us against Them mentality? Discuss amongst yourselves. (a little "Coffee Talk" humor there for you Saturday Night Live fans) Oye Vey..................... Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Lucy Brooks" 8/31/2004 6:10:52 PM >>> Joe!! Speak your mind!!! The Histonet is a powerful thing!! ----- Original Message ----- From: "Joe Nocito" To: "Histonet" Sent: Tuesday, August 31, 2004 4:01 PM Subject: [Histonet] I have a funny > Hello all, > for once, I have, what I think is funny, is a short story. A couple of > months ago, I purchased a piece of equipment from a vendor (who will remain > anonymous, at this time at least). I was told it was 6 weeks for delivery. > It's been over 8 weeks. I called the rep and left a voice mail asking when I > can expect my equipment. S/he called me right back. I asked when I can > expect the equipment. Then I said "Don't make me go on the Histonet". > My techs started laughing and I couldn't keep a straight face. The rep was > all apologetic and promised me I'll have my equipment in 2 weeks. Gee, I > didn't think I was getting that bad. Maybe I should settle down a little > bit. > > Joe Nocito, BS, HT(ASCP) QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> ihcworld.com Tue Aug 31 21:04:25 2004 From: info <@t> ihcworld.com (IHC World) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Factor XIII In-Reply-To: Message-ID: Dako's Factor VIII (vWF) works very well on human tissues. For detailed protocol, go to http://www.ihcworld.com/protocol_database.htm and type keyword "vwf" and search. You will find the protocol from our database. Good Luck. IHC World Online Information Center for Immunohistochemistry http:/www.ihcworld.com -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Tuesday, August 31, 2004 12:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Factor XIII Calbiotech has discont'd their Factor XIII Ab. Can anyone tell me of another supplier of this Ab? We're using it on FFPE human tissue for IHC. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet