From Dorothy.L.Webb <@t> HealthPartners.Com Tue Nov 1 06:49:50 2005 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Tue Nov 1 06:50:00 2005 Subject: [Histonet] Microwave processor Message-ID: <0E394B648E5284478A6CCB78E5AFDA2718C493@hpes1.HealthPartners.int> We purchased this year the Thermo Electron "Tissue Wave" made originally by Energy Beam Sciences and we love it. The reason I find it so nice after using a Ted Pella at another institution is that it has vacuum infiltration for the paraffin and the cost of the unit is affordable compared to Hacker's. The company, Thermo, is very helpful in setting it up in your lab according to what you want to use it for in processing, fixation, decalcification, etc. They have a "specialist" who you can call or Email at anytime for guidance. We are having excellent results with our processing of cell blocks and tissues that are 1 mm or less in size. We have protocols for 2-3 mm specimans, but, are not implementing that into our daily routine yet. I see a great future in better TAT for all tissues. The GI biopsies look like "digital" images under the microscope when done in this microwave process! Any questions, I would be glad to answer or help in any way! Just Email me at Dorothy.L.Webb@Healthpartners.com ________________________________________ 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 jqb7 <@t> cdc.gov Tue Nov 1 07:02:56 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Tue Nov 1 07:03:14 2005 Subject: [Histonet] floor mats Message-ID: Hi everyone: We are about to move into our gorgeous new lab and I need some help. Until now we have had the luxury of keeping our embedding centers in a room separate from the main lab. In the new lab this will no longer be the case. Since there is occasionally some dripping of paraffin during the embedding process I am trying to find a mat that I can place in the embedding area to protect the lab floor. What is everyone else out there using? I'd really appreciate some suggestions. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 (404) 639-3590 From algranth <@t> u.arizona.edu Tue Nov 1 07:46:21 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Tue Nov 1 07:46:34 2005 Subject: [Histonet] floor mats In-Reply-To: Message-ID: <4.3.2.7.2.20051101063909.00ccc430@algranth.inbox.email.arizona.edu> I had to replace our floor mats recently and I got new mats from Market Lab. 800-237-3604 or http://www.marketlabinc.com/ The mats are featured in their catalog and on the web site as designed for histology labs - No Slip Histology Floor Mat - 3' x 10' (ML9803). You can get them in custom sizes also. We just vacuum them periodically and they are great. Andi At 08:02 AM 11/1/2005 -0500, Bartlett, Jeanine wrote: >Hi everyone: > >We are about to move into our gorgeous new lab and I need some help. >Until now we have had the luxury of keeping our embedding centers in a >room separate from the main lab. In the new lab this will no longer be >the case. Since there is occasionally some dripping of paraffin during >the embedding process I am trying to find a mat that I can place in the >embedding area to protect the lab floor. What is everyone else out >there using? I'd really appreciate some suggestions. > >Thanks, > >Jeanine Bartlett, HT(ASCP) >Centers for Disease Control and Prevention >1600 Clifton Road, MS/G-32 >Atlanta, GA 30333 >(404) 639-3590 > >_______________________________________________ >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 jqb7 <@t> cdc.gov Tue Nov 1 07:55:37 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Tue Nov 1 08:00:15 2005 Subject: [Histonet] floor mats Message-ID: So they would hold up well if someone dripped paraffin on them? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Andrea Grantham Sent: Tuesday, November 01, 2005 8:46 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] floor mats I had to replace our floor mats recently and I got new mats from Market Lab. 800-237-3604 or http://www.marketlabinc.com/ The mats are featured in their catalog and on the web site as designed for histology labs - No Slip Histology Floor Mat - 3' x 10' (ML9803). You can get them in custom sizes also. We just vacuum them periodically and they are great. Andi At 08:02 AM 11/1/2005 -0500, Bartlett, Jeanine wrote: >Hi everyone: > >We are about to move into our gorgeous new lab and I need some help. >Until now we have had the luxury of keeping our embedding centers in a >room separate from the main lab. In the new lab this will no longer be >the case. Since there is occasionally some dripping of paraffin during >the embedding process I am trying to find a mat that I can place in the >embedding area to protect the lab floor. What is everyone else out >there using? I'd really appreciate some suggestions. > >Thanks, > >Jeanine Bartlett, HT(ASCP) >Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 >Atlanta, GA 30333 >(404) 639-3590 > >_______________________________________________ >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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BlazekL <@t> childrensdayton.org Tue Nov 1 09:12:17 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Tue Nov 1 09:13:57 2005 Subject: [Histonet] LIS job description Message-ID: Does anyone have a job description of a pathology LIS person that is not an administrator but someone that does general updates such as testing and adding new procedures or reference files, writing procedures, training staff and generally maintaining the system? Also is this a separate position or is a supervisor or tech doing this task? Any kind of input will be appreciated. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org From mcauliff <@t> umdnj.edu Tue Nov 1 09:22:18 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Tue Nov 1 09:24:22 2005 Subject: [Histonet] Salivary gland tissue fetal pig problems In-Reply-To: References: Message-ID: <4367882A.6050704@umdnj.edu> I agree with the other person who responded, you have embalmed material and it will never have good morphology. Also, 30 minutes in formalin won't provide much fixation. If you want well-fixed salivary glands, you need fresh material that you can perfuse or remove, slice into thin pieces, and fix yourself in NBF or Bouin's fixative (for 48 hours or longer). Geoff MVaughan4@ucok.edu wrote: >Hello again, >I want to get salivary gland tissue (parotid, submandibular, sublingual) >for the students. I thought I would try using the glands from the fetal >pigs (sent from Carolina Bio) we use for student dissection. I post-fixed >them briefly (~30 minutes) in 10%NBF, followed by >dehydration/infiltration. After sectioning, the connective tissue is >mostly degenerated and the gland tissue doesn't look that good either. The >good archival gland tissue we have is yellowish-colored, maybe it is fixed >in something different than NBF. Any suggestions? Thanks. >Mel >Melville B. Vaughan, Ph. D. >Assistant Professor >Department of Biology >University of Central Oklahoma >Edmond, OK 73034 >_______________________________________________ >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 int09018 <@t> alphahunt.com Tue Nov 1 10:35:29 2005 From: int09018 <@t> alphahunt.com (HCS) Date: Tue Nov 1 10:39:55 2005 Subject: [Histonet] need help with Shandon Embedding center repair Message-ID: <000901c5df02$4968f0f0$6601a8c0@hp> Hi. I am repairing my Shandon embedding center 11. It is overheating and will not respond to setting the temp adjustment. With a similar problem, I replaced a component many years ago and need to do this again, but have no manual or instructions. Would someone have a service manual (or just a copy of the appropriate pages to guide me through this. I am also wondering about the exact part number for the module that is to be replaced. ( I believe it is BC332X14A). Any help would be greatly appreciated. LeRoy Brown HT(ASCP) HTL Histology Consultation Services PO Box 770 207 N Harkness St Everson, WA 98247 www.histocs.com 360-966-7300 fax : 360-543-5626 From krat18 <@t> aol.com Tue Nov 1 11:51:14 2005 From: krat18 <@t> aol.com (krat18@aol.com) Date: Tue Nov 1 11:51:34 2005 Subject: [Histonet] floor mats In-Reply-To: <4.3.2.7.2.20051101063909.00ccc430@algranth.inbox.email.arizona.edu> Message-ID: <8C7AD23FCFDAE36-41C-1F458@MBLK-M26.sysops.aol.com> Our lab is now using Sticky Mats from Cole Static Control Inc. You get 30 or 60 layers (peel-off) that sticks to the paraffin and keeps it from tracking onto the floor. MarketLab also has these but at more than double the price. We've been happy with them, although you do need to peel off the layers just about daily. Phone # for the company: 800-915-3535. I hope this is what you need! Karen Raterman Histology Dept. St. Mary's Health Center St. Louis, MO 63117 krat18@aol.com -----Original Message----- From: Andrea Grantham To: histonet@lists.utsouthwestern.edu Sent: Tue, 01 Nov 2005 06:46:21 -0700 Subject: Re: [Histonet] floor mats I had to replace our floor mats recently and I got new mats from Market Lab. 800-237-3604 or http://www.marketlabinc.com/ The mats are featured in their catalog and on the web site as designed for histology labs - No Slip Histology Floor Mat - 3' x 10' (ML9803). You can get them in custom sizes also. We just vacuum them periodically and they are great. Andi At 08:02 AM 11/1/2005 -0500, Bartlett, Jeanine wrote: >Hi everyone: > >We are about to move into our gorgeous new lab and I need some help. >Until now we have had the luxury of keeping our embedding centers in a >room separate from the main lab. In the new lab this will no longer be >the case. Since there is occasionally some dripping of paraffin during >the embedding process I am trying to find a mat that I can place in the >embedding area to protect the lab floor. What is everyone else out >there using? I'd really appreciate some suggestions. > >Thanks, > >Jeanine Bartlett, HT(ASCP) >Centers for Disease Control and Prevention >1600 Clifton Road, MS/G-32 >Atlanta, GA 30333 >(404) 639-3590 > >_______________________________________________ >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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sandosis <@t> uia.net Tue Nov 1 12:33:16 2005 From: sandosis <@t> uia.net (sandosis@uia.net) Date: Tue Nov 1 12:33:27 2005 Subject: [Histonet] Ventana HER-2/neu rabbit monoclonal Message-ID: <200511011833.jA1IXHr27971@smtp3.uia.net> Has anyone used the new HER-2/neu rabbit monoclonal, clone 4B5 from Ventana? They are discontinuing the CB11 clone. Are there any out there that are IVD and not ASR? Thanks, Sandy So-Cal --------------------------------------------- This message was sent using the UIA Web Mail Server. ULTIMATE Internet Access, Inc http://www.uia.net/ From caron_fournier <@t> yahoo.ca Tue Nov 1 13:18:39 2005 From: caron_fournier <@t> yahoo.ca (caron fournier) Date: Tue Nov 1 13:18:49 2005 Subject: [Histonet] re: EXAKT system Message-ID: <20051101191839.55329.qmail@web36312.mail.mud.yahoo.com> Hello: I have just started a position where I am using the EXAKT cutting and grinding system for bone sectioning. Does anyone have a protocal for grinding which includes paper grades and rough times to get me started as I do not have the time to spend "reinventing the wheel" as the position has been vacant for some time and there are projects that must get done soon. Any help would be greatly appreciated. Caron --------------------------------- Find your next car at Yahoo! Canada Autos From JMahoney <@t> alegent.org Tue Nov 1 13:43:21 2005 From: JMahoney <@t> alegent.org (Janice A Mahoney) Date: Tue Nov 1 13:43:55 2005 Subject: [Histonet] AFB Question Message-ID: Hello Netters, I have a question that may seem very silly to most of you but I'm asking to make a point and to see what feedback I get. On special stains our practice is to place the control tissue on the same slide as the patient tissue. We are automated and the cost of performing the stain is much less this way. Has anyone ever heard of the bacteria from an AFB control section washing off and ending up "on" the patient tissue on the same slide? Any and all feedback is appreciated. Jan Mahoney Omaha Nebraska From LINDA.MARGRAF <@t> childrens.com Tue Nov 1 14:15:06 2005 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Tue Nov 1 14:15:26 2005 Subject: [Histonet] rabbit femurs Message-ID: Dear Histonetters: Here's a message from Virginia she was having trouble posting. Please respond to her not me. Thanks We are wondering if anyone can give us a quote on embedding 48 rabbit femurs in MMA? How much for 100u sections of those femurs with a titanium implant? If you have a contact for us we would greatly appreciate the information. I just want an idea of cost since we have experienced medical problems with our technicians who handle MMA. We are in Canada. Virginia.ross@nrc-cnrc.gc.ca Virginia Ross Parathyroid Hormone Group, Institute for Biological Sciences, National Research Council From Jackie.O'Connor <@t> abbott.com Tue Nov 1 14:18:24 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Tue Nov 1 14:19:04 2005 Subject: [Histonet] AFB Question Message-ID: Yes - organisms can migrate to the patient tissue rendering false positives. Additionally, if you are staining manually in coplin jars - floater organisms can also transfer to patient tissue from the solution. Fresh Carbol Fuchsin should always be dropped onto individual slides. 2 cents from Jackie O'. "Janice A Mahoney" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/01/2005 01:43 PM To: cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] AFB Question Hello Netters, I have a question that may seem very silly to most of you but I'm asking to make a point and to see what feedback I get. On special stains our practice is to place the control tissue on the same slide as the patient tissue. We are automated and the cost of performing the stain is much less this way. Has anyone ever heard of the bacteria from an AFB control section washing off and ending up "on" the patient tissue on the same slide? Any and all feedback is appreciated. Jan Mahoney Omaha Nebraska _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BGapinski <@t> pathgroup.com Tue Nov 1 14:31:39 2005 From: BGapinski <@t> pathgroup.com (Bruce Gapinski) Date: Tue Nov 1 14:31:54 2005 Subject: [Histonet] Microwave tissue processing Message-ID: We're buying a microwave tissue processor, and I'm wondering if any of you have any advice, and or tricks to share? Peace & Thanks Bruce --------------------------------------------------------------------- Important Notice: This e-mail is intended for the use of the person to whom it is addressed and may contain information that is privileged and confidential. If you are not the intended recipient, any disclosure, copying, distribution, or use of the contents of this message is strictly prohibited. If you have received this e-mail in error, please destroy this message and contact the Security Officer at PathGroup, Inc. immediately at 615-221-4431. From vazquezr <@t> ohsu.edu Tue Nov 1 14:38:43 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Tue Nov 1 14:39:15 2005 Subject: [Histonet] AFB Question Message-ID: Never heard of it... Robyn OHSU From sharon.osborn <@t> dnax.org Tue Nov 1 15:33:07 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Tue Nov 1 15:33:46 2005 Subject: [Histonet] RE: Microwave Processor Message-ID: <29B25753F6B1D51196110002A589D444032C4888@PALMSG30.us.schp.com> Muhammad, Please look at the Milestone Microwave Tissue Processor. It is built by an Italian company and marketed worldwide. I first saw it in Salt Lake at the 1998 NSH meeting then again in San Jose, CA 1005 California State meeting. I have used the Pella microwave and watched the Sakura rapid processing equipment for small specimens do the processing. From my research, I would strongly consider the Milestone's latest edition processor for it is almost hands free and produces great specimens ready for embedding. sharon osborn DNAX, ScheringPlough BioPharma Palo Alto, CA Date: Sat, 29 Oct 2005 09:31:21 +1300 From: "Muhammad Tahseen" Subject: [Histonet] Purcase a microwave tissue processor To: Cc: histo@skm.org.pk Message-ID: <015d01c5dbfe$9073a000$972bfea9@m7c0y4> Content-Type: text/plain; charset="iso-8859-1" We are looking to purchase a microwave tissue processor for surgical specimens. What has been the experience of users in Histoland as to preferred manufacturers? Muhammad Tahseen Histology Supervisor SKMCH & RC Lahore Pakistan. ********************************************************************* 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 Tue Nov 1 16:22:40 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Nov 1 16:22:55 2005 Subject: [Histonet] re: EXAKT system In-Reply-To: <20051101191839.55329.qmail@web36312.mail.mud.yahoo.com> References: <20051101191839.55329.qmail@web36312.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20051101152052.01b1e8f8@gemini.msu.montana.edu> Contact the EXAKT rep, herself - Linda Durbin can help you. . She is a very nice lady. At 12:18 PM 11/1/2005, you wrote: >Hello: >I have just started a position where I am using the EXAKT cutting and >grinding system for bone sectioning. Does anyone have a protocal for >grinding which includes paper grades and rough times to get me started as >I do not have the time to spend "reinventing the wheel" as the position >has been vacant for some time and there are projects that must get done >soon. Any help would be greatly appreciated. >Caron > > >--------------------------------- >Find your next car at Yahoo! Canada Autos >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From themagoos <@t> rushmore.com Tue Nov 1 17:18:27 2005 From: themagoos <@t> rushmore.com (Jason and Heather) Date: Tue Nov 1 17:18:56 2005 Subject: [Histonet] AFB Question References: Message-ID: <001a01c5df3a$91bcbd90$b3fc9fd1@magoo> Yes. We place the patient tissue on a seperate slide because of this. We also perform AFB's on an automated stainer. Jason McGough HT Clinical Laboratory of the Black Hills Rapid City SD 57701 From ree3 <@t> leicester.ac.uk Wed Nov 2 04:22:04 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Wed Nov 2 04:22:26 2005 Subject: [Histonet] keratin antibodies Message-ID: Looking for antibodies to cytokeratins, all types for the moment, that work on formalin fixed paraffin processed mouse tissues. Many thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER...U.K.... From BMolinari <@t> heart.thi.tmc.edu Wed Nov 2 05:37:48 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Wed Nov 2 05:40:14 2005 Subject: [Histonet] re: EXAKT system Message-ID: Hi Caron, We have recently started using the Exakt system. I would have to say that it really depends on what you are grinding. As far as I can tell there are no hard and fast protocols for papers and time. I know there are more people on the Histonet with a heck of a lot more experience than I do in this procedure and I am sure they will speak up. Also you could try to contact Linda Durbin. She is the owner of Exakt (1-800-866-7172). Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of caron fournier Sent: Tuesday, November 01, 2005 1:19 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] re: EXAKT system Hello: I have just started a position where I am using the EXAKT cutting and grinding system for bone sectioning. Does anyone have a protocal for grinding which includes paper grades and rough times to get me started as I do not have the time to spend "reinventing the wheel" as the position has been vacant for some time and there are projects that must get done soon. Any help would be greatly appreciated. Caron --------------------------------- Find your next car at Yahoo! Canada Autos _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From failm <@t> musc.edu Wed Nov 2 06:42:48 2005 From: failm <@t> musc.edu (Mildred Fail) Date: Wed Nov 2 06:43:17 2005 Subject: [Histonet] AFB Question Message-ID: YES!! We don't even run the control and patient through the same hydration/ dehydration set-up Rena Fail >>> "Janice A Mahoney" 11/01/05 02:43PM >>> Hello Netters, I have a question that may seem very silly to most of you but I'm asking to make a point and to see what feedback I get. On special stains our practice is to place the control tissue on the same slide as the patient tissue. We are automated and the cost of performing the stain is much less this way. Has anyone ever heard of the bacteria from an AFB control section washing off and ending up "on" the patient tissue on the same slide? Any and all feedback is appreciated. Jan Mahoney Omaha Nebraska _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Nov 2 07:23:36 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 2 07:23:45 2005 Subject: [Histonet] questions about rhodanine-copper stain In-Reply-To: Message-ID: <20051102132336.24497.qmail@web61225.mail.yahoo.com> I gave up rodamine many years ago because of weak staining and problematic. I switched to Timm's sulfur oxide methods, very reliable and fast (although a little "smelling"). Rene J. Mildred Fail wrote: i keep the rhodanine stock a year. Our protocol is very simple Rena fail RHODANINE SATURATED SOLUTION (STOCK) Absolute Alcohol 100.0 ml 5 p-dimethylaminobenzylidene* 0.2 gm RHODANINE SOLUTION (WORKING) Rhodanine Saturated Solution 3.0 ml Distilled Water 47.0 ml Shake stock before measuring and mixing solutions. 1. Deparaffinize and hydrate to distilled water. 2. Using plastic Coplin Jars; preheat the Rhodanine solution in 1200 watt microwave on half power 30 seconds then incubate for 5 minutes. 3. Wash well in several changes of distilled water. 4. Mayer's Hematoxylin for 1 minute. 5. Rinse with distilled water. 6. A quick rinse in 0.5% Sodium Borax. 7. Rinse well with distilled water 8. Deparaffinize, clear, and mount It will fade if not cover slipped right away udrun Lang" 10/26/05 08:23AM >>> I am dealing with the rhodanine stain, because we shall establish it in our histolab. I've read three different protocols (AFIP, Churukian, webpath). AFIP and Churukian require sodiumacetat solutions to mix with the stocksolution, the others require just the dilution of the stocksolution in Aqua dest. Why? Do both versions work? Churukian demands coverslipping with aqueous medium, the others don't. Is the staining result soluble in ethanol? The stability of the 0,2% rhodanine/ethanol stocksolution is described between one day and three months. What is the right shelf live? I hope someone can explain it to me. Gudrun Lang General hospital Linz, Austria _______________________________________________ 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 --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From rjbuesa <@t> yahoo.com Wed Nov 2 07:37:37 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 2 07:37:46 2005 Subject: [Histonet] Expired Stains In-Reply-To: Message-ID: <20051102133737.87765.qmail@web61217.mail.yahoo.com> Two different issues: 1- with regards to stains solutions JCAH consider they should be replaced (cannot be used) even if they still work well after the expiration date. Costs cuts are a concern and that is why I always prepared my staining solutions to the amounts I was going to need for 2-3 months. That is the most economical way of doing special stains. 2- regarding dye powders they are extremely stable. I remember using in 1960 some anilin dyes prepared by Merck in Darmstad (Germany) just after the "Great War" (WW I) and they were fantastic. Now you should add a label to the bottle with the remark:"No expiration date" (as I use to do). Rene J. "Frank, Matthew" wrote: The Biological Stain Commission has performed a stability study on dye powders and found that dye powders kept in closed containers stained well after over 30 years of storage. Solutions of course are a different story. -----Original Message----- From: Heckford, Karen - SMMC-SF [mailto:Karen.Heckford@CHW.edu] Sent: Wednesday, October 26, 2005 1:57 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Expired Stains Hi Everyone, I was wondering if anyone knows how JCAHO views expired Special Stains. I have some expired Special Stains. We do low volume, trying to use up some stains before they expire can be difficult. I can still get the stains to work and the Pathologists like them. Should I throw them away and order new? As you know the stains can be costly. Thanks for your input, Karen Heckford St. Mary's Medical Center San Francisco, California _______________________________________________ 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 --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From pex0220 <@t> yahoo.com.cn Wed Nov 2 07:51:36 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Wed Nov 2 07:51:47 2005 Subject: [Histonet] decalcification Message-ID: <20051102135136.99694.qmail@web15510.mail.cnb.yahoo.com> Dear all, I have a question about decalcification. I have got some bone tissues (such as femur) from newborn pups, I would like to know if they need to be decalcified before paraffin embedding. Thanks a lot. Guofeng --------------------------------- ÑÅ»¢Ãâ·ÑGÓÊÏ䣭ÖйúµÚÒ»¾øÎÞÀ¬»øÓʼþɧÈų¬´óÓÊÏä ÑÅ»¢ÖúÊÖ¡§DËÑË÷¡¢É±¶¾¡¢·ÀɧÈÅ From rjbuesa <@t> yahoo.com Wed Nov 2 07:56:51 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 2 07:57:02 2005 Subject: [Histonet] decalcification In-Reply-To: <20051102135136.99694.qmail@web15510.mail.cnb.yahoo.com> Message-ID: <20051102135651.29502.qmail@web61225.mail.yahoo.com> Yes, they need to be decalcified, gently though! If you use an acid decalcifier (like "RDO") you may want to dilute it in half and check the bones for flexibility more often than usual. Rene J. pex wrote: Dear all, I have a question about decalcification. I have got some bone tissues (such as femur) from newborn pups, I would like to know if they need to be decalcified before paraffin embedding. Thanks a lot. Guofeng --------------------------------- ????????G?????­???????????????????????????????? ??????????D?????????????????? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From pex0220 <@t> yahoo.com.cn Wed Nov 2 07:57:34 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Wed Nov 2 07:57:51 2005 Subject: [Histonet] antibody Message-ID: <20051102135734.16180.qmail@web15509.mail.cnb.yahoo.com> Dear all, I have ordered two primary antibodies, their isotypes are IgG1 and IgG3 repectively. In our lab, there are some secondary antibodies, their isotypes are IgG (H+L), so I wonder if I use these secondary antibodies for immunofluorescence. Thanks. Guofeng --------------------------------- ÑÅ»¢Ãâ·ÑGÓÊÏ䣭ÖйúµÚÒ»¾øÎÞÀ¬»øÓʼþɧÈų¬´óÓÊÏä ÑÅ»¢ÖúÊÖ¡§DËÑË÷¡¢É±¶¾¡¢·ÀɧÈÅ From Malcolm.McCallum <@t> tamut.edu Wed Nov 2 08:15:15 2005 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Wed Nov 2 08:15:24 2005 Subject: [Histonet] teaching histology newbee Message-ID: Hi, I will be teaching an undergraduate histology this spring. For those of you who teach this, what fraction of the course do you devote to techniques versus anatomy? Do you include pathology, if so, how extensively? How extensively do you require them to understand the differences among stains, and how do you structure your laboratories? Anything else I should consider? Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html From John.Sheppard <@t> Health-Partners.org Wed Nov 2 08:34:27 2005 From: John.Sheppard <@t> Health-Partners.org (John.Sheppard@Health-Partners.org) Date: Wed Nov 2 08:30:01 2005 Subject: [Histonet] Histotechnician productivity Message-ID: Hello Everyone, I have been assigned a "mission" by our medical director. He wants me to find out what would be a reasonable number of blocks per day for a histotech. I have tried searching the archieves for this information because I know it seems to be discussed quite often. However most of the old threads I found seemed to end up with histology professionals discussing that they would not work in a "factory like" atmosphere where they are slaves to productivity and turn around times. Which to be honest I agree with, histology should be a quality oriented product. I have also looked at the ASCP and NSH websites where this information is not readily accessible. I will probably e-mail them to see if they have any research or insight on this topic. The numbers that I am most familiar with, is 50 to 60 blocks per day per histotech, when one is including embedding, cutting, manual H&E and Pap staining(gyn & non-gyn), manual coverslipping, manual special stains, and frozen sections. I am just not sure how accurate those numbers are for staffing a histology lab. This information is needed because we will be consolidating two separate small community hospital histology labs onto one campus. Apparently this will be used to justify the current staff of both locations. Unfortunately, I believe this information will ultimately be used to eliminate a position or two. Thank you for your time John Sheppard HT(ASCP) CONFIDENTIALITY NOTICE: This message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From Charles.Embrey <@t> carle.com Wed Nov 2 08:36:13 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Wed Nov 2 08:36:34 2005 Subject: [Histonet] labeling regulations Message-ID: Years ago, the CAP checklist for histology specified what information was required on the specimen container and even the request form. The newest checklists seem to have completely thrown this out. Is specimen container labeling regulated anywhere else? Is there a requirement anywhere that specimen type/site be noted on the container? Thanks, Charles Embrey PA(ASCP) From mcauliff <@t> umdnj.edu Wed Nov 2 09:00:47 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Wed Nov 2 09:01:06 2005 Subject: [Histonet] teaching histology newbee In-Reply-To: References: Message-ID: <4368D49F.7010400@umdnj.edu> Hi Malcolm: I would give a short (15-30 minute) talk about how tissue is prepared (fix, dehydrate, clear, embed, section, mount, stain) and show a few artifacts (folds, tears, etc). I do think you should briefly explain H&E and maybe trichromes and PAS. Definitely explain, with illustrations, how immunostaining works. In the old days (when I was a student) there might be some discussion of how different fixatives can give different 'looks' but now that everything is done in buffered formalin (or some secret, commercial mixture of who knows what) there seems to be no point in this. Save that for teaching microtechnique. Good luck! Geoff Malcolm McCallum wrote: >Hi, >I will be teaching an undergraduate histology this spring. For those of you who teach this, what fraction of the course do you devote to techniques versus anatomy? Do you include pathology, if so, how extensively? How extensively do you require them to understand the differences among stains, and how do you structure your laboratories? Anything else I should consider? > >Malcolm L. McCallum >Assistant Professor >Department of Biological Sciences >Texas A&M University Texarkana >2600 Robison Rd. >Texarkana, TX 75501 >O: 1-903-233-3134 >H: 1-903-791-3843 >Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.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 vazquezr <@t> ohsu.edu Wed Nov 2 09:16:54 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Wed Nov 2 09:17:19 2005 Subject: [Histonet] Histotechnician productivity Message-ID: I used to do an average 150-170 a day. Robyn OHSU From gcallis <@t> montana.edu Wed Nov 2 09:57:50 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 2 09:58:05 2005 Subject: [Histonet] RE: EXAKT system In-Reply-To: References: Message-ID: <6.0.0.22.1.20051102085632.01b43510@gemini.msu.montana.edu> Linda Jenkins is an expert with Exakt system, you can contact her for help. jlinda@ces.clemson.edu At 04:37 AM 11/2/2005, you wrote: >Hi Caron, >We have recently started using the Exakt system. I would have to say >that it really depends on what you are grinding. As far as I can tell >there are no hard and fast protocols for papers and time. I know there >are more people on the Histonet with a heck of a lot more experience >than I do in this procedure and I am sure they will speak up. Also you >could try to contact Linda Durbin. She is the owner of Exakt >(1-800-866-7172). > > >Betsy Molinari HT (ASCP) >Texas Heart Institute >Cardiovascular Pathology >6770 Bertner Ave. >Houston,TX 77030 >832-355-6524 >832-355-6812 (fax) > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of caron >fournier >Sent: Tuesday, November 01, 2005 1:19 PM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] re: EXAKT system > >Hello: >I have just started a position where I am using the EXAKT cutting and >grinding system for bone sectioning. Does anyone have a protocal for >grinding which includes paper grades and rough times to get me started >as I do not have the time to spend "reinventing the wheel" as the >position has been vacant for some time and there are projects that must >get done soon. Any help would be greatly appreciated. >Caron > > >--------------------------------- >Find your next car at Yahoo! Canada Autos >_______________________________________________ >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 Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From osteffe <@t> online.no Wed Nov 2 10:03:38 2005 From: osteffe <@t> online.no (ole) Date: Wed Nov 2 10:03:53 2005 Subject: [Histonet] Anti human; P2X7, TP1, Podocalyxin Message-ID: <003001c5dfc6$fd2646f0$0100000a@ole> Histonetters; First; thanks for the suggestions for vendors of anti-alpha(s) collagen (IV) - antibodies (last week). Im looking for some anti-human antibodies that works on formalin fixed parrafin embedded tissue. Anti P2X7 and anti-TP1 ( antibodies wich is useful differentiating keratoacantoma from squamous cell carcinoma) And; Anti-Podocalyxin (i have found a podocalyxin ab that looks promising, but are interested if you have any recommandations) regards, Ole Johnny Steffensen Dep of Pathology Aalesund Norway From gcallis <@t> montana.edu Wed Nov 2 10:11:16 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 2 10:11:30 2005 Subject: [Histonet] EXAKT system Message-ID: <6.0.0.22.1.20051102090929.01b42c68@gemini.msu.montana.edu> It is always a good idea to try and get training on this system and Linda Durbin, who sells it for EXAKT is available to do this, but it may cost a bit. In the long run, her training will save you money in the long run. Nothing like some good hands on training rather than trying to piece it together with email messaging. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From PMonfils <@t> Lifespan.org Wed Nov 2 10:23:08 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Wed Nov 2 10:23:19 2005 Subject: [Histonet] Specimen Discs for Cryocut 1800 Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717604@lsexch.lsmaster.lifespan.org> Where can I purchase specimen discs for a Reichert-Jung Cryocut 1800 cryostat? Or - if you have some you don't need, I'll be glad to purchase them from you. Used is fine. From cking <@t> rallansci.com Wed Nov 2 10:39:51 2005 From: cking <@t> rallansci.com (King, Curtis - RAS) Date: Wed Nov 2 10:40:03 2005 Subject: [Histonet] EXAKT system Message-ID: <490C1EC04BA10F4891494D0ED756AC93034241CB@usmi0012k03.rallansci.com> I agree with Gayle. Linda trained me on the Exakt and I was batching within 2 weeks. Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Wednesday, November 02, 2005 11:11 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] EXAKT system It is always a good idea to try and get training on this system and Linda Durbin, who sells it for EXAKT is available to do this, but it may cost a bit. In the long run, her training will save you money in the long run. Nothing like some good hands on training rather than trying to piece it together with email messaging. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mari.ann.mailhiot <@t> leica-microsystems.com Wed Nov 2 10:55:07 2005 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Wed Nov 2 10:55:21 2005 Subject: [Histonet] Specimen Discs for Cryocut 1800 In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE01717604@lsexch.lsmaster.lifespan.org> Message-ID: Paul You can order the discs direct from Leica. Give me a call and I can give you the information. Best 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 "Monfils, Paul" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/02/2005 10:23 AM To "'histonet@lists.utsouthwestern.edu'" cc Subject [Histonet] Specimen Discs for Cryocut 1800 Where can I purchase specimen discs for a Reichert-Jung Cryocut 1800 cryostat? Or - if you have some you don't need, I'll be glad to purchase them from you. Used is fine. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com From Eric <@t> ategra.com Wed Nov 2 11:09:30 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Wed Nov 2 11:00:18 2005 Subject: [Histonet] Immediate opportunities for Histotechnologists( follow up) Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. (I have a few travel/temp HistoTech positions as well - see below). Most of my Histo Techs positions are clinical, although I do have a few Research Histo Tech positions as well. Here are some of my Hottest jobs: 1. Iowa Openings for a HistoTech(Bench) No weekends, No call, and Top Dollar. 2. Ohio(Dayton Area) HistoTech Manager No Weekends, No call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 3. Colorado (Greater Denver area) Opening for Several Bench Positions(Night Shift) No weekends, No call, Top Dollar 4. Georgia Openings for Histotech(Temp to Perm) No Weekends, No Call, Top Dollar, Excellent Benefits if you are interested, please call me today at 1-800-466-9919 ext 223 5. Pennsylvania(Philadelphia area) Openings for HistoTech(Bench) No Weekends, No Call, Top Dollar 6.Rhode Island Openings for Histotech(Bench) No weekends, No call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 7. California( Southern) HistoTech Supervisor(2nd shift) Openings for Histotechs(Bench,2nd shift) Great Location, No weekends, No call, Top Dollar 8. California(Southern) Openings for HistoTechs(Bench) Great Location, No weekends, No call, Top Dollar 9. Pennsylvania(Multiple jobs in greater Pittsburgh area) HistoTech opening(Bench) if you are interested, please call me today at 1-800-466-9919 ext 223 10. Illinois(Multiple jobs in Greater Illinois area) Opening for Histotech(Bench) 11. Michigan(Multiple jobs in Greater Michigan area) Openings for Histotechs(Bench) 12. Washington State(Eastern) Opening for Lead HistoTech(Bench) No weekends, No call if you are interested, please call me today at 1-800-466-9919 ext 223 13. Oklahoma Opening for Histotech(Bench) No weekends, No call. 14. Massachusetts (Several Opportunities in the Boston Area) Part time Histo Tech(Permanent) No weekends, No call 15. New Mexico openings for Supervisor and Bench Techs No Weekends, No Call, Excellent Benefits.. 16. Nebraska Openings for Histo Techs(Bench) No Weekends, No Call, Top Dollar 17. Virginia(Western Virginia) Opening for Bench Histotech No weekends, No call if you are interested, please call me today at 1-800-466-9919 ext 223 18. Ohio( Southern) Opening for Bench HistoTech No weekends, No call 19. Missouri(Greater Missouri area) Opening for Bench Histotech No weekends, No call 20. Wisconsin(Eastern area) Opening for a Bench Histotech No weekends, No call 21. Florida(Greater Florida area) Openings for Bench Histotech No weekends, No call The clients are currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 From lh2 <@t> sanger.ac.uk Wed Nov 2 11:00:20 2005 From: lh2 <@t> sanger.ac.uk (Lindsay Hall) Date: Wed Nov 2 11:00:25 2005 Subject: [Histonet] Antibodies for frozen mouse sections Message-ID: <131DF14ECE564A4D8F35376FD346517207CB7B42@EXCHSRV1.internal.sanger.ac.uk> Hi, I am staining some frozen mouse NALT and cervical lymph node sections with a number of antibodies, CD4, CD8, B220, CD11c (all from Pharmingen) and F4/80 (Serotec) and the staining is working fine (thanks to Gayle Callis!!). However I want to start staining for some more cell markers and I was wondering if anybody could give me details of primary antibodies (dilutions etc) and where you get them from, that are good for staining natural killer (NK) cells, ICAM-1 (CD54), CD5, MadCAM-1, VCAM-1, PNAd and a neutrophil marker, i.e. Ly-6G. Thanks very much in advance. Lindsay Hall Wellcome Trust Sanger Institute Wellcome Trust Genome Campus Hinxton Cambridge CB10 1SA UK From vazquezr <@t> ohsu.edu Wed Nov 2 11:30:47 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Wed Nov 2 11:31:15 2005 Subject: [Histonet] Specimen Discs for Cryocut 1800 Message-ID: You can get them from Internation Medical Equipment cat #370085 30mm dia $33.00 and #370086 45mm dia $43.00 each OR you can get tem from Thermo Electron Corp cat#8002 29mm dia for apprx $22.00 (they are alittle long, but you can cut them down with a hacksaw). I had a fellow in facililties do it for me. Robyn OHSU From TJasper <@t> smdc.org Wed Nov 2 11:39:52 2005 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Wed Nov 2 11:39:58 2005 Subject: [Histonet] Histotechnician productivity Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA07D390AE@harrier> John, I am quite certain that you'll get a variety of opinions on this topic. In approaching this "mission" I think you need to be clear about 2 things. 1) What is the scope of your service? and 2) Where are you going? You do mention a 50-60 block per day figure and basic clinical histology duties performed in general manually. Does this currently work well for you? Are your pathologists happy with the turnaround time and the quality of the work they receive? Are you trying to expand your business? Are your volumes naturally increasing and/or are they projected to increase? Do you want to develop technically, i.e. Immunohistochemistry, In Situ Hybridization? Are you looking at automating some of your current processes, i.e. special stains and coverslipping? Holy buckets! That's a lot of questions right out of the gate! But, you need to answer questions like these to make an accurate assessment and good decisions. One thing I'm willing to bet is that your organization is looking to save money. That's very understandable in this day and age, however you've got to be real careful in doing it. Evaluate your service very carefully as this will reveal true costs and true savings. This may not apply to you, but for instance, take estrogen and progesterone receptors. I'm guessing that you probably send those tests out somewhere. How much does that cost? Do you have the talent in-house to take it on yourself? If you do and your volumes warrant it, you may save money. Money that's easily demonstrated up front and savings in other ways such as faster turnaround times. This in turn increases the overall value of your service and makes the techs more valuable as well. Again, this may not apply to you I just created a scenario to make a point. This leads me to your last statement which is concerning, and that's the comment about losing a position or two. I firmly believe that people are the greatest asset of an organization. You need to hold on to positions with tenacity. Everyone wants to do more with less, and that's great. I like the modification of doing more with what you've got, especially if you've got good people. Even if the people aren't that good you can try to get others as long as you still have positions to fill. Good histotechs are hard to find, enough said there. Geez, I really haven't given you any hard numbers and I guess I'm loathe to do so considering all the unknowns and variables. I can tell you this, years ago when I worked in a contract lab (non-clinical, sweatshop) we were expected to cut 200 blocks in 8 hours with no more than 15 recuts. I did learn to cut pretty fast there but I wouldn't recommend it. I track hours and production here very closely but comparisons are not going to be apples to apples. I value quality over quantity and I expect that people will do their best day in and day out. Techs are people and some are stronger in different areas than others. We average 200-250 blocks per day, with 2 techs embedding and 3-4 techs cutting. We try to finish cutting in about 5 to 5.5 hours. Again, this is fraught with variables. To me, if someone here is cutting 25 to 30 blocks an hour I think that's pretty good. Please excuse my rambling, but this is a tough nut to crack as the world is filled with shades of grey. Good luck to you. tj -----Original Message----- From: John.Sheppard@Health-Partners.org [mailto:John.Sheppard@Health-Partners.org] Sent: Wednesday, November 02, 2005 8:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histotechnician productivity Hello Everyone, I have been assigned a "mission" by our medical director. He wants me to find out what would be a reasonable number of blocks per day for a histotech. I have tried searching the archieves for this information because I know it seems to be discussed quite often. However most of the old threads I found seemed to end up with histology professionals discussing that they would not work in a "factory like" atmosphere where they are slaves to productivity and turn around times. Which to be honest I agree with, histology should be a quality oriented product. I have also looked at the ASCP and NSH websites where this information is not readily accessible. I will probably e-mail them to see if they have any research or insight on this topic. The numbers that I am most familiar with, is 50 to 60 blocks per day per histotech, when one is including embedding, cutting, manual H&E and Pap staining(gyn & non-gyn), manual coverslipping, manual special stains, and frozen sections. I am just not sure how accurate those numbers are for staffing a histology lab. This information is needed because we will be consolidating two separate small community hospital histology labs onto one campus. Apparently this will be used to justify the current staff of both locations. Unfortunately, I believe this information will ultimately be used to eliminate a position or two. Thank you for your time John Sheppard HT(ASCP) CONFIDENTIALITY NOTICE: This message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. From gcallis <@t> montana.edu Wed Nov 2 12:40:22 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 2 12:40:37 2005 Subject: [Histonet] Specimen Discs for Cryocut 1800 In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE01717604@lsexch.lsmaster.l ifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE01717604@lsexch.lsmaster.lifespan.org> Message-ID: <6.0.0.22.1.20051102113659.01b37958@gemini.msu.montana.edu> You can purchase them from Leica Microsystems, they are the same disks as for the 1850. We found it more economical to buy the waffle weave style metal disks from either ThermoElectron or Electron Microscopy Services, and cut off the stem to match the length for the disks that have the circles. You can get a dozen of these inexpensively but the stems must be shortened. I guess one could call these the old fashioned metal chucks used for paraffin blocks way back when! you wrote: >Where can I purchase specimen discs for a Reichert-Jung Cryocut 1800 >cryostat? > >Or - if you have some you don't need, I'll be glad to purchase them from >you. Used is fine. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From Karen.Heckford <@t> CHW.edu Wed Nov 2 13:02:47 2005 From: Karen.Heckford <@t> CHW.edu (Heckford, Karen - SMMC-SF) Date: Wed Nov 2 13:04:49 2005 Subject: [Histonet] Microtome Message-ID: I was just wondering if anyone has a Microm 335 E microtome. I was thinking about purchasing one. I am a little scared to face into a block without manual manipulation. Do I have to worry about this? Please let me know your input? Karen Heckford St. Mary's Medical Center San Francisco, Ca. From nienhuis <@t> ucla.edu Wed Nov 2 13:09:01 2005 From: nienhuis <@t> ucla.edu (nienhuis@ucla.edu) Date: Wed Nov 2 13:09:10 2005 Subject: [Histonet] Antigen retrieval on very old tissue In-Reply-To: <6.0.0.22.1.20051102113659.01b37958@gemini.msu.montana.edu> References: <09C945920A6B654199F7A58A1D7D1FDE01717604@lsexch.lsmaster.lifespan.org> <6.0.0.22.1.20051102113659.01b37958@gemini.msu.montana.edu> Message-ID: <20051102110901.9exojjmpkw4c0c8s@mail.ucla.edu> What is your recommendation for doing AR on very old brain tissue, while minimizing damage? I have a brain that has been floating in formalin for 20 years, and need to do immuno staining on it. Bob Nienhuis Neurobiology Research UCLA / VA Medical Center Los Angeles, CA From gcallis <@t> montana.edu Wed Nov 2 13:10:54 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 2 13:11:53 2005 Subject: [Histonet] Antibodies for frozen mouse sections In-Reply-To: <131DF14ECE564A4D8F35376FD346517207CB7B42@EXCHSRV1.internal .sanger.ac.uk> References: <131DF14ECE564A4D8F35376FD346517207CB7B42@EXCHSRV1.internal.sanger.ac.uk> Message-ID: <6.0.0.22.1.20051102120508.01b643c8@gemini.msu.montana.edu> Once again, you can buy these antibodies from BD Pharmingen or SEROTEC - however you need to determine the working concentration of your primaries using a dilution panel starting at 10 ug/ml - no different than for CD4, CD8, etc. and do this in your own laboratory working conditions/buffers, etc. We double immunofluorescence with MadCAM and PNAd so we can see colocalization of these markers in NALT, and lymph nodes located in neck/head region. Mesenteric lymph nodes is the positive control for these two antibodies. We do NOT use FFPE for any of these antibodies - frozen sections fixed with an acetone/alcohol fixation is wonderful for MadCAM, PNAd and probably the other markers as well. At 10:00 AM 11/2/2005, you wrote: >Hi, I am staining some frozen mouse NALT and cervical lymph node >sections with a number of antibodies, CD4, CD8, B220, CD11c (all from >Pharmingen) and F4/80 (Serotec) and the staining is working fine (thanks >to Gayle Callis!!). However I want to start staining for some more cell >markers and I was wondering if anybody could give me details of primary >antibodies (dilutions etc) and where you get them from, that are good >for staining natural killer (NK) cells, ICAM-1 (CD54), CD5, MadCAM-1, >VCAM-1, PNAd and a neutrophil marker, i.e. Ly-6G. > >Thanks very much in advance. > >Lindsay Hall >Wellcome Trust Sanger Institute >Wellcome Trust Genome Campus >Hinxton >Cambridge >CB10 1SA >UK >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From bjdewe <@t> aol.com Wed Nov 2 13:14:39 2005 From: bjdewe <@t> aol.com (bjdewe@aol.com) Date: Wed Nov 2 13:14:51 2005 Subject: [Histonet] Microtome In-Reply-To: References: Message-ID: <8C7ADF8CE192792-1E4C-1BC8@MBLK-M15.sysops.aol.com> I do...I would be happy to answer your questions... Lorie ******************************************************************* Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s)and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ******************************************************************* -----Original Message----- From: Heckford, Karen - SMMC-SF To: Histonet (E-mail) Sent: Wed, 2 Nov 2005 12:02:47 -0700 Subject: [Histonet] Microtome I was just wondering if anyone has a Microm 335 E microtome. I was thinking about purchasing one. I am a little scared to face into a block without manual manipulation. Do I have to worry about this? Please let me know your input? Karen Heckford St. Mary's Medical Center San Francisco, Ca. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RRA <@t> Stowers-Institute.org Wed Nov 2 13:18:18 2005 From: RRA <@t> Stowers-Institute.org (Allen, Rhonda) Date: Wed Nov 2 13:18:48 2005 Subject: [Histonet] decalcification Message-ID: Hello,=20 I wanted to put my 2 cents worth in about decacifying newborn mouse = femurs. At this age there should not be any calcium yet formed in the = bones and do not need decalcifying. I have done mouse heads at the PO = age and I do not have any problem cutting them, and they do not need = decalcified. Rhonda Allen BA HT(ASCP)HTL, QIHC Stowers Institute Histotechnology Specialist II =09 1000 E. 50th street Kansas City, Missouri 64110 rra@stowers-institute.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu = [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pex Sent: Wednesday, November 02, 2005 7:52 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] decalcification Dear all, =20 I have a question about decalcification. I have got some bone tissues=20 (such as femur) from newborn pups, I would like to know if they need to=20 be decalcified before paraffin embedding. =20 Thanks a lot. =20 Guofeng =09 --------------------------------- = =D1=C5=BB=A2=C3=E2=B7=D1G=D3=CA=CF=E4=A3=AD=D6=D0=B9=FA=B5=DA=D2=BB=BE=F8= =CE=DE=C0=AC=BB=F8=D3=CA=BC=FE=C9=A7=C8=C5=B3=AC=B4=F3=D3=CA=CF=E4 = =D1=C5=BB=A2=D6=FA=CA=D6=A1=A7D=CB=D1=CB=F7=A1=A2=C9=B1=B6=BE=A1=A2=B7=C0= =C9=A7=C8=C5 =20 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu = http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Tracey.Lenek <@t> CLS.ab.ca Wed Nov 2 13:30:30 2005 From: Tracey.Lenek <@t> CLS.ab.ca (Tracey.Lenek@CLS.ab.ca) Date: Wed Nov 2 13:33:46 2005 Subject: [Histonet] c4d Message-ID: <4BC300747AF87A48BCDF8E48BC2885CE010E10DE@mail1.calgary.com> Could anyone recommend a monoclonal C4d antibody for FFPE human tissue? Thanks Tracey CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify the sender immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. From liz <@t> premierlab.com Wed Nov 2 13:44:54 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Wed Nov 2 13:45:14 2005 Subject: [Histonet] Microtome In-Reply-To: Message-ID: <000501c5dfe5$e5d5dc30$a7d48a80@AMY> Karen We just finished a demo on the micron the rough trimming feature was nice. I did not see any problems with it at all, in fact we really liked it. We have to face in a lot of bone (mouse and rat joints) the automated trim feature was nice since we could adjust the thickness and speed that we faced the block and if anyone who cuts a lot of bone knows rough trimming is very important because you can pop out portions of the joint if done too aggressively. It normally takes us longer to rough trim our blocks than to cut them. When you have to trim in 100's of blocks it puts lot of wear and tear on your wrists, elbows and shoulders. With the automated rough trim all we had to do was push buttons and let the microtome do the work for us, it was great. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heckford, Karen - SMMC-SF Sent: Wednesday, November 02, 2005 12:03 PM To: Histonet (E-mail) Subject: [Histonet] Microtome I was just wondering if anyone has a Microm 335 E microtome. I was thinking about purchasing one. I am a little scared to face into a block without manual manipulation. Do I have to worry about this? Please let me know your input? Karen Heckford St. Mary's Medical Center San Francisco, Ca. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jennifer.l.hofecker <@t> Vanderbilt.Edu Wed Nov 2 14:01:06 2005 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Wed Nov 2 14:02:34 2005 Subject: [Histonet] trying to get Bill Morris from Biotherm Message-ID: <898D946569A27444B65667A49C074052075500@mailbe06.mc.vanderbilt.edu> Hi, Does anybody have Bill's email address? I have tried to send him two messages at the old address and they keep coming back. Thanks & have a great rest of the week. Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax From gcallis <@t> montana.edu Wed Nov 2 14:31:29 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 2 14:31:44 2005 Subject: [Histonet] automated versus manual microtomy In-Reply-To: <000501c5dfe5$e5d5dc30$a7d48a80@AMY> References: <000501c5dfe5$e5d5dc30$a7d48a80@AMY> Message-ID: <6.0.0.22.1.20051102131839.01b32760@gemini.msu.montana.edu> I agree with Liz totally, and will not be purchasing a microtome unless trimming and even sectioning can be done automatically. The wrists, hands, fingerjoints, etc have taken sush of a beating over the years with manual operation that automated is going to be welcomed into this laboratory asap. Can't wait for the manual machine to die of mechanical failure - trying to think of a way to hasten its demise but it isn't happening, maybe a sledge hammer!?? Another device that is helpful is the heated Paratrimmer from ThermoElectron in order to remove excess paraffin from sides and back of blocks. Osteoarthritic finger joints can no longer stand the pain of even holding cold, metal old scalpel blades or whatever for doing this. Trimming and even sectioning with automation is a necessity for preserving longevity of histotechs appendages and adjacent joints. All one has to do is take the time to learn how to use the automated features to get rid of repetitive motion stresses, it is not a worry factor!! Your hands will thank you! Liz wrote: It normally takes us longer to rough >trim our blocks than to cut them. When you have to trim in 100's of >blocks it puts lot of wear and tear on your wrists, elbows and >shoulders. With the automated rough trim all we had to do was push >buttons and let the microtome do the work for us, it was great. > > >-----Original Message----- >Heckford, Karen - SMMC-SF >Sent: Wednesday, November 02, 2005 12:03 PM >To: Histonet (E-mail) >Subject: [Histonet] Microtome > >I was just wondering if anyone has a Microm 335 E microtome. I was >thinking about purchasing one. I am a little scared to face into a block >without manual manipulation. Do I have to worry about this? Please let >me know >your input? > >Karen Heckford >St. Mary's Medical Center >San Francisco, 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 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From anh2006 <@t> med.cornell.edu Wed Nov 2 15:03:38 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Wed Nov 2 14:58:49 2005 Subject: [Histonet] Antibodies for frozen mouse sections In-Reply-To: <6.0.0.22.1.20051102120508.01b643c8@gemini.msu.montana.edu> References: <131DF14ECE564A4D8F35376FD346517207CB7B42@EXCHSRV1.internal.sanger.ac.uk> <6.0.0.22.1.20051102120508.01b643c8@gemini.msu.montana.edu> Message-ID: I use the neutrophil marker Gr-1 from BD Pharmingen with fabulous success on frozen sections. It is actually a granulocyte marker not just neutrophils. Clone RB6-8C5. Use spleen as a positive control. In my hands it works great anywhere from 1-10 ug/ml depending on my detection system. I have not tried it on paraffin. - Andrea >At 10:00 AM 11/2/2005, you wrote: >>Hi, I am staining some frozen mouse NALT and cervical lymph node >>sections with a number of antibodies, CD4, CD8, B220, CD11c (all from >>Pharmingen) and F4/80 (Serotec) and the staining is working fine (thanks >>to Gayle Callis!!). However I want to start staining for some more cell >>markers and I was wondering if anybody could give me details of primary >>antibodies (dilutions etc) and where you get them from, that are good >>for staining natural killer (NK) cells, ICAM-1 (CD54), CD5, MadCAM-1, >>VCAM-1, PNAd and a neutrophil marker, i.e. Ly-6G. >> >>Thanks very much in advance. >> >>Lindsay Hall >>Wellcome Trust Sanger Institute >>Wellcome Trust Genome Campus >>Hinxton >>Cambridge >>CB10 1SA >>UK -- From anh2006 <@t> med.cornell.edu Wed Nov 2 15:11:51 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Wed Nov 2 15:07:04 2005 Subject: [Histonet] antibody In-Reply-To: <20051102135734.16180.qmail@web15509.mail.cnb.yahoo.com> References: <20051102135734.16180.qmail@web15509.mail.cnb.yahoo.com> Message-ID: What do you mean the secondary antibody's isotype is IgG (H+L)? You mean they are anti-IgG (H+L)?? "Anti-IgG (H+L)" means anti-IgG (all isotypes) but reacts with both heavy and light chains. So if I understand you correctly, then yes these antibodies will react with your IgG1 and IgG3. If at all unsure I strongly recommend you talk to the technical center of the company who made the antibody or get the specification sheet for your secondary. >Dear all, > >I have ordered two primary antibodies, their isotypes are IgG1 and >IgG3 repectively. In our lab, there are some secondary antibodies, >their isotypes are IgG (H+L), so I wonder if I use these secondary >antibodies for immunofluorescence. > >Thanks. > >Guofeng > > -- From drgrant <@t> cmh.edu Wed Nov 2 15:05:01 2005 From: drgrant <@t> cmh.edu (Grant, Debra, R) Date: Wed Nov 2 15:09:01 2005 Subject: [Histonet] BK Virus controls Message-ID: <6F434E27D5DE944B9E898984CADDD14904395F@exchmail.CMH.Internal> Hi all, I am looking for a company that sells BK virus control slides that I may use for our SV-40 antibody. Thanks in advance! Debby R. Grant HT(ASCP) Histology Department The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 From godsgirlnow <@t> msn.com Wed Nov 2 15:45:12 2005 From: godsgirlnow <@t> msn.com (Roxanne Soto) Date: Wed Nov 2 15:45:28 2005 Subject: [Histonet] teaching histology newbee In-Reply-To: <4368D49F.7010400@umdnj.edu> Message-ID: If this undergrad will ever gross tissue in PLEASE express the importance of thickness of the tissue to be processed...... Roxanne ______________________________________________________________ From: Geoff McAuliffe To: Malcolm McCallum ,Histonet Subject: Re: [Histonet] teaching histology newbee Date: Wed, 02 Nov 2005 10:00:47 -0500 >Hi Malcolm: > > I would give a short (15-30 minute) talk about how tissue is >prepared (fix, dehydrate, clear, embed, section, mount, stain) and >show a few artifacts (folds, tears, etc). I do think you should >briefly explain H&E and maybe trichromes and PAS. Definitely >explain, with illustrations, how immunostaining works. > In the old days (when I was a student) there might be some >discussion of how different fixatives can give different 'looks' but >now that everything is done in buffered formalin (or some secret, >commercial mixture of who knows what) there seems to be no point in >this. Save that for teaching microtechnique. > Good luck! > >Geoff > > >Malcolm McCallum wrote: > >>Hi, >>I will be teaching an undergraduate histology this spring. For >>those of you who teach this, what fraction of the course do you >>devote to techniques versus anatomy? Do you include pathology, if >>so, how extensively? How extensively do you require them to >>understand the differences among stains, and how do you structure >>your laboratories? Anything else I should consider? >> >>Malcolm L. McCallum >>Assistant Professor >>Department of Biological Sciences >>Texas A&M University Texarkana >>2600 Robison Rd. >>Texarkana, TX 75501 >>O: 1-903-233-3134 >>H: 1-903-791-3843 >>Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.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 >********************************************** > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From malek.j <@t> ghc.org Wed Nov 2 16:59:19 2005 From: malek.j <@t> ghc.org (Malek, Jack M) Date: Wed Nov 2 16:59:35 2005 Subject: [Histonet] Glass Coverslipper Message-ID: Trying to get a feel for how many folks are using glass cover slips and coverslippers versus plastic film coverslips? We're deliberating the purchase of a coupled stainer-coverslipper... Leica versus Sakura. Anyone have experience with the Leica glass coverslipper? Cheers, Jack From caron_fournier <@t> yahoo.ca Wed Nov 2 17:34:06 2005 From: caron_fournier <@t> yahoo.ca (caron fournier) Date: Wed Nov 2 17:34:15 2005 Subject: [Histonet] re: exakt system Message-ID: <20051102233407.71406.qmail@web36302.mail.mud.yahoo.com> thank you everyone who responded to my inquiry. I have gotten enough information and contacted Linda at Exakt so things are looking good. thank you again. Caron Caron Fournier, BSc, R.T. Department of Orthopaedics, Division of Orthopaedic Engineering Research, U.B.C. --------------------------------- Find your next car at Yahoo! Canada Autos From Linresearch <@t> aol.com Wed Nov 2 17:44:57 2005 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Wed Nov 2 17:45:11 2005 Subject: [Histonet] Tunnel Kit Message-ID: <19c.3fc8e709.309aa979@aol.com> Hello, Does anyone know of a good working Tunnel Kit for FFPE rodent tissues? Could share the protocol and source? Thanks, Lin From cmalc <@t> unimelb.edu.au Wed Nov 2 20:31:18 2005 From: cmalc <@t> unimelb.edu.au (Cathy Malcontenti-Wilson) Date: Wed Nov 2 20:31:42 2005 Subject: [Histonet] Hypoxyprobe Message-ID: <6.2.1.2.2.20051103132604.02bbf008@mail.staff.unimelb.edu.au> Hi all, Has anyone tried using the Hypoxyprobe Plus kit for immunostaining hypoxic cells in mouse tissue? The kit we have is a CHEMICON kit and it has a primary antibody conjugated with FITC and then a secondary antibody to the FITC which is conjugated with HRP. They say this was developed so that the mouse monoclonal antibody could be used as a mouse on mouse. We have tried it according to the manufacturers instructions and have got a lot of background staining. I am interested in finding out if anyone else has experience with this and/or has a different method they use for this marker. Thanks in advance. Regards, Cathy From jkiernan <@t> uwo.ca Wed Nov 2 23:57:08 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Wed Nov 2 23:57:16 2005 Subject: [Histonet] Antigen retrieval on very old tissue References: <09C945920A6B654199F7A58A1D7D1FDE01717604@lsexch.lsmaster.lifespan.org> <6.0.0.22.1.20051102113659.01b37958@gemini.msu.montana.edu> <20051102110901.9exojjmpkw4c0c8s@mail.ucla.edu> Message-ID: <4369A6B4.C75804A4@uwo.ca> Ask for specimens from known negative and positive control brains. Try various antigen retrieval methods. Process and immunostain + and - controls alongside sections of your "very old" material. The antigen of interest may not be retrievable after 20 years in formalin. Just a few thoughts! John Kiernan London, Canada. ____________________________________________ nienhuis@ucla.edu wrote: > > What is your recommendation for doing AR on very old brain tissue, while > minimizing damage? > > I have a brain that has been floating in formalin for 20 years, > and need to do immuno staining on it. > > Bob Nienhuis > Neurobiology Research > UCLA / VA Medical Center > Los Angeles, CA > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From M.Prideaux <@t> sheffield.ac.uk Thu Nov 3 04:04:02 2005 From: M.Prideaux <@t> sheffield.ac.uk (Matt Prideaux) Date: Thu Nov 3 04:04:13 2005 Subject: [Histonet] decalcification In-Reply-To: <20051102135136.99694.qmail@web15510.mail.cnb.yahoo.com> References: <20051102135136.99694.qmail@web15510.mail.cnb.yahoo.com> Message-ID: <1131012242.4369e092f06a3@webmail.shef.ac.uk> We decalcify our mouse bones in EDTA, usually for about 4 weeks before processing and embedding. It's a fairly slow process but it seems to preserve the enzymes and antigens in the tissue far better than any of the acid decalcifying methods. If you're going to be doing a lot of bone histology you might want to check out the 'Handbook of Histology Methods for Bone and Cartilage' by Y.H. An and K.L. Martin. We've found it invaluable so far. Matt -- Matt Prideaux Academic Unit of Bone Biology Division of Clinical Sciences (South) D Floor (DU20) Medical School Henry Wellcome Laboratories for Medical Research University of Sheffield Beech Hill Road Sheffield S10 2RX Quoting pex : > Dear all, > > I have a question about decalcification. I have got some bone tissues > (such as femur) from newborn pups, I would like to know if they need to > be decalcified before paraffin embedding. > > Thanks a lot. > > Guofeng > > > > --------------------------------- > ????????G?????????????????????????????????????? > ??????????D?????????????????? > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From rjbuesa <@t> yahoo.com Thu Nov 3 05:59:37 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 3 05:59:49 2005 Subject: [Histonet] Glass Coverslipper In-Reply-To: Message-ID: <20051103115937.32504.qmail@web61217.mail.yahoo.com> The Leica glass coverslippers I knew sometimes presented problems with sticking coverslips. I have been told that now they are better, and it will also depend on the way you store the coverslips. Regarding film coverslippers, Sakura is the best, but some pathologists don't like them because they say the photomicrographs are not the same, but this is not true. If you are going to take photomicrographs with apochromatic objectives with coverslip thickness correction collars (something very specialized to take a maximum advantage of the objective numerical aperture) the film coverslip will not do, you will need glass coverslips No.1 thick. For routine work, the everyday diagnostic work, film coverslips are OK and will even be adequate for photomicrography. The only thing I found was that long time stored slides covered with plastic coverslips tend to have bubbles and this is caused by a not well regulated xylene drop delivery when originally coverslipped. Rene J. "Malek, Jack M" wrote: Trying to get a feel for how many folks are using glass cover slips and coverslippers versus plastic film coverslips? We're deliberating the purchase of a coupled stainer-coverslipper... Leica versus Sakura. Anyone have experience with the Leica glass coverslipper? Cheers, Jack _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From rjbuesa <@t> yahoo.com Thu Nov 3 06:06:42 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 3 06:06:52 2005 Subject: [Histonet] Histotechnician productivity In-Reply-To: <1DB04B57B04C5747B87C3B39F3E605AA07D390AE@harrier> Message-ID: <20051103120642.8727.qmail@web61214.mail.yahoo.com> John: The is a paper dealing with this specific topic published in the December,2004 issue of the Journal of Histotechnology. This paper present tha AVERAGE productivity for each task in histology; its only drawback is that it does not include the RANGE productivity for each. Rene J. "Jasper, Thomas G." wrote: John, I am quite certain that you'll get a variety of opinions on this topic. In approaching this "mission" I think you need to be clear about 2 things. 1) What is the scope of your service? and 2) Where are you going? You do mention a 50-60 block per day figure and basic clinical histology duties performed in general manually. Does this currently work well for you? Are your pathologists happy with the turnaround time and the quality of the work they receive? Are you trying to expand your business? Are your volumes naturally increasing and/or are they projected to increase? Do you want to develop technically, i.e. Immunohistochemistry, In Situ Hybridization? Are you looking at automating some of your current processes, i.e. special stains and coverslipping? Holy buckets! That's a lot of questions right out of the gate! But, you need to answer questions like these to make an accurate assessment and good decisions. One thing I'm willing to bet is that your organization is looking to save money. That's very understandable in this day and age, however you've got to be real careful in doing it. Evaluate your service very carefully as this will reveal true costs and true savings. This may not apply to you, but for instance, take estrogen and progesterone receptors. I'm guessing that you probably send those tests out somewhere. How much does that cost? Do you have the talent in-house to take it on yourself? If you do and your volumes warrant it, you may save money. Money that's easily demonstrated up front and savings in other ways such as faster turnaround times. This in turn increases the overall value of your service and makes the techs more valuable as well. Again, this may not apply to you I just created a scenario to make a point. This leads me to your last statement which is concerning, and that's the comment about losing a position or two. I firmly believe that people are the greatest asset of an organization. You need to hold on to positions with tenacity. Everyone wants to do more with less, and that's great. I like the modification of doing more with what you've got, especially if you've got good people. Even if the people aren't that good you can try to get others as long as you still have positions to fill. Good histotechs are hard to find, enough said there. Geez, I really haven't given you any hard numbers and I guess I'm loathe to do so considering all the unknowns and variables. I can tell you this, years ago when I worked in a contract lab (non-clinical, sweatshop) we were expected to cut 200 blocks in 8 hours with no more than 15 recuts. I did learn to cut pretty fast there but I wouldn't recommend it. I track hours and production here very closely but comparisons are not going to be apples to apples. I value quality over quantity and I expect that people will do their best day in and day out. Techs are people and some are stronger in different areas than others. We average 200-250 blocks per day, with 2 techs embedding and 3-4 techs cutting. We try to finish cutting in about 5 to 5.5 hours. Again, this is fraught with variables. To me, if someone here is cutting 25 to 30 blocks an hour I think that's pretty good. Please excuse my rambling, but this is a tough nut to crack as the world is filled with shades of grey. Good luck to you. tj -----Original Message----- From: John.Sheppard@Health-Partners.org [mailto:John.Sheppard@Health-Partners.org] Sent: Wednesday, November 02, 2005 8:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histotechnician productivity Hello Everyone, I have been assigned a "mission" by our medical director. He wants me to find out what would be a reasonable number of blocks per day for a histotech. I have tried searching the archieves for this information because I know it seems to be discussed quite often. However most of the old threads I found seemed to end up with histology professionals discussing that they would not work in a "factory like" atmosphere where they are slaves to productivity and turn around times. Which to be honest I agree with, histology should be a quality oriented product. I have also looked at the ASCP and NSH websites where this information is not readily accessible. I will probably e-mail them to see if they have any research or insight on this topic. The numbers that I am most familiar with, is 50 to 60 blocks per day per histotech, when one is including embedding, cutting, manual H&E and Pap staining(gyn & non-gyn), manual coverslipping, manual special stains, and frozen sections. I am just not sure how accurate those numbers are for staffing a histology lab. This information is needed because we will be consolidating two separate small community hospital histology labs onto one campus. Apparently this will be used to justify the current staff of both locations. Unfortunately, I believe this information will ultimately be used to eliminate a position or two. Thank you for your time John Sheppard HT(ASCP) CONFIDENTIALITY NOTICE: This message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From rjbuesa <@t> yahoo.com Thu Nov 3 06:28:35 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 3 06:28:44 2005 Subject: [Histonet] labeling regulations In-Reply-To: Message-ID: <20051103122836.48994.qmail@web61224.mail.yahoo.com> In Florida we have to have a label in each specimen bottle with all the information you are asking about (date, name, type, site, etc.). Rene J. "Charles.Embrey" wrote: Years ago, the CAP checklist for histology specified what information was required on the specimen container and even the request form. The newest checklists seem to have completely thrown this out. Is specimen container labeling regulated anywhere else? Is there a requirement anywhere that specimen type/site be noted on the container? Thanks, Charles Embrey PA(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From c.ingles <@t> hosp.wisc.edu Thu Nov 3 08:10:35 2005 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Thu Nov 3 08:12:11 2005 Subject: [Histonet] Glass Coverslipper Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE6A@uwhis-xchng2.hosp.wisc.edu> I have used both types. I believe the glass one is the best, but I have only seen the Leica one used. It is a bit quirky, but I have heard the tape system starts to peel after a period of time. I also don't know if you have to use xylene with the tape systems or not, but we use Propar with the Leica system. I believe it is also expensive as all get out for those tape rolls. We haven't gotten the stainer yet, so I don't know how smoothly the combo system works. Claire Ingles UW Mohs Clinic Madison WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Malek, Jack M Sent: Wed 11/2/2005 4:59 PM To: histonet@pathology.swmed.edu Cc: Subject: [Histonet] Glass Coverslipper Trying to get a feel for how many folks are using glass cover slips and coverslippers versus plastic film coverslips? We're deliberating the purchase of a coupled stainer-coverslipper... Leica versus Sakura. Anyone have experience with the Leica glass coverslipper? Cheers, Jack _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ree3 <@t> leicester.ac.uk Thu Nov 3 08:30:15 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Thu Nov 3 08:30:29 2005 Subject: [Histonet] keratin antibodies Message-ID: Looking for antibodies to cytokeratins, all types for the moment, that work on formalin fixed paraffin processed mouse tissues. Many thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER...U.K.... From barbara.bublava <@t> meduniwien.ac.at Thu Nov 3 08:57:08 2005 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Thu Nov 3 08:59:49 2005 Subject: [Histonet] Histotechnician productivity References: <20051103120642.8727.qmail@web61214.mail.yahoo.com> Message-ID: <02a001c5e086$dcbdc170$dd01a8c0@GERICHTS9XOZZ8> I would be very interested in this article, but I don?t have acsess to this paper. Could someone send it to me please? Babara Bublava ----- Original Message ----- From: "Rene J Buesa" To: "Jasper, Thomas G." ; Cc: Sent: Thursday, November 03, 2005 1:06 PM Subject: RE: [Histonet] Histotechnician productivity > John: > The is a paper dealing with this specific topic published in the December,2004 issue of the Journal of Histotechnology. > This paper present tha AVERAGE productivity for each task in histology; its only drawback is that it does not include the RANGE productivity for each. > Rene J. > > "Jasper, Thomas G." wrote: > John, > > I am quite certain that you'll get a variety of opinions on this topic. In > approaching this "mission" I think you need to be clear about 2 things. 1) > What is the scope of your service? and 2) Where are you going? > You do mention a 50-60 block per day figure and basic clinical histology > duties performed in general manually. Does this currently work well for > you? Are your pathologists happy with the turnaround time and the quality > of the work they receive? Are you trying to expand your business? Are your > volumes naturally increasing and/or are they projected to increase? Do you > want to develop technically, i.e. Immunohistochemistry, In Situ > Hybridization? Are you looking at automating some of your current > processes, i.e. special stains and coverslipping? > Holy buckets! That's a lot of questions right out of the gate! But, you > need to answer questions like these to make an accurate assessment and good > decisions. One thing I'm willing to bet is that your organization is > looking to save money. That's very understandable in this day and age, > however you've got to be real careful in doing it. Evaluate your service > very carefully as this will reveal true costs and true savings. This may > not apply to you, but for instance, take estrogen and progesterone > receptors. I'm guessing that you probably send those tests out somewhere. > How much does that cost? Do you have the talent in-house to take it on > yourself? If you do and your volumes warrant it, you may save money. Money > that's easily demonstrated up front and savings in other ways such as faster > turnaround times. This in turn increases the overall value of your service > and makes the techs more valuable as well. Again, this may not apply to you > I just created a scenario to make a point. > This leads me to your last statement which is concerning, and that's the > comment about losing a position or two. I firmly believe that people are > the greatest asset of an organization. You need to hold on to positions > with tenacity. Everyone wants to do more with less, and that's great. I > like the modification of doing more with what you've got, especially if > you've got good people. Even if the people aren't that good you can try to > get others as long as you still have positions to fill. Good histotechs are > hard to find, enough said there. > Geez, I really haven't given you any hard numbers and I guess I'm loathe to > do so considering all the unknowns and variables. I can tell you this, years > ago when I worked in a contract lab (non-clinical, sweatshop) we were > expected to cut 200 blocks in 8 hours with no more than 15 recuts. I did > learn to cut pretty fast there but I wouldn't recommend it. I track hours > and production here very closely but comparisons are not going to be apples > to apples. I value quality over quantity and I expect that people will do > their best day in and day out. Techs are people and some are stronger in > different areas than others. We average 200-250 blocks per day, with 2 > techs embedding and 3-4 techs cutting. We try to finish cutting in about 5 > to 5.5 hours. Again, this is fraught with variables. To me, if someone > here is cutting 25 to 30 blocks an hour I think that's pretty good. Please > excuse my rambling, but this is a tough nut to crack as the world is filled > with shades of grey. > Good luck to you. > tj > > -----Original Message----- > From: John.Sheppard@Health-Partners.org > [mailto:John.Sheppard@Health-Partners.org] > Sent: Wednesday, November 02, 2005 8:34 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histotechnician productivity > > > Hello Everyone, > I have been assigned a "mission" by our medical director. He > wants me to find out what would be a reasonable number of blocks per day > for a histotech. I have tried searching the archieves for this information > because I know it seems to be discussed quite often. However most of the > old threads I found seemed to end up with histology professionals > discussing that they would not work in a "factory like" atmosphere where > they are slaves to productivity and turn around times. Which to be honest > I agree with, histology should be a quality oriented product. > I have also looked at the ASCP and NSH websites where this > information is not readily accessible. I will probably e-mail them to see > if they have any research or insight on this topic. > The numbers that I am most familiar with, is 50 to 60 blocks per day > per histotech, when one is including embedding, cutting, manual H&E and Pap > staining(gyn & non-gyn), manual coverslipping, manual special stains, and > frozen sections. I am just not sure how accurate those numbers are for > staffing a histology lab. > This information is needed because we will be consolidating two > separate small community hospital histology labs onto one campus. > Apparently this will be used to justify the current staff of both > locations. Unfortunately, I believe this information will ultimately be > used to eliminate a position or two. > Thank you for your time > John Sheppard HT(ASCP) > > CONFIDENTIALITY NOTICE: This message, including any attachments, is for the > sole use of the intended recipient(s) and may contain confidential and > privileged information. Any unauthorized review, use, disclosure or > distribution is prohibited. If you are not the intended recipient, please > contact the sender by reply e-mail and destroy all copies of the original > message. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > --------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DeBrosse_Beatrice <@t> Allergan.com Thu Nov 3 09:02:07 2005 From: DeBrosse_Beatrice <@t> Allergan.com (DeBrosse_Beatrice) Date: Thu Nov 3 09:03:01 2005 Subject: [Histonet] Glass Cover slipper Message-ID: We are using the Sakura glass cover slipper. It seems it has its good days and bad days, with other words, it can be very quirky. Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malek, Jack M Sent: Wednesday, November 02, 2005 2:59 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Glass Coverslipper Trying to get a feel for how many folks are using glass cover slips and coverslippers versus plastic film coverslips? We're deliberating the purchase of a coupled stainer-coverslipper... Leica versus Sakura. Anyone have experience with the Leica glass coverslipper? Cheers, Jack _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Thu Nov 3 09:06:20 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Nov 3 09:06:48 2005 Subject: [Histonet] Glass Cover slipper Message-ID: I use Fisher Scientific and never had any problems. Robyn OHSU From tracy.bergeron <@t> crl.com Thu Nov 3 09:13:44 2005 From: tracy.bergeron <@t> crl.com (tracy.bergeron@crl.com) Date: Thu Nov 3 09:12:42 2005 Subject: [Histonet] Glass Cover slipper In-Reply-To: Message-ID: We have been using the Sakura glass coverslipper for several years now and really like it. Not that we haven't had any issues with it, but they have been few and far between Tracy E. Bergeron, BS, HT (ASCP) Histotechnician Charles River Laboratories Wilmington, MA 978-658-6000 x 1229 "DeBrosse_Beatrice" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/03/2005 10:02 AM To "Malek, Jack M" , cc Subject RE: [Histonet] Glass Cover slipper We are using the Sakura glass cover slipper. It seems it has its good days and bad days, with other words, it can be very quirky. Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malek, Jack M Sent: Wednesday, November 02, 2005 2:59 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Glass Coverslipper Trying to get a feel for how many folks are using glass cover slips and coverslippers versus plastic film coverslips? We're deliberating the purchase of a coupled stainer-coverslipper... Leica versus Sakura. Anyone have experience with the Leica glass coverslipper? Cheers, Jack _______________________________________________ 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 Jacqueline.Malam <@t> rli.mbht.nhs.uk Thu Nov 3 09:10:56 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Thu Nov 3 09:14:04 2005 Subject: [Histonet] PCNA Message-ID: Has anyone out there had any experience in using PCNA on the Ventana Benchmark and, if so, is there any particular clone that performs best Regards Jacqui Malam Royal Lancaster Infirmary England 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 Mary.Judd <@t> suht.swest.nhs.uk Thu Nov 3 09:15:46 2005 From: Mary.Judd <@t> suht.swest.nhs.uk (Mary Judd) Date: Thu Nov 3 09:16:13 2005 Subject: [Histonet] Re: Re glass coverslippers. Histonet Digest, Vol 24, Issue 4 Message-ID: We have had a Leica CV5030 coverslipper for about a year now and like it very much. The only weak point is the little tin tray that catches any reject coverslips. You have to make certain that dried mountant doesn't build up on it or the dispensing nozzle will catch and bend. Mary Judd Immunohistochemistry Lab. Cellular Pathology Southampton General Hospital UK 02380795144 D I S C L A I M E R 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. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Trust unless explicitly stated otherwise. If you have received this e-mail in error please delete the e-mail and contact the Southampton University Hospitals NHS Trust Helpdesk on:- 023 80796000 The information contained in this e-mail may be subject to public disclosure under the Freedom of Information Act 2000. Unless the Information is legally exempt from disclosure, the confidentiality of this e-mail and your reply cannot be guaranteed. This footnote also confirms that this email message has been swept by MailMarshal for the presence of computer viruses. Please visit our website at http://www.suht.nhs.uk From jqb7 <@t> cdc.gov Thu Nov 3 09:37:22 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Nov 3 10:23:28 2005 Subject: [Histonet] Re: Re glass coverslippers. Histonet Digest, Vol 24, Issue 4 Message-ID: We recently demo'd this coverslipper and loved it! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 (404) 639-3590 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mary Judd Sent: Thursday, November 03, 2005 10:16 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Re glass coverslippers. Histonet Digest, Vol 24,Issue 4 We have had a Leica CV5030 coverslipper for about a year now and like it very much. The only weak point is the little tin tray that catches any reject coverslips. You have to make certain that dried mountant doesn't build up on it or the dispensing nozzle will catch and bend. Mary Judd Immunohistochemistry Lab. Cellular Pathology Southampton General Hospital UK 02380795144 D I S C L A I M E R 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. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Trust unless explicitly stated otherwise. If you have received this e-mail in error please delete the e-mail and contact the Southampton University Hospitals NHS Trust Helpdesk on:- 023 80796000 The information contained in this e-mail may be subject to public disclosure under the Freedom of Information Act 2000. Unless the Information is legally exempt from disclosure, the confidentiality of this e-mail and your reply cannot be guaranteed. This footnote also confirms that this email message has been swept by MailMarshal for the presence of computer viruses. Please visit our website at http://www.suht.nhs.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joycefr <@t> frontiernet.net Thu Nov 3 11:11:55 2005 From: joycefr <@t> frontiernet.net (Joyce Friedland) Date: Thu Nov 3 10:56:02 2005 Subject: [Histonet] Re glass coverslippers. In-Reply-To: References: Message-ID: Our lab has been using the Sakura Glass Coverslipper since May and we adore it. The few problems we have had have been operator error. It is important to use coverglass designed specifically for these machines. Joyce Friedland From sharon.osborn <@t> dnax.org Thu Nov 3 11:12:37 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Thu Nov 3 11:15:47 2005 Subject: [Histonet] RE: glass coverslippers Message-ID: <29B25753F6B1D51196110002A589D444032C48A2@PALMSG30.us.schp.com> Jack, I have had experience with both the Sakura plastic film coverslipper and the Leica Glass coverslipper. Hands down I will go for the Leica everytime. Glass does not produce wavy artifact that has to be constantly adjusted out by the microscopist. The plastic film can lift over time (even though that is supposed to be corrected, there is still some occurrence). When it lifts, the tissue may be on the plastic film rather than on the glass slide. For speed, the plastic film is still the faster; however, for quality, longevity and photo shoots, glass is still superior. If you do much photography, glass is definitely better for the camera resolution. Please compare slides covered with the plastic film and glass coverslips both grossly and particularly with the microscope. In using the microscope you will find yourself contantly making adjustments with the film to get clear focus. This can contribute to repetitive motion problems due to the fine motor skills required in making the microscope adjustments. sharon osborn DNAX Palo Alto, CA Date: Wed, 2 Nov 2005 14:59:19 -0800 From: "Malek, Jack M" Subject: [Histonet] Glass Coverslipper To: Message-ID: Content-Type: text/plain; charset="us-ascii" Trying to get a feel for how many folks are using glass cover slips and coverslippers versus plastic film coverslips? We're deliberating the purchase of a coupled stainer-coverslipper... Leica versus Sakura. Anyone have experience with the Leica glass coverslipper? Cheers, Jack ********************************************************************* 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 sharon.osborn <@t> dnax.org Thu Nov 3 11:27:53 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Thu Nov 3 11:28:15 2005 Subject: [Histonet] RE: coverslippers Message-ID: <29B25753F6B1D51196110002A589D444032C48A3@PALMSG30.us.schp.com> Jack, Additional note: If you consider a glass coverslipper that is stand alone and not hooked into a staining system such as the Leica combo, please look at the Hacker coverslipper or the Meisei glass coverslipper. Hacker makes excellent coverslippers. Years ago I used the Meisei (then under Hacker now through TBS) and loved it. It was easy to regulate and repair since we were miles from an easy service call. I learned to maintain it including taking it apart to do the PM. Yes, Leica coverslippers have improved the glass pickup process. And it does behoove the tech loading in the glass coverslips(in any coverslipper!) to have clean, dry hands or gloved clean dry hands in handling the coverslips. Coverslips that are not stuck together are necessary; there are recommended brands that work better than others. We currently purchase our coverslips for the Leica through StatLab in Lewisville, TX. The coverslip is called Anapath. They are very reasonable, loose(not sticky) and work well in the Leica coverslipper. sharon osborn DNAX Palo Alto, CA ********************************************************************* 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 tracy.bergeron <@t> crl.com Thu Nov 3 11:40:57 2005 From: tracy.bergeron <@t> crl.com (tracy.bergeron@crl.com) Date: Thu Nov 3 11:40:03 2005 Subject: [Histonet] Re glass coverslippers. In-Reply-To: Message-ID: Interesting... Over the past 2 yrs we have used coverslips from Mercedes medical, Med supply partners, and Surgipath, and have had no issues with any of those brands with our Sakura glass coverslipper. Tracy E. Bergeron, BS, HT (ASCP) Histotechnician Charles River Laboratories Wilmington, MA 978-658-6000 x 1229 Joyce Friedland Sent by: histonet-bounces@lists.utsouthwestern.edu 11/03/2005 12:11 PM To Histonet@lists.utsouthwestern.edu cc Joyce Friedland Subject [Histonet] Re glass coverslippers. Our lab has been using the Sakura Glass Coverslipper since May and we adore it. The few problems we have had have been operator error. It is important to use coverglass designed specifically for these machines. Joyce Friedland _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From STEGTM <@t> samcstl.org Thu Nov 3 12:34:50 2005 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Thu Nov 3 12:35:30 2005 Subject: [Histonet] Patient identifiers Message-ID: Greetings! I have been asked by our lab director to "put the word out" regarding identifiers on our pathology cases. At present, we label our blocks with the accession number and that is all. We have heard that some places use the accession number, and also write the patient name on the side of the cassette. Of course we check corporate number, name and date of birth on each pathology requisition when accessioning, but, as stated previously, we only label cassettes with the accession number. Should we do more for identity? Oh, by the way, I play bassoon, started in 7th grade. My dad made a belt cup for the bottom of the instrument, I sat on the strap. I always soak the reed in my mouth for a while before playing. Anyone who says that they split reeds on a daily basis is either not getting them wet/pliabe enough, or chewing on them. There is no such thing as playing all night and wearing them out. After about three hours concert play I can't even pucker or purse my lips. Peace, Terre From rjbuesa <@t> yahoo.com Thu Nov 3 12:59:40 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 3 12:59:48 2005 Subject: [Histonet] Solvent Recyclers In-Reply-To: <000501c5db54$49652b70$d86d140a@ITP36079> Message-ID: <20051103185940.69265.qmail@web61221.mail.yahoo.com> I used a xylene recicler for 12 years and it paid itself after 36 months (in disposal and buying savings). The recycled xylene was used for everything (tissue processing and staining). When we began using mineral oil for tissue processing, we cut our expenses even more and then the recycled xylene was used to clean the tissue processors. I had only wished to have been recycling earlier! Rene J. Rod Coombe wrote: I am interested in comments from people who have used solvent recylers. Is the recylced xylene suitable to use in processing machines? Are you recycling alcohol from the processing machines and have you had problems with xylene contamination in the alcohol through the processing machine? How long does it take to recycle 20 litres of alcohol? Is anybody recycling formalin in preference to having it commercially disposed of and if so how long does it take to recycle 20 litres of formalin? Which are the best recyclers? Thankyou Rod Coombe Manager Tissue Pathology Institute of Medical and Veterinary Science Frome Road, Adelaide, South Australia Phone +61 8 8222 3201 Fax +61 8 82223204 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From maria <@t> ski.org Thu Nov 3 14:43:15 2005 From: maria <@t> ski.org (Maria Mejia) Date: Thu Nov 3 14:43:26 2005 Subject: [Histonet] Antigen retrieval on very old tissue References: <09C945920A6B654199F7A58A1D7D1FDE01717604@lsexch.lsmaster.lifespan.org> <6.0.0.22.1.20051102113659.01b37958@gemini.msu.montana.edu> <20051102110901.9exojjmpkw4c0c8s@mail.ucla.edu> Message-ID: <436A7663.4090601@ski.org> Dear Bob, You realize that an extended fixation times in formalin are damaging to many antigens, secondary to the induction of molecular cross-links in proteins. Since your (archival) specimen has been in formalin for 20 years your level of difficulty has increased dramatically since now you need to do IHC on this critical specimen. However, I don't think problem is unusual. I'm sure there are others doing IHC on archival tissues for various studies. You didn't mention if this was a whole brain and from what species? Do you know if the fixative is really (10%) NBF? Has the fixative ever been changed during this period? What type of IHC do you plan to do? Free-floating sections or paraffin sections? Even if you don't know the answers to all these question, I would start by removing the brain from the formalin and place in a container with tap water and wash the specimen in running water for a long period of time. I would start with a 3-4 week washing or longer (Puchtler & Meloan 1985 "On the Chemistry of Formaldehydr Fixative & its Effects on IHC Reactions, Histochemistry 82: 201-204. Also Julies Elias, The Journal of Histotechnology Vol.24, No. 3 2001.) This is the simpliest non-heating method of AR. Elias 1990 adopted this method routinely by storing de-paraffinized sections in 10% sucrose in PBS at 4C for 16 hours (before) immunostaining. Subsequently, it's shown that some chemicals, such as urea, acid, or borohydride in water, may improve the retrieval of masked antigens (Larson 1988, Costa et al, 1986, Hausen & Dreyer 1982.) In your case your will have to do a "test battery" with different times. In fact, I would suggest that you do a "test battery" on every critical level of IHC. These conditions will give you an optimal protocol for your IHC (and) even if nothing works you'll know you tried a whole gamut of things to make it work! I would try different AR solutions: citrate buffer, Tris-HCl buffers or even a good commerical at pH 1-4, pH 6-8, pH 10-11. Try different heating methods: microwave (MW) alone, MW & pressure cooker...maybe low heating or shorter heating times. As you know crosslinkages are complicated processes that depend on a variety of conditions: pH, temp, conditions of tissue & fixation that lead to a variety of protein alteration. Then you need to think about which detection system is best for your needs and the factors that deal with this system. I hope this helps some. Yours Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 nienhuis@ucla.edu wrote: > > What is your recommendation for doing AR on very old brain tissue, while > minimizing damage? > > I have a brain that has been floating in formalin for 20 years, > and need to do immuno staining on it. > > Bob Nienhuis > Neurobiology Research > UCLA / VA Medical Center > Los Angeles, CA > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cbass <@t> bidmc.harvard.edu Thu Nov 3 16:51:54 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Thu Nov 3 16:52:03 2005 Subject: [Histonet] how to get this cryostat working? Message-ID: <7C09AE29-381D-448C-A28F-2F9B21A73F14@bidmc.harvard.edu> Hello, I have located a cryostat that someone is willing to let me use. It was inherited from another lab and has been moved to a new building and has not been powered up for several months. Does anyone have advice on how I should proceed with getting this cryostat in working order? It appears to be fairly clean inside and was decontaminated before it was left to sit. It is a Shandon Cryotome Motorised Electronic cryostat (no. 77210160 GB). I will probably use the cryostat initially about once a week. Should the cryostat be powered on constantly? Or should I turn it on prior to each use? Should it be kept at a particular temperature between uses? The cryostat has a waste bottle that collects water from the cooling coils and also debris from washing the chamber. It is suggested that this bottle should always contain 10% formalin, I assume to serve as a decontaminant. Does this always have to be in use, and should I change the bottle often? Where can I find the correct type of disposable blades to use with this cryostat model. I plan to section brain, spinal cord and liver. Is there a particular brand or type of disposable blade I should use for this purpose? How often should the blades be changed? I have used a cryostat before but I haven't been responsible for its upkeep beyond regular cleaning after use, so if there is anything that I am missing, or any suggestions, please let me know. By the way, as far as cryostats go, how does this model stack up? Any and all suggestions are appreciated. Thanks, Caroline Bass From yang5yc <@t> yahoo.com Thu Nov 3 17:00:41 2005 From: yang5yc <@t> yahoo.com (Kelly Yang) Date: Thu Nov 3 17:00:51 2005 Subject: [Histonet] double staining in frozen tissue Message-ID: <20051103230041.23717.qmail@web33905.mail.mud.yahoo.com> Dear all, Could someone be so kind to share or advise me on a protocol for double staining immunofluorescence on frozen human bladder tissue? 1)Would it cause problem if I use FITC donkey anti-goat IgG and Alexa 647 goat anti-rabbit together as my secondary antibody? 2)Which fixation is better? I had tried 4% fornaldehyde (at room temp) for 5 min and icy acetone (-20) for5 min, and found with acetone-fixation, the morphology of nucleus is very poor. Any suggestion? Any help will be highly appreciated, Kelly Yang Graduate student Department of Epidemiology School of Public Heath University of California, Los Angeles 310-409-9179 ext. 57795 yangyc@ucla.edu __________________________________ Yahoo! Mail - PC Magazine Editors' Choice 2005 http://mail.yahoo.com From ROrr <@t> enh.org Fri Nov 4 07:41:23 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Nov 4 07:41:32 2005 Subject: [Histonet] Recording Benchmark Temperatures Message-ID: Hello everyone, In preparing for my first CAP inspection in 10 years, I have a temperature question. What do all of you Benchmark users do in reference to the temps during the Cell Conditioning phase? Do you record it? Since the Benchmark will alarm if it goes out of range, and the temps on the Benchmark are factory preset...I'm thinking I don't have to record, as long as I make a statement about the temps used in the procedure manual. I also use a Biocare Decloaker and I do record these temps. Opinions please! Many Thanks and Happy Friday Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From rjbuesa <@t> yahoo.com Fri Nov 4 07:49:00 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 4 07:49:08 2005 Subject: [Histonet] Colon mets - cytokeratin IHC In-Reply-To: Message-ID: <20051104134900.85153.qmail@web61214.mail.yahoo.com> Hi Andrea: I think that you should try to get a paper by Moll, Franke & Schiller (The Catalog of Human Cytokeratins) it is fantastic review (a little old though) that may answer your questions. It appeared published in Cell, vol.31, pp.11-24, Nov.,1982. >From the "practical" point of view, for colon I always used CK20 (DAKO; Mo. 1:50, HIER pH6); for skin AE1/AE3 (DAKO; Mo; 1:50 HIER pH6) and CK 5/6 ( BoeringhenManhein Mo, 1:25 HIER pH6); for prostate CK7 (Dako;Mo; 1:25, HIER pH6) and CK34BE12 (DAKO, Mo, 1:75, HIER pH6). Hope this will help you! Rene J. "Andrea T. Hooper" wrote: Hi All, I am looking for the best possible IHC stain to use on colon met samples to distinguish single met cells amongst normal adjacent liver. I was thinking AE1/AE3 or some CK20 antibody or any others the experts may suggest. I would love to get suggestions of best clones and sources for the antibodies as well as staining dilutions and protocols if possible. Thanks, Andrea -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From rjbuesa <@t> yahoo.com Fri Nov 4 09:10:41 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 4 09:10:51 2005 Subject: [Histonet] teaching histology newbee In-Reply-To: Message-ID: <20051104151041.76692.qmail@web61220.mail.yahoo.com> Hi Malcom: Between 1991 and 1996 I taught 10 times a Histology Review course aimed at histotechs that were going to take the state certification examination. What I am going to do is to give you the general scope of the printed course and the number of pages dedicated to each subject: 1- Historical account with an overview of the types of techniques-----6 pp. 2- Laboratory mathematics (preparation of all types of solutions/dilutions)---16 pp. 3- Fixation/fixatives types/uses/advantages/disadvantages----11 pp. 4- Dehydrating/clearing agents -- 2pp. 5- Cutting and extremely difficult specimens to cut---18 pp. 6- Decalcification procedures ---4 pp. 7- Summary of electron microscopy---2 pp. 8- Principles of frozen sectioning, celloidin, carbowax and double embedding--- 3pp. 9- Staining of some special tissue components ---- 18 pp. 10- Use of the microwave oven in the histology lab./ oven calibrations --- 7 pp 11- Microscopy--- 5pp. 12- The fundamental importance of cellular surface in biology--- 5pp. 13- "DOS" and "DON'TS" in the histology laboratory--- 6pp. 14- Safety in the histology laboratory ---4 pp 15- Quality control ---5 pp 16- Fundaments of immunohistochemistry procedures --- 17 pp 17- Assorted information (some things I considered would help the students to know regarding the histology lab/procedures)--- 6 pp 18- Summary of other special procedures --- 6 pp. This review was given during 8 hours (lunch break), earned the attendants 8 CEU and this text was the guideline to be "seasoned" with 2-3 "carrousels" full of slides (specially photos of the IHC procedures and photomicrographs of all the "special stains" done at my lab). Perhaps this will help you to "chery-pick" what you want to present at your course. Rene J. Malcolm McCallum wrote: Hi, I will be teaching an undergraduate histology this spring. For those of you who teach this, what fraction of the course do you devote to techniques versus anatomy? Do you include pathology, if so, how extensively? How extensively do you require them to understand the differences among stains, and how do you structure your laboratories? Anything else I should consider? Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From lvalenta <@t> firstsourcelab.com Fri Nov 4 09:56:18 2005 From: lvalenta <@t> firstsourcelab.com (Valenta, Laura) Date: Fri Nov 4 09:55:27 2005 Subject: [Histonet] Histology Position Opening Message-ID: <365FCDF2F3FC4745916F6034F4B4CC0A07481F@fsfiles.FirstSource.local> FirstSource Laboratory Solutions, a full reference laboratory located near Indianapolis, Indiana has an opening for a Histotechnologist. All interested applicants should respond to this email address or fax a resume to 317.566.9847 Attn: HR Department. The job description and qualifications are listed below. HISTOTECHNOLOGIST The Histotechnologist is qualified to provide preparation of Histology and Cytology specimens under the supervision of the Pathologist Medical Director. RESPONSIBILITIES Competently identify, record, measure, and gross specimens. Processes tissue in processor, embeds tissue in paraffin cassette blocks, cuts tissue on microtome and transfers to water bath/slides. Stains and coverslips finished slides to present to Pathologist for examination. Maintains and operates all equipment in Histology and Cytology laboratory. Prepares cytology specimens and presents slides to Cytotechnologists for examination. Performs assignments from Pathologists as directed. From esther.peters <@t> verizon.net Fri Nov 4 10:24:56 2005 From: esther.peters <@t> verizon.net (Esther Peters) Date: Fri Nov 4 10:26:51 2005 Subject: [Histonet] teaching histology newbee References: <20051104151041.76692.qmail@web61220.mail.yahoo.com> Message-ID: <436B8B58.4020104@verizon.net> Dear Malcolm, I taught undergraduate histology for three years. The basic slide preparation equipment was available and I thought it would be important for students to have an appreciation of all the work that goes into making slides even if they never had to do this again when they went to medical or dental school (and I had supervised and worked in research histology labs), or they might decide to go into histology as a career or use it in research. I had contacted some other histology professors and learned that they only did slide reading during their labs. I stuck to just normal histology, although I am also trained in pathology. (One of the histology profs I contacted said he had never had any pathology experience, so only used "normal" prepared slides to teach.) The course was 4 credits, one 2 1/2-hour lecture and one 2 1/2-hour lab each week for one semester. Lectures followed the selected textbook (Wheater's or Gartner and Hiatt). I found it useful to review the previous lecture at the beginning of the next lecture (after the lab that had used those organs/tissues). I prepared brief bulleted notes for both lectures and labs and provided them to the students on what I thought they should really know as undergrads (realizing that if they did go on to med or dental school the subject will be taught in much more depth). The lab lectures touched on most all of the topics Rene mentioned. The lab was roughly half techniques and half slide reading based on the lecture topic. For the techniques part we obtained a beheaded rat from the psychology department (which uses the brain for nervous system studies) and dissected it in class, and also a fish (from a field fisheries biologist), and clams (from grocery store), and students could bring in their own research animal if they wished (we had corals and insects). Each was dissected and representative portions of organs and tissues were fixed in NBF and Bouin's (for freshwater and terrestrial organisms), or seawater-formalin and Helly's made with zinc chloride (for marine organisms). Students then took turns trimming tissues into cassettes, embedding, sectioning (to make 4 slides from each block), and stained with Harris's H&E, Cason's (5-min aniline blue procedure), and alcian blue/PAS with and without diastase. For the final lab report, students were given 5 "mystery blocks" and the slides for each of those (usually a mix, for example liver from rat fixed with Bouin's and NBF, liver from fish 2 fixatives, whole clam), and then had to identify which organs/tissues were present, also explain the quality of the staining and why the fixation might have mattered and what was wrong with the histoslides. (We did not have time to work on perfecting sections, so just went with what was produced in one or two lab sessions.) The final three labs 1 hour was devoted to working with the students as they studied their histoslides and providing clues to help them identify what they had (and pointing out why). If we found a pathologic change, parasite, or pathogen, I pointed it out. It was a bit hectic at times.... Of course, I had students who never understood that they should attend the lectures, really WORK on slide reading (they were given worksheets with directions for which slides to use, what to identify, and sketching and received points for completing them), and thought the slide prep was a waste of time. I also had students who "got the message" and realized that the "mystery blocks" exercise gave them a taste of what a pathologist would encounter when facing a new histoslide for diagnosis. I wish you good luck in this endeavor! There are several good Web sites these days for additional histology study. Let me know if you'd like more information. Esther Peters, Ph.D. Affiliate Professor George Mason University Rene J Buesa wrote: > Hi Malcom: > Between 1991 and 1996 I taught 10 times a Histology Review course aimed at histotechs > that were going to take the state certification examination. > What I am going to do is to give you the general scope of the printed course and the number > of pages dedicated to each subject: > 1- Historical account with an overview of the types of techniques-----6 pp. > 2- Laboratory mathematics (preparation of all types of solutions/dilutions)---16 pp. > 3- Fixation/fixatives types/uses/advantages/disadvantages----11 pp. > 4- Dehydrating/clearing agents -- 2pp. > 5- Cutting and extremely difficult specimens to cut---18 pp. > 6- Decalcification procedures ---4 pp. > 7- Summary of electron microscopy---2 pp. > 8- Principles of frozen sectioning, celloidin, carbowax and double embedding--- 3pp. > 9- Staining of some special tissue components ---- 18 pp. > 10- Use of the microwave oven in the histology lab./ oven calibrations --- 7 pp > 11- Microscopy--- 5pp. > 12- The fundamental importance of cellular surface in biology--- 5pp. > 13- "DOS" and "DON'TS" in the histology laboratory--- 6pp. > 14- Safety in the histology laboratory ---4 pp > 15- Quality control ---5 pp > 16- Fundaments of immunohistochemistry procedures --- 17 pp > 17- Assorted information (some things I considered would help the students to know > regarding the histology lab/procedures)--- 6 pp > 18- Summary of other special procedures --- 6 pp. > > This review was given during 8 hours (lunch break), earned the attendants 8 CEU and this text was the guideline to be > "seasoned" with 2-3 "carrousels" full of slides (specially photos of the IHC procedures and > photomicrographs of all the "special stains" done at my lab). > Perhaps this will help you to "chery-pick" what you want to present at your course. > Rene J. > > Malcolm McCallum wrote: > Hi, > I will be teaching an undergraduate histology this spring. For those of you who teach this, what fraction of the course do you devote to techniques versus anatomy? Do you include pathology, if so, how extensively? How extensively do you require them to understand the differences among stains, and how do you structure your laboratories? Anything else I should consider? > > Malcolm L. McCallum > Assistant Professor > Department of Biological Sciences > Texas A&M University Texarkana > 2600 Robison Rd. > Texarkana, TX 75501 > O: 1-903-233-3134 > H: 1-903-791-3843 > Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > --------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From sjchtascp <@t> yahoo.com Fri Nov 4 10:31:45 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Nov 4 10:31:56 2005 Subject: [Histonet] CD83 Message-ID: <20051104163146.15022.qmail@web90207.mail.scd.yahoo.com> Has anyone had any experiance using this antibody on mouse tissue. We are trying to work up the "eBioscience" CD83 for IF. Steve --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From ree3 <@t> leicester.ac.uk Fri Nov 4 10:39:14 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Nov 4 10:39:25 2005 Subject: [Histonet] keratin antibodies Message-ID: Looking for antibodies to cytokeratins, all types for the moment, that work on formalin fixed paraffin processed mouse tissues. Many thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER...U.K.... From dford <@t> deltapathology.com Fri Nov 4 11:33:28 2005 From: dford <@t> deltapathology.com (D. Ford) Date: Fri Nov 4 11:43:09 2005 Subject: [Histonet] Water Source for Routine H&E Staining Message-ID: I was recently told that there has been a regulatory change regarding the water used for H&E staining. I am told that this change requires that our stainers be attached to a filtration system. Has anyone else heard of this regulation? Darlene Ford, B.S. CT, HT (ASCP) Technical Supervisor 2915 Missouri Avenue Shreveport, LA 71109 318-621-8820 phone 318-671-5922 fax From jgemmanual <@t> wlgore.com Fri Nov 4 11:47:47 2005 From: jgemmanual <@t> wlgore.com (Jeannie G Emmanuel) Date: Fri Nov 4 11:47:51 2005 Subject: [Histonet] sectioning JB-4 or GMA blocks Message-ID: Hi! If you have worked with GMA-based embedded specimen I would like to know how you obtained beautiful (wrinkle free) 3-5 micron thick sections without using chloroform. Did everything the tech support person said to do but it didn't help. My sample/block size is at least 1" x 1". From liz <@t> premierlab.com Fri Nov 4 12:08:04 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Nov 4 12:08:19 2005 Subject: [Histonet] large glass slides and coverslips Message-ID: <000001c5e16a$b53a3180$a7d48a80@AMY> I know this question has been asked before, but I've tried the histonet search and other search engines, but who's the supplier for larger glass slides and coverglass. Is it Brain Research Laboratories? I can't seem to locate the web page. These are for whole mount guinea pig lung specimens. Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From PMonfils <@t> Lifespan.org Fri Nov 4 12:44:31 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Nov 4 12:44:46 2005 Subject: [Histonet] large glass slides and coverslips Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171760C@lsexch.lsmaster.lifespan.org> Yes, Brain Research Laboratories offers large slides and coverglass. http://www.brainresearchlab.com/ > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Elizabeth Chlipala > Sent: Friday, November 4, 2005 10:08 AM > To: 'Histonet' > Subject: [Histonet] large glass slides and coverslips > > I know this question has been asked before, but I've tried the histonet > search and other search engines, but who's the supplier for larger glass > slides and coverglass. Is it Brain Research Laboratories? I can't seem > to locate the web page. These are for whole mount guinea pig lung > specimens. > > Thanks in advance > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > Manager > Premier Laboratory, LLC > P.O. Box 18592 > Boulder, Colorado 80308 > Office: (303) 735-5001 > Fax: (303) 735-3540 > liz@premierlab.com > www.premierlab.com > > Ship to Address: > Premier Laboratory > University of Colorado > MCDB, Room A3B40 > Boulder, Colorado 80309 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From LChen <@t> mednet.ucla.edu Fri Nov 4 12:59:07 2005 From: LChen <@t> mednet.ucla.edu (Chen, Leslie) Date: Fri Nov 4 12:59:22 2005 Subject: [Histonet] Patient Identifiers Message-ID: <9F8AE7E7B303F44DB0CAB587F1E96C3F02A67737@admedmail3.ad.medctr.ucla.edu> Hi Terre, Here in California we have HIPAA/Patient Privacy regulations that prevent us from using identifiers. You should check what privacy regulations you have there before including patient name on the blocks. Here at ucla we only use the accession number. UCLA Lung Spore Program Phone: x49461 Pager: 94317 Message: 1 Date: Thu, 03 Nov 2005 12:34:50 -0600 From: "Therersa Stegall" Subject: [Histonet] Patient identifiers To: Message-ID: Content-Type: text/plain; charset=US-ASCII Greetings! I have been asked by our lab director to "put the word out" regarding identifiers on our pathology cases. At present, we label our blocks with the accession number and that is all. We have heard that some places use the accession number, and also write the patient name on the side of the cassette. Of course we check corporate number, name and date of birth on each pathology requisition when accessioning, but, as stated previously, we only label cassettes with the accession number. Should we do more for identity? Oh, by the way, I play bassoon, started in 7th grade. My dad made a belt cup for the bottom of the instrument, I sat on the strap. I always soak the reed in my mouth for a while before playing. Anyone who says that they split reeds on a daily basis is either not getting them wet/pliabe enough, or chewing on them. There is no such thing as playing all night and wearing them out. After about three hours concert play I can't even pucker or purse my lips. Peace, Terre ---------------------------------------------------------- IMPORTANT WARNING: This email (and any attachments) is only intended for the use of the person or entity to which it is addressed, and may contain information that is privileged and confidential. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Unauthorized redisclosure or failure to maintain confidentiality may subject you to federal and state penalties. If you are not the intended recipient, please immediately notify us by return email, and delete this message from your computer. ---------------------------------------------------------- From histomjans <@t> yahoo.com Fri Nov 4 13:06:53 2005 From: histomjans <@t> yahoo.com (Melissa Jans) Date: Fri Nov 4 13:07:02 2005 Subject: [Histonet] pneumatic tube for specimen transport Message-ID: <20051104190654.39074.qmail@web35611.mail.mud.yahoo.com> Posted on behalf of Sue Lewis: I would like to know if there are work areas or labs using a pneumatic tube system to transport wet tissue, FFPE blocks and/or slides between work areas. If anyone were using a tube system, would you share any challenges, successes or recommendations? Thank you, Sue Lewis Sue E. Lewis, BS, HTL (ASCP) Assistant Laboratory Manager Anatomic Pathology, 5216B RCP University of Iowa Hospitals and Clinics Iowa City, IA 52242-1011 319-353-8661 sue-lewis@uiowa.edu --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From TJJ <@t> Stowers-Institute.org Fri Nov 4 13:27:41 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Nov 4 13:28:04 2005 Subject: [Histonet] Re: double staining in frozen tissue Message-ID: Kelly, Answers to your questions: 1)Would it cause problem if I use FITC donkey anti-goat IgG and Alexa 647 goat anti-rabbit together as my secondary antibody? Yes, it would cause a lot of problems because the FITC anti-goat secondary would bind with the goat anti-rabbit. I would use donkey-anti rabbit instead. I recommend anybody doing multiple staining methods purchase Chris van der Loos' book. You can find it on Amazon.com...it's an excellent reference for these types of questions and problems. 2)Which fixation is better? I had tried 4% fornaldehyde (at room temp) for 5 min and icy acetone (-20) for5 min, and found with acetone-fixation, the morphology of nucleus is very poor. Any suggestion? It depends on what fixation does to the epitope you're trying to visualize. Formalin can cause a lot of autofluorescence and can, in some cases, mask the antigenicity. In other cases, it provides excellent fixation to the proteins you're staining, and of course provides excellent morphology. Try it and see if it works. Many antibodies are successfully used on acetone-fixed cryosections. However you have found the limitation of using acetone alone. Do a search on the histonet (www.histosearch.com/histonet) and look for Gayle Callis' recipe and instructions on using acetone and absolute alcohol mixture. In many cases, it retains antigenicity and preserves morphology very well. Do not let the slides air dry after acetone/alcohol fixation, but rather go into your buffer solution and continue your protocol from there. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From Tiffany.L.Sheffield <@t> uth.tmc.edu Fri Nov 4 14:04:34 2005 From: Tiffany.L.Sheffield <@t> uth.tmc.edu (Sheffield, Tiffany L) Date: Fri Nov 4 14:04:37 2005 Subject: [Histonet] Fixative Question Message-ID: Hello fellow Histonetters! Could someone recommend a fixative for EM/TEM for bone. I am familiar with a few techniques from the Handbook of Histology Methods for Bone and Cartilage by An & Martin. I am wanting to preserve synovial and bone/cartilage tissue with attached antibodies for electron microscopy without destroying the antibody-tissue binding? I was just wondering if any of you had any experience with these techniques or could recommend a fixative?Any help would greatly be appreciated. Thanks! Tiffany Sheffield-Lopez, HT(ASCP) Supervisor, Bone Histomorphometry & Biomaterials Lab Department of Orthopaedic Surgery UTHSC-Houston Medical School 6431 Fannin, Suite 6.144 MSB Houston , TX 77030 713-500-6803 WK 713-500-0729 Fax From MDiCarlo <@t> KaleidaHealth.Org Fri Nov 4 14:12:55 2005 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Nov 4 14:11:18 2005 Subject: [Histonet] vacuum oven Message-ID: <139141F8BAF4A642A945ECC528511AF001F5ED43@kalmb02.kaleidahealth.org> Hello histonetters, I am in need of a vacuum oven that has similar dimensions to the one I have been using. The inside dimensions are 8 inches wide by 8 inches high by 12 inches deep. The entire outside of the oven measures 13 inches wide by 14 inches high by 15 inches deep. Can anyone suggest a company that sells vacuum ovens around this same size? I would appreciate any information you have to offer. Thank you. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 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 rjbuesa <@t> yahoo.com Fri Nov 4 14:33:25 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 4 14:33:38 2005 Subject: [Histonet] large glass slides and coverslips In-Reply-To: <000001c5e16a$b53a3180$a7d48a80@AMY> Message-ID: <20051104203325.7966.qmail@web61215.mail.yahoo.com> You could try the following: www.brainresearchlab.com www.thomasscientific.com www.fishersci.com www.proscitech.com www.surgipath.com Rene J. Elizabeth Chlipala wrote: I know this question has been asked before, but I've tried the histonet search and other search engines, but who's the supplier for larger glass slides and coverglass. Is it Brain Research Laboratories? I can't seem to locate the web page. These are for whole mount guinea pig lung specimens. Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From BlazekL <@t> childrensdayton.org Fri Nov 4 14:34:55 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Nov 4 14:36:31 2005 Subject: [Histonet] pneumatic tube for specimen transport Message-ID: We don't use the tube system for transporting wet tissue at all. We have used it for blocks and slides though. - Linda Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Melissa Jans" 11/04/2005 2:06:53 PM >>> Posted on behalf of Sue Lewis: I would like to know if there are work areas or labs using a pneumatic tube system to transport wet tissue, FFPE blocks and/or slides between work areas. If anyone were using a tube system, would you share any challenges, successes or recommendations? Thank you, Sue Lewis Sue E. Lewis, BS, HTL (ASCP) Assistant Laboratory Manager Anatomic Pathology, 5216B RCP University of Iowa Hospitals and Clinics Iowa City, IA 52242-1011 319-353-8661 sue-lewis@uiowa.edu --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Thomas.Crowell <@t> biogenidec.com Fri Nov 4 14:48:09 2005 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Fri Nov 4 14:48:13 2005 Subject: [Histonet] CD80 In-Reply-To: Message-ID: Has anyone worked out a protocol to detect CD80 in FFPE human samples? Tom Crowell BiogenIdec Cambridge, MA From gcallis <@t> montana.edu Fri Nov 4 15:08:29 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Nov 4 15:08:41 2005 Subject: [Histonet] CD83 In-Reply-To: <20051104163146.15022.qmail@web90207.mail.scd.yahoo.com> References: <20051104163146.15022.qmail@web90207.mail.scd.yahoo.com> Message-ID: <6.0.0.22.1.20051104140752.01b678a0@gemini.msu.montana.edu> More details would help? Are you having problems? At 09:31 AM 11/4/2005, you wrote: >Has anyone had any experiance using this antibody on mouse tissue. We are >trying to work up the "eBioscience" CD83 for IF. > >Steve > > >--------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From daniel.cadolle <@t> bluewin.ch Sat Nov 5 04:13:44 2005 From: daniel.cadolle <@t> bluewin.ch (daniel.cadolle@bluewin.ch) Date: Sat Nov 5 04:13:57 2005 Subject: [Histonet] please unsubscribe from mailing list Message-ID: <200511051013.jA5ADiRj007277@sun5.netoxygen.ch> PLEASE UNSUBSCRIBE FROM MAILING LIST BECAUSE OF SUBSCRIBER'S DEATH. THANK YOU - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - YellowSwiss, l'annuaire des entreprises suisses - www.yellowswiss.ch - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - From daniel.cadolle <@t> bluewin.ch Sat Nov 5 04:16:09 2005 From: daniel.cadolle <@t> bluewin.ch (daniel.cadolle@bluewin.ch) Date: Sat Nov 5 04:16:23 2005 Subject: [Histonet] (no subject) Message-ID: <200511051016.jA5AG9fD007435@sun5.netoxygen.ch> PLEASE UNSUBSCRIBE FROM MAILING LIST BECAUSE OF SUBSCRIBER'S DEATH. THANK YOU - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - YellowSwiss, l'annuaire des entreprises suisses - www.yellowswiss.ch - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - From daniel.cadolle <@t> bluewin.ch Sat Nov 5 04:38:39 2005 From: daniel.cadolle <@t> bluewin.ch (daniel.cadolle@bluewin.ch) Date: Sat Nov 5 04:38:48 2005 Subject: [Histonet] UNSUBSCRIPTION Message-ID: <200511051038.jA5Acdc6008140@sun5.netoxygen.ch> PLEASE UNSUBSCRIBE FROM MAILING LIST BECAUSE OF SUBSCRIBER'S DEATH. THANK YOU - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - YellowSwiss, l'annuaire des entreprises suisses - www.yellowswiss.ch - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - From rjbuesa <@t> yahoo.com Sat Nov 5 11:04:33 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Nov 5 11:04:46 2005 Subject: [Histonet] TOT: third (and final) update summary. Message-ID: <20051105170433.57580.qmail@web61214.mail.yahoo.com> To all Histonet subscribers: After being disconnected for @ 10 days (no electricy/telephone) due to hurricane WILMA I am now in the position of posting the third update about the received TOTs. Since 17 Oct./95 I received just 4 additional: 23- use 10% KOH during 20-45 min at 45C to soften toe nails; 24- use plus slides for toe nails sections; 25-use sulfuric acid x 20 secs.,rinse with water and follow with 0.5% aq. toluidine blue for "thick" (0.5-1 um) epoxy sections for EM (more details will be offered when final the version is used); 26- soak toe nails blocks in soapy water before cutting. Althogh I am certain that most of you have some TOTs to share with all of us, the response has been weaker than I initially thought it would be, so it is not worth it to keep posting byweekly summaries. This will be the last. Nevertheless you could keep sending me your TOTs and they will be summarized by the end of the year. To all of you that have contributed to the common good by sharing your TOTs, I thank you. Sincerely, Rene J. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From rmeyers <@t> ctcsurge.com Sat Nov 5 12:17:37 2005 From: rmeyers <@t> ctcsurge.com (Roger Meyers) Date: Sat Nov 5 12:21:03 2005 Subject: [Histonet] automated response Message-ID: <10511051317.AA11097@mail1.ctcsurge.com> I will be out of the office from Thursday, November 3, 2005 through Friday, November 4, 2005. If the matter is urgent, please call Computer Trust Corporation at 617-557-9264 for immediate assistance. From gu.lang <@t> gmx.at Sat Nov 5 12:42:40 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat Nov 5 12:42:47 2005 Subject: AW: [Histonet] pneumatic tube for specimen transport In-Reply-To: <20051104190654.39074.qmail@web35611.mail.mud.yahoo.com> Message-ID: We use it as fast transportsystem between the surgery and the histolab for frozen sections. It works fine, because the nurses wrap the wet tissue in paper or plastic bags. Or they put it in small, tight containers. So, until now the system was kept clean. The other specimen come with a "little train" cruising through the whole house. Gudrun Lang Akh Linz Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Melissa Jans Gesendet: Freitag, 04. November 2005 20:07 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] pneumatic tube for specimen transport Posted on behalf of Sue Lewis: I would like to know if there are work areas or labs using a pneumatic tube system to transport wet tissue, FFPE blocks and/or slides between work areas. If anyone were using a tube system, would you share any challenges, successes or recommendations? Thank you, Sue Lewis Sue E. Lewis, BS, HTL (ASCP) Assistant Laboratory Manager Anatomic Pathology, 5216B RCP University of Iowa Hospitals and Clinics Iowa City, IA 52242-1011 319-353-8661 sue-lewis@uiowa.edu --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Sat Nov 5 13:41:54 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Nov 5 13:42:03 2005 Subject: [Histonet] Toe Nail TOTs summary. Message-ID: <20051105194154.30086.qmail@web61222.mail.yahoo.com> Dear Histonet subscribers: I finished editing the summary I announced on Sun 16 Oct./05 In case you are interested in its contents, please let me know and I will send you a copy. Rene J. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From MVaughan4 <@t> ucok.edu Sat Nov 5 19:01:03 2005 From: MVaughan4 <@t> ucok.edu (MVaughan4@ucok.edu) Date: Sat Nov 5 19:01:25 2005 Subject: [Histonet] Thanks salivary gland questions Message-ID: Thanks to all who responded. I have another class where I have th students prepare and stain slides, then I use these slides for Histology vet school close by allow me to harvest tiss return the favor. Mel Melville B. Vaughan, Ph. D. Assistant&nb Department of Biology University of Edmond, OK 73034 From fivetoads <@t> charter.net Mon Nov 7 01:10:01 2005 From: fivetoads <@t> charter.net (fivetoads@charter.net) Date: Mon Nov 7 01:10:12 2005 Subject: [Histonet] Please Unsubscribe Message-ID: <4enjo0$1kqh9cg@mxip16a.cluster1.charter.net> Please delete my email address from the mailing list. Thanks From mikael.niku <@t> helsinki.fi Mon Nov 7 01:39:28 2005 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Mon Nov 7 01:39:40 2005 Subject: [Histonet] Bone marrow unspecific staining Message-ID: <436F04B0.2020809@helsinki.fi> Dear Histonetters, I'm trying to do some immunos with bone marrow sections, but have problems with unspecific staining. It seems a large proportion of the BM cells is unspecifically binding any antibody, including my secondaries. These are paraffin-embedded sections of paraformaldehyde-fixed material, either decalcified tissue samples or agar-embedded cells. I have tried to improve blocking, for example by increasing the serum concentration in the pretreatments, and by adding serum to the antibody solutions, but it doesn't help at all. I have also tried different secondaries (biotinylated and fluorescent) but the results are always the same. I need to do HIER in an acid buffer for my most important primary in this project, but it doesn't seem to be the cause of the problem. Any ideas, anyone? With best regards, Mikael -- //////////////////////////////////////////////////////////// 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 //////////////////////////////////////////////////////////// From John.Sheppard <@t> Health-Partners.org Mon Nov 7 04:20:40 2005 From: John.Sheppard <@t> Health-Partners.org (John.Sheppard@Health-Partners.org) Date: Mon Nov 7 04:16:06 2005 Subject: [Histonet] Re: Histotechnician productivity Message-ID: Hello again, I just wanted to say "thank you" to everyone who sent me their insight on my previous question. I think that it has given me even more questions to figure out than answers. But, thats why I love Histonet so much! Some people had questions about our current scope of practice, and productivity. We do basic histology and special stains - no immuno's. We currently do about 8000 cases a year (approx. 20,000 slides) with two HT's a gross assistant/autopsy tech and a part-time cytotech who knows enough histology to cover the HT's sick days and vacations. We probably average 80 blocks a day. There has been times we have had 150 or more blocks too. We staff histology from 5am to 3pm and pathology from 9am to 5pm. When we merge the two labs we will have 4 more people but only about 6000 more cases, also our autopsies per year will more than double. So we will see what happens. I am going to give all of the responses to my supervisor and my medical director. Thanks again John Sheppard HT(ASCP) CONFIDENTIALITY NOTICE: This message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From MDiCarlo <@t> KaleidaHealth.Org Mon Nov 7 07:25:18 2005 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Mon Nov 7 07:23:43 2005 Subject: [Histonet] large glass slides and coverslips Message-ID: <139141F8BAF4A642A945ECC528511AF001F5ED44@kalmb02.kaleidahealth.org> Yes, Brain Research Laboratories is a supplier of large glass slides and coverglass. Their address is : Waban P.O. Box 88 Newton, MA 02468 Phone (617)965-5544 Fax (617) 965-6220 Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rene J Buesa Sent: Friday, November 04, 2005 15:33 To: Elizabeth Chlipala; 'Histonet' Subject: Re: [Histonet] large glass slides and coverslips You could try the following: www.brainresearchlab.com www.thomasscientific.com www.fishersci.com www.proscitech.com www.surgipath.com Rene J. Elizabeth Chlipala wrote: I know this question has been asked before, but I've tried the histonet search and other search engines, but who's the supplier for larger glass slides and coverglass. Is it Brain Research Laboratories? I can't seem to locate the web page. These are for whole mount guinea pig lung specimens. Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ 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 steem21 <@t> hotmail.com Mon Nov 7 08:05:08 2005 From: steem21 <@t> hotmail.com (Steve Leung) Date: Mon Nov 7 08:05:21 2005 Subject: [Histonet] IHC for markers of chronic inflammation Message-ID: Dear All, I wish to use IHC for the detection of chronic inflammation in human tissues - specifically within the prostate. I am using formalin fixed sections of prostate. The list of possible markers for chronic inflammation out there is rather overwhelming. I was wondering if anyone had a small panel of tried and tested antibodies that would cover most of the chronic inflammatory cells or give a good representative picture of chronic inflammation within the tissue. Or even suggest a few antibodies with which I could start with. Many thanks for attention, Steve Leung Clinical Research Fellow University of Edinburgh U.K. From niloof <@t> yahoo.com Mon Nov 7 09:00:09 2005 From: niloof <@t> yahoo.com (Niloufar Fozouni) Date: Mon Nov 7 09:00:37 2005 Subject: [Histonet] measuring collagen Message-ID: <20051107150019.64420.qmail@web35602.mail.mud.yahoo.com> Hello, Does anybody know, how I can measure collagen content in cartilage? Thank you. Niloo __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From rjbuesa <@t> yahoo.com Mon Nov 7 11:05:36 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 7 11:05:47 2005 Subject: [Histonet] Bone marrow unspecific staining In-Reply-To: <436F04B0.2020809@helsinki.fi> Message-ID: <20051107170536.8447.qmail@web61216.mail.yahoo.com> Mikael: In the description of your steps I don't see you blocking the endogenous peroxidase; do you do that step (5 minutes with 3% hydrogen peroxide after HIER)? Rene J. Mikael Niku wrote: Dear Histonetters, I'm trying to do some immunos with bone marrow sections, but have problems with unspecific staining. It seems a large proportion of the BM cells is unspecifically binding any antibody, including my secondaries. These are paraffin-embedded sections of paraformaldehyde-fixed material, either decalcified tissue samples or agar-embedded cells. I have tried to improve blocking, for example by increasing the serum concentration in the pretreatments, and by adding serum to the antibody solutions, but it doesn't help at all. I have also tried different secondaries (biotinylated and fluorescent) but the results are always the same. I need to do HIER in an acid buffer for my most important primary in this project, but it doesn't seem to be the cause of the problem. Any ideas, anyone? With best regards, Mikael -- //////////////////////////////////////////////////////////// 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 //////////////////////////////////////////////////////////// _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From anh2006 <@t> med.cornell.edu Mon Nov 7 11:10:14 2005 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Mon Nov 7 11:10:40 2005 Subject: [Histonet] Bone marrow unspecific staining Message-ID: <9badb0bbc8ae.436f4426@med.cornell.edu> Hi, Can you provide more details of your staining protocol? Without more info it is a little hard to guess what the problem may be. But on first glance these are my comments: (1) Are your secondaries cross-adsorbed against whatever species you are working with? So for example if you are staining mouse tissue with rat primaries, your secondary needs to be anti-rat IgG, no cross to mouse. (2) BM is a tricky tissue with hard to quench endogenous peroxidases. If you are using HRP based protocols, you may need a special addendum protocol for peroxidase blocking. [Doubtful though since you are using FFPE sections.] (3) What antigen are you staining for? Are you sure your antibody is "good"? Do you have a positive control which is not bone? What concentration are you using primaries and secondaries at? Did you titrate them? (4) Finally, what were your tissues decalcified with? I would suggest you run the following controls if you haven't done so already: (a) No primary, no secondary, no tertiary - only DAB - check for endogenous peroxidases (b) No primary, no secondary, only Strep-HRP and DAB - check for endogenous biotin (c) No primary, only secondary, tertiary, and DAB - check for secondary antibody non-specific binding (d)? Isotype control of primary, then secondary, tertiary, and DAB - check for non-specific binding associated with isotype of primary Good luck! Andrea At 9:39 AM +0200 11/7/05, Mikael Niku wrote: Dear Histonetters, I'm trying to do some immunos with bone marrow sections, but have problems with unspecific staining. It seems a large proportion of the BM cells is unspecifically binding any antibody, including my secondaries. These are paraffin-embedded sections of paraformaldehyde-fixed material, either decalcified tissue samples or agar-embedded cells. I have tried to improve blocking, for example by increasing the serum concentration in the pretreatments, and by adding serum to the antibody solutions, but it doesn't help at all. I have also tried different secondaries (biotinylated and fluorescent) but the results are always the same. I need to do HIER in an acid buffer for my most important primary in this project, but it doesn't seem to be the cause of the problem. Any ideas, anyone? With best regards, Mikael -- //////////////////////////////////////////////////////////// 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 //////////////////////////////////////////////////////////// _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From boycea <@t> mail.nih.gov Mon Nov 7 12:18:26 2005 From: boycea <@t> mail.nih.gov (Boyce, Amanda (NIH/NIAMS)) Date: Mon Nov 7 12:18:40 2005 Subject: [Histonet] RE: measuring collagen Message-ID: <0E3E7E8F6E23DF4C8127A063568356B510B98802@nihexchange12.nih.gov> Our lab uses a hydroxyproline assay. I've never used it, so I can't troubleshoot it for you, but here's the protocol: Acid hydrolysis * Papain digest and 12N HCL 1:1 * 110 overnight in sealed glass ampules * dry sample 24-48 h in vacuum-pump with NaOH * resuspend in 1 ml assay buffer, vortex, overnight at 4, vortex again * store at -20 or assay Hydroxyproline assay Reagents: * stock buffer: 50 g citric acid monohydrate, 12 ml glacial acetic acid , 120 g sodium acetate trihydrate, 34 g sodium hydroxide ad 1 liter, pH 6.0, store at 4 * assay buffer: 1:10 dilution of stock buffer * chloramin-T-reagent: 0.3525g Chloramine T dissolved in 5.175 ml water, add 6.5 ml m-propanol and 13.325 ml stock buffer, make fresh * Dimethylaminobenzaldehyde reagent: 3.75 g dimethylaminobenzaldehyde in 15 ml n-propanol, add 6.5 ml perchloric acid, make fresh * Standard: 1 mg/ml hydroyxproline, store -20 Method: 1. standard dilutions: 0, 0 , 0.2, 0.4, 0.6, 0.8, 1, 2, 4, 6, 8 ,10 ?g/ml 2. sample dilutions: 1:5, 1:10, 1:20, 1:40, 1:80,1: 100 3. 200 ?l sample/standard in eppendorf 4. add 100 ?l Chloramine T, pipett mix 20 min room temp 5. add 100 ?l dimethylaminobenzaldehyde, pipet mix, 15 min 60 water bath 6. 300 ?l in 96 well plate and read immediately 540 nm -------------------------- Amanda Taylor Boyce, Ph.D. Postdoctoral IRTA Fellow Cartilage Biology and Orthopaedics Branch National Institute of Arthritis and Musculoskeletal and Skin Diseases National Institutes of Health Bldg. 13, Rm. 3W17 Bethesda, MD 20892-5755 Phone: 301-451-6860 Fax: 301-480-4315 Email: boycea@mail.nih.gov > Message: 6 > Date: Mon, 7 Nov 2005 07:00:09 -0800 (PST) > From: Niloufar Fozouni > Subject: [Histonet] measuring collagen > To: histonet@lists.utsouthwestern.edu > Message-ID: <20051107150019.64420.qmail@web35602.mail.mud.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > Hello, > > Does anybody know, how I can measure collagen content > in cartilage? > Thank you. > Niloo > > From GauchV <@t> mail.amc.edu Mon Nov 7 12:24:25 2005 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Mon Nov 7 12:24:58 2005 Subject: [Histonet] H. Pylori Controls Message-ID: Does anyone know of a good source for H. Pylori Controls?? We normally order from Newcomer Supply but they are experiencing a large backorder right now and we need something to hold us over until they arrive. Any help would be greatly appreciated... Thanks so much, Vicki Gauch AMCH Albany, NY ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. From rjbuesa <@t> yahoo.com Mon Nov 7 13:30:08 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 7 13:30:19 2005 Subject: [Histonet] H. Pylori Controls In-Reply-To: Message-ID: <20051107193008.74020.qmail@web61212.mail.yahoo.com> We used positive cases that were selected by the pathologists, but there is another approarch. Usually pathologists want to be able to have a positive control of exactly what they are looking for; if this is the case with your pathologists you will need controls comntaining H. pylori. Now, the whole idea with the control is to make sure that the procedure worked, and this you can obtain with any tissue containing abundant number of bacteria. After a long "convincing" process with our pathologists I was able to make them aware of this fact. We used the Steiner method (with phosphotungstic acid instead of the radiactive uranyl nitrate) and for control we used appendix (and you will not find a more easy to find tissue with more bacteria than an appendix). Perhaps you would like to try to talk about this approach with your pathologist. Just a thought! Rene J. Vicki Gauch wrote: Does anyone know of a good source for H. Pylori Controls?? We normally order from Newcomer Supply but they are experiencing a large backorder right now and we need something to hold us over until they arrive. Any help would be greatly appreciated... Thanks so much, Vicki Gauch AMCH Albany, NY ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From LINDA.MARGRAF <@t> childrens.com Mon Nov 7 13:49:23 2005 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Mon Nov 7 13:49:48 2005 Subject: [Histonet] histologist/pathologist ratio Message-ID: Here's a message Beatrice is having trouble posting to the list. Please email her or send your reply to the entire list (and not me directly). Thanks Linda M (Histonet administrator) "DeBrosse_Beatrice" Hi! I would like to know what the ratio of how many histologists to a Pathologist is. Does the ratio differ from hospitals to reference labs to pharmaceutical labs? Your help is greatly appreciated. Sincerely, Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 From JGordon <@t> cellmarque.com Mon Nov 7 14:16:27 2005 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Mon Nov 7 14:16:38 2005 Subject: [Histonet] H. Pylori Controls Message-ID: Cell Marque offers H. pylori positive controls for IHC, as well as controls for every other antibody that we carry. Jeff Gordon Cell Marque Corp. http://www.cellmarque.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Vicki Gauch Sent: Mon 11/7/2005 12:24 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] H. Pylori Controls Does anyone know of a good source for H. Pylori Controls?? We normally order from Newcomer Supply but they are experiencing a large backorder right now and we need something to hold us over until they arrive. Any help would be greatly appreciated... Thanks so much, Vicki Gauch AMCH Albany, NY ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From yun.li <@t> utsouthwestern.edu Mon Nov 7 14:21:17 2005 From: yun.li <@t> utsouthwestern.edu (Yun Li) Date: Mon Nov 7 14:19:57 2005 Subject: [Histonet] Sheep anti BrdU Message-ID: <436FB73D.3090606@utsouthwestern.edu> Greetings everyone, I am trying to do some double immuno-staining for BrdU and other antigens. Since the antibodies for my "other" antigens are mostly from mouse, I am looking for a non-mouse (and preferably non-rat) antibody against BrdU. I came across this sheep anti brdU antibody in the literature, it seems that several different companies offer it, including maine biotech, abcam, novus and one called genetex. So my question is has anyone here used it before, what type of sections was used, how did it work and which company was it purchased from? Thanks a lot! yun From rjbuesa <@t> yahoo.com Mon Nov 7 14:43:36 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 7 14:43:44 2005 Subject: [Histonet] histologist/pathologist ratio In-Reply-To: Message-ID: <20051107204336.58622.qmail@web61216.mail.yahoo.com> Hi Beatrice: Some months ago I reviewed different sources (US Dept Labor; US Dept. Commerce;ASCP) and got to the figure of about 1.4 to 2.0 histotechs per pathologist. This, as any other average, is "not written in stone" and has to vary between laboratories. Of the laboratories I personally know the averages have been from 1 to 1, to 2.2 to 1 These figures will also depend on the type of laboratory: laboratories in small hospitals will probably will be closer to 1:1, in large reference laboratories, although will have many more pathologists, the huge sample volume will require the larger side of the scale (2 or 2:2 histotechs / pathologist). Rene J. LINDA MARGRAF wrote: Here's a message Beatrice is having trouble posting to the list. Please email her or send your reply to the entire list (and not me directly). Thanks Linda M (Histonet administrator) "DeBrosse_Beatrice" Hi! I would like to know what the ratio of how many histologists to a Pathologist is. Does the ratio differ from hospitals to reference labs to pharmaceutical labs? Your help is greatly appreciated. Sincerely, Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From dholmes <@t> anatomy.umsmed.edu Mon Nov 7 15:34:24 2005 From: dholmes <@t> anatomy.umsmed.edu (Dianne Holmes) Date: Mon Nov 7 15:34:51 2005 Subject: [Histonet] Sihler's Stain Message-ID: Is anyone familiar with this process to counterstain nerve fibers within a relatively transparent muscle? I need a materials and method list. Can anyone help me? From mplhisto <@t> aol.com Mon Nov 7 15:56:31 2005 From: mplhisto <@t> aol.com (mplhisto@aol.com) Date: Mon Nov 7 15:56:45 2005 Subject: [Histonet] (no subject) In-Reply-To: References: Message-ID: <8C7B1FD3F626F95-B64-AA33@FWM-R21.sysops.aol.com> Can anyone tell me the name of a good computer safety training program ? we are looking to update our current program ? Thanks Meredith Hale HT (ASCP) Histology Supervisor Pathology Partners Irving, Texas From rjbuesa <@t> yahoo.com Mon Nov 7 16:03:50 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 7 16:03:58 2005 Subject: [Histonet] Sihler's Stain In-Reply-To: Message-ID: <20051107220350.18981.qmail@web61225.mail.yahoo.com> Hi Dianne: Sihler's method was published by Bohm and Oppel in 1907 and is used in pieces of fesh muscle. The solutions are: A- water 75 mL + glycerol 12 mL+ acetic acid 12 mL + chloral hydrate 0.75 g B- water 75 ml + Ehrlich hematoxylin (1886 formula) 12 mL + glycerol 12 mL + chloral hydrate 0.75 g C- 0.1% acetic acid in glycerol. Method: pieces of fresh muscle in Sol.A for 18 hours; transfer to Sol.B for 3 to 10 days; transfer to Sol.C untill differentiated, mount in glycerol, seal coverslip with wax. Hope this will help! Rene J. Dianne Holmes wrote: Is anyone familiar with this process to counterstain nerve fibers within a relatively transparent muscle? I need a materials and method list. Can anyone help me? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From jcolclefa <@t> aol.com Mon Nov 7 16:23:10 2005 From: jcolclefa <@t> aol.com (John PJ Coleman) Date: Mon Nov 7 16:23:30 2005 Subject: [Histonet] Inflammation In-Reply-To: <200511071300.8d436f965416b@rly-xb02.mx.aol.com> References: <200511071300.8d436f965416b@rly-xb02.mx.aol.com> Message-ID: For inflammation (?) I personally would start trying to label the leucocytes. What about CD15 for Polys, CD117 for mast cells, and CD30 or CD68 for Histiocytes? On Nov 7, 2005, at 1:01 PM, histonet-request@lists.utsouthwestern.edu wrote: Dear All, I wish to use IHC for the detection of chronic inflammation in human tissues - specifically within the prostate. I am using formalin fixed sections of prostate. The list of possible markers for chronic inflammation out there is rather overwhelming. I was wondering if anyone had a small panel of tried and tested antibodies that would cover most of the chronic inflammatory cells or give a good representative picture of chronic inflammation within the tissue. Or even suggest a few antibodies with which I could start with. Many thanks for attention, Steve Leung Clinical Research Fellow University of Edinburgh U.K. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 24, Issue 9 *************************************** JPJC-757 335-2159 http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ From Linresearch <@t> aol.com Mon Nov 7 16:24:57 2005 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Mon Nov 7 16:25:13 2005 Subject: [Histonet] Histo Online Program Message-ID: <252.72dde0.30a12e39@aol.com> Hello, Does anyone know of a current online program for Histology? Lin From vargasb <@t> nova.edu Mon Nov 7 17:17:21 2005 From: vargasb <@t> nova.edu (Bernardo Vargas Angel) Date: Mon Nov 7 17:17:33 2005 Subject: [Histonet] Pleaes unsuscribe Message-ID: <1131405441.436fe08192c3b@mail2.nova.edu> Please unsuscribe Bernardo Vargas Angel Ph.D From caron_fournier <@t> yahoo.ca Mon Nov 7 17:54:26 2005 From: caron_fournier <@t> yahoo.ca (caron fournier) Date: Mon Nov 7 17:54:34 2005 Subject: [Histonet] re: grinding bone sections Message-ID: <20051107235426.70464.qmail@web36313.mail.mud.yahoo.com> Hi All: what is the finest paper that you have used on an exakt system for grinding bone samples (undecalcified). We were told that the 2500 was the highest they had but there is literature that notes 4000. Caron Fournier, BSc, R.T. Department of Orthopaedics, Division of Orthopaedic Engineering Research, U.B.C. --------------------------------- Find your next car at Yahoo! Canada Autos From liz <@t> premierlab.com Mon Nov 7 18:36:36 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Mon Nov 7 18:36:50 2005 Subject: [Histonet] cytospin or cytek machine Message-ID: <000001c5e3fc$79e12870$a7d48a80@AMY> Hello Histonetters I have a unique request. I have a client that is in Emeryville California and they need access to a cytospin or a Thin Prep machine, they are willing to pay for time, etc. If there is anyone out there that is willing to do this can they e-mail me back with contact information so I can forward it to them. Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From jkiernan <@t> uwo.ca Mon Nov 7 23:15:03 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Nov 7 23:15:13 2005 Subject: [Histonet] Sihler's Stain References: <20051107220350.18981.qmail@web61225.mail.yahoo.com> Message-ID: <43703457.F66DB4E3@uwo.ca> Before going ahead with Sihler's haemalum method (for peripheral nerves in cleared whole mounts) consult recent publications: Wu BL, Sanders I (1994) The Sihler stain: A technique for studying peripheral nerves in whole-mount specimens. Neuroscience Protocols Module 4: 83-93. Wu B, Saunders I (1994) The Sihler stain: a technique for studying peripheral nerves in whole-mount specimens. In: FG Wouterlood, Neuroscience Protocols. Elsevier, Amsterdam. pp. 94-050-07-01 to 94-050-07-10. Mu LC, Sanders I (1998) Neuromuscular organization of the human upper esophageal sphincter. Annals of Otology Rhinology and Laryngology 107: 370-377. Mu LC, Sanders I (1999) Neuromuscular organization of the canine tongue. Anatomical Record 256: 412-424. Gozil R, Kadioglu D, Calguner E, Erdogan D, Bahcelioglu M, Elmas C (2002) Branching patterns of rabbit oculomotor and trochlear nerves demonstrated by Sihler's stain technique. Biotechnic & Histochemistry 77: 21-25. This is an important method for examining the "gross" anatomy of small nerves. Rene: do you have a copy of Bohm & Appel 1907 or of any of Sihler's papers? If so, I'd be really grateful for bibliographic details! John Kiernan Anatomy, UWO London, Canada. ______________________________________________________________ Rene J Buesa wrote: > > Hi Dianne: > Sihler's method was published by Bohm and Oppel in 1907 and is used in pieces of fesh muscle. The solutions are: > A- water 75 mL + glycerol 12 mL+ acetic acid 12 mL + chloral hydrate 0.75 g > B- water 75 ml + Ehrlich hematoxylin (1886 formula) 12 mL + glycerol 12 mL + chloral hydrate 0.75 g > C- 0.1% acetic acid in glycerol. > Method: pieces of fresh muscle in Sol.A for 18 hours; transfer to Sol.B for 3 to 10 days; transfer to Sol.C untill differentiated, mount in glycerol, seal coverslip with wax. > Hope this will help! > Rene J. > > Dianne Holmes wrote: > Is anyone familiar with this process to counterstain nerve fibers within a relatively transparent muscle? I need a materials and method list. Can anyone help me? > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > --------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From louise.renton <@t> gmail.com Tue Nov 8 00:55:14 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Tue Nov 8 00:55:22 2005 Subject: [Histonet] re: grinding bone sections In-Reply-To: <20051107235426.70464.qmail@web36313.mail.mud.yahoo.com> References: <20051107235426.70464.qmail@web36313.mail.mud.yahoo.com> Message-ID: Yep, we GRIND on 2500, but POLISH on 4000. This help?? On 11/8/05, caron fournier wrote: > Hi All: > what is the finest paper that you have used on an exakt system for grinding bone samples (undecalcified). We were told that the 2500 was the highest they had but there is literature that notes 4000. > > > Caron Fournier, BSc, R.T. > Department of Orthopaedics, > Division of Orthopaedic Engineering Research, > U.B.C. > > > > --------------------------------- > Find your next car at Yahoo! Canada Autos > _______________________________________________ > 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 "....onwards through the fog!" From mikael.niku <@t> helsinki.fi Tue Nov 8 02:08:19 2005 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Tue Nov 8 02:08:32 2005 Subject: [Histonet] Bone marrow unspecific staining In-Reply-To: <9badb0bbc8ae.436f4426@med.cornell.edu> References: <9badb0bbc8ae.436f4426@med.cornell.edu> Message-ID: <43705CF3.4010806@helsinki.fi> Andrea and Rene, thanks for your comments. I'm sorry for being a bit unspecific with my unspecificity question :) Some more details on the problem: - Seems to be caused by unspecific antibody binding, but not a specific property of this primary ab - Primary antibody works very well with other tissues - Also other primaries (and secondary alone) show unspecific staining with BM - Not caused by endogenous biotin, peroxidase, or autofluorescence (done enough controls) And some more details on the protocol: - Bovine BM tissue samples decalcified with EDTA, or cell samples embedded in agar - Paraformaldehyde fixation - HIER (glycine-HCl pH 3) + Tween-20 permeabilization + mild protease digestion - For peroxidase-based protocol, biotin block (tried also peroxidase block, doesn't help) - Serum block - Rabbit primary antibody (overnight +4C) - Sheep secondary antibody (biotinylated or fluorescent) - For biotinylated secondary, tyramide amplification & DAB reaction, hematoxylin counterstain - For fluorescent secondary, Sudan Black B for autofluorescence suppression - Embedding with Faramount -- //////////////////////////////////////////////////////////// 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 //////////////////////////////////////////////////////////// From peter_bannister <@t> hotmail.co.uk Tue Nov 8 06:07:52 2005 From: peter_bannister <@t> hotmail.co.uk (Peter Bannister) Date: Tue Nov 8 06:08:01 2005 Subject: [Histonet] Thermal polymerisation of LR gold resin for electron microscopy Message-ID: Hello Histonetters, Does anyone have a method for the thermal polymerisation of LR gold resin for electron microscopy? I am trying to produce blocks of cultured cell pellets fixed in para/glut, post fixed in Osmium tetroxide, dehydrated in ethanol, infiltrated with the resin and cured. I have tried polymerisation using light with mixed results - non osmium treated blocks polymerise okay and are just about suitable for sectioning. However, osmium treated blocks will only polymerise down as far as the cell pellet (I understand that the dark colour absorbs light and therefore interferes with the polymerisation). I have tried to keep exposure to oxygen during polymerisation to a minimum by using closed BEEM capsules or snap fit gelatin capsules. I would therefore like to try the more classical thermal polymerisation at +60°C. The problem is that when I add the initiator (benzoyl peroxide) to the pure resin I end up with a solid mass of resin even before the peroxide is fully dissolved, at room temperature -so i can't use it on my samples. If anyone can help me out with a protocol that will ensure the resin only polymerises quickly when at heated to +60°C it would be very much appreciated. Many thanks for your time, Peter Bannister. Dr. Peter Bannister, Thrombosis Research Institute, Histopathology, Emmanuel Kaye Building, Manresa Road, London SW3 6LR. UK. _________________________________________________________________ Be the first to hear what's new at MSN - sign up to our free newsletters! http://www.msn.co.uk/newsletters From sjchtascp <@t> yahoo.com Tue Nov 8 07:47:03 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Tue Nov 8 07:47:12 2005 Subject: [Histonet] Red marker Message-ID: <20051108134703.44353.qmail@web90207.mail.scd.yahoo.com> Does anyone know of a company that makes permanant red markers that resist organic solvents? Thanks, Steve --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From germckeon <@t> excite.com Tue Nov 8 07:51:01 2005 From: germckeon <@t> excite.com (germckeon@excite.com) Date: Tue Nov 8 07:51:10 2005 Subject: [Histonet] Mounting Gelatine Embedded Cryostat CNS Sections Message-ID: <20051108135101.79C1B3E01@xprdmailfe6.nwk.excite.com> Hello, I am currently working on spinal cord and am having trouble with sections floating off. I have embedded with gelatine, am performing cryostat sectioning and will be doing some of the more structural stains eg Kultschitsky's stain for myelin. I have been using Superfrost slides. Can anyone advise me regarding choices of slides, coatings, methodologies, etc which would maximise adherence? In gratitude for any advice received, Gerald McKeon _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! From c.ingles <@t> hosp.wisc.edu Tue Nov 8 08:00:50 2005 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Tue Nov 8 08:01:00 2005 Subject: [Histonet] Red marker Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE6D@uwhis-xchng2.hosp.wisc.edu> I believe I just saw some new ones in a Market lab catalogue yesterday. 1-800-237-3604 or www.marketlabinc.com. :) Claire Ingles UW Mohs Clinic Lab Madison Wisconsin -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Steven Coakley Sent: Tue 11/8/2005 7:47 AM To: Histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Red marker Does anyone know of a company that makes permanant red markers that resist organic solvents? Thanks, Steve --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RJLevier <@t> LancasterGeneral.org Tue Nov 8 08:38:41 2005 From: RJLevier <@t> LancasterGeneral.org (LeVier, Rebecca J) Date: Tue Nov 8 08:38:55 2005 Subject: [Histonet] Reagent Grade Alcohol Message-ID: <0FDBF29200C637468CCC1F9E7E85A3D25E2214@MAIL-LR.lha.org> Hello Histonetters, I was just wondering what everyone was using for alcohol. Does anyone see any differences in Reagent Grade Alcohol compared to LCB regulated alcohol? ...... Any information on this topic would be greatly appreciated. Thanks in advance. Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From rjbuesa <@t> yahoo.com Tue Nov 8 08:50:28 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 8 08:50:40 2005 Subject: [Histonet] Bone marrow unspecific staining In-Reply-To: <43705CF3.4010806@helsinki.fi> Message-ID: <20051108145028.81939.qmail@web61218.mail.yahoo.com> Although I cannot be absolutely sure, for me it seems that perhaps the problem may reside in the paraformaldehyde fixation. Do you have to use it? Could you use another fixative in another parallel sample just to try to find out? Just a thoght! Rene J. Mikael Niku wrote: Andrea and Rene, thanks for your comments. I'm sorry for being a bit unspecific with my unspecificity question :) Some more details on the problem: - Seems to be caused by unspecific antibody binding, but not a specific property of this primary ab - Primary antibody works very well with other tissues - Also other primaries (and secondary alone) show unspecific staining with BM - Not caused by endogenous biotin, peroxidase, or autofluorescence (done enough controls) And some more details on the protocol: - Bovine BM tissue samples decalcified with EDTA, or cell samples embedded in agar - Paraformaldehyde fixation - HIER (glycine-HCl pH 3) + Tween-20 permeabilization + mild protease digestion - For peroxidase-based protocol, biotin block (tried also peroxidase block, doesn't help) - Serum block - Rabbit primary antibody (overnight +4C) - Sheep secondary antibody (biotinylated or fluorescent) - For biotinylated secondary, tyramide amplification & DAB reaction, hematoxylin counterstain - For fluorescent secondary, Sudan Black B for autofluorescence suppression - Embedding with Faramount -- //////////////////////////////////////////////////////////// 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 //////////////////////////////////////////////////////////// _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From HornHV <@t> archildrens.org Tue Nov 8 08:56:38 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Tue Nov 8 08:56:19 2005 Subject: [Histonet] histologist/pathologist ratio Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDE9A@EMAIL.archildrens.org> We have 5 anatomic pathologists and 3 histotechs. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Monday, November 07, 2005 2:44 PM To: LINDA MARGRAF; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] histologist/pathologist ratio Hi Beatrice: Some months ago I reviewed different sources (US Dept Labor; US Dept. Commerce;ASCP) and got to the figure of about 1.4 to 2.0 histotechs per pathologist. This, as any other average, is "not written in stone" and has to vary between laboratories. Of the laboratories I personally know the averages have been from 1 to 1, to 2.2 to 1 These figures will also depend on the type of laboratory: laboratories in small hospitals will probably will be closer to 1:1, in large reference laboratories, although will have many more pathologists, the huge sample volume will require the larger side of the scale (2 or 2:2 histotechs / pathologist). Rene J. LINDA MARGRAF wrote: Here's a message Beatrice is having trouble posting to the list. Please email her or send your reply to the entire list (and not me directly). Thanks Linda M (Histonet administrator) "DeBrosse_Beatrice" Hi! I would like to know what the ratio of how many histologists to a Pathologist is. Does the ratio differ from hospitals to reference labs to pharmaceutical labs? Your help is greatly appreciated. Sincerely, Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ 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 rjbuesa <@t> yahoo.com Tue Nov 8 08:59:22 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 8 08:59:31 2005 Subject: [Histonet] Reagent Grade Alcohol In-Reply-To: <0FDBF29200C637468CCC1F9E7E85A3D25E2214@MAIL-LR.lha.org> Message-ID: <20051108145923.84304.qmail@web61212.mail.yahoo.com> I always used 100% (= "200 grade") ,bought in bulk tax exempted, ethanol. Anything you need to do with "pure" ethanol will be simpler to calculate (solutions, mixtures, etc.) with 100% ethanol. Besides that, the "denatured" ethanol contains substances other than ethanol. Now, "reagent grade" sold by chemical companies with "impurities certificates", are much more expensive and offer no advantage over the type of "200 grade" ethanol I used to use. Rene J. "LeVier, Rebecca J" wrote: Hello Histonetters, I was just wondering what everyone was using for alcohol. Does anyone see any differences in Reagent Grade Alcohol compared to LCB regulated alcohol? ...... Any information on this topic would be greatly appreciated. Thanks in advance. Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From GauchV <@t> mail.amc.edu Tue Nov 8 09:00:22 2005 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Tue Nov 8 09:01:02 2005 Subject: [Histonet] Thank you Message-ID: Thank you to everyone who responded to my query for H. Pylori Controls...we are looking into them now....You really are GREAT !!! Vicki AMCH Albany ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. From rjbuesa <@t> yahoo.com Tue Nov 8 09:02:52 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 8 09:03:01 2005 Subject: [Histonet] histologist/pathologist ratio In-Reply-To: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDE9A@EMAIL.archildrens.org> Message-ID: <20051108150252.25624.qmail@web61217.mail.yahoo.com> Question to all subscribers: why don't you post the histologists/pathologists ratios you have in each of your own laboratories? It will be like a survey from Histonet that for sure will usefull for all. Yesterday I posted the ratois I was aware of; now is your turns! Rene J. "Horn, Hazel V" wrote: We have 5 anatomic pathologists and 3 histotechs. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Monday, November 07, 2005 2:44 PM To: LINDA MARGRAF; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] histologist/pathologist ratio Hi Beatrice: Some months ago I reviewed different sources (US Dept Labor; US Dept. Commerce;ASCP) and got to the figure of about 1.4 to 2.0 histotechs per pathologist. This, as any other average, is "not written in stone" and has to vary between laboratories. Of the laboratories I personally know the averages have been from 1 to 1, to 2.2 to 1 These figures will also depend on the type of laboratory: laboratories in small hospitals will probably will be closer to 1:1, in large reference laboratories, although will have many more pathologists, the huge sample volume will require the larger side of the scale (2 or 2:2 histotechs / pathologist). Rene J. LINDA MARGRAF wrote: Here's a message Beatrice is having trouble posting to the list. Please email her or send your reply to the entire list (and not me directly). Thanks Linda M (Histonet administrator) "DeBrosse_Beatrice" Hi! I would like to know what the ratio of how many histologists to a Pathologist is. Does the ratio differ from hospitals to reference labs to pharmaceutical labs? Your help is greatly appreciated. Sincerely, Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ 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. ============================================================================== --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From pmarcum <@t> vet.upenn.edu Tue Nov 8 09:16:18 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Tue Nov 8 09:16:39 2005 Subject: [Histonet] Bone marrow unspecific staining In-Reply-To: <20051108145028.81939.qmail@web61218.mail.yahoo.com> References: <43705CF3.4010806@helsinki.fi> <20051108145028.81939.qmail@web61218.mail.yahoo.com> Message-ID: <6.1.1.1.2.20051108100701.019a08b0@mail.vet.upenn.edu> Paraformaldehyde is just freshly made formaldehyde solution from powder in a buffer. It should not make a difference in the staining or background. As this is a bovine sample in origin you may need to change your blocking serum. You do not give the type of serum so here I can not really make a suggestion. Animal tissues often requires making adjustments not suggested in standard protocols. Pamela A Marcum UPENN Vet School Kennett Square, PA 610-925-6278 At 09:50 AM 11/8/2005, Rene J Buesa wrote: >Although I cannot be absolutely sure, for me it seems that perhaps the >problem may reside in the paraformaldehyde fixation. Do you have to use >it? Could you use another fixative in another parallel sample just to try >to find out? >Just a thoght! >Rene J. > >Mikael Niku wrote: >Andrea and Rene, thanks for your comments. I'm sorry for being a bit >unspecific with my unspecificity question :) > >Some more details on the problem: > >- Seems to be caused by unspecific antibody binding, but not a specific >property of this primary ab >- Primary antibody works very well with other tissues >- Also other primaries (and secondary alone) show unspecific staining >with BM >- Not caused by endogenous biotin, peroxidase, or autofluorescence (done >enough controls) > >And some more details on the protocol: > >- Bovine BM tissue samples decalcified with EDTA, or cell samples >embedded in agar >- Paraformaldehyde fixation >- HIER (glycine-HCl pH 3) + Tween-20 permeabilization + mild protease >digestion >- For peroxidase-based protocol, biotin block (tried also peroxidase >block, doesn't help) >- Serum block >- Rabbit primary antibody (overnight +4C) >- Sheep secondary antibody (biotinylated or fluorescent) >- For biotinylated secondary, tyramide amplification & DAB reaction, >hematoxylin counterstain >- For fluorescent secondary, Sudan Black B for autofluorescence suppression >- Embedding with Faramount > >-- >//////////////////////////////////////////////////////////// > >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 > >//////////////////////////////////////////////////////////// > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >--------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sbreeden <@t> nmda.nmsu.edu Tue Nov 8 09:23:32 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Tue Nov 8 09:23:42 2005 Subject: [Histonet] HT/Path Ratio Message-ID: For your survey: I'm the only HT and I have 3 pathologists. Workload this year will be about 12,500 blocks. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From pmarcum <@t> vet.upenn.edu Tue Nov 8 09:26:39 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Tue Nov 8 09:26:56 2005 Subject: [Histonet] Reagent Grade Alcohol In-Reply-To: <20051108145923.84304.qmail@web61212.mail.yahoo.com> References: <0FDBF29200C637468CCC1F9E7E85A3D25E2214@MAIL-LR.lha.org> <20051108145923.84304.qmail@web61212.mail.yahoo.com> Message-ID: <6.1.1.1.2.20051108101656.019b99f8@mail.vet.upenn.edu> At 09:59 AM 11/8/2005, Rene J Buesa wrote: >I always used 100% (= "200 grade") ,bought in bulk tax exempted, ethanol. >Anything you need to do with "pure" ethanol will be simpler to calculate >(solutions, mixtures, etc.) with 100% ethanol. Besides that, the >"denatured" ethanol contains substances other than ethanol. >Now, "reagent grade" sold by chemical companies with "impurities >certificates", are much more expensive and offer no advantage over the >type of "200 grade" ethanol I used to use. >Rene J. > >"LeVier, Rebecca J" wrote: >Hello Histonetters, > >I was just wondering what everyone was using for alcohol. Does anyone >see any differences in Reagent Grade Alcohol compared to LCB regulated >alcohol? ...... Any information on this topic would be greatly >appreciated. > >Thanks in advance. > > >Confidentiality Notice: This e-mail message, including any >attachments, is for the sole use of intended recipient(s) and may >contain confidential and privileged information. Any unauthorized >review, use, disclosure or distribution is prohibited. If you are not >the intended recipient, please contact the sender by reply e-mail and >destroy all copies of the original message. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >--------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Sorry those comments are misleading. The 200 proof and 190 proof alcohols are taxed at $13.50 per proof (100 proof = 1 proof taxable) so a gallon of 200 proof ethanol is given a tax of $27.00 before you even get the price of the actual product. Only tax exempt or non-profit organizations can afford to use these routinely as we do not pay this tax. Reagent alcohols can be purchased that work very well however, you must read the MSDS as the original formula of 90% ethanol, 5% methanol and 5% isopropanol is now being sold with varying percentages of isopropanol and methanol. These formula changes are legal so check the MSDS and if it is not a stable formula do not use it as it can cause many issues in your lab. Example: 90% EtOH with 1 to 5% isopropanol and 1 to 5% methanol. You would not know from lot to lot how much of the last two you have in the solution and it can effect your staining and processing when these are varied. If you go to the internet and type in Reagent Alcohol you will be able to see a number of MSDS sheets for various companies and gauge which have stable formulas and which do not. ALWAYS LOOK AT THE MSDS FOR ANY PRODUCT YOU PURCHASE TO ASSURE YOU ARE ALWAYS GETTING THE SAME PRODUCT FORMULA. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From naje1972 <@t> yahoo.com Tue Nov 8 09:32:10 2005 From: naje1972 <@t> yahoo.com (cynthia haynes) Date: Tue Nov 8 09:32:17 2005 Subject: [Histonet] Thank you Message-ID: <20051108153210.97531.qmail@web33005.mail.mud.yahoo.com> Good Morning fellower Histonetter.I would like to thank those who responded to my post for help with brain/20 micron sections. All the advise really helped me to get good sections. Thank you again. Cynthia Haynes H.T. From J.Kane <@t> wriwindber.org Tue Nov 8 09:43:42 2005 From: J.Kane <@t> wriwindber.org (Jennifer Kane) Date: Tue Nov 8 09:45:13 2005 Subject: [Histonet] CryoJane Message-ID: <7690673B79E70D429A120579193392231D0B2B@wri-xchng.WRIWINDBER.ORG> Hi, Does anyone have experience with the CryoJane tape transfer system from Intrumedics? Unfortunately we can not demo the product and would like to hear some feedback good or bad. Thank you, Jennifer From gentras <@t> vetmed.auburn.edu Tue Nov 8 09:48:01 2005 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Tue Nov 8 09:48:17 2005 Subject: [Histonet] HT/Path ratio Message-ID: <6.0.1.1.0.20051108094408.01ad6f40@mailhost.vetmed.auburn.edu> 1 histotech / 1 pathologist; workload varies since we're primarily research and our lab is involved with other disciplines of medical research in addition to histology as required. Thanks, ASG From anh2006 <@t> med.cornell.edu Tue Nov 8 09:56:19 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Tue Nov 8 09:51:23 2005 Subject: [Histonet] Bone marrow unspecific staining In-Reply-To: <43705CF3.4010806@helsinki.fi> References: <9badb0bbc8ae.436f4426@med.cornell.edu> <43705CF3.4010806@helsinki.fi> Message-ID: Bone is FAMOUS for being tricky to work with so everything must be done with patience :) Here are my new batch of thoughts based on your problems: - Do you have a picture of the background??? Is it specific or diffuse? - Is your rabbit primary an antiserum or a affini-pure antibody? - Is your sheep anti-rabbit cross adsorbed against cow tissue? - What species was your anti-rabbit secondary made in? - Bone marrow is filled with cells of hematopoietic origin which have a lot of endogenous peroxidases as well as Fc receptors which may bind to your primary antibody (or secondary for that matter). This is why isotype controls and a serum block is so necessary. So, what serum have you been using for blocking? - Why are you using tyramide? Is this necessary for your antigen? - Have you titrated the primary at different dilutions and at different times for incubation? I doubt that the PFA is the problem as I routinely work with PFA fixed murine BM samples and don't have any issues but how long are you fixing for? You do the fixation *after* EDTA decal correct? >Andrea and Rene, thanks for your comments. I'm >sorry for being a bit unspecific with my >unspecificity question :) > >Some more details on the problem: > >- Seems to be caused by unspecific antibody >binding, but not a specific property of this >primary ab >- Primary antibody works very well with other tissues >- Also other primaries (and secondary alone) show unspecific staining with BM >- Not caused by endogenous biotin, peroxidase, >or autofluorescence (done enough controls) > >And some more details on the protocol: > >- Bovine BM tissue samples decalcified with >EDTA, or cell samples embedded in agar >- Paraformaldehyde fixation >- HIER (glycine-HCl pH 3) + Tween-20 >permeabilization + mild protease digestion >- For peroxidase-based protocol, biotin block >(tried also peroxidase block, doesn't help) >- Serum block >- Rabbit primary antibody (overnight +4C) >- Sheep secondary antibody (biotinylated or fluorescent) >- For biotinylated secondary, tyramide >amplification & DAB reaction, hematoxylin >counterstain >- For fluorescent secondary, Sudan Black B for autofluorescence suppression >- Embedding with Faramount > >-- >//////////////////////////////////////////////////////////// > > 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 > >//////////////////////////////////////////////////////////// > -- From ldibernardo <@t> synecor.com Tue Nov 8 10:11:28 2005 From: ldibernardo <@t> synecor.com (Louis DiBernardo) Date: Tue Nov 8 10:11:43 2005 Subject: [Histonet] MMA and stents (yup again) Message-ID: working through the process for embedding and cutting coronary stents using MMA. would welcome any advice or protocols that are not closely guarded! any helpful hints on your infiltration and embedding, anti-bubble techniques and cutting /fold reduction would be much appreciated! Thanks From ree3 <@t> leicester.ac.uk Tue Nov 8 10:32:45 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Tue Nov 8 10:32:57 2005 Subject: [Histonet] : beta-2- microglobulin Message-ID: Looking for an antibody to the above that works, as ever, on paraffin processed mouse tissues... Many thanks Richard Edwards MRC TOX UNIT LEICESTER...U.K.... _ From TJJ <@t> Stowers-Institute.org Tue Nov 8 10:46:58 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Tue Nov 8 10:47:36 2005 Subject: [Histonet] Re: Sheep anti BrdU Message-ID: Yun, Do you currently have an anti-BrdU made in mouse that is working for you? And are you using chromogenic or fluorescent techniques? If you are using chromogenic, consider biotinylating one of your mouse monoclonals to do the double staining. Do a sequential immunostain technique using your 1st mouse antibody, anti-mouse secondary, DAB (or other stable) chromogen, then do the biotinylated antibody, ABC/streptavidin or avidin biotin/LSAB Alk Phos label, and chromogen of choice after that. If you are using fluorescent techniques, it might be best to use the sheep anti-BrdU in a cocktail and do your double staining that way. Another thing to consider when doing BrdU doublestaining is what effect the denaturing step will have on the antigenicity of your other proteins of interest. We had some challenges trying to do BrdU doublestaining because of this. Most often when working up antibodies, you don't usually include this step. Therefore trying to just plug in an antibody that has previously worked ok in the single staining techniques may or may not work well when combined with the protocols needed for BrdU staining. Good luck, and let us know how the sheep antibody works if you decide to go that route. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From mclarke <@t> allsaintshealthcare.org Tue Nov 8 11:05:52 2005 From: mclarke <@t> allsaintshealthcare.org (Clarke, Mary) Date: Tue Nov 8 11:06:04 2005 Subject: [Histonet] Histologist ratio Message-ID: <6FE2A453DA437F4D96FAAF363F20ABB2FBBC94@WFEXBE04.wfsi.priv> We have 4 pathologists and 4 histotechs, one who is registry eligible and 3 who are registered. Terri Clarke Histology Supervisor All Saints Laboratory 3801 Spring Street Racine, Wi 53405 mclarke@allsaintshealthcare.org 262-687-6609 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, November 08, 2005 9:06 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 24, Issue 10 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: measuring collagen (Boyce, Amanda (NIH/NIAMS)) 2. H. Pylori Controls (Vicki Gauch) 3. Re: H. Pylori Controls (Rene J Buesa) 4. histologist/pathologist ratio (LINDA MARGRAF) 5. RE: H. Pylori Controls (Jeff Gordon) 6. Sheep anti BrdU (Yun Li) 7. Re: histologist/pathologist ratio (Rene J Buesa) 8. Sihler's Stain (Dianne Holmes) 9. (no subject) (mplhisto@aol.com) 10. Re: Sihler's Stain (Rene J Buesa) 11. Inflammation (John PJ Coleman) 12. Histo Online Program (Linresearch@aol.com) 13. Pleaes unsuscribe (Bernardo Vargas Angel) 14. re: grinding bone sections (caron fournier) 15. cytospin or cytek machine (Elizabeth Chlipala) 16. Re: Sihler's Stain (John Kiernan) 17. Re: re: grinding bone sections (louise renton) 18. Re: Bone marrow unspecific staining (Mikael Niku) 19. Thermal polymerisation of LR gold resin for electron microscopy (Peter Bannister) 20. Red marker (Steven Coakley) 21. Mounting Gelatine Embedded Cryostat CNS Sections (germckeon@excite.com) 22. RE: Red marker (Ingles Claire) 23. Reagent Grade Alcohol (LeVier, Rebecca J) 24. Re: Bone marrow unspecific staining (Rene J Buesa) 25. RE: histologist/pathologist ratio (Horn, Hazel V) 26. Re: Reagent Grade Alcohol (Rene J Buesa) 27. Thank you (Vicki Gauch) 28. RE: histologist/pathologist ratio (Rene J Buesa) ---------------------------------------------------------------------- Message: 1 Date: Mon, 7 Nov 2005 13:18:26 -0500 From: "Boyce, Amanda \(NIH/NIAMS\)" Subject: [Histonet] RE: measuring collagen To: Message-ID: <0E3E7E8F6E23DF4C8127A063568356B510B98802@nihexchange12.nih.gov> Content-Type: text/plain; charset="iso-8859-1" Our lab uses a hydroxyproline assay. I've never used it, so I can't troubleshoot it for you, but here's the protocol: Acid hydrolysis * Papain digest and 12N HCL 1:1 * 110 overnight in sealed glass ampules * dry sample 24-48 h in vacuum-pump with NaOH * resuspend in 1 ml assay buffer, vortex, overnight at 4, vortex again * store at -20 or assay Hydroxyproline assay Reagents: * stock buffer: 50 g citric acid monohydrate, 12 ml glacial acetic acid , 120 g sodium acetate trihydrate, 34 g sodium hydroxide ad 1 liter, pH 6.0, store at 4 * assay buffer: 1:10 dilution of stock buffer * chloramin-T-reagent: 0.3525g Chloramine T dissolved in 5.175 ml water, add 6.5 ml m-propanol and 13.325 ml stock buffer, make fresh * Dimethylaminobenzaldehyde reagent: 3.75 g dimethylaminobenzaldehyde in 15 ml n-propanol, add 6.5 ml perchloric acid, make fresh * Standard: 1 mg/ml hydroyxproline, store -20 Method: 1. standard dilutions: 0, 0 , 0.2, 0.4, 0.6, 0.8, 1, 2, 4, 6, 8 ,10 ?g/ml 2. sample dilutions: 1:5, 1:10, 1:20, 1:40, 1:80,1: 100 3. 200 ?l sample/standard in eppendorf 4. add 100 ?l Chloramine T, pipett mix 20 min room temp 5. add 100 ?l dimethylaminobenzaldehyde, pipet mix, 15 min 60 water bath 6. 300 ?l in 96 well plate and read immediately 540 nm -------------------------- Amanda Taylor Boyce, Ph.D. Postdoctoral IRTA Fellow Cartilage Biology and Orthopaedics Branch National Institute of Arthritis and Musculoskeletal and Skin Diseases National Institutes of Health Bldg. 13, Rm. 3W17 Bethesda, MD 20892-5755 Phone: 301-451-6860 Fax: 301-480-4315 Email: boycea@mail.nih.gov > Message: 6 > Date: Mon, 7 Nov 2005 07:00:09 -0800 (PST) > From: Niloufar Fozouni > Subject: [Histonet] measuring collagen > To: histonet@lists.utsouthwestern.edu > Message-ID: <20051107150019.64420.qmail@web35602.mail.mud.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > Hello, > > Does anybody know, how I can measure collagen content > in cartilage? > Thank you. > Niloo > > ------------------------------ Message: 2 Date: Mon, 07 Nov 2005 13:24:25 -0500 From: "Vicki Gauch" Subject: [Histonet] H. Pylori Controls To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Does anyone know of a good source for H. Pylori Controls?? We normally order from Newcomer Supply but they are experiencing a large backorder right now and we need something to hold us over until they arrive. Any help would be greatly appreciated... Thanks so much, Vicki Gauch AMCH Albany, NY ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. ------------------------------ Message: 3 Date: Mon, 7 Nov 2005 11:30:08 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] H. Pylori Controls To: Vicki Gauch , histonet@lists.utsouthwestern.edu Message-ID: <20051107193008.74020.qmail@web61212.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 We used positive cases that were selected by the pathologists, but there is another approarch. Usually pathologists want to be able to have a positive control of exactly what they are looking for; if this is the case with your pathologists you will need controls comntaining H. pylori. Now, the whole idea with the control is to make sure that the procedure worked, and this you can obtain with any tissue containing abundant number of bacteria. After a long "convincing" process with our pathologists I was able to make them aware of this fact. We used the Steiner method (with phosphotungstic acid instead of the radiactive uranyl nitrate) and for control we used appendix (and you will not find a more easy to find tissue with more bacteria than an appendix). Perhaps you would like to try to talk about this approach with your pathologist. Just a thought! Rene J. Vicki Gauch wrote: Does anyone know of a good source for H. Pylori Controls?? We normally order from Newcomer Supply but they are experiencing a large backorder right now and we need something to hold us over until they arrive. Any help would be greatly appreciated... Thanks so much, Vicki Gauch AMCH Albany, NY ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 4 Date: Mon, 07 Nov 2005 13:49:23 -0600 From: "LINDA MARGRAF" Subject: [Histonet] histologist/pathologist ratio To: Message-ID: Content-Type: text/plain; charset=US-ASCII Here's a message Beatrice is having trouble posting to the list. Please email her or send your reply to the entire list (and not me directly). Thanks Linda M (Histonet administrator) "DeBrosse_Beatrice" Hi! I would like to know what the ratio of how many histologists to a Pathologist is. Does the ratio differ from hospitals to reference labs to pharmaceutical labs? Your help is greatly appreciated. Sincerely, Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 ------------------------------ Message: 5 Date: Mon, 7 Nov 2005 14:16:27 -0600 From: "Jeff Gordon" Subject: RE: [Histonet] H. Pylori Controls To: "Vicki Gauch" , Message-ID: Content-Type: text/plain; charset="utf-8" Cell Marque offers H. pylori positive controls for IHC, as well as controls for every other antibody that we carry. Jeff Gordon Cell Marque Corp. http://www.cellmarque.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Vicki Gauch Sent: Mon 11/7/2005 12:24 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] H. Pylori Controls Does anyone know of a good source for H. Pylori Controls?? We normally order from Newcomer Supply but they are experiencing a large backorder right now and we need something to hold us over until they arrive. Any help would be greatly appreciated... Thanks so much, Vicki Gauch AMCH Albany, NY ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 07 Nov 2005 14:21:17 -0600 From: Yun Li Subject: [Histonet] Sheep anti BrdU To: histonet@lists.utsouthwestern.edu Message-ID: <436FB73D.3090606@utsouthwestern.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Greetings everyone, I am trying to do some double immuno-staining for BrdU and other antigens. Since the antibodies for my "other" antigens are mostly from mouse, I am looking for a non-mouse (and preferably non-rat) antibody against BrdU. I came across this sheep anti brdU antibody in the literature, it seems that several different companies offer it, including maine biotech, abcam, novus and one called genetex. So my question is has anyone here used it before, what type of sections was used, how did it work and which company was it purchased from? Thanks a lot! yun ------------------------------ Message: 7 Date: Mon, 7 Nov 2005 12:43:36 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] histologist/pathologist ratio To: LINDA MARGRAF , Histonet@lists.utsouthwestern.edu Message-ID: <20051107204336.58622.qmail@web61216.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Beatrice: Some months ago I reviewed different sources (US Dept Labor; US Dept. Commerce;ASCP) and got to the figure of about 1.4 to 2.0 histotechs per pathologist. This, as any other average, is "not written in stone" and has to vary between laboratories. Of the laboratories I personally know the averages have been from 1 to 1, to 2.2 to 1 These figures will also depend on the type of laboratory: laboratories in small hospitals will probably will be closer to 1:1, in large reference laboratories, although will have many more pathologists, the huge sample volume will require the larger side of the scale (2 or 2:2 histotechs / pathologist). Rene J. LINDA MARGRAF wrote: Here's a message Beatrice is having trouble posting to the list. Please email her or send your reply to the entire list (and not me directly). Thanks Linda M (Histonet administrator) "DeBrosse_Beatrice" Hi! I would like to know what the ratio of how many histologists to a Pathologist is. Does the ratio differ from hospitals to reference labs to pharmaceutical labs? Your help is greatly appreciated. Sincerely, Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 8 Date: Mon, 07 Nov 2005 15:34:24 -0600 From: "Dianne Holmes" Subject: [Histonet] Sihler's Stain To: Message-ID: Content-Type: text/plain; charset=US-ASCII Is anyone familiar with this process to counterstain nerve fibers within a relatively transparent muscle? I need a materials and method list. Can anyone help me? ------------------------------ Message: 9 Date: Mon, 07 Nov 2005 16:56:31 -0500 From: mplhisto@aol.com Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Message-ID: <8C7B1FD3F626F95-B64-AA33@FWM-R21.sysops.aol.com> Content-Type: text/plain; charset="us-ascii" Can anyone tell me the name of a good computer safety training program ? we are looking to update our current program ? Thanks Meredith Hale HT (ASCP) Histology Supervisor Pathology Partners Irving, Texas ------------------------------ Message: 10 Date: Mon, 7 Nov 2005 14:03:50 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Sihler's Stain To: Dianne Holmes , Histonet@lists.utsouthwestern.edu Message-ID: <20051107220350.18981.qmail@web61225.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Dianne: Sihler's method was published by Bohm and Oppel in 1907 and is used in pieces of fesh muscle. The solutions are: A- water 75 mL + glycerol 12 mL+ acetic acid 12 mL + chloral hydrate 0.75 g B- water 75 ml + Ehrlich hematoxylin (1886 formula) 12 mL + glycerol 12 mL + chloral hydrate 0.75 g C- 0.1% acetic acid in glycerol. Method: pieces of fresh muscle in Sol.A for 18 hours; transfer to Sol.B for 3 to 10 days; transfer to Sol.C untill differentiated, mount in glycerol, seal coverslip with wax. Hope this will help! Rene J. Dianne Holmes wrote: Is anyone familiar with this process to counterstain nerve fibers within a relatively transparent muscle? I need a materials and method list. Can anyone help me? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 11 Date: Mon, 7 Nov 2005 17:23:10 -0500 From: John PJ Coleman Subject: [Histonet] Inflammation To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII; format=flowed For inflammation (?) I personally would start trying to label the leucocytes. What about CD15 for Polys, CD117 for mast cells, and CD30 or CD68 for Histiocytes? On Nov 7, 2005, at 1:01 PM, histonet-request@lists.utsouthwestern.edu wrote: Dear All, I wish to use IHC for the detection of chronic inflammation in human tissues - specifically within the prostate. I am using formalin fixed sections of prostate. The list of possible markers for chronic inflammation out there is rather overwhelming. I was wondering if anyone had a small panel of tried and tested antibodies that would cover most of the chronic inflammatory cells or give a good representative picture of chronic inflammation within the tissue. Or even suggest a few antibodies with which I could start with. Many thanks for attention, Steve Leung Clinical Research Fellow University of Edinburgh U.K. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 24, Issue 9 *************************************** JPJC-757 335-2159 http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ ------------------------------ Message: 12 Date: Mon, 7 Nov 2005 17:24:57 EST From: Linresearch@aol.com Subject: [Histonet] Histo Online Program To: histonet@pathology.swmed.edu Message-ID: <252.72dde0.30a12e39@aol.com> Content-Type: text/plain; charset="US-ASCII" Hello, Does anyone know of a current online program for Histology? Lin ------------------------------ Message: 13 Date: Mon, 7 Nov 2005 18:17:21 -0500 From: Bernardo Vargas Angel Subject: [Histonet] Pleaes unsuscribe To: Histonet@lists.utsouthwestern.edu Message-ID: <1131405441.436fe08192c3b@mail2.nova.edu> Content-Type: text/plain Please unsuscribe Bernardo Vargas Angel Ph.D ------------------------------ Message: 14 Date: Mon, 7 Nov 2005 18:54:26 -0500 (EST) From: caron fournier Subject: [Histonet] re: grinding bone sections To: histonet@lists.utsouthwestern.edu Message-ID: <20051107235426.70464.qmail@web36313.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi All: what is the finest paper that you have used on an exakt system for grinding bone samples (undecalcified). We were told that the 2500 was the highest they had but there is literature that notes 4000. Caron Fournier, BSc, R.T. Department of Orthopaedics, Division of Orthopaedic Engineering Research, U.B.C. --------------------------------- Find your next car at Yahoo! Canada Autos ------------------------------ Message: 15 Date: Mon, 7 Nov 2005 17:36:36 -0700 From: "Elizabeth Chlipala" Subject: [Histonet] cytospin or cytek machine To: "'Histonet'" Message-ID: <000001c5e3fc$79e12870$a7d48a80@AMY> Content-Type: text/plain; charset="us-ascii" Hello Histonetters I have a unique request. I have a client that is in Emeryville California and they need access to a cytospin or a Thin Prep machine, they are willing to pay for time, etc. If there is anyone out there that is willing to do this can they e-mail me back with contact information so I can forward it to them. Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 ------------------------------ Message: 16 Date: Tue, 08 Nov 2005 00:15:03 -0500 From: John Kiernan Subject: Re: [Histonet] Sihler's Stain To: Rene J Buesa Cc: Histonet@lists.utsouthwestern.edu Message-ID: <43703457.F66DB4E3@uwo.ca> Content-Type: text/plain; charset=us-ascii Before going ahead with Sihler's haemalum method (for peripheral nerves in cleared whole mounts) consult recent publications: Wu BL, Sanders I (1994) The Sihler stain: A technique for studying peripheral nerves in whole-mount specimens. Neuroscience Protocols Module 4: 83-93. Wu B, Saunders I (1994) The Sihler stain: a technique for studying peripheral nerves in whole-mount specimens. In: FG Wouterlood, Neuroscience Protocols. Elsevier, Amsterdam. pp. 94-050-07-01 to 94-050-07-10. Mu LC, Sanders I (1998) Neuromuscular organization of the human upper esophageal sphincter. Annals of Otology Rhinology and Laryngology 107: 370-377. Mu LC, Sanders I (1999) Neuromuscular organization of the canine tongue. Anatomical Record 256: 412-424. Gozil R, Kadioglu D, Calguner E, Erdogan D, Bahcelioglu M, Elmas C (2002) Branching patterns of rabbit oculomotor and trochlear nerves demonstrated by Sihler's stain technique. Biotechnic & Histochemistry 77: 21-25. This is an important method for examining the "gross" anatomy of small nerves. Rene: do you have a copy of Bohm & Appel 1907 or of any of Sihler's papers? If so, I'd be really grateful for bibliographic details! John Kiernan Anatomy, UWO London, Canada. ______________________________________________________________ Rene J Buesa wrote: > > Hi Dianne: > Sihler's method was published by Bohm and Oppel in 1907 and is used in > pieces of fesh muscle. The solutions are: > A- water 75 mL + glycerol 12 mL+ acetic acid 12 mL + chloral hydrate 0.75 g > B- water 75 ml + Ehrlich hematoxylin (1886 formula) 12 mL + glycerol 12 mL + chloral hydrate 0.75 g > C- 0.1% acetic acid in glycerol. > Method: pieces of fresh muscle in Sol.A for 18 hours; transfer to Sol.B for 3 to 10 days; transfer to Sol.C untill differentiated, mount in glycerol, seal coverslip with wax. > Hope this will help! > Rene J. > > Dianne Holmes wrote: > Is anyone familiar with this process to counterstain nerve fibers > within a relatively transparent muscle? I need a materials and method > list. Can anyone help me? > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > --------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Tue, 8 Nov 2005 08:55:14 +0200 From: louise renton Subject: Re: [Histonet] re: grinding bone sections To: caron fournier , Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Yep, we GRIND on 2500, but POLISH on 4000. This help?? On 11/8/05, caron fournier wrote: > Hi All: > what is the finest paper that you have used on an exakt system for > grinding bone samples (undecalcified). We were told that the 2500 was > the highest they had but there is literature that notes 4000. > > > Caron Fournier, BSc, R.T. > Department of Orthopaedics, > Division of Orthopaedic Engineering Research, > U.B.C. > > > > --------------------------------- > Find your next car at Yahoo! Canada Autos > _______________________________________________ > 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 "....onwards through the fog!" ------------------------------ Message: 18 Date: Tue, 08 Nov 2005 10:08:19 +0200 From: Mikael Niku Subject: Re: [Histonet] Bone marrow unspecific staining To: Histonet@lists.utsouthwestern.edu Message-ID: <43705CF3.4010806@helsinki.fi> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Andrea and Rene, thanks for your comments. I'm sorry for being a bit unspecific with my unspecificity question :) Some more details on the problem: - Seems to be caused by unspecific antibody binding, but not a specific property of this primary ab - Primary antibody works very well with other tissues - Also other primaries (and secondary alone) show unspecific staining with BM - Not caused by endogenous biotin, peroxidase, or autofluorescence (done enough controls) And some more details on the protocol: - Bovine BM tissue samples decalcified with EDTA, or cell samples embedded in agar - Paraformaldehyde fixation - HIER (glycine-HCl pH 3) + Tween-20 permeabilization + mild protease digestion - For peroxidase-based protocol, biotin block (tried also peroxidase block, doesn't help) - Serum block - Rabbit primary antibody (overnight +4C) - Sheep secondary antibody (biotinylated or fluorescent) - For biotinylated secondary, tyramide amplification & DAB reaction, hematoxylin counterstain - For fluorescent secondary, Sudan Black B for autofluorescence suppression - Embedding with Faramount -- //////////////////////////////////////////////////////////// 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 //////////////////////////////////////////////////////////// ------------------------------ Message: 19 Date: Tue, 08 Nov 2005 12:07:52 +0000 From: "Peter Bannister" Subject: [Histonet] Thermal polymerisation of LR gold resin for electron microscopy To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hello Histonetters, Does anyone have a method for the thermal polymerisation of LR gold resin for electron microscopy? I am trying to produce blocks of cultured cell pellets fixed in para/glut, post fixed in Osmium tetroxide, dehydrated in ethanol, infiltrated with the resin and cured. I have tried polymerisation using light with mixed results - non osmium treated blocks polymerise okay and are just about suitable for sectioning. However, osmium treated blocks will only polymerise down as far as the cell pellet (I understand that the dark colour absorbs light and therefore interferes with the polymerisation). I have tried to keep exposure to oxygen during polymerisation to a minimum by using closed BEEM capsules or snap fit gelatin capsules. I would therefore like to try the more classical thermal polymerisation at +60?C. The problem is that when I add the initiator (benzoyl peroxide) to the pure resin I end up with a solid mass of resin even before the peroxide is fully dissolved, at room temperature -so i can't use it on my samples. If anyone can help me out with a protocol that will ensure the resin only polymerises quickly when at heated to +60?C it would be very much appreciated. Many thanks for your time, Peter Bannister. Dr. Peter Bannister, Thrombosis Research Institute, Histopathology, Emmanuel Kaye Building, Manresa Road, London SW3 6LR. UK. _________________________________________________________________ Be the first to hear what's new at MSN - sign up to our free newsletters! http://www.msn.co.uk/newsletters ------------------------------ Message: 20 Date: Tue, 8 Nov 2005 05:47:03 -0800 (PST) From: Steven Coakley Subject: [Histonet] Red marker To: Histonet@lists.utsouthwestern.edu Message-ID: <20051108134703.44353.qmail@web90207.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Does anyone know of a company that makes permanant red markers that resist organic solvents? Thanks, Steve --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 21 Date: Tue, 8 Nov 2005 08:51:01 -0500 (EST) From: "germckeon@excite.com" Subject: [Histonet] Mounting Gelatine Embedded Cryostat CNS Sections To: histonet@lists.utsouthwestern.edu Message-ID: <20051108135101.79C1B3E01@xprdmailfe6.nwk.excite.com> Content-Type: text/plain; charset="us-ascii" Hello, I am currently working on spinal cord and am having trouble with sections floating off. I have embedded with gelatine, am performing cryostat sectioning and will be doing some of the more structural stains eg Kultschitsky's stain for myelin. I have been using Superfrost slides. Can anyone advise me regarding choices of slides, coatings, methodologies, etc which would maximise adherence? In gratitude for any advice received, Gerald McKeon _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! ------------------------------ Message: 22 Date: Tue, 8 Nov 2005 08:00:50 -0600 From: "Ingles Claire" Subject: RE: [Histonet] Red marker To: "Steven Coakley" , Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE6D@uwhis-xchng2.hosp.wisc.edu> Content-Type: text/plain; charset="utf-8" I believe I just saw some new ones in a Market lab catalogue yesterday. 1-800-237-3604 or www.marketlabinc.com. :) Claire Ingles UW Mohs Clinic Lab Madison Wisconsin -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Steven Coakley Sent: Tue 11/8/2005 7:47 AM To: Histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Red marker Does anyone know of a company that makes permanant red markers that resist organic solvents? Thanks, Steve --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 23 Date: Tue, 8 Nov 2005 09:38:41 -0500 From: "LeVier, Rebecca J" Subject: [Histonet] Reagent Grade Alcohol To: Message-ID: <0FDBF29200C637468CCC1F9E7E85A3D25E2214@MAIL-LR.lha.org> Content-Type: text/plain; charset="us-ascii" Hello Histonetters, I was just wondering what everyone was using for alcohol. Does anyone see any differences in Reagent Grade Alcohol compared to LCB regulated alcohol? ...... Any information on this topic would be greatly appreciated. Thanks in advance. Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 24 Date: Tue, 8 Nov 2005 06:50:28 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Bone marrow unspecific staining To: Mikael Niku , Histonet@lists.utsouthwestern.edu Message-ID: <20051108145028.81939.qmail@web61218.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Although I cannot be absolutely sure, for me it seems that perhaps the problem may reside in the paraformaldehyde fixation. Do you have to use it? Could you use another fixative in another parallel sample just to try to find out? Just a thoght! Rene J. Mikael Niku wrote: Andrea and Rene, thanks for your comments. I'm sorry for being a bit unspecific with my unspecificity question :) Some more details on the problem: - Seems to be caused by unspecific antibody binding, but not a specific property of this primary ab - Primary antibody works very well with other tissues - Also other primaries (and secondary alone) show unspecific staining with BM - Not caused by endogenous biotin, peroxidase, or autofluorescence (done enough controls) And some more details on the protocol: - Bovine BM tissue samples decalcified with EDTA, or cell samples embedded in agar - Paraformaldehyde fixation - HIER (glycine-HCl pH 3) + Tween-20 permeabilization + mild protease digestion - For peroxidase-based protocol, biotin block (tried also peroxidase block, doesn't help) - Serum block - Rabbit primary antibody (overnight +4C) - Sheep secondary antibody (biotinylated or fluorescent) - For biotinylated secondary, tyramide amplification & DAB reaction, hematoxylin counterstain - For fluorescent secondary, Sudan Black B for autofluorescence suppression - Embedding with Faramount -- //////////////////////////////////////////////////////////// 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 //////////////////////////////////////////////////////////// _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 25 Date: Tue, 8 Nov 2005 08:56:38 -0600 From: "Horn, Hazel V" Subject: RE: [Histonet] histologist/pathologist ratio To: "Rene J Buesa" , "LINDA MARGRAF" , Histonet@lists.utsouthwestern.edu Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDE9A@EMAIL.archildrens.org> Content-Type: text/plain; charset=us-ascii We have 5 anatomic pathologists and 3 histotechs. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Monday, November 07, 2005 2:44 PM To: LINDA MARGRAF; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] histologist/pathologist ratio Hi Beatrice: Some months ago I reviewed different sources (US Dept Labor; US Dept. Commerce;ASCP) and got to the figure of about 1.4 to 2.0 histotechs per pathologist. This, as any other average, is "not written in stone" and has to vary between laboratories. Of the laboratories I personally know the averages have been from 1 to 1, to 2.2 to 1 These figures will also depend on the type of laboratory: laboratories in small hospitals will probably will be closer to 1:1, in large reference laboratories, although will have many more pathologists, the huge sample volume will require the larger side of the scale (2 or 2:2 histotechs / pathologist). Rene J. LINDA MARGRAF wrote: Here's a message Beatrice is having trouble posting to the list. Please email her or send your reply to the entire list (and not me directly). Thanks Linda M (Histonet administrator) "DeBrosse_Beatrice" Hi! I would like to know what the ratio of how many histologists to a Pathologist is. Does the ratio differ from hospitals to reference labs to pharmaceutical labs? Your help is greatly appreciated. Sincerely, Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ 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. ============================================================================== ------------------------------ Message: 26 Date: Tue, 8 Nov 2005 06:59:22 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Reagent Grade Alcohol To: "LeVier, Rebecca J" , Histonet@lists.utsouthwestern.edu Message-ID: <20051108145923.84304.qmail@web61212.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I always used 100% (= "200 grade") ,bought in bulk tax exempted, ethanol. Anything you need to do with "pure" ethanol will be simpler to calculate (solutions, mixtures, etc.) with 100% ethanol. Besides that, the "denatured" ethanol contains substances other than ethanol. Now, "reagent grade" sold by chemical companies with "impurities certificates", are much more expensive and offer no advantage over the type of "200 grade" ethanol I used to use. Rene J. "LeVier, Rebecca J" wrote: Hello Histonetters, I was just wondering what everyone was using for alcohol. Does anyone see any differences in Reagent Grade Alcohol compared to LCB regulated alcohol? ...... Any information on this topic would be greatly appreciated. Thanks in advance. Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 27 Date: Tue, 08 Nov 2005 10:00:22 -0500 From: "Vicki Gauch" Subject: [Histonet] Thank you To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Thank you to everyone who responded to my query for H. Pylori Controls...we are looking into them now....You really are GREAT !!! Vicki AMCH Albany ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. ------------------------------ Message: 28 Date: Tue, 8 Nov 2005 07:02:52 -0800 (PST) From: Rene J Buesa Subject: RE: [Histonet] histologist/pathologist ratio To: "Horn, Hazel V" , LINDA MARGRAF , Histonet@lists.utsouthwestern.edu Message-ID: <20051108150252.25624.qmail@web61217.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Question to all subscribers: why don't you post the histologists/pathologists ratios you have in each of your own laboratories? It will be like a survey from Histonet that for sure will usefull for all. Yesterday I posted the ratois I was aware of; now is your turns! Rene J. "Horn, Hazel V" wrote: We have 5 anatomic pathologists and 3 histotechs. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Monday, November 07, 2005 2:44 PM To: LINDA MARGRAF; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] histologist/pathologist ratio Hi Beatrice: Some months ago I reviewed different sources (US Dept Labor; US Dept. Commerce;ASCP) and got to the figure of about 1.4 to 2.0 histotechs per pathologist. This, as any other average, is "not written in stone" and has to vary between laboratories. Of the laboratories I personally know the averages have been from 1 to 1, to 2.2 to 1 These figures will also depend on the type of laboratory: laboratories in small hospitals will probably will be closer to 1:1, in large reference laboratories, although will have many more pathologists, the huge sample volume will require the larger side of the scale (2 or 2:2 histotechs / pathologist). Rene J. LINDA MARGRAF wrote: Here's a message Beatrice is having trouble posting to the list. Please email her or send your reply to the entire list (and not me directly). Thanks Linda M (Histonet administrator) "DeBrosse_Beatrice" Hi! I would like to know what the ratio of how many histologists to a Pathologist is. Does the ratio differ from hospitals to reference labs to pharmaceutical labs? Your help is greatly appreciated. Sincerely, Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ 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. 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From rjr6 <@t> psu.edu Tue Nov 8 11:29:55 2005 From: rjr6 <@t> psu.edu (Roberta Horner) Date: Tue Nov 8 11:30:04 2005 Subject: [Histonet] Histologist ratio Message-ID: We have 1 Histologist and 2 Avian pathologists and 3 mammalian pathologists. Roberta Horner HT/HTL Penn State University Animal Diagnostic Lab From TJJ <@t> Stowers-Institute.org Tue Nov 8 11:34:32 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Tue Nov 8 11:35:02 2005 Subject: [Histonet] Re: Bone marrow unspecific staining Message-ID: Thanks for the additional information. Only you know what you've tried and haven't, so I'll give you some things to consider: ~you indicate the antibodies are working well in "other tissues." And you are currently using "Bovine BM tissue samples decalcified with EDTA, or cell samples embedded in agar". You may have to use a different pretreatment protocol to get these same antibodies to work in decalcified bone and whole cell preparations. It could be that the pretreatments you are using in these samples require a different concentration of primary or secondary antibody. ~Try pH 6 or pH 8 HIER solutions with and without enzyme and see what happens. ~Any time I see non-specific nuclear staining, I immediately suspect the retrieval method combined with antibody concentration. Just because you haven't seen this in other tissues, doesn't mean you won't see it due to differences in specimen types (cells v. tissues) and handling (decal) prior to staining. Do you see any specific staining at all in either? If so, take the dilutions of the primary and secondary antibodies down to reduce nonspecific background. If you're not seeing any specific signal at all, I wouldn't bother trying to abolish background staining, especially if the answer lies in further antibody dilution. ~If you are seeing this effect only in the tyramide amplified samples, again check the concentration of your antibodies. It could be you need to dilute them out further. Or try using a streptavidin conjugated enzyme or fluorophore and see if you get signal with less background and go from there. ~You may never get some antibodies to stain well in decalcified aldehyde-fixed bone marrow. We had this issue with some markers and ended up using non-fixed, non-decalcified frozen sections using the Cryo-Jane tape transfer system. This takes time and practice to use on these difficult samples, but it may be another option. ~if you've got cells to stain, why not just fix and stain the cell cultures rather than paraffin processing them? Some antibodies work better in ICC protocols (permeabilization only and no digestion or retrieval) than for paraffin-embedded IHC protocols. Good luck! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From jcline <@t> wchsys.org Tue Nov 8 11:34:47 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Tue Nov 8 11:35:03 2005 Subject: [Histonet] Path/Tech ratio Message-ID: <001201c5e48a$b8d713d0$1d2a14ac@wchsys.org> We have four pathologists and if fully staffed I have 5 histotechs including myself. Our caseload per year is around 19,000. ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From mbryhan <@t> hotmail.com Tue Nov 8 12:42:22 2005 From: mbryhan <@t> hotmail.com (Mary Bryhan (Quandt)) Date: Tue Nov 8 12:42:34 2005 Subject: [Histonet] histologist/pathologist ratio Message-ID: Does anyone know if there are any regulations regarding this issue? Could it be used in either Workman's compenstation or malpractice litigation? For example, histology labs that are run with insufficient staffing, overworked, and stressed staff making mistakes which lead or contribute to litigation. Mary Bryhan HT (ASCP) >From: "LINDA MARGRAF" >To: >Subject: [Histonet] histologist/pathologist ratio >Date: Mon, 07 Nov 2005 13:49:23 -0600 > >Here's a message Beatrice is having trouble posting to the list. >Please email her or send your reply to the entire list (and not me >directly). > Thanks Linda M (Histonet administrator) > >"DeBrosse_Beatrice" >Hi! > > I would like to know what the ratio of how many histologists to a >Pathologist is. Does the ratio differ from hospitals to reference labs >to pharmaceutical labs? Your help is greatly appreciated. > > Sincerely, > >Beatrice DeBrosse-Serra >HT(ASCP)QIHC >Allergan, Inc. >2525 Dupont Drive RD-2A >Irvine, CA 92612 >714-246-5116 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dahmed <@t> mdanderson.org Tue Nov 8 13:09:48 2005 From: dahmed <@t> mdanderson.org (dahmed@mdanderson.org) Date: Tue Nov 8 13:09:58 2005 Subject: [Histonet] Job Opportunities- M.D. Anderson Cancer Center, Houston, Texas Message-ID: M.D. Anderson Cancer Center in Houston, Texas has two full-time histotech positions, one in frozen section and one in the main histology lab. We also have one part-time position available. If anyone is interested, please contact Dianna Menard in Human Resources at 713-745-6184. Thank you. David S. Ahmed, HT(ASCP) Supervisor, Clinical Histology Laboratory Department of Pathology M.D. Anderson Cancer Center From shive003 <@t> umn.edu Tue Nov 8 13:21:48 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Tue Nov 8 13:21:58 2005 Subject: Fw: [Histonet] histologist/pathologist ratio Message-ID: <00c501c5e499$aaae0e30$41065486@auxs.umn.edu> We have 12 pathologists, 3 histotechs, and 3 IHC techs. Jan Shivers U of MN Vet Diag Lab ----- Original Message ----- From: "LINDA MARGRAF" To: Sent: Monday, November 07, 2005 1:49 PM Subject: [Histonet] histologist/pathologist ratio Here's a message Beatrice is having trouble posting to the list. Please email her or send your reply to the entire list (and not me directly). Thanks Linda M (Histonet administrator) "DeBrosse_Beatrice" Hi! I would like to know what the ratio of how many histologists to a Pathologist is. Does the ratio differ from hospitals to reference labs to pharmaceutical labs? Your help is greatly appreciated. Sincerely, Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jennifer.l.hofecker <@t> Vanderbilt.Edu Tue Nov 8 13:50:03 2005 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Tue Nov 8 13:50:11 2005 Subject: [Histonet] Tennessee Society Meeting...Call for Abstracts Message-ID: <898D946569A27444B65667A49C074052075510@mailbe06.mc.vanderbilt.edu> Hi everyone, We are now accepting abstracts from anybody interested in presenting at our annual meeting. The meeting is later next year because the region III meeting is scheduled for March. Our meeting will be June 1-3rd in Chattanooga. If you would be interested in presenting, please contact me. We are interested in 3hr workshops as well as shorter lectures. Thank you for your interest and please let me know if you need more information. Jennifer Hofecker TSH President Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax From dmarsha3 <@t> utmem.edu Tue Nov 8 14:37:32 2005 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Tue Nov 8 14:43:24 2005 Subject: [Histonet] crumbly femurs Message-ID: <008a01c5e4a5$0b478360$f623c084@DanaM> Hi everyone, I am trying to section some mouse legs. The bones were fixed/decalcified for one week then quick frozen in OCT and put at -20C where they have been for about 2 months. Regardless of how thick I cut the section (5-20 microns), whether I cut free hand, use our cryojane or the old fashioned scotch tape type of tape transfer, there is little/no visible bone in the section. Just OCT and an empty space where bone would be. If I cut without tape I can see the bone crumble as I cut. This seems to be a catastrophic failure of some sort and may be beyond any type of help at this point but if anyone has any ideas as to why this is happening I would be happy to hear from you. Thanks, Dana From rjbuesa <@t> yahoo.com Tue Nov 8 14:49:17 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 8 14:49:25 2005 Subject: [Histonet] histologist/pathologist ratio In-Reply-To: Message-ID: <20051108204917.30891.qmail@web61221.mail.yahoo.com> No, there is no regulation that relates to the ratio histotechs/pathologists. That is always an internal laboratory issue. Rene J. "Mary Bryhan (Quandt)" wrote: Does anyone know if there are any regulations regarding this issue? Could it be used in either Workman's compenstation or malpractice litigation? For example, histology labs that are run with insufficient staffing, overworked, and stressed staff making mistakes which lead or contribute to litigation. Mary Bryhan HT (ASCP) >From: "LINDA MARGRAF" >To: >Subject: [Histonet] histologist/pathologist ratio >Date: Mon, 07 Nov 2005 13:49:23 -0600 > >Here's a message Beatrice is having trouble posting to the list. >Please email her or send your reply to the entire list (and not me >directly). > Thanks Linda M (Histonet administrator) > >"DeBrosse_Beatrice" >Hi! > > I would like to know what the ratio of how many histologists to a >Pathologist is. Does the ratio differ from hospitals to reference labs >to pharmaceutical labs? Your help is greatly appreciated. > > Sincerely, > >Beatrice DeBrosse-Serra >HT(ASCP)QIHC >Allergan, Inc. >2525 Dupont Drive RD-2A >Irvine, CA 92612 >714-246-5116 > > > >_______________________________________________ >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 --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From portera <@t> msu.edu Tue Nov 8 14:51:05 2005 From: portera <@t> msu.edu (Amy Porter) Date: Tue Nov 8 14:52:02 2005 Subject: [Histonet] crumbly femurs References: <008a01c5e4a5$0b478360$f623c084@DanaM> Message-ID: <002d01c5e4a6$23531a40$8e7a0923@HistoJJ> Potentially lack of rinsing after decal....we always rinse our decal specimens in running tap water before proceeding with any other steps. Maybe you needed to rinse and then cryoprotect with sucrose prior to flash freezing. Just a thought. ----- Original Message ----- From: "Dana Marshall" To: Sent: Tuesday, November 08, 2005 3:37 PM Subject: [Histonet] crumbly femurs Hi everyone, I am trying to section some mouse legs. The bones were fixed/decalcified for one week then quick frozen in OCT and put at -20C where they have been for about 2 months. Regardless of how thick I cut the section (5-20 microns), whether I cut free hand, use our cryojane or the old fashioned scotch tape type of tape transfer, there is little/no visible bone in the section. Just OCT and an empty space where bone would be. If I cut without tape I can see the bone crumble as I cut. This seems to be a catastrophic failure of some sort and may be beyond any type of help at this point but if anyone has any ideas as to why this is happening I would be happy to hear from you. Thanks, Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Tue Nov 8 14:55:09 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Tue Nov 8 14:55:19 2005 Subject: [Histonet] histologist/pathologist ratio Message-ID: <110820052055.12934.437110AC000DDDFA000032862200763692CECE030E9D0C9A03@comcast.net> Mary and Rene, Your point is extemely taken and understood however, I have never heard of any regulations for histology regarding number of techs. It is frightening when one thinks about it. In my years it was always handled as an internal matter. Hopefully if there is a hard and fast rule some one will give the reference for us. Pam Marcum > No, there is no regulation that relates to the ratio histotechs/pathologists. > That is always an internal laboratory issue. > Rene J. > > "Mary Bryhan (Quandt)" wrote: > Does anyone know if there are any regulations regarding this issue? Could > it be used in either Workman's compenstation or malpractice litigation? For > example, histology labs that are run with insufficient staffing, overworked, > and stressed staff making mistakes which lead or contribute to litigation. > > Mary Bryhan HT (ASCP) > > > >From: "LINDA MARGRAF" > > >To: > >Subject: [Histonet] histologist/pathologist ratio > >Date: Mon, 07 Nov 2005 13:49:23 -0600 > > > >Here's a message Beatrice is having trouble posting to the list. > >Please email her or send your reply to the entire list (and not me > >directly). > > Thanks Linda M (Histonet administrator) > > > >"DeBrosse_Beatrice" > >Hi! > > > > I would like to know what the ratio of how many histologists to a > >Pathologist is. Does the ratio differ from hospitals to reference labs > >to pharmaceutical labs? Your help is greatly appreciated. > > > > Sincerely, > > > >Beatrice DeBrosse-Serra > >HT(ASCP)QIHC > >Allergan, Inc. > >2525 Dupont Drive RD-2A > >Irvine, CA 92612 > >714-246-5116 > > > > > > > >_______________________________________________ > >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 > > > > > --------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Tue Nov 8 14:57:17 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 8 14:57:26 2005 Subject: [Histonet] histologist/pathologist ratio In-Reply-To: <110820052055.12934.437110AC000DDDFA000032862200763692CECE030E9D0C9A03@comcast.net> Message-ID: <20051108205717.63976.qmail@web61212.mail.yahoo.com> Today as well. How many histotechs are in a laboratory should (and ususally is) dictated by the work load not by the number of pathologists being served by the laboratory. It is just interesting to find out how are things around. Rene J. Pam Marcum wrote: Mary and Rene, Your point is extemely taken and understood however, I have never heard of any regulations for histology regarding number of techs. It is frightening when one thinks about it. In my years it was always handled as an internal matter. Hopefully if there is a hard and fast rule some one will give the reference for us. Pam Marcum > No, there is no regulation that relates to the ratio histotechs/pathologists. > That is always an internal laboratory issue. > Rene J. > > "Mary Bryhan (Quandt)" wrote: > Does anyone know if there are any regulations regarding this issue? Could > it be used in either Workman's compenstation or malpractice litigation? For > example, histology labs that are run with insufficient staffing, overworked, > and stressed staff making mistakes which lead or contribute to litigation. > > Mary Bryhan HT (ASCP) > > > >From: "LINDA MARGRAF" > > >To: > >Subject: [Histonet] histologist/pathologist ratio > >Date: Mon, 07 Nov 2005 13:49:23 -0600 > > > >Here's a message Beatrice is having trouble posting to the list. > >Please email her or send your reply to the entire list (and not me > >directly). > > Thanks Linda M (Histonet administrator) > > > >"DeBrosse_Beatrice" > >Hi! > > > > I would like to know what the ratio of how many histologists to a > >Pathologist is. Does the ratio differ from hospitals to reference labs > >to pharmaceutical labs? Your help is greatly appreciated. > > > > Sincerely, > > > >Beatrice DeBrosse-Serra > >HT(ASCP)QIHC > >Allergan, Inc. > >2525 Dupont Drive RD-2A > >Irvine, CA 92612 > >714-246-5116 > > > > > > > >_______________________________________________ > >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 > > > > > --------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From anh2006 <@t> med.cornell.edu Tue Nov 8 15:20:35 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Tue Nov 8 15:15:39 2005 Subject: [Histonet] Santa Cruz anti-CD31 Message-ID: Has there been any advancements in knowledge on whether Santa Cruz has produced a CD31 from the new goat or whatnot that works in IHC? I know there was some trouble as the animal was no longer producing batches that were good for IHC and someone mentioned that they were going to come out with something new/more reliable late this year? Anyone know what I am talking about? Thanks, Andrea -- From dusko.trajkovic <@t> pfizer.com Tue Nov 8 15:35:10 2005 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Tue Nov 8 15:35:32 2005 Subject: [Histonet] crumbly femurs Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B201E90C5A@lajamrexm01.amer.pfizer.com> I had a similar experience a few months ago, and it was corrected by minimally cutting into the surface of the bone or in your case, the entire side of the leg and placing it in 30% sucrose at 4C for at least 24 hours, then transfer the legs to OCT and let sit for 24 hours at 4C, before re-freezing in OCT. Since I was doing an Oil Red O on the sections, and morphology was not an issue, I melted the blocks down, trimmed the bone as I mentioned above and placed it in sucrose overnight and then in OCT at 4C. After re-freezing in OCT, bones were sectioning beautifully, and my ORO worked as well. Thanks Dusko Trajkovic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dana Marshall Sent: Tuesday, November 08, 2005 12:38 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] crumbly femurs Hi everyone, I am trying to section some mouse legs. The bones were fixed/decalcified for one week then quick frozen in OCT and put at -20C where they have been for about 2 months. Regardless of how thick I cut the section (5-20 microns), whether I cut free hand, use our cryojane or the old fashioned scotch tape type of tape transfer, there is little/no visible bone in the section. Just OCT and an empty space where bone would be. If I cut without tape I can see the bone crumble as I cut. This seems to be a catastrophic failure of some sort and may be beyond any type of help at this point but if anyone has any ideas as to why this is happening I would be happy to hear from you. Thanks, Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Tue Nov 8 16:05:15 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Tue Nov 8 16:00:23 2005 Subject: [Histonet] REPOST: Type I and Type II pneumocytes Message-ID: Any takers on my question below? I have received no responses yet :( >Date: Wed, 26 Oct 2005 23:13:00 -0500 >To: Histonet >From: "Andrea T. Hooper" >Subject: Type I and Type II pneumocytes >Cc: >Bcc: >X-Attachments: > >I am looking for markers against Type I and Type II pneumocytes in >human and mouse tissues for IHC. Any suggestions? Markers, clones, >catalog #s, protocols - all are welcome - even from vendors. > >Thanks, >Andrea -- From tgoodpas <@t> fhcrc.org Tue Nov 8 16:30:13 2005 From: tgoodpas <@t> fhcrc.org (Goodpaster, Tracy A) Date: Tue Nov 8 16:30:22 2005 Subject: [Histonet] CryoJane Message-ID: <8BD501B158A381409FE54DB421F82FCA012BB3D3@groucho.fhcrc.org> We have the CryoJane tape transfer system and we don't use this system for all of our routine frozen slides because of the cost, we just use it for specimens that are difficult. It has allowed us to get good sections when we otherwise have difficulty, for example for mouse femurs. The morphology is very good. Tracy Goodpaster ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jennifer Kane Sent: Tue 11/8/2005 7:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CryoJane Hi, Does anyone have experience with the CryoJane tape transfer system from Intrumedics? Unfortunately we can not demo the product and would like to hear some feedback good or bad. Thank you, Jennifer _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From d.gianotti <@t> comcast.net Tue Nov 8 17:25:05 2005 From: d.gianotti <@t> comcast.net (Dennis Gianotti) Date: Tue Nov 8 17:25:27 2005 Subject: [Histonet] fatty breast processing Message-ID: <437133D1.3010004@comcast.net> We have a sakura VIP processor and need to process breast tissue that comes in between 4 & 5 pm and have it out the next day. What would be a good protocal to use on the processor using formalin, zylene and alcohols. The end time could be as late as 10 am the next day. thanks From CrochiereSteve <@t> aol.com Tue Nov 8 20:35:11 2005 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Tue Nov 8 20:35:25 2005 Subject: [Histonet] HT/Path ratio Message-ID: <256.9312bd.30a2ba5f@aol.com> I have 4 techs (counting myself) and 6 pathologists. Our volume is near 20,000 cases per year. Some days we cut 800 slides while others only 100 or so. Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 From tahseen <@t> brain.net.pk Tue Nov 8 15:06:09 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Tue Nov 8 23:09:14 2005 Subject: [Histonet] histologist/pathologist ratio References: Message-ID: <003201c5e4a8$3f4ee1a0$972bfea9@m7c0y4> We are six Pathologist and ten histologists including supervisor.Our workload is 20,000case/year, 300blocks/day,45slidesIHC/day and 15sp stains/day. Muhammad Tahseen Histology Supervisor SKMCH & RC Lahore Pakistan. ----- Original Message ----- From: Mary Bryhan (Quandt) To: ; Sent: Wednesday, November 09, 2005 7:42 AM Subject: RE: [Histonet] histologist/pathologist ratio > Does anyone know if there are any regulations regarding this issue? Could > it be used in either Workman's compenstation or malpractice litigation? For > example, histology labs that are run with insufficient staffing, overworked, > and stressed staff making mistakes which lead or contribute to litigation. > > Mary Bryhan HT (ASCP) > > > >From: "LINDA MARGRAF" > >To: > >Subject: [Histonet] histologist/pathologist ratio > >Date: Mon, 07 Nov 2005 13:49:23 -0600 > > > >Here's a message Beatrice is having trouble posting to the list. > >Please email her or send your reply to the entire list (and not me > >directly). > > Thanks Linda M (Histonet administrator) > > > >"DeBrosse_Beatrice" > >Hi! > > > > I would like to know what the ratio of how many histologists to a > >Pathologist is. Does the ratio differ from hospitals to reference labs > >to pharmaceutical labs? Your help is greatly appreciated. > > > > Sincerely, > > > >Beatrice DeBrosse-Serra > >HT(ASCP)QIHC > >Allergan, Inc. > >2525 Dupont Drive RD-2A > >Irvine, CA 92612 > >714-246-5116 > > > > > > > >_______________________________________________ > >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 vince86 <@t> hotmail.com Wed Nov 9 00:32:36 2005 From: vince86 <@t> hotmail.com (Steven Van Cleave) Date: Wed Nov 9 00:32:44 2005 Subject: [Histonet] Obtaining PA exam requirements Message-ID: I am currently in search of an institution or hospital willing to accept a an intern student for a short period of time (~2wks). Compensation to this institution and/or trainer would be without question. I am in need of obtaining the post-mordem requirements for the pathologist assistant route 2 ASCP qualification for examination. I have all requirements except for 10 autopsies, 2 of which must include brain. My name, or initials would have to be on report and I would have to be involved in FAD, clinical summary and the like. If anyone has any suggestions that may be helpful, I have open ears. I would really like to get this cert. before it's too late and would have to go back to school somewhere up north and have to give up my job as lab supervisor. I have HT and HTL certification and have been in Histology since 1996. I graduated from Abilene Christian University with a degree in Biology in 2002. I am 25. My Pathologists have trained me exhaustively in gross examination and many other areas, but we only do a couple of autopsies per year and I must apply for examination by Dec of 2007. We are an independent lab. I am in Abilene, TX at Clinical Pathology Associates. I really just don't feel comfortable stopping at my BS, when this cert. is out there now. Thanks for your time. -Vince Van Cleave, HT, BS, HTL(ASCP) P.S. If anyone is interested and would like some info, I am doing an in-house study with the CAMVIR-1 monoclonal antibody against major capsid protein L1 which has been said to react strongly against HPV type 16 and 33. I am testing it with ThinPreps and testing it on more high risk types. So far, it looks like the cross reactivity with squamous cells would make this a very technically and professionally difficult test for routine screening of HPV. Hopefully our Paths will want to give some antibodies against MCMs (minichromosomal maintenance proteins) and Topoisomerase II-alpha which are overexpressed in abnormal cervical cells. Anyone else have any neat findings with either of these???? It looks like TriPath has a neat ab cocktail like this out now??? -Vince Van Cleave From BMolinari <@t> heart.thi.tmc.edu Wed Nov 9 06:01:16 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Wed Nov 9 06:04:00 2005 Subject: [Histonet] MMA and stents (yup again) Message-ID: Hi Louis, I float my sections on a DH2O bath set at 25C. I do not have any additives in the water. I cut at 2-3 microns on a tungsten carbide D profile knife. I pick the sections off the knife with forceps or a small brush, being careful to pick up a small edge of the section. If the section curls I gently blow on it (of course only works if forceps is being used :). I hold the section about 6-8 inches above the water bath and let it float down into the bath. Yes, sometimes it folds but more often than not the section lands flat. I do not have a problem with bubbles. As far as processing and embedding, we have had to develop our own protocol due to size, amount of tissue attached and humidity. I am still working on a VVG stain, if you or anyone out there has a protocol they would like to share..... Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Louis DiBernardo Sent: Tuesday, November 08, 2005 10:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] MMA and stents (yup again) working through the process for embedding and cutting coronary stents using MMA. would welcome any advice or protocols that are not closely guarded! any helpful hints on your infiltration and embedding, anti-bubble techniques and cutting /fold reduction would be much appreciated! Thanks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From petepath <@t> yahoo.com Wed Nov 9 06:29:28 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Wed Nov 9 06:29:37 2005 Subject: [Histonet] fatty breast processing Message-ID: <20051109122929.23707.qmail@web30410.mail.mud.yahoo.com> I'll leave the protocols to the histologists. Better make sure your pathologists and PA's are cutting there sections less than 3 mm thick across the full section. No pillows with 3 mm thick edges and 4mm thick centers or your pathologists are going to be wondering where the middle of their sections went no matter what protocol you are using. From Barbara_Lentz <@t> dahlchase.com Wed Nov 9 06:59:55 2005 From: Barbara_Lentz <@t> dahlchase.com (Barbara Lentz) Date: Wed Nov 9 07:03:31 2005 Subject: [Histonet] HT/Path ratio Message-ID: We have 13 pathologists, 15 techs with a workload about 47,000 cases per year. As an independent lab, we also do Flow Cytometry and Cytology. Barb From angela.mcnabola.b <@t> bayer.com Wed Nov 9 07:09:41 2005 From: angela.mcnabola.b <@t> bayer.com (Angela McNabola) Date: Wed Nov 9 07:09:31 2005 Subject: [Histonet] Santa Cruz anti-CD31 In-Reply-To: Message-ID: Hi, I do know what your talking about...I think I might have been the one who originally posted the first note. I placed a call to the person at SC who I had been discussing this with, just this past week. They were out of the office until I believe yesterday, so the plan is to give him a call today. We have several lots of the CD31that is are not working in IHC. The new goat's antibody, I have been told, will work in westerns, but not in FFPE tissues. I hope that they get it corrected quickly, it has become, for us, one of our routine IHC's in mouse xenografts. I'll post a note if I hear anything further. Angela Angela McNabola MS, HT(ASCP)SLS, QIHC Scientist Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 203-812-5001 fax# 203-812-5820 angela.mcnabola.b@bayer.com "Andrea T. Hooper" To: Histonet Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Santa Cruz anti-CD31 western.edu 11/08/2005 04:20 PM Has there been any advancements in knowledge on whether Santa Cruz has produced a CD31 from the new goat or whatnot that works in IHC? I know there was some trouble as the animal was no longer producing batches that were good for IHC and someone mentioned that they were going to come out with something new/more reliable late this year? Anyone know what I am talking about? Thanks, Andrea -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Nov 9 07:33:45 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 9 07:33:53 2005 Subject: [Histonet] fatty breast processing In-Reply-To: <437133D1.3010004@comcast.net> Message-ID: <20051109133345.90958.qmail@web61211.mail.yahoo.com> Dennis: First of all, more important than the actual protocol is the fixation of the breast tissue, and in order to get good fixation, you have to start with THIN slices of breast, 3 mm thick OR LESS if at all possible. After this thickness factor is obtained you will need a good fixation time. Lets say that you get the tissues between 4&5 pm; the should be described preferentially, meaning, immediately and put in cassettes into the NBF at 5 or 6 PM. If you have a VIP and your end time (I gather that you mean the end time for the processing protocol) is at 10AM, you could start processing at midnight which will mean that the tissues will be in the retort in formalin from 5 or 6 PM until midnight and IF the sections are THIN enough, they will be fixed by then. Start your ptotocol in the first alcohol station, with the following sequence: 60%EthOlX50 min; 80%EthOlX50 min; 95%EthOlX50 min; 100%EthOlX1 hour;100%EthOlX45 min; 100%EthOlX45 in; XyleneX45 min; XyleneX45 min; Paraf.1X45 min; Paraf2 X50 min; Paraf 3X 55 min and Paraf 4 X1hour This protocol will give 10 hours (without secondary fixation since it will skip station 2 and the tissues will be in the retort with the formalin from station 1). This protocol will give you a (clearing+infiltration)/dehydration ratio of 0.97 which is a good ratio to avoid excessive dryness. Really we stop having problems with breast (and any other difficult tissue) when we abandoned xylene and start using mineral oil. Hope this will help you! Rene J. Dennis Gianotti wrote: We have a sakura VIP processor and need to process breast tissue that comes in between 4 & 5 pm and have it out the next day. What would be a good protocal to use on the processor using formalin, zylene and alcohols. The end time could be as late as 10 am the next day. thanks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From tyler-wellington <@t> northwestern.edu Wed Nov 9 07:40:50 2005 From: tyler-wellington <@t> northwestern.edu (Tyler W.) Date: Wed Nov 9 07:40:58 2005 Subject: [Histonet] Mouse Ovary Processing Message-ID: <20051109134050.882C99E845@merle.it.northwestern.edu> Does anyone have a good and reliable processing protocol for 6 month old mouse ovaries? Curently the processing program is set up at 45 minutes per station through 70%, 80%, 3 stations of 95%, 3 stations of 100%, 2 stations xylene and 3 stations of paraffin (surgipath). Does this sound right? Any modifications or helpful suggestions? Thank you so much for everything, as always. This list is fantastic. From rjbuesa <@t> yahoo.com Wed Nov 9 07:53:40 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 9 07:53:49 2005 Subject: [Histonet] Obtaining PA exam requirements In-Reply-To: Message-ID: <20051109135340.11171.qmail@web61223.mail.yahoo.com> Try the Ohio State University, they run a very reputable and recognized PA program. Rene J. Steven Van Cleave wrote: I am currently in search of an institution or hospital willing to accept a an intern student for a short period of time (~2wks). Compensation to this institution and/or trainer would be without question. I am in need of obtaining the post-mordem requirements for the pathologist assistant route 2 ASCP qualification for examination. I have all requirements except for 10 autopsies, 2 of which must include brain. My name, or initials would have to be on report and I would have to be involved in FAD, clinical summary and the like. If anyone has any suggestions that may be helpful, I have open ears. I would really like to get this cert. before it's too late and would have to go back to school somewhere up north and have to give up my job as lab supervisor. I have HT and HTL certification and have been in Histology since 1996. I graduated from Abilene Christian University with a degree in Biology in 2002. I am 25. My Pathologists have trained me exhaustively in gross examination and many other areas, but we only do a couple of autopsies per year and I must apply for examination by Dec of 2007. We are an independent lab. I am in Abilene, TX at Clinical Pathology Associates. I really just don't feel comfortable stopping at my BS, when this cert. is out there now. Thanks for your time. -Vince Van Cleave, HT, BS, HTL(ASCP) P.S. If anyone is interested and would like some info, I am doing an in-house study with the CAMVIR-1 monoclonal antibody against major capsid protein L1 which has been said to react strongly against HPV type 16 and 33. I am testing it with ThinPreps and testing it on more high risk types. So far, it looks like the cross reactivity with squamous cells would make this a very technically and professionally difficult test for routine screening of HPV. Hopefully our Paths will want to give some antibodies against MCMs (minichromosomal maintenance proteins) and Topoisomerase II-alpha which are overexpressed in abnormal cervical cells. Anyone else have any neat findings with either of these???? It looks like TriPath has a neat ab cocktail like this out now??? -Vince Van Cleave _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From gcallis <@t> montana.edu Wed Nov 9 09:44:14 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 9 09:44:26 2005 Subject: [Histonet] crumbly femurs In-Reply-To: <008a01c5e4a5$0b478360$f623c084@DanaM> References: <008a01c5e4a5$0b478360$f623c084@DanaM> Message-ID: <6.0.0.22.1.20051109083338.01b7c530@gemini.msu.montana.edu> Dana, You did not say that you cryoprotected the fixed/decalcified mouse bones in 30% sucrose at 4C for a few days. If not, you don't get very good frozen sections. Then you should section at -26C with Cryojane, even with decalcified/cryoprotected. I'm a bit surprised that Cryojane is not helping you. It also helps to use a high profile blade or even use a c profile tungsten carbide for doing decalcified free hand bone FS, much sturdier knives/blades for denser bone. Did you do endpoint determination during decalcification? The crumbly bone may be in indication your bones are NOT completely decalcified - a disaster for frozen sections. If you are doing frozen sections with Cryojane in the first place, why do you need to have bones fixed and decalcified? I suggest you try undecalcified, snap frozen FRESH bones instead but make sure the tungsten carbide knife d profile is really sharp. Some use c profile, we prefer d profile. If you did EDTA decalcification, they bones sometimes take more than a week to be totally calcium free. At 01:37 PM 11/8/2005, you wrote: >Hi everyone, >I am trying to section some mouse legs. The bones were fixed/decalcified >for one week then quick frozen in OCT and put at -20C where they have been >for about 2 months. Regardless of how thick I cut the section (5-20 >microns), whether I cut free hand, use our cryojane or the old fashioned >scotch tape type of tape transfer, there is little/no visible bone in the >section. Just OCT and an empty space where bone would be. If I cut >without tape I can see the bone crumble as I cut. This seems to be a >catastrophic failure of some sort and may be beyond any type of help at >this point but if anyone has any ideas as to why this is happening I would >be happy to hear from you. >Thanks, >Dana >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From ploykasek <@t> phenopath.com Wed Nov 9 09:55:10 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Wed Nov 9 09:55:36 2005 Subject: [Histonet] Tech to doc ratio Message-ID: We have 7 full time IHC techs, 1 full time & 2 part-time histologists, 1 lab aide, 3 full time pathologists, and 1 fellow. We run 300-400 immuno slides a day, and 10-20+ FISH cases. We are a reference lab. The last hospital based 'routine' histo lab that I worked in had 1 lab aide, 5 full time techs, 1 IHC tech and 6- 7 pathologists. We did about 250 blocks a day. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From livieira <@t> ualg.pt Wed Nov 9 10:01:12 2005 From: livieira <@t> ualg.pt (Lina Vieira) Date: Wed Nov 9 10:04:48 2005 Subject: [Histonet] bone fish samples embedded in Historesin Plus Message-ID: <006f01c5e546$ce935280$2914100a@labhistologia> Can anyone offer some advice on cutting bone fish samples embedded in Historesin Plus using rigid disposable microtome blades? We are using the same blades for cut samples embedded in Historesin without problems. But now we have bone fish sample without decalcification embedded in Historesin Plus and we can?t get good sections. Tanks in advance Lina Vieira Universidade do Algarve From abright <@t> brightinstruments.com Wed Nov 9 10:15:47 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Wed Nov 9 10:08:57 2005 Subject: [Histonet] crumbly femurs Message-ID: Dear Dana, Looks to me that your specimen has freeze dried due to being stored @ -20?C for 2 months. I agree that you should have no problems sectioning fresh frozen undecalcified mouse legs with a tungsten carbide tipped knife, but do not see the need for using any tape removal system, just a good cryostat and anti roll plate sectioning @ -30?C with a slow cutting speed. 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: Gayle Callis [mailto:gcallis@montana.edu] Sent: 09 November 2005 15:44 To: Dana Marshall; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] crumbly femurs Dana, You did not say that you cryoprotected the fixed/decalcified mouse bones in 30% sucrose at 4C for a few days. If not, you don't get very good frozen sections. Then you should section at -26C with Cryojane, even with decalcified/cryoprotected. I'm a bit surprised that Cryojane is not helping you. It also helps to use a high profile blade or even use a c profile tungsten carbide for doing decalcified free hand bone FS, much sturdier knives/blades for denser bone. Did you do endpoint determination during decalcification? The crumbly bone may be in indication your bones are NOT completely decalcified - a disaster for frozen sections. If you are doing frozen sections with Cryojane in the first place, why do you need to have bones fixed and decalcified? I suggest you try undecalcified, snap frozen FRESH bones instead but make sure the tungsten carbide knife d profile is really sharp. Some use c profile, we prefer d profile. If you did EDTA decalcification, they bones sometimes take more than a week to be totally calcium free. At 01:37 PM 11/8/2005, you wrote: >Hi everyone, >I am trying to section some mouse legs. The bones were >fixed/decalcified >for one week then quick frozen in OCT and put at -20C where they have been >for about 2 months. Regardless of how thick I cut the section (5-20 >microns), whether I cut free hand, use our cryojane or the old fashioned >scotch tape type of tape transfer, there is little/no visible bone in the >section. Just OCT and an empty space where bone would be. If I cut >without tape I can see the bone crumble as I cut. This seems to be a >catastrophic failure of some sort and may be beyond any type of help at >this point but if anyone has any ideas as to why this is happening I would >be happy to hear from you. >Thanks, >Dana >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From petepath <@t> yahoo.com Wed Nov 9 10:23:40 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Wed Nov 9 10:23:50 2005 Subject: [Histonet] Tech to doc ratio Message-ID: <20051109162340.90620.qmail@web30410.mail.mud.yahoo.com> This tech to doc ratio is not going to yield very practical info because of the great variability of practice and specimen types. All of these entrys could actually yield some valid data if we look at blocks per histotechs and not include immunotechs. The immunotechs will vary depending on the volumes. We cut about 500 blocks /day with 10 techs and a section head.. We do about 80 immunos/ day and have a dedicated immunotech. We have seven surgical pathologists reading it. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From japoteete <@t> saintfrancis.com Wed Nov 9 10:35:56 2005 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Wed Nov 9 10:37:11 2005 Subject: [Histonet] Tech to doc ratio Message-ID: A note from Patti's last hospital. We now have 9 pathologists, 4 full time and 1 part time histologists, 3 lab aides, do 300+ blocks per day, and 1 IHC Tech who does 40-80 slides per day. I think they are thinking about adding another pathologist, plus CISH testing. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Lab Saint Francis Hospital, Tulsa, OK -----Original Message----- From: Patti Loykasek [mailto:ploykasek@phenopath.com] Sent: Wednesday, November 09, 2005 9:55 AM To: histonet Subject: [Histonet] Tech to doc ratio We have 7 full time IHC techs, 1 full time & 2 part-time histologists, 1 lab aide, 3 full time pathologists, and 1 fellow. We run 300-400 immuno slides a day, and 10-20+ FISH cases. We are a reference lab. The last hospital based 'routine' histo lab that I worked in had 1 lab aide, 5 full time techs, 1 IHC tech and 6- 7 pathologists. We did about 250 blocks a day. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From livieira <@t> ualg.pt Wed Nov 9 11:28:41 2005 From: livieira <@t> ualg.pt (Lina Vieira) Date: Wed Nov 9 11:32:17 2005 Subject: [Histonet] bone fish samples embedded in Historesin Plus References: <006f01c5e546$ce935280$2914100a@labhistologia> <6.0.0.22.1.20051109095751.01b920d8@gemini.msu.montana.edu> Message-ID: <007a01c5e553$072504c0$2914100a@labhistologia> We are using samples without decalcification because we are studing the bone fish development. Historesin is glycol metacrylate based (manufactured by Leica). ----- Original Message ----- From: "Gayle Callis" To: "Lina Vieira" Sent: Wednesday, November 09, 2005 4:58 PM Subject: Re: [Histonet] bone fish samples embedded in Historesin Plus What is Historesin Plus, a plastic? Which one? At 09:01 AM 11/9/2005, you wrote: >Can anyone offer some advice on cutting bone fish samples embedded in >Historesin Plus using rigid disposable microtome blades? > >We are using the same blades for cut samples embedded in Historesin >without problems. > >But now we have bone fish sample without decalcification embedded in >Historesin Plus and we can?t get good sections. > >Tanks in advance > >Lina Vieira >Universidade do Algarve >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dmarsha3 <@t> utmem.edu Wed Nov 9 11:43:13 2005 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Wed Nov 9 11:43:22 2005 Subject: [Histonet] crumbly femurs References: <3AD0BD3142459B4E9B12CBEAFF2B89B201E90C5A@lajamrexm01.amer.pfizer.com> Message-ID: <000f01c5e555$0f2e7190$f623c084@DanaM> Thanks to those of you have have made suggestions re: my crumbly femurs. I will try to salvage those that i have using these suggestions and be a bit more proactive in the future. Since we do have a cryojane we don't need to do the decal so i'll try to get the experiments planned so that people are doing that first. I am happy to hear any experiences with bone so please continue to write if you have thoughts on crumbly femurs or good ones! Thanks, Dana ----- Original Message ----- From: "Trajkovic, Dusko" To: "Dana Marshall" ; Sent: Tuesday, November 08, 2005 3:35 PM Subject: RE: [Histonet] crumbly femurs I had a similar experience a few months ago, and it was corrected by minimally cutting into the surface of the bone or in your case, the entire side of the leg and placing it in 30% sucrose at 4C for at least 24 hours, then transfer the legs to OCT and let sit for 24 hours at 4C, before re-freezing in OCT. Since I was doing an Oil Red O on the sections, and morphology was not an issue, I melted the blocks down, trimmed the bone as I mentioned above and placed it in sucrose overnight and then in OCT at 4C. After re-freezing in OCT, bones were sectioning beautifully, and my ORO worked as well. Thanks Dusko Trajkovic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dana Marshall Sent: Tuesday, November 08, 2005 12:38 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] crumbly femurs Hi everyone, I am trying to section some mouse legs. The bones were fixed/decalcified for one week then quick frozen in OCT and put at -20C where they have been for about 2 months. Regardless of how thick I cut the section (5-20 microns), whether I cut free hand, use our cryojane or the old fashioned scotch tape type of tape transfer, there is little/no visible bone in the section. Just OCT and an empty space where bone would be. If I cut without tape I can see the bone crumble as I cut. This seems to be a catastrophic failure of some sort and may be beyond any type of help at this point but if anyone has any ideas as to why this is happening I would be happy to hear from you. Thanks, Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From maria <@t> ski.org Wed Nov 9 11:53:00 2005 From: maria <@t> ski.org (Maria Mejia) Date: Wed Nov 9 11:53:17 2005 Subject: [Histonet] Sihler's Stain References: Message-ID: <4372377C.7050001@ski.org> Hello Dianne, I have done this stain in the past (and) if interested in the protocol I will send it to your email address. Just let me know. Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco CA, 94103 Email: maria@ski.org Phone: (415)-345-2185 Dianne Holmes wrote: >Is anyone familiar with this process to counterstain nerve fibers within a relatively transparent muscle? I need a materials and method list. Can anyone help me? > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From MadaryJ <@t> MedImmune.com Wed Nov 9 11:54:06 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Wed Nov 9 11:56:05 2005 Subject: [Histonet] path/tech ratio Message-ID: <746FDB897740814EA52BDDCB5ED1DDBC02B61571@medimmune4.medimmune.com> Doesn't it depend on the type of pathologist one has? New or insecure pathologists tend to over order, where as the minimalist pathologists(we love them don't we?) order what they need to get the job done, make mission, go fishin. For the record we have two experienced pathologists and 4 experienced histotechs who perform everything. I think PA kind of throw a curveball in the mix too since some are more pathologistlike and others are more histotechlike. Nick Madary, HT/HTL(ASCP)QIHC Medimmune Histology Laboratory One Medimmune Way Gaithersburg, MD 20878 ph 301.398.4745/6113/6141 fx 301.398.9745 From gcallis <@t> montana.edu Wed Nov 9 12:06:46 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 9 12:07:00 2005 Subject: [Histonet] bone fish samples embedded in Historesin Plus In-Reply-To: <007a01c5e553$072504c0$2914100a@labhistologia> References: <006f01c5e546$ce935280$2914100a@labhistologia> <6.0.0.22.1.20051109095751.01b920d8@gemini.msu.montana.edu> <007a01c5e553$072504c0$2914100a@labhistologia> Message-ID: <6.0.0.22.1.20051109110151.01b3c4e8@gemini.msu.montana.edu> It may be better to do the sectioning of these samples using a tungsten carbide tipped knife , d profile should work. A regular steel c profile knife will not cut through undecalcified bones very well. It take the extremely hard TC knife for this purpose. Since Historesin Plus is GMA - you may want to enter into some discussion about this plastic and sectioning with "Patsy Ruegg" as she has experience using this kind of knife with undecalcified bone embedded in GMA. At 10:28 AM 11/9/2005, you wrote: >We are using samples without decalcification because we are studing the bone >fish development. > >Historesin is glycol metacrylate based (manufactured by Leica). > Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From jcline <@t> wchsys.org Wed Nov 9 12:29:48 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Wed Nov 9 12:30:09 2005 Subject: [Histonet] susan/ postition open Message-ID: <000501c5e55b$92a022c0$1d2a14ac@wchsys.org> I currently have a full time 6:00-3:30 varying shift, one Saturday a month. Embedding, cutting, specials & IHC. We are located in Hagerstown, Md about 70 miles from Baltimore and Wash. D.C. My lab is clean and roomy. I have a VIP E300, Sakura DRS stainer, microwave processor, Ventana NeXes equipment and Leica 2135 microtomes. We average about 19.000 cases a year with about 82,000 slides. Please contact personnel at 301-665-4500. ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From RJLevier <@t> LancasterGeneral.org Wed Nov 9 12:40:38 2005 From: RJLevier <@t> LancasterGeneral.org (LeVier, Rebecca J) Date: Wed Nov 9 12:40:53 2005 Subject: [Histonet] Cassette Labels Message-ID: <0FDBF29200C637468CCC1F9E7E85A3D25E221E@MAIL-LR.lha.org> Hello Histonetters, I was wondering if there was anyone that can give me some feedback on any of the automated cassette label systems. We are looking at a few and any feedback would be greatly appreciated. Thank you in advance. Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From rjbuesa <@t> yahoo.com Wed Nov 9 12:46:58 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 9 12:47:08 2005 Subject: [Histonet] path/tech ratio In-Reply-To: <746FDB897740814EA52BDDCB5ED1DDBC02B61571@medimmune4.medimmune.com> Message-ID: <20051109184659.24370.qmail@web61224.mail.yahoo.com> I think that the ratio really depends on economics/management rather than in the pathologist's profficiency. If you have a large volume of work you will have to take care of it in timely manner no matter how the pathologist feels about his diagnosis. An insecure pathologist will only make things worse (from the workload point of view) but it will also be a matter of management to "reign in" some "overacting" pathologist (something that will be the task of the chief pathologist if s/he considers the action as adequate). Thank you for your data. Rene J. "Madary, Joseph" wrote: Doesn't it depend on the type of pathologist one has? New or insecure pathologists tend to over order, where as the minimalist pathologists(we love them don't we?) order what they need to get the job done, make mission, go fishin. For the record we have two experienced pathologists and 4 experienced histotechs who perform everything. I think PA kind of throw a curveball in the mix too since some are more pathologistlike and others are more histotechlike. Nick Madary, HT/HTL(ASCP)QIHC Medimmune Histology Laboratory One Medimmune Way Gaithersburg, MD 20878 ph 301.398.4745/6113/6141 fx 301.398.9745 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From maria <@t> ski.org Wed Nov 9 12:55:53 2005 From: maria <@t> ski.org (Maria Mejia) Date: Wed Nov 9 12:56:07 2005 Subject: [Histonet] Mounting Gelatine Embedded Cryostat CNS Sections References: <20051108135101.79C1B3E01@xprdmailfe6.nwk.excite.com> Message-ID: <43724639.8070709@ski.org> Dear Gerald, For large brain specimens, I always embed them in gelatin (my recipe) and to section them I use the sliding microtome and section 30um to 50ums. These sections are free-floated in 0.1M phosphate buffer and are mounted on gelatin sub slides. As for in-house sub slides there are numerous recipes and each differs slightly in each lab. To gel sub slides use UNCOATED glass slides and clean...again (even though it says on the box - clean slides). I place the slides in a chromerge acid solution overnight (using hood). I can provide this recipe if you are interested. Next day, the slides are removed from the acid solution and washed in running tap water for 1-2 hours with final rinse in distilled water. Then I gelatin coat using Sigma's 300 bloom gelatin. Gelatin Subbing Solution distilled H20 - (cold) - 500ml gelatin (300 bloom) - 6gms Stir to mix well with heat (58 degrees C - don't heat solution above 58C or it will denature the gelatin. Add chromium potassium sulfate (chrom alum) - 0.2gms and mix again. Cool abit and filter using #4 paper. When coating slides make sure the solution is NOT cool - makes for better even coating. Air dry slides well and store in slide boxes. I keep these gel sub slide for no longer than 6 months, then use a freshly made batch. Yours Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 germckeon@excite.com wrote: > >Hello, > >I am currently working on spinal cord and am having trouble with sections floating off. I have embedded with gelatine, am performing cryostat sectioning and will be doing some of the more structural stains eg Kultschitsky's stain for myelin. I have been using Superfrost slides. > >Can anyone advise me regarding choices of slides, coatings, methodologies, etc which would maximise adherence? > > > >In gratitude for any advice received, > >Gerald McKeon > > > > > > > >_______________________________________________ >Join Excite! - http://www.excite.com >The most personalized portal on the Web! > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From TillRenee <@t> uams.edu Wed Nov 9 13:13:41 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Wed Nov 9 13:14:03 2005 Subject: [Histonet] any suggestions? Message-ID: Another tech who does not have much experience with histology came to me with questions about his immunos. They are doing IHC with various cd markers on frozen sections of mouse aorta. He has encountered particularly strong background(or so he's been told, he thinks it is actual staining) with one of the antibodies that was made in rat. He is using a goat anti-rat F(ab')2 from Jackson as the secondary. It is not absorbed against mouse. I have asked all about his dilutions and incubations times, but he doesn't seem to think that is the problem. I gave him an avidin/biotin block to try and see if that helps. Any other ideas? I am not familiar with cd markers myself. The only problem I could find just in talking to him was that he was blocking with rabbit serum? I told him you normally match your block with the host of the secondary, but would that make that big a difference as far as background is concerned? Would the fact he is using frozen sections have anything to do with it? Or could it just be the stain? I know they are doing cd54, but I'm not sure if this is the one he is having a problem with. I know this is not much informations, but I would still appreciate any input Thanks, Renee' ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ From maxim_71 <@t> mail.ru Wed Nov 9 13:52:20 2005 From: maxim_71 <@t> mail.ru (=?koi8-r?Q?=F0=C5=DB=CB=CF=D7=20=ED=C1=CB=D3=C9=CD?=) Date: Wed Nov 9 13:52:33 2005 Subject: [Histonet] histologist/pathologist Message-ID: Good Day! We have 7 pathologists and 11 techs. Workload is 70.000 byposies, 600 autopsies, special stains 2000, frozen secton 300, cytology (FNA) 5000/year or so. We did about 400-500 blocks a day. Peshkov Maxim Russia Taganrog mailto:maxim_71@mail.ru From gentras <@t> vetmed.auburn.edu Wed Nov 9 14:30:24 2005 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Wed Nov 9 14:30:40 2005 Subject: [Histonet] CryoJane Message-ID: <6.0.1.1.0.20051108100801.01aba020@mailhost.vetmed.auburn.edu> Jennifer, I have been using the Instrumedics CryoJane Tape Transfer Method for approximately 5 years and I'm very pleased with it. It allows one to render great sections of those hard to cut frozen sections (i.e. brain) even at lower than usual micron thickness. Although, it is expensive to use it for routine sectioning ( $1.00) per slide it works miracles on brain and other ordinarily problematic tissues. Best wishes. Atoska From tbourm <@t> olympicmedical.org Wed Nov 9 14:40:35 2005 From: tbourm <@t> olympicmedical.org (Tasha Bourm) Date: Wed Nov 9 14:40:45 2005 Subject: [Histonet] Banjo Adesuyi Message-ID: <7B6A91693097B34CBB897045D45E4E2B02A0DCE6@is-210s.olympicmedical.local> I'm looking for any information on where Banjo Adesuyi might be. Please e-mail me back if you have any info. From gcallis <@t> montana.edu Wed Nov 9 15:13:01 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 9 15:13:19 2005 Subject: Murine CD marker woes Re: [Histonet] any suggestions? In-Reply-To: References: Message-ID: <6.0.0.22.1.20051109135020.01b4fd10@gemini.msu.montana.edu> REnee, It would be nice to know which CD marker and know exactly how he is doing his staining???? Including the buffer and if he uses any detergent, protein in buffer - serum or BSA? A lot of murine CD IHC background woes are not just the secondary. He can adsorb his secondary by simply using from 1 to 5% mouse serum in the diluent of secondary and in the normal serum block, i.e. 10% goat with 1 to 5% mouse serum for 30 minutes before the primary. I have, on occasion added mouse serum to primary diluent, and do this routinely when diluting a biotinylated Rat antiMouse primary for a CD marker followed by Strepavidin-HRP. You were correct in advising to match the blocking serum to the host of the secondary. If he wants to use a serum other than goat, swine may be a better choice than rabbit. No, frozen sections are not going to be a factor and he may not be able to use FFPE for the CD markers anyway, crosslinking by NBF will ruin the antigens to the point of NO retrieval, ever!!! What do you mean by the stain? Chromogen? Which one? Hopefully he did a dilution panel starting with a target concentration of 10 ug/ml for primary, and the secondary should be good at 1:250 or approx 2 to 5 ug/ml - Does he control the development of chromogen with a microscope? Is he using HRP or AP? Blocking? Tween 20 in buffer? Protein additive to buffer? How are frozen sections fixed? Cold acetone? Is he using an isotype matched IgG control for the antibody used at the same concentration? More details would help - so I ask a lot of questions!! At 12:13 PM 11/9/2005, you wrote: >Another tech who does not have much experience with histology came to me >with questions about his immunos. They are doing IHC with various cd >markers on frozen sections of mouse aorta. He has encountered >particularly strong background(or so he's been told, he thinks it is >actual staining) with one of the antibodies that was made in rat. He is >using a goat anti-rat F(ab')2 from Jackson as the secondary. It is not >absorbed against mouse. I have asked all about his dilutions and >incubations times, but he doesn't seem to think that is the problem. I >gave him an avidin/biotin block to try and see if that helps. Any other >ideas? I am not familiar with cd markers myself. The only problem I >could find just in talking to him was that he was blocking with rabbit >serum? I told him you normally match your block with the host of the >secondary, but would that make that big a difference as far as >background is concerned? Would the fact he is using frozen sections >have anything to do with it? Or could it just be the stain? I know they >are doing cd54, but I'm not sure if this is the one he is having a >problem with. > >I know this is not much informations, but I would still appreciate any >input > >Thanks, > >Renee' > > >================================================================================================ > >Confidentiality Notice: This e-mail message, including any attachments, is >for the sole use of the intended recipient(s) and may contain confidential >and privileged information. Any unauthorized review, use, disclosure or >distribution is prohibited. If you are not the intended recipient, please >contact the sender by reply e-mail and destroy all copies of the original >message. >================================================================================================ >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From rjbuesa <@t> yahoo.com Wed Nov 9 15:28:08 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 9 15:28:17 2005 Subject: [Histonet] any suggestions? In-Reply-To: Message-ID: <20051109212808.47118.qmail@web61212.mail.yahoo.com> With all due respect I think that your colleague's procedure is a royal mess. On the other hand it seems that he is not very willing to change his ways, so the only thing I have to tell you (for him) is: good luck! He should be the one doing the asking, and not you! Rene J. "Till, Renee" wrote: Another tech who does not have much experience with histology came to me with questions about his immunos. They are doing IHC with various cd markers on frozen sections of mouse aorta. He has encountered particularly strong background(or so he's been told, he thinks it is actual staining) with one of the antibodies that was made in rat. He is using a goat anti-rat F(ab')2 from Jackson as the secondary. It is not absorbed against mouse. I have asked all about his dilutions and incubations times, but he doesn't seem to think that is the problem. I gave him an avidin/biotin block to try and see if that helps. Any other ideas? I am not familiar with cd markers myself. The only problem I could find just in talking to him was that he was blocking with rabbit serum? I told him you normally match your block with the host of the secondary, but would that make that big a difference as far as background is concerned? Would the fact he is using frozen sections have anything to do with it? Or could it just be the stain? I know they are doing cd54, but I'm not sure if this is the one he is having a problem with. I know this is not much informations, but I would still appreciate any input Thanks, Renee' ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From jcresor <@t> lcpath.com Wed Nov 9 15:56:50 2005 From: jcresor <@t> lcpath.com (Jennifer N. Cresor) Date: Wed Nov 9 15:57:07 2005 Subject: [Histonet] Clean glassware education Message-ID: <200511092156.jA9LuvD20115@plus34.host4u.net> Hello all, I would like to do a presentation about the importance of clean glassware. Does anyone have pictures of what stains look like when glassware is not properly cleaned? Maybe a web site? I would really like to show examples. Thank you, Jennifer jcresor@icpath.com From gcallis <@t> montana.edu Wed Nov 9 16:33:29 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 9 16:33:43 2005 Subject: [Histonet] any suggestions? In-Reply-To: <20051109212808.47118.qmail@web61212.mail.yahoo.com> References: <20051109212808.47118.qmail@web61212.mail.yahoo.com> Message-ID: <6.0.0.22.1.20051109150749.01b10198@gemini.msu.montana.edu> Rene, Sorry to disagree, and I certainly did not understand what YOU meant by "changing his ways"? I couldn't see many changes here, other than some refinement of his IHC method. Many people working with murine CD marker IHC often need a little help with murine animal model IHC. Since our lab works with murine CD marker IHC approx. 95% of the time, on frozen sections etc per the inquiry, we are happy to sleuth problems. The questions were valid ones for learning/refining of his techniques and being a mousie person, I didn't have a problem with the inquiry. Some people (including a lot of experts in our field) do NOT want to be on Histonet and second hand messaging certainly doesn't bother me as long as more details are provided. I frequently ask questions on Histonet to access information for others in my department - that's part of my job. Your colleague is welcome to email privately if you or they wish. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) - At 02:28 PM 11/9/2005, you wrote: >With all due respect I think that your colleague's procedure is a royal mess. >On the other hand it seems that he is not very willing to change his ways, >so the only thing I have to tell you (for him) is: good luck! He should be >the one doing the asking, and not you! >Rene J. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From llewllew <@t> shaw.ca Wed Nov 9 16:49:21 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Wed Nov 9 16:50:49 2005 Subject: [Histonet] Van Cleave Hematoxylin Message-ID: <000501c5e57f$d31d7ea0$690a4246@yourlk4rlmsu> Some months ago I came across a reference to Van Cleave's hematoxylin. I have searched since then but have not found any more references. It was to: Pritchard, M. H., and Kruse, G. O. W., (1982) The collection and preservation of animal parasites, Technical Bulletin No. 1. University of Nebraska Press, Lincoln, Nebraska I have gone to the U. Nebraska web site but could not order the publication as it was not listed. If anyone has information about the solution or a copy of the bulletin and could let me know what it says about the hematoxylin, I would appreciate it. Bryan Llewellyn From rjbuesa <@t> yahoo.com Wed Nov 9 17:03:30 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 9 17:03:39 2005 Subject: [Histonet] any suggestions? In-Reply-To: <6.0.0.22.1.20051109150749.01b10198@gemini.msu.montana.edu> Message-ID: <20051109230330.79818.qmail@web61224.mail.yahoo.com> Gayle: Reading the posting it seems that the histotech has not given Renee the information she wanted to have in order to help him, that is what I meant with not willing to change his ways.Sorry that you disagree with me but that is how I feel. He should be the one asking for advise not Renee Rene J. Gayle Callis wrote: Rene, Sorry to disagree, and I certainly did not understand what YOU meant by "changing his ways"? I couldn't see many changes here, other than some refinement of his IHC method. Many people working with murine CD marker IHC often need a little help with murine animal model IHC. Since our lab works with murine CD marker IHC approx. 95% of the time, on frozen sections etc per the inquiry, we are happy to sleuth problems. The questions were valid ones for learning/refining of his techniques and being a mousie person, I didn't have a problem with the inquiry. Some people (including a lot of experts in our field) do NOT want to be on Histonet and second hand messaging certainly doesn't bother me as long as more details are provided. I frequently ask questions on Histonet to access information for others in my department - that's part of my job. Your colleague is welcome to email privately if you or they wish. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) - At 02:28 PM 11/9/2005, you wrote: >With all due respect I think that your colleague's procedure is a royal mess. >On the other hand it seems that he is not very willing to change his ways, >so the only thing I have to tell you (for him) is: good luck! He should be >the one doing the asking, and not you! >Rene J. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From chesarato <@t> hotmail.com Wed Nov 9 21:57:30 2005 From: chesarato <@t> hotmail.com (Cesar Francisco Romero) Date: Wed Nov 9 21:57:39 2005 Subject: [Histonet] Santa Cruz Antibodies ! Message-ID: Dear People, this is my question: Does Anybody know what does Santa Cruz people mean when they say " DO NOT FREEZE " in their Data sheets ? Can I Freeze aliquotes only once for storage ? They say that the antibody is good only for one year. I think is a big waste of money to buy it only for a year. Thank You in advance Cesar Romero Buenos Aires Argentina _________________________________________________________________ Nuevo MSN Messenger [1]Una forma r?pida y divertida de enviar mensajes References 1. http://g.msn.com/8HMAESAR/2749??PS=47575 From mikael.niku <@t> helsinki.fi Thu Nov 10 01:24:19 2005 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Thu Nov 10 01:24:35 2005 Subject: [Histonet] Re: Bone marrow unspecific staining In-Reply-To: References: Message-ID: <4372F5A3.2070502@helsinki.fi> Thanks for your very valuable comments, everyone! With best regards, Mikael -- //////////////////////////////////////////////////////////// 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 //////////////////////////////////////////////////////////// From drosini <@t> comcast.net Thu Nov 10 05:53:56 2005 From: drosini <@t> comcast.net (drosini@comcast.net) Date: Thu Nov 10 05:54:05 2005 Subject: [Histonet] (no subject) Message-ID: <111020051153.26883.437334D400063D3F0000690322007358340702079C019D0B@comcast.net> please take my name off the histonet list From rjbuesa <@t> yahoo.com Thu Nov 10 07:25:52 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 10 07:26:00 2005 Subject: [Histonet] Santa Cruz Antibodies ! In-Reply-To: Message-ID: <20051110132552.78235.qmail@web61215.mail.yahoo.com> Cesar: Santa Cruz, as many other antibodies manufacturers, usually place a warning label of "do not freeze". They are referring to avoid a cycle of "freeze-thaw-freeze and thaw again and again". This procedure will affect the proteins and should be avoided. Other manufacturers, like Novocastra sell their antibodies in lyophilized form and have to be reconstituted (usually with deionized water because the preservatives also come with the anibody) and they can be aliquoted and frozen until their expiration date. We use to to than routinely with many antibodies and all of our FITC conjugated for immunofluorescence. The temperature at which they are frozen should be the lowest possible; we used -80Celsius and kept our aliquoted antibodies with the speciments in our tumours bank. So, answering to your question more directly: no, you cannot freeze-thaw-freeze-thaw.... your antibodies. Yes, you can aliquote them and freeze the aliquotes and, yes, they could be used after their expiration date; there are a few publications concluding that they can be used (I just don't have the reference now, perhaps one of the readers of this posting can forward them to you). Rene J. Cesar Francisco Romero wrote: Dear People, this is my question: Does Anybody know what does Santa Cruz people mean when they say " DO NOT FREEZE " in their Data sheets ? Can I Freeze aliquotes only once for storage ? They say that the antibody is good only for one year. I think is a big waste of money to buy it only for a year. Thank You in advance Cesar Romero Buenos Aires Argentina _________________________________________________________________ Nuevo MSN Messenger [1]Una forma r?pida y divertida de enviar mensajes References 1. http://g.msn.com/8HMAESAR/2749??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From KevinMcGovern <@t> catholichealth.net Thu Nov 10 08:22:22 2005 From: KevinMcGovern <@t> catholichealth.net (McGovern, Kevin) Date: Thu Nov 10 08:22:36 2005 Subject: [Histonet] Old Style Bacti-Cinerator Element Message-ID: Hello Histoneteurs, I work with a great Histo tech who needs to be able to sterilize forceps in his old Bacti-Cinerator. The newer models have a ceramic collar that requires he get his forceps in deep to get them heated properly, which is proving a bit rough on his fingertips. The older type of element heated pretty much right up to the front of the unit, and his has gasped out its last. Do any of you have an old element (it's pretty wide--about 5cm, compared to the new elements) you'd like to sell, or do you have a source for them to which you could direct moi? Our tech plays jazz sax, and I hate to see him give up a promising career, simply because he has burned fingers... Thanks very much for your help! Kevin Kevin S. McGovern, BMETII Good Samaritan Hospital Clinical Engineering Dept. 10 East 31st Street Kearney, NE 68836 Phone: (308) 865-7051 Fax: (308) 865-2914 e-Mail: kevinmcgovern@catholichealth.net Confidentiality Notice: This email, including any attachments, contains CONFIDENTIAL information. The information is intended only for the use of the individual (s) or entity name above. If you are not the intended recipient, you are notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this email is not permissible. If you have received this email in error, please notify the sender immediately by reply mail, and destroy all copies of the original message. From pmarcum <@t> vet.upenn.edu Thu Nov 10 08:27:29 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Thu Nov 10 08:27:43 2005 Subject: [Histonet] any suggestions? In-Reply-To: <6.0.0.22.1.20051109150749.01b10198@gemini.msu.montana.edu> References: <20051109212808.47118.qmail@web61212.mail.yahoo.com> <6.0.0.22.1.20051109150749.01b10198@gemini.msu.montana.edu> Message-ID: <6.1.1.1.2.20051110092057.019acc78@mail.vet.upenn.edu> Thank You Gayle!! It is sometimes easy to forget not everyone has access to the Internet or a way to even place a message on HistoNet. We are attempting to assist people who are learning and while some may seem to be using HistoNet as way to avoid looking anything up the majority are asking questions that are not readily accessible or in an area like murine animal models that is more difficult. I have no problem with this and will answer most questions off line if it is a more esoteric question. I am surprised that the person with the issue about Rene's assistance for her friend is also the same Rene who has been asking all of us to contribute to Tricks of the Trade. Pamela A Marcum Special Procedures Manager Bone Histology UPENN Vet School At 05:33 PM 11/9/2005, Gayle Callis wrote: >Rene, > >Sorry to disagree, and I certainly did not understand what YOU meant by >"changing his ways"? I couldn't see many changes here, other than some >refinement of his IHC method. Many people working with murine CD marker >IHC often need a little help with murine animal model IHC. Since our >lab works with murine CD marker IHC approx. 95% of the time, on frozen >sections etc per the inquiry, we are happy to sleuth problems. The >questions were valid ones for learning/refining of his techniques and >being a mousie person, I didn't have a problem with the inquiry. > >Some people (including a lot of experts in our field) do NOT want to be on >Histonet and second hand messaging certainly doesn't bother me as long as >more details are provided. I frequently ask questions on Histonet to >access information for others in my department - that's part of my job. >Your colleague is welcome to email privately if you or they wish. > >Gayle Callis >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 >406 994-4303 (FAX) > > > > > > >- At 02:28 PM 11/9/2005, you wrote: >>With all due respect I think that your colleague's procedure is a royal mess. >>On the other hand it seems that he is not very willing to change his >>ways, so the only thing I have to tell you (for him) is: good luck! He >>should be the one doing the asking, and not you! >>Rene J. > >Gayle Callis >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From j.w.degroot <@t> med.uu.nl Thu Nov 10 08:59:51 2005 From: j.w.degroot <@t> med.uu.nl (Jan Willem de Groot) Date: Thu Nov 10 09:00:04 2005 Subject: [Histonet] Pmv cryomicrotome Message-ID: Hi Histonetters, I was wondering if there are laboratories that are working with the PMV 450mp large cryomicrotome (Palmstiernas Instruments Stockholm) from Leica or another company. Using the PMV 450mp, mammals like mature rabbits,rats and mice can be sectioned. If so I would like to exchange information about you experience with this cryomcrotome e.g. in what way you embed and collect sections of these large specimens etc. Thank you, Jan Willem JanWillem de Groot University Medical Centre Utrecht Dept. of Functional Anatomy P.O. Box 85060 3508 AB Utrecht, the Netherlands tel.+31 30 2538323 fax +31 30 2539030 e-mail: j.w.degroot@med.uu.nl From TillRenee <@t> uams.edu Thu Nov 10 09:18:58 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Thu Nov 10 09:19:21 2005 Subject: [Histonet] Her2/Neu Message-ID: I am looking for a Her2/Neu antibody to use on rat tissue. I keep coming across ErbB-2 and C-erbB-2 in relation to it. Is this the same thing? I have found far more under the latter 2 name than under Her2 or Neu. Thanks! Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ From rjbuesa <@t> yahoo.com Thu Nov 10 09:40:22 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 10 09:40:32 2005 Subject: [Histonet] Her2/Neu In-Reply-To: Message-ID: <20051110154022.13153.qmail@web61223.mail.yahoo.com> Dear Namesake: Her2/Neu is a trade mark of DakoCytomation and it related to the human HER-2 gene , also known as NEU or ERBB2 that encodes a protein often referred to as HER2 protein or p185 The c-erbB-2 oncoproten is closely related in structure to the epidermal growth factor and is a member of a large family of cell surface growth factor receptors. C-erbB-2 oncoprotein is present in some breast and adenocarcinomas and in some transitional cell carcinomas. This Ab is provided by Novocastra and presents the same reaction aspect as Her2/Neu. Her2/Neu is an FDA diagnostic approved protocol as DAKO Hercept Test measured in 3 levels of reaction intensity with treatment prognostic value. They are essentially the same but commercialized under different names. Rene J. "Till, Renee" wrote: I am looking for a Her2/Neu antibody to use on rat tissue. I keep coming across ErbB-2 and C-erbB-2 in relation to it. Is this the same thing? I have found far more under the latter 2 name than under Her2 or Neu. Thanks! Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From BlazekL <@t> childrensdayton.org Thu Nov 10 09:54:21 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Thu Nov 10 09:56:39 2005 Subject: [Histonet] tonsils Message-ID: I need some information on how other facilities are handling tonsils. Are they gross only and what criteria is used to have microscopic performed on them? Thanks for any info. - Linda Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org From DavidH <@t> marketlabinc.com Thu Nov 10 10:02:03 2005 From: DavidH <@t> marketlabinc.com (David Haagsma) Date: Thu Nov 10 10:02:14 2005 Subject: [Histonet] Old Style Bacti-Cinerator Element Message-ID: <19E3602A16438E48B51A4250CA04B5F638B556@exchange.marketlab.com> Kevin, You need to protect those fingers. We still have a few (6) of the older model (model II and III) heating elements available at MarketLab. (ML0633) Just a little recent history on the Bacti-Cinerator - It was a Sherwood/Monoject item which was owned by Tyco Healthcare. Tyco has been selling off a lot of their healthcare products including the line of Bacti-Cinerators. They are now made by McCormick. In the transition, there have been supply problems and price increases. If you're feeling lucky, there are some new in box units available at Eglin AFB on the military surplus auction site: http://cgi.govliquidation.com/auction/view?id=717138 There are deals on there sometimes, they have sold off a pallet of Leica microtomes for $850.00 a piece, embedding centers and other items of interest for histology labs. Regards, Dave Haagsma MT MarketLab Inc. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of McGovern, Kevin Sent: Thursday, November 10, 2005 9:22 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Old Style Bacti-Cinerator Element Hello Histoneteurs, I work with a great Histo tech who needs to be able to sterilize forceps in his old Bacti-Cinerator. The newer models have a ceramic collar that requires he get his forceps in deep to get them heated properly, which is proving a bit rough on his fingertips. The older type of element heated pretty much right up to the front of the unit, and his has gasped out its last. Do any of you have an old element (it's pretty wide--about 5cm, compared to the new elements) you'd like to sell, or do you have a source for them to which you could direct moi? Our tech plays jazz sax, and I hate to see him give up a promising career, simply because he has burned fingers... Thanks very much for your help! Kevin Kevin S. McGovern, BMETII Good Samaritan Hospital Clinical Engineering Dept. 10 East 31st Street Kearney, NE 68836 Phone: (308) 865-7051 Fax: (308) 865-2914 e-Mail: kevinmcgovern@catholichealth.net Confidentiality Notice: This email, including any attachments, contains CONFIDENTIAL information. The information is intended only for the use of the individual (s) or entity name above. If you are not the intended recipient, you are notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this email is not permissible. If you have received this email in error, please notify the sender immediately by reply mail, and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jennifer.l.hofecker <@t> Vanderbilt.Edu Thu Nov 10 10:03:33 2005 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Thu Nov 10 10:03:43 2005 Subject: [Histonet] any suggestions? References: <20051109212808.47118.qmail@web61212.mail.yahoo.com><6.0.0.22.1.20051109150749.01b10198@gemini.msu.montana.edu> <6.1.1.1.2.20051110092057.019acc78@mail.vet.upenn.edu> Message-ID: <898D946569A27444B65667A49C07405207551D@mailbe06.mc.vanderbilt.edu> Well said Pam and Gayle! I know that every time I have a problem, my first thought is "ask histonet" not because I am lazy or because I can't look it up. I think first of histonet because of the diversity of our listmembers and their experience. Just because a publication lists the steps of a protocol, doesn't mean it will work. I would rather get my info from the ones who make these protocols work. On the other hand, every time I am about to post a query, I have to read it 6 times to make sure there isn't anything in the post that I will be chastised for. While I agree that we should not "post with reckless abandon", I do not think we should be terrified of backlash for asking how to do something we've never done before, especially if it affects patient care, whether directly or indirectly. Well, I am going to put on my bullet proof vest! Thanks for listening, Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax Thank You Gayle!! It is sometimes easy to forget not everyone has access to the Internet or a way to even place a message on HistoNet. We are attempting to assist people who are learning and while some may seem to be using HistoNet as way to avoid looking anything up the majority are asking questions that are not readily accessible or in an area like murine animal models that is more difficult. I have no problem with this and will answer most questions off line if it is a more esoteric question. I am surprised that the person with the issue about Rene's assistance for her friend is also the same Rene who has been asking all of us to contribute to Tricks of the Trade. Pamela A Marcum Special Procedures Manager Bone Histology UPENN Vet School At 05:33 PM 11/9/2005, Gayle Callis wrote: >Rene, > >Sorry to disagree, and I certainly did not understand what YOU meant by >"changing his ways"? I couldn't see many changes here, other than some >refinement of his IHC method. Many people working with murine CD marker >IHC often need a little help with murine animal model IHC. Since our >lab works with murine CD marker IHC approx. 95% of the time, on frozen >sections etc per the inquiry, we are happy to sleuth problems. The >questions were valid ones for learning/refining of his techniques and >being a mousie person, I didn't have a problem with the inquiry. > >Some people (including a lot of experts in our field) do NOT want to be on >Histonet and second hand messaging certainly doesn't bother me as long as >more details are provided. I frequently ask questions on Histonet to >access information for others in my department - that's part of my job. >Your colleague is welcome to email privately if you or they wish. > >Gayle Callis >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 >406 994-4303 (FAX) > > > > > > >- At 02:28 PM 11/9/2005, you wrote: >>With all due respect I think that your colleague's procedure is a royal mess. >>On the other hand it seems that he is not very willing to change his >>ways, so the only thing I have to tell you (for him) is: good luck! He >>should be the one doing the asking, and not you! >>Rene J. > >Gayle Callis >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 >406 994-4303 (FAX) > > > >_______________________________________________ >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 rjbuesa <@t> yahoo.com Thu Nov 10 10:19:49 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 10 10:19:57 2005 Subject: [Histonet] tonsils In-Reply-To: Message-ID: <20051110161949.42007.qmail@web61216.mail.yahoo.com> We used to gross them only but the PA (usually at requests from the surgeon or the medical practitioner) could ask for a microscopic evaluation sometimes as results of a request by the insurance company!. In that case we used to process them and do just H&E. Essentially you will have to ask your pathologist what to do about them. Rene J. Linda Blazek wrote: I need some information on how other facilities are handling tonsils. Are they gross only and what criteria is used to have microscopic performed on them? Thanks for any info. - Linda Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 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 --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From pruegg <@t> ihctech.net Thu Nov 10 10:23:37 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Nov 10 10:23:51 2005 Subject: [Histonet] any suggestions? In-Reply-To: <20051109212808.47118.qmail@web61212.mail.yahoo.com> Message-ID: <200511101623.jAAGNbxT021885@chip.viawest.net> Renee the IHC question asker. I too, like Gayle think that your question was appropriate for this list and I thought that you gave a lot of information. In my opinion yes he should be using goat serum block to match up with his secondary host. There may also be a too close species with his anti-rat primary on ms tissue causing non specific binding of ms serum IgG just because ms and rat are so closely related. Frozen sections can be very active with antigen since they don't go thru all the processing related to ffpe tissue, so in my experience you need to carefully titer your IHC antibody reagents often diluting much more than you would for ffpe tissue. Also much of the endogenous issues, such as biotin, peroxidase and alk.phos. Are inhanced in frozens, or preserved more than in ffpe tissue. AB block, fc receptor block, serum block before the primary and before the secondary are all things I consider with frozen IHC, or I try to avoid the biotin issue altogether by using a non biotin labeled polymer detection. Maybe this person is inexperienced but we all started somewhere, so probably with just a little help we can make a big difference for this person, that is what we are here for, right? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Wednesday, November 09, 2005 2:28 PM To: Till, Renee; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] any suggestions? With all due respect I think that your colleague's procedure is a royal mess. On the other hand it seems that he is not very willing to change his ways, so the only thing I have to tell you (for him) is: good luck! He should be the one doing the asking, and not you! Rene J. "Till, Renee" wrote: Another tech who does not have much experience with histology came to me with questions about his immunos. They are doing IHC with various cd markers on frozen sections of mouse aorta. He has encountered particularly strong background(or so he's been told, he thinks it is actual staining) with one of the antibodies that was made in rat. He is using a goat anti-rat F(ab')2 from Jackson as the secondary. It is not absorbed against mouse. I have asked all about his dilutions and incubations times, but he doesn't seem to think that is the problem. I gave him an avidin/biotin block to try and see if that helps. Any other ideas? I am not familiar with cd markers myself. The only problem I could find just in talking to him was that he was blocking with rabbit serum? I told him you normally match your block with the host of the secondary, but would that make that big a difference as far as background is concerned? Would the fact he is using frozen sections have anything to do with it? Or could it just be the stain? I know they are doing cd54, but I'm not sure if this is the one he is having a problem with. I know this is not much informations, but I would still appreciate any input Thanks, Renee' ============================================================================ ==================== Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ============================================================================ ==================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Thu Nov 10 10:23:46 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Thu Nov 10 10:24:03 2005 Subject: [Histonet] any suggestions? In-Reply-To: <898D946569A27444B65667A49C07405207551D@mailbe06.mc.vanderb ilt.edu> References: <20051109212808.47118.qmail@web61212.mail.yahoo.com> <6.0.0.22.1.20051109150749.01b10198@gemini.msu.montana.edu> <6.1.1.1.2.20051110092057.019acc78@mail.vet.upenn.edu> <898D946569A27444B65667A49C07405207551D@mailbe06.mc.vanderbilt.edu> Message-ID: <6.1.1.1.2.20051110112330.019a6c60@mail.vet.upenn.edu> Thanks, Jennifer. Pam At 11:03 AM 11/10/2005, Hofecker, Jennifer L wrote: >Well said Pam and Gayle! >I know that every time I have a problem, my first thought is "ask >histonet" not because I am lazy or because I can't look it up. I think >first of histonet because of the diversity of our listmembers and their >experience. Just because a publication lists the steps of a protocol, >doesn't mean it will work. I would rather get my info from the ones who >make these protocols work. >On the other hand, every time I am about to post a query, I have to read >it 6 times to make sure there isn't anything in the post that I will be >chastised for. While I agree that we should not "post with reckless >abandon", I do not think we should be terrified of backlash for asking how >to do something we've never done before, especially if it affects patient >care, whether directly or indirectly. > >Well, I am going to put on my bullet proof vest! >Thanks for listening, >Jennifer > > > >Jennifer Hofecker, HT (ASCP) >Vanderbilt University Medical Center >Division of Neuropathology >(615) 343-0083 >(615) 343-7089 fax > >Thank You Gayle!! It is sometimes easy to forget not everyone has access >to the Internet or a way to even place a message on HistoNet. We are >attempting to assist people who are learning and while some may seem to be >using HistoNet as way to avoid looking anything up the majority are asking >questions that are not readily accessible or in an area like murine animal >models that is more difficult. I have no problem with this and will answer >most questions off line if it is a more esoteric question. I am surprised >that the person with the issue about Rene's assistance for her friend is >also the same Rene who has been asking all of us to contribute to Tricks of >the Trade. > >Pamela A Marcum >Special Procedures Manager >Bone Histology >UPENN Vet School > > >At 05:33 PM 11/9/2005, Gayle Callis wrote: > >Rene, > > > >Sorry to disagree, and I certainly did not understand what YOU meant by > >"changing his ways"? I couldn't see many changes here, other than some > >refinement of his IHC method. Many people working with murine CD marker > >IHC often need a little help with murine animal model IHC. Since our > >lab works with murine CD marker IHC approx. 95% of the time, on frozen > >sections etc per the inquiry, we are happy to sleuth problems. The > >questions were valid ones for learning/refining of his techniques and > >being a mousie person, I didn't have a problem with the inquiry. > > > >Some people (including a lot of experts in our field) do NOT want to be on > >Histonet and second hand messaging certainly doesn't bother me as long as > >more details are provided. I frequently ask questions on Histonet to > >access information for others in my department - that's part of my job. > >Your colleague is welcome to email privately if you or they wish. > > > >Gayle Callis > >Research Histopathology Supervisor > >Veterinary Molecular Biology > >Montana State University - Bozeman > >PO Box 173610 > >Bozeman MT 59717-3610 > >406 994-6367 > >406 994-4303 (FAX) > > > > > > > > > > > > > >- At 02:28 PM 11/9/2005, you wrote: > >>With all due respect I think that your colleague's procedure is a royal > mess. > >>On the other hand it seems that he is not very willing to change his > >>ways, so the only thing I have to tell you (for him) is: good luck! He > >>should be the one doing the asking, and not you! > >>Rene J. > > > >Gayle Callis > >Research Histopathology Supervisor > >Veterinary Molecular Biology > >Montana State University - Bozeman > >PO Box 173610 > >Bozeman MT 59717-3610 > >406 994-6367 > >406 994-4303 (FAX) > > > > > > > >_______________________________________________ > >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 Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From John.Sheppard <@t> Health-Partners.org Thu Nov 10 10:56:13 2005 From: John.Sheppard <@t> Health-Partners.org (John.Sheppard@Health-Partners.org) Date: Thu Nov 10 10:51:35 2005 Subject: [Histonet] Process improvement monitors Message-ID: Hello "Histonetters", Once again I am on a "mission" for my medical director. I am hoping to find out if anyone out there has been looking at quality management / process improvement areas in histology / pathology labs in their community hospitals. We are currently reworking our P.I. program, and we are looking at some of the most obvious errors we find. It seems to us that many mistakes come to our lab from doctors offices, the O.R. and our birthing center. These errors come in the form of incomplete or no requisition, discrepancies between specimen labels and requisitions, and no or incomplete label on specimens. These are the issues that are of particular interest to my director. We are trying to establish a reasonable range to strive for. Ideally we would like to have all specimens labelled correctly with the correct reqistion and full clinical history / diagnosis coming in to our lab, but realistically we are not sure if that is possible. If anyone has reviewed these monitors and does not mind sharing their information, we would greatly appreciate using it for comparitive data purposes. We have been monitoring this for a couple of years and we do not have a sky high rate, but it is higher than we believe it should be. Thanks again John Sheppard HT(ASCP) P.S. If anyone has ideas on how to improve this performance, short of firing nuses everytime they mess up please let me know ? (this is a joke) CONFIDENTIALITY NOTICE: This message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From PMonfils <@t> Lifespan.org Thu Nov 10 11:18:36 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Thu Nov 10 11:18:48 2005 Subject: [Histonet] tonsils Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717610@lsexch.lsmaster.lifespan.org> Years ago, when I worked in a clinical histology lab (I am now in research), we processed and sectioned all tonsils. Since no serious pathology had ever been discovered in any of the thousands of subadult tonsils we had processed, the department was considering a "gross only" policy for tonsils from patients under 16 years of age. Don't you know, that very month they found an undiagnosed lymphoma in the tonsils of a 12 year old girl. We continued processing all tonsils. As I said, that was quite a few years ago, and they may have changed the policy since then. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Linda > Blazek > Sent: Thursday, November 10, 2005 7:54 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] tonsils > > I need some information on how other facilities are handling tonsils. Are > they gross only and what criteria is used to have microscopic performed on > them? Thanks for any info. - Linda > > > Linda Blazek, HT (ASCP) > Department of Pathology > Children's Medical Center > 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 > > From Siobhan.O'Reilly <@t> nyumc.org Thu Nov 10 11:20:00 2005 From: Siobhan.O'Reilly <@t> nyumc.org (O'Reilly, Siobhan) Date: Thu Nov 10 11:20:35 2005 Subject: [Histonet] (no subject) Message-ID: <4BCB18390D553D41A3FFEB77D5A3BA97012DB631@excnyuebw2k63.nyumc.org> Please remove mine as well. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of drosini@comcast.net Sent: Thursday, November 10, 2005 6:54 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) please take my name off the histonet list _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------- This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email. ============================================================================== From bernaweston <@t> hotmail.com Thu Nov 10 11:48:12 2005 From: bernaweston <@t> hotmail.com (Bernadette Weston) Date: Thu Nov 10 11:48:21 2005 Subject: [Histonet] reproducing microwave temperatures Message-ID: A few years back I attended a convention taht did a workshop on how to check your microwave for reproducible tempertures. I remember having a worksheet for that seminar, does anyone have a method they like to share for this monitoring? Bernadette Weston HT Barberton Citizens Hospital From rjbuesa <@t> yahoo.com Thu Nov 10 12:05:06 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 10 12:05:13 2005 Subject: [Histonet] reproducing microwave temperatures In-Reply-To: Message-ID: <20051110180506.65450.qmail@web61220.mail.yahoo.com> Bernadette: If you are talking about a "home cooking" microwaves oven (MWO) temperatures are difficult to reproduce unless you calibrate your MWO. I published a paper on the subject but I don't have an electronic version of it. If you are interested in a copye you can e-mail me your FAX number I will send you a copy. Now if you are talking about a MWO of the recent breed for tissue processing, that is a a given since they have thermo probes connected to the magnetron tube in a way that you set a desied operation temperature and the MWO will reach and keep it. Now, even in this category there are MWO with variable wattage output that will deliver the energy in a continuous way (these are to be preferred) and others just deliver wattage in "bursts" meaning that the magnetron tube will be off and on during alternate periods of time resulting in a average energy to be correlated to certain temperature. Rene J. Bernadette Weston wrote: A few years back I attended a convention taht did a workshop on how to check your microwave for reproducible tempertures. I remember having a worksheet for that seminar, does anyone have a method they like to share for this monitoring? Bernadette Weston HT Barberton Citizens Hospital _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From ja.mitchell <@t> hosp.wisc.edu Thu Nov 10 12:54:41 2005 From: ja.mitchell <@t> hosp.wisc.edu (Mitchell Jean A.) Date: Thu Nov 10 12:54:50 2005 Subject: [Histonet] Disinfectant Solutions Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3147CF832@uwhis-xchng2.hosp.wisc.edu> What disinfectant solutions or products do any of your laboratories use for forceps, bench tops, etc, that come in contact with blood and or surgical tissues? Thanks!! Jean Mitchell University of Wisconsin Hospital & Clinics 600 Highland Ave. Neuromuscular Laboratory, H6/547 Madison, WI 53792 From cgfields <@t> lexhealth.org Thu Nov 10 13:04:21 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Thu Nov 10 13:02:05 2005 Subject: [Histonet] Disinfectant Solutions Message-ID: Amphyl....it is strong so you have to dilute. Most hospitals carry it inhouse. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 -----Original Message----- From: Mitchell Jean A. [mailto:ja.mitchell@hosp.wisc.edu] Sent: Thursday, November 10, 2005 1:55 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Disinfectant Solutions What disinfectant solutions or products do any of your laboratories use for forceps, bench tops, etc, that come in contact with blood and or surgical tissues? Thanks!! Jean Mitchell University of Wisconsin Hospital & Clinics 600 Highland Ave. Neuromuscular Laboratory, H6/547 Madison, WI 53792 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________ 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 som <@t> kvl.dk Thu Nov 10 08:07:32 2005 From: som <@t> kvl.dk (Sophia Moesgaard) Date: Thu Nov 10 13:03:15 2005 Subject: [Histonet] eNOS in paraffin sections from dogs and pigs? Message-ID: Hi! I am looking for antibodies that detect eNOS on formalin-paraffin embedded cardiac tissue from dogs and/or pigs on immunohistochemistry. From the list it appears that transduction laboratories produces a monoclonal antibody (610296/30020) that works well in paraffin sections - but does anybody have experience with this or any other eNOS antibody in pigs or dogs? Any help or comment will be greatly appreciated! Best Regards Sophia Moesgaard Sophia Gry Moesgaard DVM, Ph.D. student Department of Basic Animal and Veterinary Sciences The Royal Veterinary and Agricultural University (KVL) Dyrl?gevej 100, lok. E-09 1870 Frederiksberg Denmark Phone: +45-35283884 Fax: +45-35282525 e-mail: som@kvl.dk From Michele_Marggi <@t> ssmhc.com Thu Nov 10 13:08:10 2005 From: Michele_Marggi <@t> ssmhc.com (Michele_Marggi@ssmhc.com) Date: Thu Nov 10 13:09:48 2005 Subject: [Histonet] tonsils In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE01717610@lsexch.lsmaster.lifespan.org> Message-ID: Our lab does a Gross-only for tonsils and adenoids for those under 12 years of age. (Unless ordered specifically by the ordering physician or a pathologist feels a micro is warranted). Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 "Monfils, Paul" To Sent by: "'histonet@lists.utsouthwestern.edu histonet-bounces@ '" lists.utsouthwest ern.edu cc Subject 11/10/2005 11:18 RE: [Histonet] tonsils AM Years ago, when I worked in a clinical histology lab (I am now in research), we processed and sectioned all tonsils. Since no serious pathology had ever been discovered in any of the thousands of subadult tonsils we had processed, the department was considering a "gross only" policy for tonsils from patients under 16 years of age. Don't you know, that very month they found an undiagnosed lymphoma in the tonsils of a 12 year old girl. We continued processing all tonsils. As I said, that was quite a few years ago, and they may have changed the policy since then. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Linda > Blazek > Sent: Thursday, November 10, 2005 7:54 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] tonsils > > I need some information on how other facilities are handling tonsils. Are > they gross only and what criteria is used to have microscopic performed on > them? Thanks for any info. - Linda > > > Linda Blazek, HT (ASCP) > Department of Pathology > Children's Medical Center > 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 > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. From pruegg <@t> ihctech.net Thu Nov 10 13:20:43 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Nov 10 13:20:56 2005 Subject: [Histonet] SERCA-2 Message-ID: <200511101920.jAAJKhxT013389@chip.viawest.net> We are interested in a new protein - SERCA2. We are interested in > looking at its expression in human lung and sheep (and possibly later > in baboon). We have the antibody to this protein working in westerns. Has anyone experience with this antibody to this protein in tissue by IHC methods. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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 MICHAEL.OWEN <@t> fda.gov Thu Nov 10 13:30:26 2005 From: MICHAEL.OWEN <@t> fda.gov (Owen, Michael P) Date: Thu Nov 10 13:30:47 2005 Subject: [Histonet] Disinfectant Solutions Message-ID: Dear Jean, A formal risk assessment of the pathogens expected to be encountered should be performed before using a chemical disinfectant. This evaluation will determine what chemical is appropriate. Although phenolics like Amphyl are effective against many types of microorganisms they will not be effective against everything. I recommend you consolt the following books and Web sites for discussions on the strengths and weaknesses of most chemical disinfectants. For example, chlorine compounds such as household bleach are very effective disinfectants but contact times may be long (depending on use) and their corrosive action will be damaging to instruments. I am not an expert on risk assessments or biological safety. On the contrary, I am a humble novice and I have learned when writing a Web-based course on biological safety this year disinfectant selection is not a simple process. Good luck! WHO Biological Safety Manual Third Edition http://www.who.int/csr/resources/publications/biosafety/WHO_CDS_CSR_LYO_2004 _11/en World Health Organization December 2004 University of Washington-Seattle Environmental Health and Safety http://www.ehs.washington.edu http://www.ehs.washington.edu/rbsbiosafe/index.shtm Biological Safety Principles and Practices Third Edition Edited by Diane O. Fleming and Debra L. Hunt American Society of Microbiology December 2000 Laboratory-acquired Infections: History, Incidence, Causes and Preventions, Fourth Edition C. H. Collins and D. A. Kennedy Arnold Publishers January 1999 Michael P. Owen, Regulatory Microbiologist U.S. Food and Drug Administration Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From psanquin <@t> lugo.usc.es Thu Nov 10 13:42:01 2005 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Thu Nov 10 13:35:43 2005 Subject: [Histonet] antibody storage Message-ID: <3.0.6.32.20051110204201.008abb80@pop.lugo.usc.es> Dear Histonetters, Lately I am having difficulties with the storage of my antibodies. I keep them at -20?C in undiluted aliquots. After few months a kind of web or lattice grows in the liquid. It is evident after unfreezening. The antibody keeps working but I am afraid this is not any pleasant. I would very greatful with your input. By the way the provider states an expiring date of only six months. Is this reasonable? Best regards from Spain Pablo Sanchez From Michael.Rice <@t> holy-cross.com Thu Nov 10 13:44:30 2005 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Thu Nov 10 13:45:16 2005 Subject: [Histonet] RE: Histonet Digest, Vol 24, Issue 15 labeling issues Message-ID: <3BC92F29BE821745AB15E04C98EE028D6D38D4@HCH2KMAIL.holy-cross.com> In response to the question regarding an acceptable range for identification errors, there can be no range. It must be 100% correct. It has always been my habit to refuse specimens that are incorrectly labeled or otherwise lacking information. In the case of biopsies or other signicant cases where we do not want to lose control of the specimen, we insist that the person who was involved in the origin of the specimen come to pathology and identify the patient or correct the error. We do not process the specimen until all problems have been rectified. after a while when the people involved get tired of your calls and their having to come to the lab, they will hopefully change their ways. We recently had an ongoing problem with the OR sending specimens down without labeling the containers. I met with the nurse manager for the OR and went over the problem with her. She was able to identify the employee who was ignoring the step of labeling the specimens. That employee was counseled and was warned that her job was at risk. It has not happened again with that employee. Drastic steps have to be taken to avoid these errors as part of meeting JCAHO standards Michael Rice CT.HT(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Thursday, November 10, 2005 1:04 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 24, Issue 15 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: tonsils (Rene J Buesa) 2. RE: any suggestions? (Patsy Ruegg) 3. RE: any suggestions? (Pamela Marcum) 4. Process improvement monitors (John.Sheppard@Health-Partners.org) 5. RE: tonsils (Monfils, Paul) 6. RE: (no subject) (O'Reilly, Siobhan) 7. reproducing microwave temperatures (Bernadette Weston) ---------------------------------------------------------------------- Message: 1 Date: Thu, 10 Nov 2005 08:19:49 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] tonsils To: Linda Blazek , histonet@lists.utsouthwestern.edu Message-ID: <20051110161949.42007.qmail@web61216.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 We used to gross them only but the PA (usually at requests from the surgeon or the medical practitioner) could ask for a microscopic evaluation sometimes as results of a request by the insurance company!. In that case we used to process them and do just H&E. Essentially you will have to ask your pathologist what to do about them. Rene J. Linda Blazek wrote: I need some information on how other facilities are handling tonsils. Are they gross only and what criteria is used to have microscopic performed on them? Thanks for any info. - Linda Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 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 --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 2 Date: Thu, 10 Nov 2005 09:23:37 -0700 From: "Patsy Ruegg" Subject: RE: [Histonet] any suggestions? To: "'Rene J Buesa'" , "'Till, Renee'" , Message-ID: <200511101623.jAAGNbxT021885@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Renee the IHC question asker. I too, like Gayle think that your question was appropriate for this list and I thought that you gave a lot of information. In my opinion yes he should be using goat serum block to match up with his secondary host. There may also be a too close species with his anti-rat primary on ms tissue causing non specific binding of ms serum IgG just because ms and rat are so closely related. Frozen sections can be very active with antigen since they don't go thru all the processing related to ffpe tissue, so in my experience you need to carefully titer your IHC antibody reagents often diluting much more than you would for ffpe tissue. Also much of the endogenous issues, such as biotin, peroxidase and alk.phos. Are inhanced in frozens, or preserved more than in ffpe tissue. AB block, fc receptor block, serum block before the primary and before the secondary are all things I consider with frozen IHC, or I try to avoid the biotin issue altogether by using a non biotin labeled polymer detection. Maybe this person is inexperienced but we all started somewhere, so probably with just a little help we can make a big difference for this person, that is what we are here for, right? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Wednesday, November 09, 2005 2:28 PM To: Till, Renee; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] any suggestions? With all due respect I think that your colleague's procedure is a royal mess. On the other hand it seems that he is not very willing to change his ways, so the only thing I have to tell you (for him) is: good luck! He should be the one doing the asking, and not you! Rene J. "Till, Renee" wrote: Another tech who does not have much experience with histology came to me with questions about his immunos. They are doing IHC with various cd markers on frozen sections of mouse aorta. He has encountered particularly strong background(or so he's been told, he thinks it is actual staining) with one of the antibodies that was made in rat. He is using a goat anti-rat F(ab')2 from Jackson as the secondary. It is not absorbed against mouse. I have asked all about his dilutions and incubations times, but he doesn't seem to think that is the problem. I gave him an avidin/biotin block to try and see if that helps. Any other ideas? I am not familiar with cd markers myself. The only problem I could find just in talking to him was that he was blocking with rabbit serum? I told him you normally match your block with the host of the secondary, but would that make that big a difference as far as background is concerned? Would the fact he is using frozen sections have anything to do with it? Or could it just be the stain? I know they are doing cd54, but I'm not sure if this is the one he is having a problem with. I know this is not much informations, but I would still appreciate any input Thanks, Renee' ============================================================================ ==================== Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ============================================================================ ==================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Thu, 10 Nov 2005 11:23:46 -0500 From: Pamela Marcum Subject: RE: [Histonet] any suggestions? To: "Hofecker, Jennifer L" , "histonet" Message-ID: <6.1.1.1.2.20051110112330.019a6c60@mail.vet.upenn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Thanks, Jennifer. Pam At 11:03 AM 11/10/2005, Hofecker, Jennifer L wrote: >Well said Pam and Gayle! >I know that every time I have a problem, my first thought is "ask >histonet" not because I am lazy or because I can't look it up. I think >first of histonet because of the diversity of our listmembers and their >experience. Just because a publication lists the steps of a protocol, >doesn't mean it will work. I would rather get my info from the ones who >make these protocols work. >On the other hand, every time I am about to post a query, I have to read >it 6 times to make sure there isn't anything in the post that I will be >chastised for. While I agree that we should not "post with reckless >abandon", I do not think we should be terrified of backlash for asking how >to do something we've never done before, especially if it affects patient >care, whether directly or indirectly. > >Well, I am going to put on my bullet proof vest! >Thanks for listening, >Jennifer > > > >Jennifer Hofecker, HT (ASCP) >Vanderbilt University Medical Center >Division of Neuropathology >(615) 343-0083 >(615) 343-7089 fax > >Thank You Gayle!! It is sometimes easy to forget not everyone has access >to the Internet or a way to even place a message on HistoNet. We are >attempting to assist people who are learning and while some may seem to be >using HistoNet as way to avoid looking anything up the majority are asking >questions that are not readily accessible or in an area like murine animal >models that is more difficult. I have no problem with this and will answer >most questions off line if it is a more esoteric question. I am surprised >that the person with the issue about Rene's assistance for her friend is >also the same Rene who has been asking all of us to contribute to Tricks of >the Trade. > >Pamela A Marcum >Special Procedures Manager >Bone Histology >UPENN Vet School > > >At 05:33 PM 11/9/2005, Gayle Callis wrote: > >Rene, > > > >Sorry to disagree, and I certainly did not understand what YOU meant by > >"changing his ways"? I couldn't see many changes here, other than some > >refinement of his IHC method. Many people working with murine CD marker > >IHC often need a little help with murine animal model IHC. Since our > >lab works with murine CD marker IHC approx. 95% of the time, on frozen > >sections etc per the inquiry, we are happy to sleuth problems. The > >questions were valid ones for learning/refining of his techniques and > >being a mousie person, I didn't have a problem with the inquiry. > > > >Some people (including a lot of experts in our field) do NOT want to be on > >Histonet and second hand messaging certainly doesn't bother me as long as > >more details are provided. I frequently ask questions on Histonet to > >access information for others in my department - that's part of my job. > >Your colleague is welcome to email privately if you or they wish. > > > >Gayle Callis > >Research Histopathology Supervisor > >Veterinary Molecular Biology > >Montana State University - Bozeman > >PO Box 173610 > >Bozeman MT 59717-3610 > >406 994-6367 > >406 994-4303 (FAX) > > > > > > > > > > > > > >- At 02:28 PM 11/9/2005, you wrote: > >>With all due respect I think that your colleague's procedure is a royal > mess. > >>On the other hand it seems that he is not very willing to change his > >>ways, so the only thing I have to tell you (for him) is: good luck! He > >>should be the one doing the asking, and not you! > >>Rene J. > > > >Gayle Callis > >Research Histopathology Supervisor > >Veterinary Molecular Biology > >Montana State University - Bozeman > >PO Box 173610 > >Bozeman MT 59717-3610 > >406 994-6367 > >406 994-4303 (FAX) > > > > > > > >_______________________________________________ > >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 Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu ------------------------------ Message: 4 Date: Thu, 10 Nov 2005 11:56:13 -0500 From: John.Sheppard@Health-Partners.org Subject: [Histonet] Process improvement monitors To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hello "Histonetters", Once again I am on a "mission" for my medical director. I am hoping to find out if anyone out there has been looking at quality management / process improvement areas in histology / pathology labs in their community hospitals. We are currently reworking our P.I. program, and we are looking at some of the most obvious errors we find. It seems to us that many mistakes come to our lab from doctors offices, the O.R. and our birthing center. These errors come in the form of incomplete or no requisition, discrepancies between specimen labels and requisitions, and no or incomplete label on specimens. These are the issues that are of particular interest to my director. We are trying to establish a reasonable range to strive for. Ideally we would like to have all specimens labelled correctly with the correct reqistion and full clinical history / diagnosis coming in to our lab, but realistically we are not sure if that is possible. If anyone has reviewed these monitors and does not mind sharing their information, we would greatly appreciate using it for comparitive data purposes. We have been monitoring this for a couple of years and we do not have a sky high rate, but it is higher than we believe it should be. Thanks again John Sheppard HT(ASCP) P.S. If anyone has ideas on how to improve this performance, short of firing nuses everytime they mess up please let me know ? (this is a joke) CONFIDENTIALITY NOTICE: This message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 5 Date: Thu, 10 Nov 2005 12:18:36 -0500 From: "Monfils, Paul" Subject: RE: [Histonet] tonsils To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717610@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" Years ago, when I worked in a clinical histology lab (I am now in research), we processed and sectioned all tonsils. Since no serious pathology had ever been discovered in any of the thousands of subadult tonsils we had processed, the department was considering a "gross only" policy for tonsils from patients under 16 years of age. Don't you know, that very month they found an undiagnosed lymphoma in the tonsils of a 12 year old girl. We continued processing all tonsils. As I said, that was quite a few years ago, and they may have changed the policy since then. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Linda > Blazek > Sent: Thursday, November 10, 2005 7:54 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] tonsils > > I need some information on how other facilities are handling tonsils. Are > they gross only and what criteria is used to have microscopic performed on > them? Thanks for any info. - Linda > > > Linda Blazek, HT (ASCP) > Department of Pathology > Children's Medical Center > 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 > > ------------------------------ Message: 6 Date: Thu, 10 Nov 2005 12:20:00 -0500 From: "O'Reilly, Siobhan" Subject: RE: [Histonet] (no subject) To: Histonet@lists.utsouthwestern.edu Message-ID: <4BCB18390D553D41A3FFEB77D5A3BA97012DB631@excnyuebw2k63.nyumc.org> Content-Type: text/plain; charset=iso-8859-1 Please remove mine as well. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of drosini@comcast.net Sent: Thursday, November 10, 2005 6:54 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) please take my name off the histonet list _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------- This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email. ============================================================================== ------------------------------ Message: 7 Date: Thu, 10 Nov 2005 12:48:12 -0500 From: "Bernadette Weston" Subject: [Histonet] reproducing microwave temperatures To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed A few years back I attended a convention taht did a workshop on how to check your microwave for reproducible tempertures. I remember having a worksheet for that seminar, does anyone have a method they like to share for this monitoring? Bernadette Weston HT Barberton Citizens Hospital ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 24, Issue 15 **************************************** ---------------------- Important news about Holy Cross email communications! Within 30 days, Holy Cross Hospital will be implementing a secure email policy. You may receive a ZixMail Secure Message with a link to view your message. To access your message follow the three easy steps below: 1. Click on the link provided in the notification email 2. Create a password 3. Click ?submit? If you need assistance in accessing your Secure message, click on the link below http://www.zixit.com/support/zixmailnethelp.htm#4.0 or contact ZixCorp Support at support@zixcorp.com General questions or concerns related to the implementation of this new policy at Holy Cross can be forwarded to ishelpdesk@holy-cross.com. ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From GoodwinD <@t> pahosp.com Thu Nov 10 13:47:27 2005 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Thu Nov 10 13:47:48 2005 Subject: [Histonet] IL-5 and Eotaxin Message-ID: <80CDD9C3FEEAFD4982B114C4A6DFD00E256EC3@uphsmbx2.UPHS.PENNHEALTH.PRV> Anyone out there using IL-5 or Eotaxin antibodies on human tissue? If so, with which detection kit/system, and at what dilution? Also, what works best as a positive control? Thanks, Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com From rjbuesa <@t> yahoo.com Thu Nov 10 14:34:01 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 10 14:34:10 2005 Subject: [Histonet] Using antibodies beyond their expiration dates. Message-ID: <20051110203401.74207.qmail@web61221.mail.yahoo.com> Cesar, Susan et alli: Digging amongst my papers I found the 2 referneces I wrote Cesar this morning: 1- Is a paper by Balaton et alli, published in Applied Immunohistochemistry & Molecular Morphology 7(3):221-225, 1999. Using standard protocols they tested 65 antibodies (Ab) that had expired 6 to 134 months previously (average of 33). Concluded that all Ab performed satisfactorily beyond the recommended expiration dates. 2- (And this is more important because it was published as part of a CAP Laboratory Improvement Program) by Tubbs et alli in Arch Pathol Lab Med, Vol 122: 1051-1052, 1998 tested a limited number of Abs, but in the test intervened221 different laboratories with the following conclusions:"The data suggest review of the Health Care Financing Administration's ruling on extending the useful reagent shelf life beyond manufacturers recommendations". Both are good news for those who want (or need) for economical reasons to extend the time frame of your Abs beyond the manufacturers' date. At least no CAP inspector I think will go against their own conclusions. So now you know! Rene J. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From rjbuesa <@t> yahoo.com Thu Nov 10 14:38:21 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 10 14:38:29 2005 Subject: [Histonet] Fwd: Using antibodies beyond their expiration dates. Message-ID: <20051110203821.9766.qmail@web61213.mail.yahoo.com> E-mail addrsses corrected! Note: forwarded message attached. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From rjbuesa <@t> yahoo.com Thu Nov 10 14:57:08 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 10 14:57:19 2005 Subject: [Histonet] antibody storage In-Reply-To: <3.0.6.32.20051110204201.008abb80@pop.lugo.usc.es> Message-ID: <20051110205709.5550.qmail@web61216.mail.yahoo.com> Pablo: I just mailed you some information I have just posted about use of Abs beyond their "expiration dates". Rene J. Pablo S?nchez Quinteiro wrote: Dear Histonetters, Lately I am having difficulties with the storage of my antibodies. I keep them at -20?C in undiluted aliquots. After few months a kind of web or lattice grows in the liquid. It is evident after unfreezening. The antibody keeps working but I am afraid this is not any pleasant. I would very greatful with your input. By the way the provider states an expiring date of only six months. Is this reasonable? Best regards from Spain Pablo Sanchez _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From ploykasek <@t> phenopath.com Thu Nov 10 15:08:04 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Thu Nov 10 15:08:30 2005 Subject: [Histonet] Expired Antibodies. Message-ID: While I have often used and believe in using expired antibodies that are performing correctly, this is no longer an option under CAP. The rule is as follows: ANP.22432 Phase II N/A YES NO Are all immunohistochemical reagents used within their indicated expiration dates? COMMENTARY: N/A REFERENCE: Department of Health and Human Services, Centers for Medicare and Medicaid Services. Clinical laboratory improvement amendments of 1988; final rule. Fed Register. 2003(Jan 24):7164 [42CFR493.1252(c)]. This can be an expensive proposition. I have often thought maybe labs in a general area or city could split ordering seldom used antibodies & share aliquots between themselves. Just a thought. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From japoteete <@t> saintfrancis.com Thu Nov 10 15:34:25 2005 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Thu Nov 10 15:35:41 2005 Subject: [Histonet] Using antibodies beyond their expiration dates. Message-ID: When I called a couple of months ago to get a clarification on this question, I was told that the question stands as printed in the April 28, 2005 Checklist, and that it was no longer open for interpretation otherwise, regardless of previous studies. I'm not going to push my luck with it, as a friend who works in a different state was given a deficiency because of an antibody which had expired less than one month previously. I understand that the inspectors are being just as strict for ANY reagent these days. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK -----Original Message----- From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Thursday, November 10, 2005 2:34 PM To: histonet@lists.utsouthwestern.edu; chesarato@htmail.com; sbryant@labpath.com Subject: [Histonet] Using antibodies beyond their expiration dates. Cesar, Susan et alli: Digging amongst my papers I found the 2 referneces I wrote Cesar this morning: 1- Is a paper by Balaton et alli, published in Applied Immunohistochemistry & Molecular Morphology 7(3):221-225, 1999. Using standard protocols they tested 65 antibodies (Ab) that had expired 6 to 134 months previously (average of 33). Concluded that all Ab performed satisfactorily beyond the recommended expiration dates. 2- (And this is more important because it was published as part of a CAP Laboratory Improvement Program) by Tubbs et alli in Arch Pathol Lab Med, Vol 122: 1051-1052, 1998 tested a limited number of Abs, but in the test intervened221 different laboratories with the following conclusions:"The data suggest review of the Health Care Financing Administration's ruling on extending the useful reagent shelf life beyond manufacturers recommendations". Both are good news for those who want (or need) for economical reasons to extend the time frame of your Abs beyond the manufacturers' date. At least no CAP inspector I think will go against their own conclusions. So now you know! Rene J. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From anh2006 <@t> med.cornell.edu Thu Nov 10 17:36:22 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Thu Nov 10 17:31:24 2005 Subject: [Histonet] any suggestions? In-Reply-To: References: Message-ID: Dear Renee, It is going to be essential that his secondary antibody be cross-adsorbed against mouse even though his primary is anti-F(ab')2. When I switched over the non-cross reacting secondaries my whole quality of staining mouse tissue improved a 1000%. In addition, I suggest he run a panel of controls if he hasn't already including: (1) no primary but with secondary and strep-HRP (non-specific secondary binding control) (2) isotype control with secondary and strep-HRP (primary FcR interaction and non-specific binding control) (3) Only strep-HRP (biotin control) (4) Only DAB (peroxidase control) Cheers, Andrea At 1:13 PM -0600 11/9/05, Till, Renee wrote: >Another tech who does not have much experience with histology came to me >with questions about his immunos. They are doing IHC with various cd >markers on frozen sections of mouse aorta. He has encountered >particularly strong background(or so he's been told, he thinks it is >actual staining) with one of the antibodies that was made in rat. He is >using a goat anti-rat F(ab')2 from Jackson as the secondary. It is not >absorbed against mouse. I have asked all about his dilutions and >incubations times, but he doesn't seem to think that is the problem. I >gave him an avidin/biotin block to try and see if that helps. Any other >ideas? I am not familiar with cd markers myself. The only problem I >could find just in talking to him was that he was blocking with rabbit >serum? I told him you normally match your block with the host of the >secondary, but would that make that big a difference as far as >background is concerned? Would the fact he is using frozen sections >have anything to do with it? Or could it just be the stain? I know they >are doing cd54, but I'm not sure if this is the one he is having a >problem with. > >I know this is not much informations, but I would still appreciate any >input > >Thanks, > >Renee' > -- From Eric <@t> ategra.com Thu Nov 10 19:40:08 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Nov 11 04:54:24 2005 Subject: [Histonet] Histo tech jobs Hiring immediately Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. This is a follow up e-mail with the newest jobs listed first(1-5) (I have a few travel/temp HistoTech positions as well - see below). Most of my Histo Techs positions are clinical, although I do have a few Research Histo Tech positions as well. Here are some of my Hottest jobs: 1. Virginia Opening for a Histotech(bench) No weekends, No Call, Top dollar 2. Florida(Greater Florida area) Openings for Bench Histotech No weekends, No call 3. Ohio Opening for Histotechs(bench, NightShift and Dayshift) No Weekends, No Call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 4. Ohio Opening for CytoTech No Weekends, No Call, Top Dollar 5. Tennessee Opening for a Histotech Great Location, No Call, No Weekends, Top Dollar 6.Iowa Openings for a HistoTech(Bench) No weekends, No call, and Top Dollar. 7. Ohio(Dayton Area) HistoTech Manager No Weekends, No call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 8. Colorado (Greater Denver area) Opening for Several Bench Positions(Night Shift) No weekends, No call, Top Dollar 9. Georgia Openings for Histotech(Temp to Perm) No Weekends, No Call, Top Dollar, Excellent Benefits if you are interested, please call me today at 1-800-466-9919 ext 223 10. Pennsylvania(Philadelphia area) Openings for HistoTech(Bench) No Weekends, No Call, Top Dollar 11.Rhode Island Openings for Histotech(Bench) No weekends, No call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 12. California( Southern) HistoTech Supervisor(2nd shift) Openings for Histotechs(Bench,2nd shift) Great Location, No weekends, No call, Top Dollar 13. California(Southern) Openings for HistoTechs(Bench) Great Location, No weekends, No call, Top Dollar 14. Pennsylvania(Multiple jobs in greater Pittsburgh area) HistoTech opening(Bench) if you are interested, please call me today at 1-800-466-9919 ext 223 15. Illinois(Multiple jobs in Greater Illinois area) Opening for Histotech(Bench) 16. Michigan(Multiple jobs in Greater Michigan area) Openings for Histotechs(Bench) 17. Washington State(Eastern) Opening for Lead HistoTech(Bench) No weekends, No call if you are interested, please call me today at 1-800-466-9919 ext 223 18. Oklahoma Opening for Histotech(Bench) No weekends, No call. 19. Massachusetts (Several Opportunities in the Boston Area) Part time Histo Tech(Permanent) No weekends, No call 20. New Mexico openings for Supervisor and Bench Techs No Weekends, No Call, Excellent Benefits.. 21. Nebraska Openings for Histo Techs(Bench) No Weekends, No Call, Top Dollar 22. Virginia(Western Virginia) Opening for Bench Histotech No weekends, No call if you are interested, please call me today at 1-800-466-9919 ext 223 23. Ohio( Southern) Opening for Bench HistoTech No weekends, No call 24. Missouri(Greater Missouri area) Opening for Bench Histotech No weekends, No call 25. Wisconsin(Eastern area) Opening for a Bench Histotech No weekends, No call 26. Florida(Greater Florida area) Openings for Bench Histotech No weekends, No call The clients are currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 From Nancy.Temple <@t> ssfhs.org Fri Nov 11 07:46:49 2005 From: Nancy.Temple <@t> ssfhs.org (Temple Nancy) Date: Fri Nov 11 07:47:01 2005 Subject: [Histonet] Bone Saw Message-ID: We are currently using a bone saw made by Mar-Med. We do a lot of bones, and are in need of a heavier duty bone saw. We have had to replace the motor twice this year on the Mar-Med. Any suggestions? Thanks, Nancy St. Francis Hospital Indianapolis, In The information contained in this e-mail and any accompanying documents is intended for the sole use of the recipient to whom it is addressed, and may contain information that is privileged, confidential, and prohibited from disclosure under applicable law. If you are not the intended recipient, or authorized to receive this on behalf of the recipient, you are hereby notified that any review, use, disclosure, copying, or distribution is prohibited. If you are not the intended recipients(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. From donna <@t> milestonemed.com Fri Nov 11 08:36:38 2005 From: donna <@t> milestonemed.com (Donna Willis) Date: Fri Nov 11 08:36:49 2005 Subject: [Histonet] Back on Line Message-ID: <1131719802_6594@osiris.serverlogistics.com> For those that did not already know, I have changed jobs. After 35 years in the lab, and 22 of them as a manager, I have moved to the vendor side of the business. I started my position with Milestone Medical the last week in October. Everyone I know will understand the move considering my passion of Microwave processing. I can be reached by e-mail at donna@milestonemed.com or by phone at 214-725-6184. My training and demo lab will be operational by the 1st of the year. If anyone has any microwave questions, please feel free to contact me. Glad to be back on the histonet. Donna Willis Milestone Medical North American Application Manager Gulf Cost Sales Rep. From donna <@t> milestonemed.com Fri Nov 11 08:40:31 2005 From: donna <@t> milestonemed.com (Donna Willis) Date: Fri Nov 11 08:40:45 2005 Subject: [Histonet] reproducing microwave temperatures In-Reply-To: Message-ID: <1131720035_6596@osiris.serverlogistics.com> Bernadette, I will be glad to send you the procedure that I have given out in the microwave workshops that I have given in the past. What type of instrument are you using, household or laboratory? If laboratory, what brand. I say this because, Milestone RHS instruments perform the wattage checks a little different that others. Let me know if you have any other questions. Donna Willis Milestone Medical North American Application Manager -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bernadette Weston Sent: Thursday, November 10, 2005 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] reproducing microwave temperatures A few years back I attended a convention taht did a workshop on how to check your microwave for reproducible tempertures. I remember having a worksheet for that seminar, does anyone have a method they like to share for this monitoring? Bernadette Weston HT Barberton Citizens Hospital _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From d.fuster <@t> ub.edu Fri Nov 11 09:26:54 2005 From: d.fuster <@t> ub.edu (Dolors Fuster) Date: Fri Nov 11 09:27:07 2005 Subject: [Histonet] Vocal Cords Message-ID: <4374B83E.3010307@ub.edu> Hello histonetters We are thinking about demonstrating slow and fast contraction muscular fibers in vocal cords. What do you think are the most advisable methods? Thanks in advance. Dolors Fuster Botella T?cnic Especialista en Anatomia Patologica i Citologia Dept. Anatomia i Embriologia Humana Universitat de Barcelona. Barcelona. Catalunya. Espa?a From MDiCarlo <@t> KaleidaHealth.Org Fri Nov 11 09:52:56 2005 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Nov 11 09:51:17 2005 Subject: [Histonet] Bone Saw Message-ID: <139141F8BAF4A642A945ECC528511AF001F5ED49@kalmb02.kaleidahealth.org> I would also be interested in a heavy duty band saw since I work with large human bones. When I looked into the Mar-Med I felt it was not sturdy or large enough to meet my lab needs. I also would appreciate any suggestions. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Temple Nancy Sent: Friday, November 11, 2005 08:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone Saw We are currently using a bone saw made by Mar-Med. We do a lot of bones, and are in need of a heavier duty bone saw. We have had to replace the motor twice this year on the Mar-Med. Any suggestions? Thanks, Nancy St. Francis Hospital Indianapolis, In The information contained in this e-mail and any accompanying documents is intended for the sole use of the recipient to whom it is addressed, and may contain information that is privileged, confidential, and prohibited from disclosure under applicable law. If you are not the intended recipient, or authorized to receive this on behalf of the recipient, you are hereby notified that any review, use, disclosure, copying, or distribution is prohibited. If you are not the intended recipients(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. _______________________________________________ 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 tyler-wellington <@t> northwestern.edu Fri Nov 11 09:53:52 2005 From: tyler-wellington <@t> northwestern.edu (Tyler W.) Date: Fri Nov 11 09:54:00 2005 Subject: [Histonet] DPT3 Message-ID: <20051111155352.505F09E847@merle.it.northwestern.edu> I am processing rat ovary samples and was wondering if anyone has a protocol for sectioning and developing slides using DPT3. Any information you have is verrrrry much apreciated. Thank you verry much--Tyler W. From juan.gutierrez <@t> christushealth.org Fri Nov 11 10:10:03 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Nov 11 10:10:14 2005 Subject: [Histonet] Bone Saw Message-ID: Try Cabelas.com they have a butcher's band saw for about $550.00. It might sound "cheap", but you have to realize it is not a scientific supply company. If it cuts elk it should work fine in humans. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DiCarlo, Margaret Sent: Friday, November 11, 2005 9:53 AM To: 'Temple Nancy'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Bone Saw I would also be interested in a heavy duty band saw since I work with large human bones. When I looked into the Mar-Med I felt it was not sturdy or large enough to meet my lab needs. I also would appreciate any suggestions. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Temple Nancy Sent: Friday, November 11, 2005 08:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone Saw We are currently using a bone saw made by Mar-Med. We do a lot of bones, and are in need of a heavier duty bone saw. We have had to replace the motor twice this year on the Mar-Med. Any suggestions? Thanks, Nancy St. Francis Hospital Indianapolis, In The information contained in this e-mail and any accompanying documents is intended for the sole use of the recipient to whom it is addressed, and may contain information that is privileged, confidential, and prohibited from disclosure under applicable law. If you are not the intended recipient, or authorized to receive this on behalf of the recipient, you are hereby notified that any review, use, disclosure, copying, or distribution is prohibited. If you are not the intended recipients(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. _______________________________________________ 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 froyer <@t> bitstream.net Fri Nov 11 10:14:40 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Nov 11 10:14:58 2005 Subject: [Histonet] Bone Saw In-Reply-To: <139141F8BAF4A642A945ECC528511AF001F5ED49@kalmb02.kaleidahealth.org> Message-ID: <005e01c5e6db$0589a420$0e00a8c0@fords> My suggestion is to check at your local Sears store. Our lab used a Sears Craftsman 10" band saw purchased off the shelf for under $150.00. It was a bench model and did not take up much room. That was 25 years ago and the last I heard, they are still using it. ~ Ford Ford M. Royer, MT(ASCP) Sales Manager, Histology Product Division Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Golden Valley, MN 55427-3601 763-542-8725 phone 888-790-9686 toll free 763-546-4830 fax email web -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DiCarlo, Margaret Sent: Friday, November 11, 2005 9:53 AM To: 'Temple Nancy'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Bone Saw I would also be interested in a heavy duty band saw since I work with large human bones. When I looked into the Mar-Med I felt it was not sturdy or large enough to meet my lab needs. I also would appreciate any suggestions. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Temple Nancy Sent: Friday, November 11, 2005 08:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone Saw We are currently using a bone saw made by Mar-Med. We do a lot of bones, and are in need of a heavier duty bone saw. We have had to replace the motor twice this year on the Mar-Med. Any suggestions? Thanks, Nancy St. Francis Hospital Indianapolis, In The information contained in this e-mail and any accompanying documents is intended for the sole use of the recipient to whom it is addressed, and may contain information that is privileged, confidential, and prohibited from disclosure under applicable law. If you are not the intended recipient, or authorized to receive this on behalf of the recipient, you are hereby notified that any review, use, disclosure, copying, or distribution is prohibited. If you are not the intended recipients(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. _______________________________________________ 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 pruegg <@t> ihctech.net Fri Nov 11 11:17:32 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Nov 11 11:17:44 2005 Subject: [Histonet] Bone Saw In-Reply-To: <139141F8BAF4A642A945ECC528511AF001F5ED49@kalmb02.kaleidahealth.org> Message-ID: <200511111717.jABHHVxT008027@chip.viawest.net> I have the Mar-med and have used it to cut some pretty large bones (human heads of femur) you can adjust the table height or take it out so that larger samples than you think can pass. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DiCarlo, Margaret Sent: Friday, November 11, 2005 8:53 AM To: 'Temple Nancy'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Bone Saw I would also be interested in a heavy duty band saw since I work with large human bones. When I looked into the Mar-Med I felt it was not sturdy or large enough to meet my lab needs. I also would appreciate any suggestions. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Temple Nancy Sent: Friday, November 11, 2005 08:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone Saw We are currently using a bone saw made by Mar-Med. We do a lot of bones, and are in need of a heavier duty bone saw. We have had to replace the motor twice this year on the Mar-Med. Any suggestions? Thanks, Nancy St. Francis Hospital Indianapolis, In The information contained in this e-mail and any accompanying documents is intended for the sole use of the recipient to whom it is addressed, and may contain information that is privileged, confidential, and prohibited from disclosure under applicable law. If you are not the intended recipient, or authorized to receive this on behalf of the recipient, you are hereby notified that any review, use, disclosure, copying, or distribution is prohibited. If you are not the intended recipients(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. _______________________________________________ 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 pruegg <@t> ihctech.net Fri Nov 11 11:45:41 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Nov 11 11:45:55 2005 Subject: [Histonet] Expired antibodies Message-ID: <200511111745.jABHjexT014530@chip.viawest.net> Please if anyone finds themselves in the position where they will be disgarding expired antibodies because of CAP rules, let me take them off your hands. I will pay for shipping. Thank you, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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 xli <@t> amgen.com Fri Nov 11 11:50:01 2005 From: xli <@t> amgen.com (Li, Xiaodong) Date: Fri Nov 11 11:50:11 2005 Subject: FW: [Histonet] (no subject) Message-ID: Please remove mine too, thanks very much. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of O'Reilly, Siobhan Sent: Thursday, November 10, 2005 9:20 AM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] (no subject) Please remove mine as well. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of drosini@comcast.net Sent: Thursday, November 10, 2005 6:54 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) please take my name off the histonet list _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- --- This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Fri Nov 11 13:54:15 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Nov 11 13:54:26 2005 Subject: [Histonet] Bone Saw Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717612@lsexch.lsmaster.lifespan.org> I have also used a 10" craftsman band saw from Sears for about 15 years, and it is still going strong. I use a fine tooth (14 teeth per inch) metal cutting blade. The wood cutting blades are too coarse. I use it not only for cutting bone but also for trimming methyl methacrylate blocks, and for innumerable other odds and ends that come up in the lab. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ford > Royer > Sent: Friday, November 11, 2005 8:14 AM > To: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Bone Saw > > My suggestion is to check at your local Sears store. Our lab used a Sears > Craftsman 10" band saw purchased off the shelf for under $150.00. It was > a > bench model and did not take up much room. That was 25 years ago and the > last I heard, they are still using it. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Sales Manager, Histology Product Division > Minnesota Medical Specialists, Inc. > 7177 Madison Ave. W. > Golden Valley, MN 55427-3601 > 763-542-8725 phone > 888-790-9686 toll free > 763-546-4830 fax > email > web > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DiCarlo, > Margaret > Sent: Friday, November 11, 2005 9:53 AM > To: 'Temple Nancy'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Bone Saw > > I would also be interested in a heavy duty band saw since I work with > large > human bones. When I looked into the Mar-Med I felt it was not sturdy or > large enough to meet my lab needs. > > I also would appreciate any suggestions. > > Peggy DiCarlo HT (ASCP) > Orthopedics Bone Lab > Buffalo General Hospital > 100 High St. > Buffalo, NY 14203 > 716-859-1293 > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Temple > Nancy > Sent: Friday, November 11, 2005 08:47 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Bone Saw > > > We are currently using a bone saw made by Mar-Med. We do a lot of bones, > and > are in need of a heavier duty bone saw. We have had to replace the motor > twice this year on the Mar-Med. Any suggestions? > Thanks, > > Nancy > St. Francis Hospital > Indianapolis, In > > > The information contained in this e-mail and any accompanying documents is > intended for the sole use of the recipient to whom it is addressed, and > may > contain information that is privileged, confidential, and prohibited from > disclosure under applicable law. If you are not the intended recipient, or > authorized to receive this on behalf of the recipient, you are hereby > notified that any review, use, disclosure, copying, or distribution is > prohibited. If you are not the intended recipients(s), please contact the > sender by e-mail and destroy all copies of the original message. Thank > you. > _______________________________________________ > 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From MVaughan4 <@t> ucok.edu Fri Nov 11 13:57:51 2005 From: MVaughan4 <@t> ucok.edu (MVaughan4@ucok.edu) Date: Fri Nov 11 13:58:20 2005 Subject: [Histonet] Antibodies beyond expiration In-Reply-To: Message-ID: I have had success using antibodies beyond their expiration point. If you have to get rid of your antibodies because of regulations, you might consider donating them to small colleges and universities in your area. There is more of a demand for undergraduate research these days at such colleges and it is difficult to find the resources to purchase many supplies. And the schools would be very appreciative of your help. Mel Melville B. Vaughan, Ph. D. Assistant Professor Department of Biology University of Central Oklahoma Edmond, OK 73034 From anh2006 <@t> med.cornell.edu Fri Nov 11 14:29:25 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Nov 11 14:24:26 2005 Subject: [Histonet] Antibodies beyond expiration In-Reply-To: References: Message-ID: I agree, great idea. Many antibodies can most definitely be used after their expiration date. Case in point, I recently used an anti-CD31 antibody clone MEC13.3 from RDI which was probably 5 years old - way past it's expiry date - and it worked as beautifully on my acetone fixed frozens as it did on the day I got it. It had been stored (as all my antibodies are) at 4 deg C. So this won't be the case for all of them, but certainly many can still be used. At 1:57 PM -0600 11/11/05, MVaughan4@ucok.edu wrote: >I have had success using antibodies beyond their expiration point. If you >have to get rid of your antibodies because of regulations, you might >consider donating them to small colleges and universities in your area. >There is more of a demand for undergraduate research these days at such >colleges and it is difficult to find the resources to purchase many >supplies. And the schools would be very appreciative of your help. >Mel > >Melville B. Vaughan, Ph. D. >Assistant Professor >Department of Biology >University of Central Oklahoma >Edmond, OK 73034 -- From jnocito <@t> pathreflab.com Fri Nov 11 14:57:02 2005 From: jnocito <@t> pathreflab.com (Joe Nocito) Date: Fri Nov 11 14:57:19 2005 Subject: [Histonet] Using antibodies beyond their expiration dates. In-Reply-To: <20051110203401.74207.qmail@web61221.mail.yahoo.com> Message-ID: Rene, Sorry to disagree with you. This January, I had my CAP inspection and the inspector told us that we couldn't use reagents beyond the expiration date. I had a written procedure to validate each antibody, plus a drawer filled with stained slides that had the test date on them. It wasn't accepted and resulted in a deficiency. 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 Rene J Buesa Sent: Thursday, November 10, 2005 2:34 PM To: histonet@lists.utsouthwestern.edu; chesarato@htmail.com; sbryant@labpath.com Subject: [Histonet] Using antibodies beyond their expiration dates. Cesar, Susan et alli: Digging amongst my papers I found the 2 referneces I wrote Cesar this morning: 1- Is a paper by Balaton et alli, published in Applied Immunohistochemistry & Molecular Morphology 7(3):221-225, 1999. Using standard protocols they tested 65 antibodies (Ab) that had expired 6 to 134 months previously (average of 33). Concluded that all Ab performed satisfactorily beyond the recommended expiration dates. 2- (And this is more important because it was published as part of a CAP Laboratory Improvement Program) by Tubbs et alli in Arch Pathol Lab Med, Vol 122: 1051-1052, 1998 tested a limited number of Abs, but in the test intervened221 different laboratories with the following conclusions:"The data suggest review of the Health Care Financing Administration's ruling on extending the useful reagent shelf life beyond manufacturers recommendations". Both are good news for those who want (or need) for economical reasons to extend the time frame of your Abs beyond the manufacturers' date. At least no CAP inspector I think will go against their own conclusions. So now you know! Rene J. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jxw387 <@t> psu.edu Fri Nov 11 15:10:32 2005 From: jxw387 <@t> psu.edu (weijianwen) Date: Fri Nov 11 15:11:28 2005 Subject: [Histonet] bone histomorphometric analysis References: <20051111155352.505F09E847@merle.it.northwestern.edu> Message-ID: <001501c5e704$5a7a3110$75ec7680@weijianwen> Dear all, I am looking for some labs or companys which can provide bone histomorphometric analysis service for mice samples. Dose anyone can give me some information? Thanks in advance! Jianwen From rjbuesa <@t> yahoo.com Fri Nov 11 15:46:35 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 11 15:46:43 2005 Subject: Fwd: RE: [Histonet] Using antibodies beyond their expiration dates. Message-ID: <20051111214635.55599.qmail@web61221.mail.yahoo.com> Note: forwarded message attached. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From ihctech2000 <@t> yahoo.com Fri Nov 11 18:59:07 2005 From: ihctech2000 <@t> yahoo.com (Sun Zhon) Date: Fri Nov 11 18:59:15 2005 Subject: [Histonet] IgG detection for mouse tissue? Message-ID: <20051112005907.47187.qmail@web50211.mail.yahoo.com> Hi All, Does any one know whether there are antibodies to detect mouse IgG on mouse tissues? Thank you for the help. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From Abizar.A.Lakdawalla <@t> appliedbiosystems.com Fri Nov 11 19:04:40 2005 From: Abizar.A.Lakdawalla <@t> appliedbiosystems.com (Abizar A Lakdawalla) Date: Fri Nov 11 19:05:05 2005 Subject: [Histonet] IgG detection for mouse tissue? In-Reply-To: <20051112005907.47187.qmail@web50211.mail.yahoo.com> Message-ID: Any Anti-mouse secondary antibody (link) in IHC kits will detect mouse IgG in mouse tissues. Abizar Sun Zhon Sent by: histonet-bounces@lists.utsouthwestern.edu 11/11/2005 04:59 PM To HistoList HistoList cc Subject [Histonet] IgG detection for mouse tissue? Hi All, Does any one know whether there are antibodies to detect mouse IgG on mouse tissues? Thank you for the help. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Fri Nov 11 19:34:16 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Nov 11 19:34:27 2005 Subject: [Histonet] IgG detection for mouse tissue? In-Reply-To: <20051112005907.47187.qmail@web50211.mail.yahoo.com> Message-ID: <000001c5e729$31a9e3a0$a7d48a80@AMY> I just did this for a client on mouse joints with arthritis. I used vectors biotinylated horse anti-mouse IgG (BA-2001) then streptavidin and then DAB, turned out great. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sun Zhon Sent: Friday, November 11, 2005 5:59 PM To: HistoList HistoList Subject: [Histonet] IgG detection for mouse tissue? Hi All, Does any one know whether there are antibodies to detect mouse IgG on mouse tissues? Thank you for the help. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ben.Shelkowsky <@t> chomp.org Sat Nov 12 16:32:30 2005 From: Ben.Shelkowsky <@t> chomp.org (Shelkowsky, Ben) Date: Sat Nov 12 16:35:13 2005 Subject: [Histonet] H. Pylori Controls References: Message-ID: <40AA34F14CA09F429945413E05DED375281F56@EXCHANGE.chomp.org> We buy from American Master Tech (1800-860 4073) 25 slides/pack ; Cat# CSHEL25 ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Vicki Gauch Sent: Mon 11/7/2005 10:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H. Pylori Controls Does anyone know of a good source for H. Pylori Controls?? We normally order from Newcomer Supply but they are experiencing a large backorder right now and we need something to hold us over until they arrive. Any help would be greatly appreciated... Thanks so much, Vicki Gauch AMCH Albany, NY ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 Ben.Shelkowsky <@t> chomp.org Sat Nov 12 16:41:52 2005 From: Ben.Shelkowsky <@t> chomp.org (Shelkowsky, Ben) Date: Sat Nov 12 16:42:02 2005 Subject: [Histonet] H. Pylori Controls References: <20051107193008.74020.qmail@web61212.mail.yahoo.com> Message-ID: <40AA34F14CA09F429945413E05DED375281F55@EXCHANGE.chomp.org> We buy them from American Mastertech Scientific (1800) 860 4073, 25/pk Item # CSHEL25 ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rene J Buesa Sent: Mon 11/7/2005 11:30 AM To: Vicki Gauch; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H. Pylori Controls We used positive cases that were selected by the pathologists, but there is another approarch. Usually pathologists want to be able to have a positive control of exactly what they are looking for; if this is the case with your pathologists you will need controls comntaining H. pylori. Now, the whole idea with the control is to make sure that the procedure worked, and this you can obtain with any tissue containing abundant number of bacteria. After a long "convincing" process with our pathologists I was able to make them aware of this fact. We used the Steiner method (with phosphotungstic acid instead of the radiactive uranyl nitrate) and for control we used appendix (and you will not find a more easy to find tissue with more bacteria than an appendix). Perhaps you would like to try to talk about this approach with your pathologist. Just a thought! Rene J. Vicki Gauch wrote: Does anyone know of a good source for H. Pylori Controls?? We normally order from Newcomer Supply but they are experiencing a large backorder right now and we need something to hold us over until they arrive. Any help would be greatly appreciated... Thanks so much, Vicki Gauch AMCH Albany, NY ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 tahseen <@t> brain.net.pk Sat Nov 12 09:40:10 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Sat Nov 12 17:43:20 2005 Subject: [Histonet] Bone Saw References: Message-ID: <006901c5e79f$5ef8b0e0$972bfea9@m7c0y4> Dear Nancy, We have used a Startrite 352S band saw for about 11 years, and it is still going strong. I use a fine tooth (14 teeth per inch) metal cutting blade. The wood cutting blades are too coarse. Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Lahore Pakistan. ----- Original Message ----- From: Temple Nancy To: Sent: Saturday, November 12, 2005 2:46 AM Subject: [Histonet] Bone Saw We are currently using a bone saw made by Mar-Med. We do a lot of bones, and are in need of a heavier duty bone saw. We have had to replace the motor twice this year on the Mar-Med. Any suggestions? Thanks, Nancy St. Francis Hospital Indianapolis, In The information contained in this e-mail and any accompanying documents is intended for the sole use of the recipient to whom it is addressed, and may contain information that is privileged, confidential, and prohibited from disclosure under applicable law. If you are not the intended recipient, or authorized to receive this on behalf of the recipient, you are hereby notified that any review, use, disclosure, copying, or distribution is prohibited. If you are not the intended recipients(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NSEARCY <@t> swmail.sw.org Mon Nov 14 08:02:56 2005 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Mon Nov 14 08:03:36 2005 Subject: [Histonet] Surgical Path TAT Message-ID: Are there any benchmarks regarding TAT for surgical pathology reporting? CAP requires that autopsy TAT and frozen section TAT be tracked but has anyone studied surgicals? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Division Manager, Anatomic Pathology 2401 S. 31st. Street 254-724-2438 Temple, Texas,76502 254-724-2438 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Anatomic Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD From KMH.02 <@t> ex.uchs.org Mon Nov 14 08:15:02 2005 From: KMH.02 <@t> ex.uchs.org (Hopkins, Karen) Date: Mon Nov 14 08:14:41 2005 Subject: [Histonet] RE: Histonet Digest, Vol 24, Issue 18 Message-ID: <640B23A5DC4B234BB065E56F2DB30596028AB108@uchex2ucmc.uchs.org> During the summer months I had submitted a request for information concerning the actual salaries of HT and supervisory level HT's from anyone who was willing to share that information. Other than one request from someone who asked if I could share the responses with her I did not receive any responses. I'm not certain if no one wanted to share this personal information or if the emails were blocked by MIS at work. I would still be interested in this information and plan to use it to see if the salaries where I work are comparable with other facilities, realizing the region where the facility is located plays into it somewhat. If anyone is willing to share how many years experience they have as a HT and what they currently make per hour I would greatly appreciate the information. Also, I would be interested in the same for supervisor. Please email me at kmh.02@ex.uchs,org instead of replying through the histonet site. I will share my information with anyone who is willing to share with me and will not include names or facilities to anyone else. Thanks for your help From rjbuesa <@t> yahoo.com Mon Nov 14 08:37:43 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 14 08:37:52 2005 Subject: [Histonet] Surgical Path TAT In-Reply-To: Message-ID: <20051114143743.89340.qmail@web61215.mail.yahoo.com> Nita: The closest to a bench mark about surgicals that I can recommend you is a paper by LaFriniere at al. published in the Journal of Histotechnology in the Dec./2004 issue (vol 27 No4, pp293-5) where the authors present the average TAT for all histology tasks, as the result of a study by the NSH Task Force on Productivity. The values are averages so maybe they will be useful for you. In the case of repporting you will have to consider the TAT for description+processing+ slides preparation+diagnosis+special stain/procedures if required+transcription time, although in this aspects we used to give a report over the telephone for urgent cases pending on the final written (hard copy) report. Later we started to link our system with that of the referring physicians and the pathologists themselves were able to e-mail the results, always also pending of a written report. Our TAT averaged (from specimen reception to report of simple cases) from 16-19 hours for regular cases and 12-14 hours for rushes. Hope this will help you. Rene J. Nita Searcy wrote: Are there any benchmarks regarding TAT for surgical pathology reporting? CAP requires that autopsy TAT and frozen section TAT be tracked but has anyone studied surgicals? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Division Manager, Anatomic Pathology 2401 S. 31st. Street 254-724-2438 Temple, Texas,76502 254-724-2438 254-724-2438 BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Anatomic Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From rjbuesa <@t> yahoo.com Mon Nov 14 08:43:45 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 14 08:43:53 2005 Subject: [Histonet] RE: Histonet Digest, Vol 24, Issue 18 In-Reply-To: <640B23A5DC4B234BB065E56F2DB30596028AB108@uchex2ucmc.uchs.org> Message-ID: <20051114144345.66625.qmail@web61221.mail.yahoo.com> Both ADVANCE and the NSH have summarized salaries ranges for different areas based on years of experience, education and position. ASCP has also summaries. You could try to contact them and get their information. Rene J. "Hopkins, Karen" wrote: During the summer months I had submitted a request for information concerning the actual salaries of HT and supervisory level HT's from anyone who was willing to share that information. Other than one request from someone who asked if I could share the responses with her I did not receive any responses. I'm not certain if no one wanted to share this personal information or if the emails were blocked by MIS at work. I would still be interested in this information and plan to use it to see if the salaries where I work are comparable with other facilities, realizing the region where the facility is located plays into it somewhat. If anyone is willing to share how many years experience they have as a HT and what they currently make per hour I would greatly appreciate the information. Also, I would be interested in the same for supervisor. Please email me at kmh.02@ex.uchs,org instead of replying through the histonet site. I will share my information with anyone who is willing to share with me and will not include names or facilities to anyone else. Thanks for your help _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From anh2006 <@t> med.cornell.edu Mon Nov 14 10:10:34 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Mon Nov 14 10:05:34 2005 Subject: [Histonet] biotin blocking for frozen sections Message-ID: For FROZEN sections of mouse tissue, what are people using to block biotin. I am having problems with the standard kits I use. Thanks, Andrea -- From MadaryJ <@t> MedImmune.com Mon Nov 14 10:11:35 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Mon Nov 14 10:11:46 2005 Subject: [Histonet] I need a quote for a tissue processor now Message-ID: <746FDB897740814EA52BDDCB5ED1DDBC02C8C22F@medimmune4.medimmune.com> All you vendors who sell tissue processors please send a quote and I will go with the best one for my needs. I NEED A QUOTE NOW! I do not want any quotes from Thermo Shandon. Also do not want pathcenters, excelsiors, citadels. Prefer a VIP, or even a Leica (not the 1050). I would consider a Ventana if the price is right. I need at least 3 paraffins, vacuum, pressure, heat, agitation, capability for a printout of the run, decent reagent managment, and the feature that can call a phone number if there is a problem during the run. I know these all exist because I have used them over the yrs. Now we are using a Citadel 1000. I had no idea that these things were still being made. Makes me think of when I learned this stuff in the 70's, the technicon. I mean the technicon actually had better features than the citadel I am using now! I will consider used as cost is a bit of an issue. Thanks folks. Nick Madary, HT/HTL(ASCP)QIHC Medimmune Histology Laboratory One Medimmune Way Gaithersburg, MD 20878 ph 301.398.4745/6113/6141 fx 301.398.9745 From vazquezr <@t> ohsu.edu Mon Nov 14 10:24:12 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Mon Nov 14 10:24:36 2005 Subject: [Histonet] Disinfectant Solutions Message-ID: We use Sanimaster IV. Look it up on internet. Broad spectrum disinfectant. Robyn OHSU From sjchtascp <@t> yahoo.com Mon Nov 14 11:21:41 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Mon Nov 14 11:21:50 2005 Subject: [Histonet] cryostat heat extractor Message-ID: <20051114172141.95661.qmail@web90202.mail.scd.yahoo.com> I am search for a used heat extractor unit to assist more rapid freezing. We have a Microm HM 505 N. But any seperate heat extractor with a plastic handle from any cryostat would suit our needs. Thanks, Steve --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From pruegg <@t> ihctech.net Mon Nov 14 11:35:30 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Nov 14 11:35:35 2005 Subject: [Histonet] I need a quote for a tissue processor now In-Reply-To: <746FDB897740814EA52BDDCB5ED1DDBC02C8C22F@medimmune4.medimmune.com> Message-ID: <200511141735.jAEHZPxT004600@chip.viawest.net> I also need a quote for a reburbished tissue processor, nothing fancy or large, 100 cassette capacity will do. I want to purchase this before the end of the year to save on taxes. I do not have time to talk to sales people on the telephone, so just email me with what you have for sale and how much it costs. I am also looking for a slide drying oven. Patsy pruegg@ihctech.net Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Monday, November 14, 2005 9:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] I need a quote for a tissue processor now All you vendors who sell tissue processors please send a quote and I will go with the best one for my needs. I NEED A QUOTE NOW! I do not want any quotes from Thermo Shandon. Also do not want pathcenters, excelsiors, citadels. Prefer a VIP, or even a Leica (not the 1050). I would consider a Ventana if the price is right. I need at least 3 paraffins, vacuum, pressure, heat, agitation, capability for a printout of the run, decent reagent managment, and the feature that can call a phone number if there is a problem during the run. I know these all exist because I have used them over the yrs. Now we are using a Citadel 1000. I had no idea that these things were still being made. Makes me think of when I learned this stuff in the 70's, the technicon. I mean the technicon actually had better features than the citadel I am using now! I will consider used as cost is a bit of an issue. Thanks folks. Nick Madary, HT/HTL(ASCP)QIHC Medimmune Histology Laboratory One Medimmune Way Gaithersburg, MD 20878 ph 301.398.4745/6113/6141 fx 301.398.9745 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Nov 14 11:39:47 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Nov 14 11:39:57 2005 Subject: [Histonet] Heavy duty bones saw In-Reply-To: <139141F8BAF4A642A945ECC528511AF001F5ED49@kalmb02.kaleidahe alth.org> References: <139141F8BAF4A642A945ECC528511AF001F5ED49@kalmb02.kaleidahealth.org> Message-ID: <6.0.0.22.1.20051114103235.01b94a08@gemini.msu.montana.edu> Some research laboratories use shop band saws and modify them for bone work. The drawback, you need to do something to contain the aerosol sprays and bone dust, plus use a device to hold the bone so YOU are not the specimen. Fixing the bone a few hours in NBF helps with sawing, you can grip bones better after a bit of fixation as this seems to make a fatty bones/tissues less slick. It is a good idea to hold big bones between two pieces of wood for griping sample and not damaging the blade. You need to have 16 tpi or 16 teeth per inch for a very fine cut or you will tear the heck out of the bone/tissue/marrow cavity. For really large bones, we also used a hand held hack saw with tungsten blade, very crude but works to get a huge bone smaller then use MarMed. Still a bit messy but containable. Check out some of the water cooled Buehler units, but these are very pricey. We had a huge floor standing meat cutters saw (Hobart) that we could water cool but this is a monster, and was located in a separate room with a drain in the floor. At 08:52 AM 11/11/2005, you wrote: >I would also be interested in a heavy duty band saw since I work with large >human bones. When I looked into the Mar-Med I felt it was not sturdy or >large enough to meet my lab needs. > >I also would appreciate any suggestions. > >Peggy DiCarlo HT (ASCP) >Orthopedics Bone Lab >Buffalo General Hospital >100 High St. >Buffalo, NY 14203 >716-859-1293 > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Temple >Nancy >Sent: Friday, November 11, 2005 08:47 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Bone Saw > > >We are currently using a bone saw made by Mar-Med. We do a lot of bones, and >are in need of a heavier duty bone saw. We have had to replace the motor >twice this year on the Mar-Med. Any suggestions? >Thanks, > >Nancy >St. Francis Hospital >Indianapolis, In > > >The information contained in this e-mail and any accompanying documents is >intended for the sole use of the recipient to whom it is addressed, and may >contain information that is privileged, confidential, and prohibited from >disclosure under applicable law. If you are not the intended recipient, or >authorized to receive this on behalf of the recipient, you are hereby >notified that any review, use, disclosure, copying, or distribution is >prohibited. If you are not the intended recipients(s), please contact the >sender by e-mail and destroy all copies of the original message. Thank you. >_______________________________________________ >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 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From RStaskiewicz <@t> agr.state.il.us Mon Nov 14 11:40:13 2005 From: RStaskiewicz <@t> agr.state.il.us (RStaskiewicz@agr.state.il.us) Date: Mon Nov 14 11:40:26 2005 Subject: [Histonet] Surgipath Cassette Writer Message-ID: Does anyone using the Surgipath Cassette Writer have it set up to accept information from a spread sheet? If so, please contact me at the number below. Rae Ann Staskiewicz Rae Ann Staskiewicz HT(ASCP) Section Head-Histology Galesburg Animal Disease Lab 2100 S. Lake Storey Rd Galesburg, IL 61401 (309) 344-2451 rstaskiewicz@agr.state.il.us From gcallis <@t> montana.edu Mon Nov 14 11:49:56 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Nov 14 11:50:03 2005 Subject: [Histonet] IgG detection for mouse tissue? In-Reply-To: <20051112005907.47187.qmail@web50211.mail.yahoo.com> References: <20051112005907.47187.qmail@web50211.mail.yahoo.com> Message-ID: <6.0.0.22.1.20051114104624.01b94c88@gemini.msu.montana.edu> Yes, but to do this, we prefer to use a monoclonal (BD Pharmingen) that is Rat antiMouse IgG. If you use a polyclonal, go to Southern Biotechnology and make sure the antibody is adsorbed to mouse tissue, and/or an F(ab')2 frag of IgG to avoid fc receptor problems. OR use an antiAt 05:59 PM 11/11/2005, you wrote: >Hi All, > Does any one know whether there are antibodies to detect mouse IgG on > mouse tissues? Thank you for the help. > > > > >--------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Nov 14 11:54:12 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Nov 14 11:54:19 2005 Subject: [Histonet] biotin blocking for frozen sections In-Reply-To: References: Message-ID: <6.0.0.22.1.20051114105109.01b9c6e0@gemini.msu.montana.edu> We have used Vectors Avidin/biotin kits. Now, however, with our Strepavidin- Biotin methods, we now use their Strepavidin/biotin blocking kit. I have heard that DAKO's avidin/biotin blocking kit is stronger, more concentrated in order to block endogenous and unwanted biotins when using the CSA or tyramide amplification method (TSA) - certainly worth a try. At 09:10 AM 11/14/2005, you wrote: >For FROZEN sections of mouse tissue, what are people using to block >biotin. I am having problems with the standard kits I use. > >Thanks, >Andrea >-- >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From pruegg <@t> ihctech.net Mon Nov 14 11:55:35 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Nov 14 11:55:42 2005 Subject: [Histonet] Heavy duty bones saw In-Reply-To: <6.0.0.22.1.20051114103235.01b94a08@gemini.msu.montana.edu> Message-ID: <200511141755.jAEHtUxT010723@chip.viawest.net> Years ago some body sold us bone people a manual bone holding and sawing system, I don't remember the manufacturer, but they still use this at the U where I retired from. It was a big metal holding rack with a V shape with teeth to hold the bone (I used to cut hard to hold round human whole heads of femur) it used a hack saw/coping saw type saw, there were spaces cut thru the holder so you could actually cut a pretty thin waffer thru the whole bone by making two cuts creating a thin waffer between the cuts, it worked well for getting really large bones down to a more managable size to work with, the thickness is the most important thing to concern yourself with, if your bone sample is more than about 3-6 mm it will take forever to infiltrate. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Monday, November 14, 2005 10:40 AM To: DiCarlo, Margaret; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Heavy duty bones saw Some research laboratories use shop band saws and modify them for bone work. The drawback, you need to do something to contain the aerosol sprays and bone dust, plus use a device to hold the bone so YOU are not the specimen. Fixing the bone a few hours in NBF helps with sawing, you can grip bones better after a bit of fixation as this seems to make a fatty bones/tissues less slick. It is a good idea to hold big bones between two pieces of wood for griping sample and not damaging the blade. You need to have 16 tpi or 16 teeth per inch for a very fine cut or you will tear the heck out of the bone/tissue/marrow cavity. For really large bones, we also used a hand held hack saw with tungsten blade, very crude but works to get a huge bone smaller then use MarMed. Still a bit messy but containable. Check out some of the water cooled Buehler units, but these are very pricey. We had a huge floor standing meat cutters saw (Hobart) that we could water cool but this is a monster, and was located in a separate room with a drain in the floor. At 08:52 AM 11/11/2005, you wrote: >I would also be interested in a heavy duty band saw since I work with >large human bones. When I looked into the Mar-Med I felt it was not >sturdy or large enough to meet my lab needs. > >I also would appreciate any suggestions. > >Peggy DiCarlo HT (ASCP) >Orthopedics Bone Lab >Buffalo General Hospital >100 High St. >Buffalo, NY 14203 >716-859-1293 > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Temple >Nancy >Sent: Friday, November 11, 2005 08:47 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Bone Saw > > >We are currently using a bone saw made by Mar-Med. We do a lot of >bones, and are in need of a heavier duty bone saw. We have had to >replace the motor twice this year on the Mar-Med. Any suggestions? >Thanks, > >Nancy >St. Francis Hospital >Indianapolis, In > > >The information contained in this e-mail and any accompanying documents >is intended for the sole use of the recipient to whom it is addressed, >and may contain information that is privileged, confidential, and >prohibited from disclosure under applicable law. If you are not the >intended recipient, or authorized to receive this on behalf of the >recipient, you are hereby notified that any review, use, disclosure, >copying, or distribution is prohibited. If you are not the intended >recipients(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. >_______________________________________________ >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 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mhorne <@t> upei.ca Mon Nov 14 12:07:49 2005 From: mhorne <@t> upei.ca (Margaret Horne) Date: Mon Nov 14 12:16:02 2005 Subject: [Histonet] crab TEM Message-ID: <43789A35.27208.1171661@localhost> Hi , I am about to process some crab Hipatopancreas and gill for TEM . I was contemplating my fish protocol but wondered if anyone has done crustaceans. Thanks, Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada From petepath <@t> yahoo.com Mon Nov 14 12:26:26 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Mon Nov 14 12:26:36 2005 Subject: [Histonet] cryostat heat extractor Message-ID: <20051114182626.879.qmail@web30407.mail.mud.yahoo.com> Steve, Are you familiar with my Precision Cryoembedding System. It will sinificantly speed your frozen section embedding while offering a high level of precision and considerably less tissue wastage. I offer a no obligation demo period and creative payment schedual so that anyone can try it and return it if not satisfied or buy it with no more than petty cash. Visit http://pathologyinnovations.com/index.html to learn more. Stephen From sjchtascp <@t> yahoo.com Mon Nov 14 13:10:46 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Mon Nov 14 13:10:55 2005 Subject: [Histonet] cyrosectioning Message-ID: <20051114191046.70513.qmail@web90201.mail.scd.yahoo.com> We do allot of cryostat sectioning of both fresh and fixed sucrose cryoprotected tissue. I was wondering what other techs have found to result in better sections, one larger piece of tissue or several smaller pieces within one section. When sectioning several pieces of smaller tissue, such as liver is orientation critical? Thanks everyone. Steve --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From gcallis <@t> montana.edu Mon Nov 14 13:50:43 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Nov 14 13:50:52 2005 Subject: [Histonet] cyrosectioning In-Reply-To: <20051114191046.70513.qmail@web90201.mail.scd.yahoo.com> References: <20051114191046.70513.qmail@web90201.mail.scd.yahoo.com> Message-ID: <6.0.0.22.1.20051114123417.01b4cdb0@gemini.msu.montana.edu> Steven, What species? Our preference is to put one slice of liver per block, or one whole spleen. If you get a lot of curling with sections right at tissue location, then you may want do one piece per block or use a whole or half lobe. As long as the tissues have the same cryosectioning characteristics at a desired cryostat temperature, then you could put more than one piece per block It has been a disaster use for to put liver with intestine. We do put murltiple tiny lymph nodes (as many at 12) or peyers patches in one block or spinal cord cut into lengths for multiple pieces destined for cross sectioning but we would never put liver in with any of these tissues those - just liver with liver. Cryosectioning liver has been best at -17C while nodes or patches works at colder, usually -20C. One can "bread slice? liver lobes, and embed on cut edges. We have done the more flat orientation of whole or partial lobe - both work, obviously the whole/half lobe gives a bigger section and more information. If you need to see the outer capsule of liver, on edge works well but you don't have a generous section. Try it both ways. As for critical orientation of slices, I would embed them in some kind sequence so you know where the slices are from any given lobe, say and outer slice, middle slice, then another outer slice - that type of thing. Marking ink helps with who belongs to who!! We keep track of where we collect spinal cord, and know which end it being cut first, usually proximal end of any given slice - we even do cervical, thoracic and lumbar portions. Bet I have confused people with this one!!! Good luck!! At 12:10 PM 11/14/2005, you wrote: > >We do allot of cryostat sectioning of both fresh and fixed sucrose >cryoprotected tissue. I was wondering what other techs have found to >result in better sections, one larger piece of tissue or several smaller >pieces within one section. When sectioning several pieces of smaller >tissue, such as liver is orientation critical? > >Thanks everyone. > >Steve Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Nov 14 14:24:55 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Nov 14 14:25:24 2005 Subject: [Histonet] Bone Saw In-Reply-To: References: Message-ID: <6.0.0.22.1.20051114132140.01b10660@gemini.msu.montana.edu> Juan, It never ceases to amaze me at the wonderful replies that come from Histonet. I looked at the Cabelas butchers saw and it is a gem!!! Best bone saw deal I've seen lately!!! Boneheads will be happy with this one!!! At 09:10 AM 11/11/2005, you wrote: >Try Cabelas.com they have a butcher's band saw for about $550.00. It >might sound "cheap", but you have to realize it is not a scientific >supply company. If it cuts elk it should work fine in humans. > >Juan C. Gutierrez, HT(ASCP) >Histology Laboratory Supervisor >(210)704-2533 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From rjbuesa <@t> yahoo.com Mon Nov 14 14:46:17 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 14 14:46:25 2005 Subject: [Histonet] crab TEM In-Reply-To: <43789A35.27208.1171661@localhost> Message-ID: <20051114204617.56107.qmail@web61218.mail.yahoo.com> In 1963 I studied the histology of the spiny lobster Panulirus argus and the hepatopancreas was one of the tissues I processed. On those days I used a procedure totally "forbidden" today, namely ethanol (60;70; 80; 95;100) followed by the carcinogen aniline oil (until tissue transparent), followed by the also carcinogen benzene before the infiltration. All steps by hand. I used the same procedure for fish tissue (Family Lutjanidae) and it worked fine, so if you have a protocol that works fine on fish I would give it a try (by the way the hepatopancreas in any decapod crustacean is quite similar). Hope this will help! Rene J. Margaret Horne wrote: Hi , I am about to process some crab Hipatopancreas and gill for TEM . I was contemplating my fish protocol but wondered if anyone has done crustaceans. Thanks, Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From CMCCOLLOUGH <@t> dnr.state.md.us Mon Nov 14 15:14:34 2005 From: CMCCOLLOUGH <@t> dnr.state.md.us (McCollough, Carol) Date: Mon Nov 14 15:14:45 2005 Subject: [Histonet] cloth yellow stain Message-ID: Greetings Histonetters: In an exhaustive inventory of laboratory chemicals we have discovered an old bottle of 'Cloth Yellow' stain powder, manufactured by Chroma. Efforts to get an MSDS from Chroma have failed. Nobody here knows what this was even used for. Would anyone care to offer any histological uses for this stain? Regards - Carol ********************** Carol B. McCollough Aquatic Animal Research Pathologist Oyster Disease Research Project Fisheries Service Maryland Department of Natural Resources Cooperative Oxford Laboratory 904 S. Morris Street Oxford, Maryland 21654 410-226-5193 x124 From BennettW <@t> pac.dfo-mpo.gc.ca Mon Nov 14 15:32:24 2005 From: BennettW <@t> pac.dfo-mpo.gc.ca (BennettW@pac.dfo-mpo.gc.ca) Date: Mon Nov 14 15:30:23 2005 Subject: [Histonet] Histology Equipment Message-ID: <7CBBD627E4E688499349A5D11D07831603130901@msgpacpbs.pac.dfo-mpo.ca> Hi Everyone, Miraculously I have money to spend on new, yes new, histology lab equipment. Does anyone have any experience with the Shandon Pathcentre enclosed tissue processor, the Shandon Histocentre 3 tissue embedding centre and the Shandon Finesse Model ME motorized electronic microtome? Are they reliable and user friendly, ergonomic etc. Thanks in advance for your time. Cheers Bill Bennett Histologist Fisheries and Oceans Canada From froyer <@t> bitstream.net Mon Nov 14 15:35:53 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Mon Nov 14 15:36:11 2005 Subject: [Histonet] Bone Saw In-Reply-To: <6.0.0.22.1.20051114132140.01b10660@gemini.msu.montana.edu> Message-ID: <000801c5e963$642e30b0$0e00a8c0@fords> Someone mentioned that you may have to have a way to control bone dust if you went the route of getting a band saw from a local home improvement center. I do not know if it would work, but Craftsman (Sears) sells a portable dust collector... This has a blower, collection bag, and a long 4" diameter hose with a large dust hood. The blower assembly is mounted on casters and can be moved around easily. ($130.00) The collection bag has a 30 micron mesh size, so I do not know if that would pass inspection by your infection control committee or not. All I know is that it works great in my basement workshop while making wooden toys for my two grandsons' Christmas presents! ;-) ~ Ford ...And NO I do not work for or represent Sears Craftsman, or do I receive any form of compensation from them... They do, however, know me by name when I go in there! ;-) Ford M. Royer, MT(ASCP) Sales Manager, Histology Product Division Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Golden Valley, MN 55427-3601 763-542-8725 phone 888-790-9686 toll free 763-546-4830 fax email web -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Monday, November 14, 2005 2:25 PM To: GUTIERREZ, JUAN; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Bone Saw Juan, It never ceases to amaze me at the wonderful replies that come from Histonet. I looked at the Cabelas butchers saw and it is a gem!!! Best bone saw deal I've seen lately!!! Boneheads will be happy with this one!!! At 09:10 AM 11/11/2005, you wrote: >Try Cabelas.com they have a butcher's band saw for about $550.00. It >might sound "cheap", but you have to realize it is not a scientific >supply company. If it cuts elk it should work fine in humans. > >Juan C. Gutierrez, HT(ASCP) >Histology Laboratory Supervisor >(210)704-2533 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jlinda <@t> ces.clemson.edu Mon Nov 14 15:47:17 2005 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Mon Nov 14 15:47:10 2005 Subject: [Histonet] More on Bone saws Message-ID: <5.2.1.1.2.20051114163826.01f66e88@mailhost.ces.clemson.edu> There are many choices to choose from when it come to choosing a bone saw. My 2 favorites are the Exakt saw and the Buehler Isomet 2000 (both of theses pieces are 15 years old and still working well!). There are also wire saws and annular saws ( try doing a web search for these). The manual saw that Patsy Ruegg referred to is still made by Mopec. From an ol' Bonehead, Good luck! Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo.htm From JMacDonald <@t> mtsac.edu Mon Nov 14 15:56:41 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Mon Nov 14 15:56:34 2005 Subject: [Histonet] California Society for Histotechnology accepting Abstracts Message-ID: The California Society for Histotechnology is accepting abstracts for our spring meeting. The meeting will be held at the Costa Mesa Hilton from May 18-21 (Orange County, Southern California). Please contact Jennifer MacDonald if interested in participating. Jennifer MacDonald jmacdonald@mtsac.edu 909-594-5611 ext 4884 From pruegg <@t> ihctech.net Mon Nov 14 16:34:48 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Nov 14 16:34:54 2005 Subject: [Histonet] Sawbones Message-ID: <200511142234.jAEMYgxT005636@chip.viawest.net> Jennifer reminded me of the name of the bone holding and sawing device, it is called Sawbones from Mopec. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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 gcallis <@t> montana.edu Mon Nov 14 17:14:01 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Nov 14 17:14:22 2005 Subject: [Histonet] cloth yellow stain In-Reply-To: References: Message-ID: <6.0.0.22.1.20051114161205.01b3ed40@gemini.msu.montana.edu> Carol, Obviously old - I found Cloth red and Cloth blue, must be related to these, but I am betting John Kiernan will come up with the answer. My old Conn's didn't do the job. At 02:14 PM 11/14/2005, you wrote: >Greetings Histonetters: > >In an exhaustive inventory of laboratory chemicals we have discovered an >old bottle of 'Cloth Yellow' stain powder, manufactured by >Chroma. Efforts to get an MSDS from Chroma have failed. Nobody here >knows what this was even used for. > >Would anyone care to offer any histological uses for this stain? > >Regards - >Carol >********************** >Carol B. McCollough >Aquatic Animal Research Pathologist >Oyster Disease Research Project >Fisheries Service >Maryland Department of Natural Resources >Cooperative Oxford Laboratory >904 S. Morris Street >Oxford, Maryland 21654 >410-226-5193 x124 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From llewllew <@t> shaw.ca Mon Nov 14 18:17:00 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Mon Nov 14 18:18:30 2005 Subject: [Histonet] cloth yellow stain References: Message-ID: <003301c5e979$e5dd08f0$690a4246@yourlk4rlmsu> Dyes have many synonyms and individual dyes have their day in the sun then fade in popularity, as it were. I suggest you go to http://www.sdc.org.uk/ (The Society of Dyers and Colourists in the UK) who publish the Colour Index and send an e-mail asking if they have any information. If anyone would know what it is (or was) they would. I do know they have responded favourably to specific requests like this in the past. If you do find out, please let us know, my curiosity is aroused. Bryan Llewellyn ----- Original Message ----- From: "McCollough, Carol" To: "Histonet (E-mail 2)" Sent: Monday, November 14, 2005 1:14 PM Subject: [Histonet] cloth yellow stain Greetings Histonetters: In an exhaustive inventory of laboratory chemicals we have discovered an old bottle of 'Cloth Yellow' stain powder, manufactured by Chroma. Efforts to get an MSDS from Chroma have failed. Nobody here knows what this was even used for. Would anyone care to offer any histological uses for this stain? Regards - Carol ********************** Carol B. McCollough Aquatic Animal Research Pathologist Oyster Disease Research Project Fisheries Service Maryland Department of Natural Resources Cooperative Oxford Laboratory 904 S. Morris Street Oxford, Maryland 21654 410-226-5193 x124 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vanann702 <@t> skmc.gov.ae Mon Nov 14 21:59:55 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Mon Nov 14 21:57:51 2005 Subject: [Histonet] Histology Equipment Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02F6D6B6@SKMCEMAIL.skmc.gov.ae> My 'two cents worth' is that you try Sakura for the closed processor (VIP5) and embedding centre and Microm for the motorized microtome. Reliable, ergonomic and VERY user friendly. AnnieinArabia -----Original Message----- From: BennettW@pac.dfo-mpo.gc.ca [mailto:BennettW@pac.dfo-mpo.gc.ca] Sent: Tuesday, November 15, 2005 1:32 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology Equipment Hi Everyone, Miraculously I have money to spend on new, yes new, histology lab equipment. Does anyone have any experience with the Shandon Pathcentre enclosed tissue processor, the Shandon Histocentre 3 tissue embedding centre and the Shandon Finesse Model ME motorized electronic microtome? Are they reliable and user friendly, ergonomic etc. Thanks in advance for your time. Cheers Bill Bennett Histologist Fisheries and Oceans Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Tue Nov 15 00:00:02 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Tue Nov 15 00:00:14 2005 Subject: [Histonet] cloth yellow stain References: Message-ID: <43797962.7E1A5558@uwo.ca> Dear Carol, As Gayle points out, cloth yellow is not in the Lillie's 9th edition of Conn's Biological Stains (Conn9). Needless to say, it's not in Conn10 (2002), from which many obsolete dyes were omitted to make room for fluorescent labels and probes, and new information about old dyes that are still used. The three "cloth" dyes in Conn9 are all anionic bis-azo dyes. The two blue ones were used, along with many other dyes, in some of many published investigations of staining by Holde Puchtler and her colleagues in the 1960s and early '70s. For the red one [which is disulphonated sudan IV] Lillie reported no published uses as a biological stain; he suggested that it might serve as a "bluish red" cytoplasmic stain. Don't ask! Bluish and yellowish red dyes exist. We should discuss them on Histonet, but not now. If the "cloth" moniker was attached to anionic bis-azo dyes, a "cloth yellow" might serve as a yellow background to contrast with blue, black or red nuclei. There's no shortage of easily available dyes for that job. If Chroma's "cloth yellow" was a dye with large hydrophilic anions, it might have served as the collagen colorant in a trichrome-type method, as does saffron in Movat's original pentachrome. Such a dye, if available, might be much less expensive than saffron. Bryan's advice to ask the Society of Dyers and Colorists is good. Follow it, and keep it up by asking more. Chroma had a reputation for selling good stains, but their bottle labels didn't always commit to the chemical identity by providing a Colour Index (CI) number/name or a Chemical Abstracts Service (CAS) number, and they did not have samples of their products certified by the Biological Stain Commission. Bryan Llewellyn would like to keep in touch with your "cloth yellow" detective work, and so would I. By way of Histonet we can all be informed! John Kiernan Anatomy, UWO London, Canada. _______________________ "McCollough, Carol" wrote: > > Greetings Histonetters: > > In an exhaustive inventory of laboratory chemicals we have discovered an old bottle of 'Cloth Yellow' stain powder, manufactured by Chroma. Efforts to get an MSDS from Chroma have failed. Nobody here knows what this was even used for. > > Would anyone care to offer any histological uses for this stain? > > Regards - > Carol > ********************** > Carol B. McCollough > Aquatic Animal Research Pathologist > Oyster Disease Research Project > Fisheries Service > Maryland Department of Natural Resources > Cooperative Oxford Laboratory > 904 S. Morris Street > Oxford, Maryland 21654 > 410-226-5193 x124 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From germckeon <@t> excite.com Tue Nov 15 07:48:16 2005 From: germckeon <@t> excite.com (germckeon@excite.com) Date: Tue Nov 15 07:48:25 2005 Subject: [Histonet] Cryostating and Kultschitsky's stain Message-ID: <20051115134816.53FDE1973F9@xprdmailfe3.nwk.excite.com> Hello everybody, I am cryostating with gelatin embedded CNS tissue. I will be staining with silver stains which I have heard is not very penetrative. I am cutting at 30um. Could I ask is this too thick for these stains? I have tried cutting thinner sections (15um) but unfortunately I am getting chucking ie it will just cut a small section out of the lower section of the block every 2nd section. Despite adjusting everything on the cryostat and changing blades nothing seems to improve this (I was getting perfect sections at 30um). Does anyone know what's wrong? Finally how long can you keep kultschitsky's stain and Iron alum? I made up a stock solution of both but the quality suddenly deteriorated. Thanking you for any help, Gerald McKeon _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! From novanity <@t> nc.rr.com Tue Nov 15 08:55:15 2005 From: novanity <@t> nc.rr.com (novanity@nc.rr.com) Date: Tue Nov 15 08:55:26 2005 Subject: [Histonet] QIHC change Message-ID: <66105b660df9.660df966105b@southeast.rr.com> I missed the change to the QIHC requirements in January. I was wondering if anyone on the histonet was involved in that change or was aware of who was involved in that change and if they could elaborate on the specific second requirement about immnunophenotyping. As far as I can tell these techniques are specific to flow cytometry/cytometry/cytometers. Techniques utilizing procedures for immunodeficiency monitors/immunoproliferative disorders and transplantation biopsies that may be using antibodies to identify certain epitopes/antigens are as far as I can tell in most cases separate sub-lab (if you will) functions. Sub-lab as in lab a part of a whole department with separate labs handling flow, immuno etc... How is a technologist supposed to actually qualify for the QIHC when most businesses are going to hire a technologist only for one of the lab areas and not both? Cross-training may be OK but that doesn't really qualify as being involved in the day to day work in both labs. Is this requirement supposed to motivate people to quit working at the one lab after the 12 month period and then work in the other lab for at least 12 months to get both experiences for the requirements to have them in the previous 5 years? Making a requirement like this on the surface seems to me encouraging people to do things like quit a job that may not be in there best interest just to qualify for the QIHC. The lucky ones(very few) will work for a place that actually does crosstrain and make the next leap by allowing the employees to rotate through the different labs. Does the ASCP really want this reality? Was this requirement thought through by someone thoroughly? I'd really like some help understanding the mentality behind this change. While the 50 question test is nice, maybe how much emphasis is placed on the employer reference submission. If they say you have been doing the other two requirements extensive immunos/and the QAQC part of the job including the routine preparation histology, then for those of us(the many) that aren't in the flow lab are we excused from that requirement? Anyone? All replies greatly appreciated in advance. From HornHV <@t> archildrens.org Tue Nov 15 09:07:52 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Tue Nov 15 09:07:07 2005 Subject: [Histonet] Surgical Path TAT Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDEA6@EMAIL.archildrens.org> I believe CAP states 2 days. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Nita Searcy Sent: Monday, November 14, 2005 8:03 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Surgical Path TAT Are there any benchmarks regarding TAT for surgical pathology reporting? CAP requires that autopsy TAT and frozen section TAT be tracked but has anyone studied surgicals? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Division Manager, Anatomic Pathology 2401 S. 31st. Street 254-724-2438 Temple, Texas,76502 254-724-2438 254-724-2438 ------------------------------------------------------------------------------ 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 jwatson <@t> gnf.org Tue Nov 15 11:56:16 2005 From: jwatson <@t> gnf.org (James Watson) Date: Tue Nov 15 11:56:29 2005 Subject: [Histonet] QIHC change Message-ID: An few additional question about the immunophenotyping requirement: 1) Are there specific antigens or cell surface markers that are required? 2) What diseases can be screened for? Only leukemia and lymphomas? 3) Does it have to be on human cells? 4) What about those of us that work in research? Can we get slides to run these antibodies, and who would then grades the results based on the number of positive vs. normal cells if you do not have access to flow cytometry. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of novanity@nc.rr.com Sent: Tuesday, November 15, 2005 6:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] QIHC change I missed the change to the QIHC requirements in January. I was wondering if anyone on the histonet was involved in that change or was aware of who was involved in that change and if they could elaborate on the specific second requirement about immnunophenotyping. As far as I can tell these techniques are specific to flow cytometry/cytometry/cytometers. Techniques utilizing procedures for immunodeficiency monitors/immunoproliferative disorders and transplantation biopsies that may be using antibodies to identify certain epitopes/antigens are as far as I can tell in most cases separate sub-lab (if you will) functions. Sub-lab as in lab a part of a whole department with separate labs handling flow, immuno etc... How is a technologist supposed to actually qualify for the QIHC when most businesses are going to hire a technologist only for one of the lab areas and not both? Cross-training may be OK but that doesn't really qualify as being involved in the day to day work in both labs. Is this requirement supposed to motivate people to quit working at the one lab after the 12 month period and then work in the other lab for at least 12 months to get both experiences for the requirements to have them in the previous 5 years? Making a requirement like this on the surface seems to me encouraging people to do things like quit a job that may not be in there best interest just to qualify for the QIHC. The lucky ones(very few) will work for a place that actually does crosstrain and make the next leap by allowing the employees to rotate through the different labs. Does the ASCP really want this reality? Was this requirement thought through by someone thoroughly? I'd really like some help understanding the mentality behind this change. While the 50 question test is nice, maybe how much emphasis is placed on the employer reference submission. If they say you have been doing the other two requirements extensive immunos/and the QAQC part of the job including the routine preparation histology, then for those of us(the many) that aren't in the flow lab are we excused from that requirement? Anyone? All replies greatly appreciated in advance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfranci <@t> rigel.com Tue Nov 15 12:25:07 2005 From: cfranci <@t> rigel.com (Christian Franci) Date: Tue Nov 15 12:24:34 2005 Subject: [Histonet] Fixing question Message-ID: <75a19107de575a993a5048fb4abe7c0b@rigel.com> dear folks, pardon my ignorance but.... It seems that pressure is applied to the processing of tissues to ensure evenness and to force out any residual alcohol and/or xylene that might over-dry one's delicate samples. I gather that depending on what machine you have you can apply pressure at each step or just at the final paraffin stage ( as in my case). Here is my question... would applying pressure during the fixation of the tissues also be beneficial? Would it speed up the process therefore minimizing the possibility of over-fixation? Do any of you fix tissue under pressure and, if so, how do you go about it? Just curious.... Cheers Chris From jcresor <@t> lcpath.com Tue Nov 15 12:38:13 2005 From: jcresor <@t> lcpath.com (Jennifer N. Cresor) Date: Tue Nov 15 12:38:30 2005 Subject: [Histonet] clean glassware Message-ID: <200511151838.jAFIcL723606@plus34.host4u.net> Hello, I would like to give a presentation on the importance of cleaning glassware. Does anyone have pictures available of what the stains look like when using dirty glassware versus clean glassware? Any info would be helpful. Thank you. Jennifer jcresor@icpath.com From sbreeden <@t> nmda.nmsu.edu Tue Nov 15 12:47:27 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Tue Nov 15 12:47:37 2005 Subject: [Histonet] Cloth Yellow Message-ID: If anyone would know about this oldie, I would put my money on Dick Dapson of Anatech. I just took his "origins of stains" workshop at NSH and found that all our histo dyes/stains were originally used for the textile industry. He's done extensive research on stains and is a wealth of information. The website is www.anatechltdusa.com. Phone 800-262-8324; 269-964-6450. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From Barry.R.Rittman <@t> uth.tmc.edu Tue Nov 15 13:01:11 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Tue Nov 15 13:01:39 2005 Subject: [Histonet] Fixing question Message-ID: Using pressure to speed up fixtion has been used by several investigators. The jury is out on this one but the fact that it has been in the literature for quite some time and never gained any real popularity is indicative that it is not very useful. Just my opinion Can send you some references if you wish. . Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Christian Franci Sent: Tue 11/15/2005 12:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fixing question dear folks, pardon my ignorance but.... It seems that pressure is applied to the processing of tissues to ensure evenness and to force out any residual alcohol and/or xylene that might over-dry one's delicate samples. I gather that depending on what machine you have you can apply pressure at each step or just at the final paraffin stage ( as in my case). Here is my question... would applying pressure during the fixation of the tissues also be beneficial? Would it speed up the process therefore minimizing the possibility of over-fixation? Do any of you fix tissue under pressure and, if so, how do you go about it? Just curious.... Cheers Chris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Tue Nov 15 14:04:44 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 15 14:04:53 2005 Subject: [Histonet] Fixing question In-Reply-To: <75a19107de575a993a5048fb4abe7c0b@rigel.com> Message-ID: <20051115200445.64231.qmail@web61219.mail.yahoo.com> Christian: There are several issues to consider including the type of fixative. I you are referring to neutral buffered formalin (NBF) overfixation occurs when tissues are left more time than necessary and this will depend on the thickness of the tissue slice to be fixed. NBF has a diffusability coefficient (K) of 0.78 mm/hour so a slice 4 mm thick will require 2.5 hours to fix (remember that the NBF will be "entering" the tissues from both sides!). Any time after that will be "theoretically" and overfixation, i.e. not needed time in NBF and this can translate into more crosslinkage of the proteins and more difficulties for immunohistochemistry (IHC) procedures. Ethanol has a K of 1.0 mm/h so the same type of slice will require only 2 hours, will fix by quagulation and theoretically will not interfere with IHC but, again, after 2 hours it is not necessary to keep the tissue in ethanol. Having said all that, in tissue processors like the Sakura VIP all stations can have vacuum/pressure only depending in how long the station is programmed to last. With thin enough slices of tissue I don't think you have to worry about overfixation as long as you keep the tissues in the fixative the time required for its penetration, whichever the fixative is. Hope this will help (although I think I gave a somewhat "convoluted" answer!). Rene J. Christian Franci wrote: dear folks, pardon my ignorance but.... It seems that pressure is applied to the processing of tissues to ensure evenness and to force out any residual alcohol and/or xylene that might over-dry one's delicate samples. I gather that depending on what machine you have you can apply pressure at each step or just at the final paraffin stage ( as in my case). Here is my question... would applying pressure during the fixation of the tissues also be beneficial? Would it speed up the process therefore minimizing the possibility of over-fixation? Do any of you fix tissue under pressure and, if so, how do you go about it? Just curious.... Cheers Chris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From Karen.Heckford <@t> CHW.edu Tue Nov 15 14:34:51 2005 From: Karen.Heckford <@t> CHW.edu (Heckford, Karen - SMMC-SF) Date: Tue Nov 15 14:34:42 2005 Subject: [Histonet] IHC and JCAHO Message-ID: Does anyone know where I can get material on what JCAHO will look for in our new IHC lab? I would greatly appreciate the help. Have a Good Day, Karen Heckford HT (ASCP) CE Lead Histology Technician St. Mary's Medical Center Histology Department 450 Stanyan St. San Francisco, Ca. 94117 1-415-668-1000 ext. 6167 email: kheckfor@chw.edu From laurie.colbert <@t> huntingtonhospital.com Tue Nov 15 14:41:44 2005 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Tue Nov 15 14:41:57 2005 Subject: [Histonet] Biohazard Labeling Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628005653880@EXCHANGE1.huntingtonhospital.com> Can anyone give me information on the labeling of fresh specimens? In the past, I was told by our infection control coordinator that fresh specimens did not need to be labeled as "Biohazard." Now, I'm being told they do need to be labeled. The new coordinator is getting me more info, but can anyone guide me to info from OSHA, CAP, JCAHO, etc. that refers to this topic? Laurie Colbert Huntington Memorial Hospital From julien_lambreydesouza <@t> uqar.qc.ca Tue Nov 15 14:58:59 2005 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien Lambrey de Souza) Date: Tue Nov 15 14:57:31 2005 Subject: [Histonet] RE: histology equipment Message-ID: <2eb6dd00c8eabb044b61404bd4a0bd1a@uqar.qc.ca> We have the Finesse ME from shandon. It does wonders!! Julien Lambrey de Souza Research assistant, Evolutionary biology University of Quebec at Rimouski Tel: (418) 723-1986 #1714 Fax: (418) 724-1849 From BGapinski <@t> pathgroup.com Tue Nov 15 15:10:53 2005 From: BGapinski <@t> pathgroup.com (Bruce Gapinski) Date: Tue Nov 15 15:11:10 2005 Subject: [Histonet] Histology schools Message-ID: Are there any Histology schools? Where do people learn how to do what we do? On the job training is too much for me anymore .I've trained about 8 now. Besides, who wants to spend their (now required) A.S. in a histo lab? Honestly, some times I wonder if we've boxed ourselves into the worst situation. Anyone ELSE having trouble finding a histologist? Or is it an histologist, I never know. Bruce Gapinski HT (ASCP) --------------------------------------------------------------------- Important Notice: This e-mail is intended for the use of the person to whom it is addressed and may contain information that is privileged and confidential. If you are not the intended recipient, any disclosure, copying, distribution, or use of the contents of this message is strictly prohibited. If you have received this e-mail in error, please destroy this message and contact the Security Officer at PathGroup, Inc. immediately at 615-221-4431. From rjbuesa <@t> yahoo.com Tue Nov 15 15:23:55 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 15 15:24:04 2005 Subject: [Histonet] Histology schools In-Reply-To: Message-ID: <20051115212355.35450.qmail@web61218.mail.yahoo.com> In South Florida we have 2 histology programs, one at Barry University and the other at Miami Dade College. Rene J. Bruce Gapinski wrote: Are there any Histology schools? Where do people learn how to do what we do? On the job training is too much for me anymore .I've trained about 8 now. Besides, who wants to spend their (now required) A.S. in a histo lab? Honestly, some times I wonder if we've boxed ourselves into the worst situation. Anyone ELSE having trouble finding a histologist? Or is it an histologist, I never know. Bruce Gapinski HT (ASCP) --------------------------------------------------------------------- Important Notice: This e-mail is intended for the use of the person to whom it is addressed and may contain information that is privileged and confidential. If you are not the intended recipient, any disclosure, copying, distribution, or use of the contents of this message is strictly prohibited. If you have received this e-mail in error, please destroy this message and contact the Security Officer at PathGroup, Inc. immediately at 615-221-4431._______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From lldewe <@t> ucdavis.edu Tue Nov 15 15:27:55 2005 From: lldewe <@t> ucdavis.edu (Loralei Dewe) Date: Tue Nov 15 15:28:04 2005 Subject: [Histonet] Histology schools In-Reply-To: References: Message-ID: Hi Bruce, On the West Coast here unfortunately there is only one program taught down at Mt San Antonio College in Southern Calif. It is fairly new but doing quite well... Best to you, Loralei Dewe On Tue, 15 Nov 2005, Bruce Gapinski wrote: > Are there any Histology schools? Where do people learn how to do what we do? On the job training is too much for me anymore .I've trained about 8 now. Besides, who wants to spend their (now required) A.S. in a histo lab? Honestly, some times I wonder if we've boxed ourselves into the worst situation. Anyone ELSE having trouble finding a histologist? Or is it an histologist, I never know. > Bruce Gapinski HT (ASCP) > > > --------------------------------------------------------------------- > Important Notice: This e-mail is intended for the use of the person > to whom it is addressed and may contain information that is > privileged and confidential. If you are not the intended recipient, > any disclosure, copying, distribution, or use of the contents of > this message is strictly prohibited. If you have received this > e-mail in error, please destroy this message and contact the > Security Officer at PathGroup, Inc. immediately at 615-221-4431. From HornHV <@t> archildrens.org Tue Nov 15 15:30:05 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Tue Nov 15 15:29:16 2005 Subject: [Histonet] Histology schools Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDEAC@EMAIL.archildrens.org> I know there is a histology school at Baptist Medical Center School for Allied Health Professions in Little Rock, AR. The trouble they only take 4-6 people a year. Which keeps all our local positions full. I guess we are lucky. Although if there were a shortage around here I think it would help salaries. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bruce Gapinski Sent: Tuesday, November 15, 2005 3:11 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology schools Are there any Histology schools? Where do people learn how to do what we do? On the job training is too much for me anymore .I've trained about 8 now. Besides, who wants to spend their (now required) A.S. in a histo lab? Honestly, some times I wonder if we've boxed ourselves into the worst situation. Anyone ELSE having trouble finding a histologist? Or is it an histologist, I never know. Bruce Gapinski HT (ASCP) --------------------------------------------------------------------- Important Notice: This e-mail is intended for the use of the person to whom it is addressed and may contain information that is privileged and confidential. If you are not the intended recipient, any disclosure, copying, distribution, or use of the contents of this message is strictly prohibited. If you have received this e-mail in error, please destroy this message and contact the Security Officer at PathGroup, Inc. immediately at 615-221-4431. ------------------------------------------------------------------------------ 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 mucram11 <@t> comcast.net Tue Nov 15 15:32:04 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Tue Nov 15 15:32:22 2005 Subject: [Histonet] Histology schools Message-ID: <111520052132.7317.437A53D400071E7800001C952200763704CECE030E9D0C9A03@comcast.net> Arizona has two at leat and I know one is at Pima Community College. Ethel Macrea is involved with it and it is quite good. Pam Marcum > I know there is a histology school at Baptist Medical Center School for > Allied Health Professions in Little Rock, AR. The trouble they only > take 4-6 people a year. Which keeps all our local positions full. > I guess we are lucky. Although if there were a shortage around here I > think it would help salaries. > > > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > 800 Marshall Mail Slot 820 > Little Rock, AR 72202 > > phone- 501.364.4240 > fax- 501.364.3912 > > visit us on the web at: www.archildrens.org > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bruce > Gapinski > Sent: Tuesday, November 15, 2005 3:11 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histology schools > > Are there any Histology schools? Where do people learn how to do what we > do? On the job training is too much for me anymore .I've trained about 8 > now. Besides, who wants to spend their (now required) A.S. in a histo > lab? Honestly, some times I wonder if we've boxed ourselves into the > worst situation. Anyone ELSE having trouble finding a histologist? Or is > it an histologist, I never know. > Bruce Gapinski HT (ASCP) > > > --------------------------------------------------------------------- > Important Notice: This e-mail is intended for the use of the person to > whom it is addressed and may contain information that is privileged and > confidential. If you are not the intended recipient, any disclosure, > copying, distribution, or use of the contents of this message is > strictly prohibited. If you have received this e-mail in error, please > destroy this message and contact the Security Officer at PathGroup, Inc. > immediately at 615-221-4431. > > ------------------------------------------------------------------------------ > 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 tpmorken <@t> labvision.com Tue Nov 15 15:58:56 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Tue Nov 15 15:59:07 2005 Subject: [Histonet] Histology schools Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D351@usca0082k08.labvision.apogent.com> I heard there is another program starting up in Seattle area. Tim Morken Lab Vision - Neomarkers www.labvision.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Loralei Dewe Sent: Tuesday, November 15, 2005 1:28 PM To: Bruce Gapinski Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology schools Hi Bruce, On the West Coast here unfortunately there is only one program taught down at Mt San Antonio College in Southern Calif. It is fairly new but doing quite well... Best to you, Loralei Dewe On Tue, 15 Nov 2005, Bruce Gapinski wrote: > Are there any Histology schools? Where do people learn how to do what > we do? On the job training is too much for me anymore .I've trained > about 8 now. Besides, who wants to spend their (now required) A.S. in > a histo lab? Honestly, some times I wonder if we've boxed ourselves > into the worst situation. Anyone ELSE having trouble finding a > histologist? Or is it an histologist, I never know. Bruce Gapinski HT > (ASCP) > > > --------------------------------------------------------------------- > Important Notice: This e-mail is intended for the use of the person to > whom it is addressed and may contain information that is privileged > and confidential. If you are not the intended recipient, any > disclosure, copying, distribution, or use of the contents of this > message is strictly prohibited. If you have received this e-mail in > error, please destroy this message and contact the Security Officer at > PathGroup, Inc. immediately at 615-221-4431. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMacDonald <@t> mtsac.edu Tue Nov 15 16:11:00 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Tue Nov 15 16:10:52 2005 Subject: [Histonet] Histology schools In-Reply-To: <111520052132.7317.437A53D400071E7800001C952200763704CECE030E9D0C9A03@comcast.net> Message-ID: This website will give you a list of NAACLS accredited Histotechnology programs in the US. http://www.nsh.org/education/schools.html mucram11@comcast.net (Pam Marcum) Sent by: histonet-bounces@lists.utsouthwestern.edu 11/15/2005 01:32 PM To "Horn, Hazel V" , "Bruce Gapinski" , histonet@lists.utsouthwestern.edu cc Subject RE: [Histonet] Histology schools Arizona has two at leat and I know one is at Pima Community College. Ethel Macrea is involved with it and it is quite good. Pam Marcum > I know there is a histology school at Baptist Medical Center School for > Allied Health Professions in Little Rock, AR. The trouble they only > take 4-6 people a year. Which keeps all our local positions full. > I guess we are lucky. Although if there were a shortage around here I > think it would help salaries. > > > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > 800 Marshall Mail Slot 820 > Little Rock, AR 72202 > > phone- 501.364.4240 > fax- 501.364.3912 > > visit us on the web at: www.archildrens.org > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bruce > Gapinski > Sent: Tuesday, November 15, 2005 3:11 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histology schools > > Are there any Histology schools? Where do people learn how to do what we > do? On the job training is too much for me anymore .I've trained about 8 > now. Besides, who wants to spend their (now required) A.S. in a histo > lab? Honestly, some times I wonder if we've boxed ourselves into the > worst situation. Anyone ELSE having trouble finding a histologist? Or is > it an histologist, I never know. > Bruce Gapinski HT (ASCP) > > > --------------------------------------------------------------------- > Important Notice: This e-mail is intended for the use of the person to > whom it is addressed and may contain information that is privileged and > confidential. If you are not the intended recipient, any disclosure, > copying, distribution, or use of the contents of this message is > strictly prohibited. If you have received this e-mail in error, please > destroy this message and contact the Security Officer at PathGroup, Inc. > immediately at 615-221-4431. > > ------------------------------------------------------------------------------ > 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Tue Nov 15 16:16:23 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Tue Nov 15 16:16:55 2005 Subject: [Histonet] Histology schools In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328D351@usca0082k08.labvision.apogent.com> Message-ID: There are 2 programs in the Seattle are that are in the 'start-up' phase. One is at Henry Cogswell College in Everett, WA (N. of Seattle) and the other at Clover Park Technical College in Lakewood, WA (near Tacoma). I would be happy to keep anyone interested informed of the situation. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA > I heard there is another program starting up in Seattle area. > > Tim Morken > Lab Vision - Neomarkers > www.labvision.com > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Loralei Dewe > Sent: Tuesday, November 15, 2005 1:28 PM > To: Bruce Gapinski > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Histology schools > > > > Hi Bruce, > > On the West Coast here unfortunately there is only one program taught down > at Mt San Antonio College in Southern Calif. It is fairly new but doing > quite well... > > Best to you, > > Loralei Dewe > > > > On Tue, 15 Nov 2005, Bruce Gapinski wrote: > >> Are there any Histology schools? Where do people learn how to do what >> we do? On the job training is too much for me anymore .I've trained >> about 8 now. Besides, who wants to spend their (now required) A.S. in >> a histo lab? Honestly, some times I wonder if we've boxed ourselves >> into the worst situation. Anyone ELSE having trouble finding a >> histologist? Or is it an histologist, I never know. Bruce Gapinski HT >> (ASCP) >> >> >> --------------------------------------------------------------------- >> Important Notice: This e-mail is intended for the use of the person to >> whom it is addressed and may contain information that is privileged >> and confidential. If you are not the intended recipient, any >> disclosure, copying, distribution, or use of the contents of this >> message is strictly prohibited. If you have received this e-mail in >> error, please destroy this message and contact the Security Officer at >> PathGroup, Inc. immediately at 615-221-4431. > > _______________________________________________ > 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 e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From ploykasek <@t> phenopath.com Tue Nov 15 16:16:23 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Tue Nov 15 16:16:56 2005 Subject: [Histonet] Histology schools In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328D351@usca0082k08.labvision.apogent.com> Message-ID: There are 2 programs in the Seattle are that are in the 'start-up' phase. One is at Henry Cogswell College in Everett, WA (N. of Seattle) and the other at Clover Park Technical College in Lakewood, WA (near Tacoma). I would be happy to keep anyone interested informed of the situation. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA > I heard there is another program starting up in Seattle area. > > Tim Morken > Lab Vision - Neomarkers > www.labvision.com > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Loralei Dewe > Sent: Tuesday, November 15, 2005 1:28 PM > To: Bruce Gapinski > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Histology schools > > > > Hi Bruce, > > On the West Coast here unfortunately there is only one program taught down > at Mt San Antonio College in Southern Calif. It is fairly new but doing > quite well... > > Best to you, > > Loralei Dewe > > > > On Tue, 15 Nov 2005, Bruce Gapinski wrote: > >> Are there any Histology schools? Where do people learn how to do what >> we do? On the job training is too much for me anymore .I've trained >> about 8 now. Besides, who wants to spend their (now required) A.S. in >> a histo lab? Honestly, some times I wonder if we've boxed ourselves >> into the worst situation. Anyone ELSE having trouble finding a >> histologist? Or is it an histologist, I never know. Bruce Gapinski HT >> (ASCP) >> >> >> --------------------------------------------------------------------- >> Important Notice: This e-mail is intended for the use of the person to >> whom it is addressed and may contain information that is privileged >> and confidential. If you are not the intended recipient, any >> disclosure, copying, distribution, or use of the contents of this >> message is strictly prohibited. If you have received this e-mail in >> error, please destroy this message and contact the Security Officer at >> PathGroup, Inc. immediately at 615-221-4431. > > _______________________________________________ > 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 e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From ngianotti <@t> g-com.com Tue Nov 15 16:20:40 2005 From: ngianotti <@t> g-com.com (Nancy) Date: Tue Nov 15 16:20:50 2005 Subject: [Fwd: [Histonet] Histology schools] Message-ID: <437A5F38.8050408@g-com.com> We are starting a histology school at UPMC hospitals in Pittsburgh, PA in January 2006 out of necessity in finding histologists. They have to have an associates and it is for 1 year. The only other school in PA is Conemaugh in Johnstown. We need to promote the field more, but who knows how?? -------- Original Message -------- Subject: [Histonet] Histology schools Date: Tue, 15 Nov 2005 15:10:53 -0600 From: Bruce Gapinski To: Are there any Histology schools? Where do people learn how to do what we do? On the job training is too much for me anymore .I've trained about 8 now. Besides, who wants to spend their (now required) A.S. in a histo lab? Honestly, some times I wonder if we've boxed ourselves into the worst situation. Anyone ELSE having trouble finding a histologist? Or is it an histologist, I never know. Bruce Gapinski HT (ASCP) --------------------------------------------------------------------- Important Notice: This e-mail is intended for the use of the person to whom it is addressed and may contain information that is privileged and confidential. If you are not the intended recipient, any disclosure, copying, distribution, or use of the contents of this message is strictly prohibited. If you have received this e-mail in error, please destroy this message and contact the Security Officer at PathGroup, Inc. immediately at 615-221-4431. -------------- next part -------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Tue Nov 15 17:06:30 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Tue Nov 15 17:06:53 2005 Subject: [Histonet] Histology schools Message-ID: Portland oregon has a program at PCC. Robyn OHSU From pathrm35 <@t> adelphia.net Tue Nov 15 17:17:27 2005 From: pathrm35 <@t> adelphia.net (Ron Martin) Date: Tue Nov 15 17:17:54 2005 Subject: [Histonet] Histology schools] References: <437A5F38.8050408@g-com.com> Message-ID: <001401c5ea3a$c05a8e10$27233418@Pathrm35> I went to a local junior college to try to set up a histology program. I offered my services as an instructor and our lab as a teaching facility. This junior college had thought about doing this in the past but never did. I was told that they didn't want to flood the local market with histotechs. Imagine that!! I tried to make my case to no avail. I have been (and still am) looking for a histotech for the past 5 months with no luck. Ron Martin, BS, HT (ASCP) HTL Palm Beach Gardens, Florida ----- Original Message ----- From: "Nancy" To: Sent: Tuesday, November 15, 2005 5:20 PM Subject: [Fwd: [Histonet] Histology schools] > We are starting a histology school at UPMC hospitals in Pittsburgh, PA > in January 2006 out of necessity in finding histologists. They have to > have an associates and it is for 1 year. The only other school in PA is > Conemaugh in Johnstown. We need to promote the field more, but who knows > how?? > > -------- Original Message -------- > Subject: [Histonet] Histology schools > Date: Tue, 15 Nov 2005 15:10:53 -0600 > From: Bruce Gapinski > To: > > > > Are there any Histology schools? Where do people learn how to do what we > do? On the job training is too much for me anymore .I've trained about 8 > now. Besides, who wants to spend their (now required) A.S. in a histo > lab? Honestly, some times I wonder if we've boxed ourselves into the worst > situation. Anyone ELSE having trouble finding a histologist? Or is it an > histologist, I never know. > Bruce Gapinski HT (ASCP) > > > --------------------------------------------------------------------- > Important Notice: This e-mail is intended for the use of the person > to whom it is addressed and may contain information that is > privileged and confidential. If you are not the intended recipient, > any disclosure, copying, distribution, or use of the contents of > this message is strictly prohibited. If you have received this > e-mail in error, please destroy this message and contact the > Security Officer at PathGroup, Inc. immediately at 615-221-4431. > > -------------------------------------------------------------------------------- > _______________________________________________ > 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 histology.bc <@t> shaw.ca Tue Nov 15 19:20:23 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Tue Nov 15 19:17:11 2005 Subject: [Histonet] cloth yellow stain References: Message-ID: <437A8957.5040009@shaw.ca> For the last couple of hours I have been racking my brain to try to recall where I have come across this dye before. But, the required neurons are not responding ... or only weakly. John Kiernan is right on target ( as usual) in suggesting that this dye is almost certainly a large molecule azo dye (actually a very long, linear molecule) anionic dye probably used in the textile industry for coloring cotton fibres. If I remember correctly, such dyes are often referred to as "milling dyes". Similar azo dyes are commonly used for dying natural fibres in industry. In histology, dyes of this type are used to show amyloid (Congo red being the classical example). Yellow dyes provide poor visibility to tissue elements, so this dyes is almost certainly a counterstain used to highlight general background structures and to contrast with a red or blue primary stain. My suggestion would be to make up a 0.5% solution of the dye in 1% acetic acid, and try it out on a few sections of tissue. Let us know what the results are ... I am sure we would all love to know for sure.. Paul Bradbury Kamloops, BC Canada McCollough, Carol wrote: >Greetings Histonetters: > >In an exhaustive inventory of laboratory chemicals we have discovered an old bottle of 'Cloth Yellow' stain powder, manufactured by Chroma. Efforts to get an MSDS from Chroma have failed. Nobody here knows what this was even used for. > >Would anyone care to offer any histological uses for this stain? > >Regards - >Carol >********************** >Carol B. McCollough >Aquatic Animal Research Pathologist >Oyster Disease Research Project >Fisheries Service >Maryland Department of Natural Resources >Cooperative Oxford Laboratory >904 S. Morris Street >Oxford, Maryland 21654 >410-226-5193 x124 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From jwell1958 <@t> msn.com Tue Nov 15 21:04:28 2005 From: jwell1958 <@t> msn.com (Janci Wellborn) Date: Tue Nov 15 21:04:37 2005 Subject: [Histonet] Neg Mouse Controls Message-ID: Does anyone know if a neg mouse control is required by CAP on every slide in a case. If it is, what is the listing. We have a new pathologist and he ordered immuno staining on 8 blocks on the same case. The tech ran a neg mouse on each block. I has been my understanding that only one negative mouse is required for a case. Janci Wellborn, BS, BSeD, HTL From Malcolm.McCallum <@t> tamut.edu Tue Nov 15 22:06:06 2005 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Tue Nov 15 22:07:00 2005 Subject: [Histonet] Histology schools] Message-ID: Now this is a new thing to me. What would a school need to do to offer a histotech certificate or degree (bs) program??? Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ron Martin Sent: Tue 11/15/2005 5:17 PM To: Nancy; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology schools] I went to a local junior college to try to set up a histology program. I offered my services as an instructor and our lab as a teaching facility. This junior college had thought about doing this in the past but never did. I was told that they didn't want to flood the local market with histotechs. Imagine that!! I tried to make my case to no avail. I have been (and still am) looking for a histotech for the past 5 months with no luck. Ron Martin, BS, HT (ASCP) HTL Palm Beach Gardens, Florida ----- Original Message ----- From: "Nancy" To: Sent: Tuesday, November 15, 2005 5:20 PM Subject: [Fwd: [Histonet] Histology schools] > We are starting a histology school at UPMC hospitals in Pittsburgh, PA > in January 2006 out of necessity in finding histologists. They have to > have an associates and it is for 1 year. The only other school in PA is > Conemaugh in Johnstown. We need to promote the field more, but who knows > how?? > > -------- Original Message -------- > Subject: [Histonet] Histology schools > Date: Tue, 15 Nov 2005 15:10:53 -0600 > From: Bruce Gapinski > To: > > > > Are there any Histology schools? Where do people learn how to do what we > do? On the job training is too much for me anymore .I've trained about 8 > now. Besides, who wants to spend their (now required) A.S. in a histo > lab? Honestly, some times I wonder if we've boxed ourselves into the worst > situation. Anyone ELSE having trouble finding a histologist? Or is it an > histologist, I never know. > Bruce Gapinski HT (ASCP) > > > --------------------------------------------------------------------- > Important Notice: This e-mail is intended for the use of the person > to whom it is addressed and may contain information that is > privileged and confidential. If you are not the intended recipient, > any disclosure, copying, distribution, or use of the contents of > this message is strictly prohibited. If you have received this > e-mail in error, please destroy this message and contact the > Security Officer at PathGroup, Inc. immediately at 615-221-4431. > > -------------------------------------------------------------------------------- > _______________________________________________ > 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 lpwenk <@t> sbcglobal.net Wed Nov 16 04:50:36 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Wed Nov 16 04:51:50 2005 Subject: [Histonet] Histology schools In-Reply-To: Message-ID: Try contacting the Indiana Univesity HT program. They have a 1 year training program, where the people stay in the lab they are in, but are taught by participating in teleconferences, emails, mailing slides in to IU, taking tests administered by their supervisor (like you), and doing a lot of studying on their own. They are NAACLS accredited HT program. They take students from about 45 sites across the country. The program started every August, if I remember correctly. 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 Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bruce Gapinski Sent: Tuesday, November 15, 2005 4:11 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology schools Are there any Histology schools? Where do people learn how to do what we do? On the job training is too much for me anymore .I've trained about 8 now. Besides, who wants to spend their (now required) A.S. in a histo lab? Honestly, some times I wonder if we've boxed ourselves into the worst situation. Anyone ELSE having trouble finding a histologist? Or is it an histologist, I never know. Bruce Gapinski HT (ASCP) --------------------------------------------------------------------- Important Notice: This e-mail is intended for the use of the person to whom it is addressed and may contain information that is privileged and confidential. If you are not the intended recipient, any disclosure, copying, distribution, or use of the contents of this message is strictly prohibited. If you have received this e-mail in error, please destroy this message and contact the Security Officer at PathGroup, Inc. immediately at 615-221-4431. From jnocito <@t> satx.rr.com Wed Nov 16 06:09:12 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Wed Nov 16 06:09:22 2005 Subject: [Histonet] Histology schools References: <20051115212355.35450.qmail@web61218.mail.yahoo.com> Message-ID: <014001c5eaa6$8ecb5c50$e8bd0b43@yourxhtr8hvc4p> Unfortunately, the only program in San Antonio, Texas has just closed. That is too bad, I hired four students out of that class too! Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Rene J Buesa" To: "Bruce Gapinski" ; Sent: Tuesday, November 15, 2005 3:23 PM Subject: Re: [Histonet] Histology schools > In South Florida we have 2 histology programs, one at Barry University and > the other at > Miami Dade College. > Rene J. > > Bruce Gapinski wrote: > Are there any Histology schools? Where do people learn how to do what we > do? On the job training is too much for me anymore .I've trained about 8 > now. Besides, who wants to spend their (now required) A.S. in a histo lab? > Honestly, some times I wonder if we've boxed ourselves into the worst > situation. Anyone ELSE having trouble finding a histologist? Or is it an > histologist, I never know. > Bruce Gapinski HT (ASCP) > > > --------------------------------------------------------------------- > Important Notice: This e-mail is intended for the use of the person > to whom it is addressed and may contain information that is > privileged and confidential. If you are not the intended recipient, > any disclosure, copying, distribution, or use of the contents of > this message is strictly prohibited. If you have received this > e-mail in error, please destroy this message and contact the > Security Officer at PathGroup, Inc. immediately at > 615-221-4431._______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > --------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Free Edition. > Version: 7.1.362 / Virus Database: 267.13.2/170 - Release Date: 11/15/2005 > From Barry.R.Rittman <@t> uth.tmc.edu Wed Nov 16 06:43:18 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Wed Nov 16 06:44:35 2005 Subject: [Histonet] cloth yellow stain Message-ID: Is it possible that this is a Procion dye (chloro- -s triazine)? these have ben used extensively to dye fabrics and still are used for this purpose. Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Paul Bradbury Sent: Tue 11/15/2005 7:20 PM To: McCollough, Carol; HistoNet Server Subject: Re: [Histonet] cloth yellow stain For the last couple of hours I have been racking my brain to try to recall where I have come across this dye before. But, the required neurons are not responding ... or only weakly. John Kiernan is right on target ( as usual) in suggesting that this dye is almost certainly a large molecule azo dye (actually a very long, linear molecule) anionic dye probably used in the textile industry for coloring cotton fibres. If I remember correctly, such dyes are often referred to as "milling dyes". Similar azo dyes are commonly used for dying natural fibres in industry. In histology, dyes of this type are used to show amyloid (Congo red being the classical example). Yellow dyes provide poor visibility to tissue elements, so this dyes is almost certainly a counterstain used to highlight general background structures and to contrast with a red or blue primary stain. My suggestion would be to make up a 0.5% solution of the dye in 1% acetic acid, and try it out on a few sections of tissue. Let us know what the results are ... I am sure we would all love to know for sure.. Paul Bradbury Kamloops, BC Canada McCollough, Carol wrote: >Greetings Histonetters: > >In an exhaustive inventory of laboratory chemicals we have discovered an old bottle of 'Cloth Yellow' stain powder, manufactured by Chroma. Efforts to get an MSDS from Chroma have failed. Nobody here knows what this was even used for. > >Would anyone care to offer any histological uses for this stain? > >Regards - >Carol >********************** >Carol B. McCollough >Aquatic Animal Research Pathologist >Oyster Disease Research Project >Fisheries Service >Maryland Department of Natural Resources >Cooperative Oxford Laboratory >904 S. Morris Street >Oxford, Maryland 21654 >410-226-5193 x124 > >_______________________________________________ >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 rjbuesa <@t> yahoo.com Wed Nov 16 07:10:35 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 16 07:10:44 2005 Subject: [Histonet] Neg Mouse Controls In-Reply-To: Message-ID: <20051116131035.77126.qmail@web61216.mail.yahoo.com> Janci: In our SOP for IHC we required 1 negative control per block and that was what we were reqiured to do by CAP. You don't have to have a negative for each antibody as we used to do way back in 1983. Now NCCLS (Document MM4-A, vol.19, No.26 of Dec./99) recommends for Inspection Requirements (heading 7.1.4; pp 20) 1 positive and 1 negative tissue or cell control with each run, although there is not included the definition of "run". Since each block can have their own characteristics with regards to how it ended being fixed or processed I always used to have 1 negative control per block and stop doing 1 negative per block per antibody. Hope this will help! Rene J. Janci Wellborn wrote: Does anyone know if a neg mouse control is required by CAP on every slide in a case. If it is, what is the listing. We have a new pathologist and he ordered immuno staining on 8 blocks on the same case. The tech ran a neg mouse on each block. I has been my understanding that only one negative mouse is required for a case. Janci Wellborn, BS, BSeD, HTL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From jr <@t> infosight.com Wed Nov 16 07:42:57 2005 From: jr <@t> infosight.com (John Robertson) Date: Wed Nov 16 07:43:09 2005 Subject: [Histonet] Slide Adhesion Message-ID: <9B614E04-8475-4F11-ABEF-EA21961E61A5@infosight.com> We are interested in seeing if a new slide coating method has advantages for the adhesion of "difficult" sections. It should work well in all applications and ultimately be less costly than existing charged slides. If you have an especially difficult adhesion problem , I would like to hear from you and offer you a few prototype samples for testing. Thanks John John Robertson P.E. Ph.D. jr@infosight.com CEO InfoSight Corporation http://www.infosight.com " We Barcode Difficult Stuff" tm P.O. Box 5000 20700 Rt. 23 Chillicothe , OH 45601 Phone (740) 642 3600 (Eastern time zone) Fax (740) 642 5001 Private Fax. (740) 642 3106 PGP Key 0x8EDD65A2 ?Never Confuse Motion with Action? Ben Franklin From algranth <@t> u.arizona.edu Wed Nov 16 07:43:48 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Wed Nov 16 07:44:00 2005 Subject: [Histonet] Histology schools In-Reply-To: References: <111520052132.7317.437A53D400071E7800001C952200763704CECE030E9D0C9A03@comcast.net> Message-ID: <4.3.2.7.2.20051116063919.024fdd20@algranth.inbox.email.arizona.edu> ARIZONA Pima Community College, West Campus Histologic Technician 2202 W. Anklam Road Tucson, AZ 85709-0270 Prgm Dir: Sandra King Tel:520-206-6031 Email: slking_htascp@cox.net Lead Faculty: Jean Lindeberg Tel: 520-206-6832 PCC is now an accredited school and is getting ready to graduate another class of registry eligible histotechs. Many of those who graduated are now working in the Tucson area but there are some who might be interested in relocating. If interested, you could contact the school with your job opportunities. Andi Grantham ..................................................................... : 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 novanity <@t> nc.rr.com Wed Nov 16 07:55:59 2005 From: novanity <@t> nc.rr.com (novanity@nc.rr.com) Date: Wed Nov 16 07:56:09 2005 Subject: [Histonet] RE: QIHC change Message-ID: <69aa13696e34.696e3469aa13@southeast.rr.com> James, to qualify for the qualification you take the 50 question test and submit an employer reference form + of course satisfy one of the three routes. There is no practical to submit anymore. I think that is what you are asking. It seems to me that it wouldn't matter about specificity antigens/markers or what diseases or human cells. There is no requirement other than what is requested on the employer reference form which you can't see the details until you order and receive your packet. Is it possible for anyone to post a copy of the employer reference form. From the ASCP website this is what it says " Qualification in Immunohistochemistry Experience requirements Applicants must have experience in the following areas * Immunohistochemical and Immunofluorescence Preparation All of the following should have been performed by the applicant o staining technique o selection of proper control material o titration of immunologic reagents * Immunophenotyping in at least one of the following applications o immunodeficiencies o immunoproliferative disorders (neoplastic and non-neoplastic disorders) o transplantation biopsies o other immunophenotyping applications please specify: ______________________ * Quality Assurance The applicant should have participated in Quality Assurance related to all of the following o specimen fixation, processing, microtomy o reagent selection, preparation, storage, disposal o method selection, validation, documentation o quality control o safety " This is the experience which I am assuming is only documented for the ASCP through the employer reference form, hence if you only do A and C and not B you can't qualify unless your employer is dishonest on the form. Because even if you crosstrain into what I assume is flow cytometry but don't actually work it day to day as part of your job you do not qualify because you have not had experience doing it for a minimun of 12 months. As for research, same thing if you do all of it every day then your good to go. If not it is a grey or is it gray area that I'm looking more information/details on. In the past you qualified your work with different immuno stains as a practical , I don't remember there being a flow requirement. Maybe I'm wrong but anyone have this info I'm looking for. G Hurlburt HT(ASCP) sunny and warm NC From juan.gutierrez <@t> christushealth.org Wed Nov 16 07:59:49 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Wed Nov 16 08:00:01 2005 Subject: [Histonet] Histology schools Message-ID: Actually Joe, the UTHSCSA still has their program running. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Wednesday, November 16, 2005 6:09 AM To: Rene J Buesa; Bruce Gapinski; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology schools Unfortunately, the only program in San Antonio, Texas has just closed. That is too bad, I hired four students out of that class too! Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Rene J Buesa" To: "Bruce Gapinski" ; Sent: Tuesday, November 15, 2005 3:23 PM Subject: Re: [Histonet] Histology schools > In South Florida we have 2 histology programs, one at Barry University and > the other at > Miami Dade College. > Rene J. > > Bruce Gapinski wrote: > Are there any Histology schools? Where do people learn how to do what we > do? On the job training is too much for me anymore .I've trained about 8 > now. Besides, who wants to spend their (now required) A.S. in a histo lab? > Honestly, some times I wonder if we've boxed ourselves into the worst > situation. Anyone ELSE having trouble finding a histologist? Or is it an > histologist, I never know. > Bruce Gapinski HT (ASCP) > > > --------------------------------------------------------------------- > Important Notice: This e-mail is intended for the use of the person > to whom it is addressed and may contain information that is > privileged and confidential. If you are not the intended recipient, > any disclosure, copying, distribution, or use of the contents of > this message is strictly prohibited. If you have received this > e-mail in error, please destroy this message and contact the > Security Officer at PathGroup, Inc. immediately at > 615-221-4431._______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > --------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Free Edition. > Version: 7.1.362 / Virus Database: 267.13.2/170 - Release Date: 11/15/2005 > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jerry.santiago <@t> jax.ufl.edu Wed Nov 16 07:59:15 2005 From: jerry.santiago <@t> jax.ufl.edu (Santiago, Jerry) Date: Wed Nov 16 08:00:05 2005 Subject: [Histonet] Histology schools Message-ID: Florida Community College in Jacksonville. This is an Associate Degree Program, which is NAACLS accredited as well as a State approved program. All the didactics are taught on-line, the students spend two weekends a semester at the campus lab. All quizzes are taken on-line and exams are proctored at their training facilities. The practicum is done with affiliated facilities near to the student home. For more information, please visit www.fccj.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bruce Gapinski Sent: Tuesday, November 15, 2005 4:11 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology schools Are there any Histology schools? Where do people learn how to do what we do? On the job training is too much for me anymore .I've trained about 8 now. Besides, who wants to spend their (now required) A.S. in a histo lab? Honestly, some times I wonder if we've boxed ourselves into the worst situation. Anyone ELSE having trouble finding a histologist? Or is it an histologist, I never know. Bruce Gapinski HT (ASCP) --------------------------------------------------------------------- Important Notice: This e-mail is intended for the use of the person to whom it is addressed and may contain information that is privileged and confidential. If you are not the intended recipient, any disclosure, copying, distribution, or use of the contents of this message is strictly prohibited. If you have received this e-mail in error, please destroy this message and contact the Security Officer at PathGroup, Inc. immediately at 615-221-4431. From JWEEMS <@t> sjha.org Wed Nov 16 08:21:58 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Nov 16 08:22:07 2005 Subject: [Histonet] Histology schools Message-ID: <83AACDB0810528418AA106F9AE9B7F7E013050FC@sjhaexc02.sjha.org> Darton Community College in Albany, GA has an online program - info at the following website. http://www.darton.edu/programs/AlliedHealth/fs.php Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrea Grantham Sent: Wednesday, November 16, 2005 8:44 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histology schools ARIZONA Pima Community College, West Campus Histologic Technician 2202 W. Anklam Road Tucson, AZ 85709-0270 Prgm Dir: Sandra King Tel:520-206-6031 Email: slking_htascp@cox.net Lead Faculty: Jean Lindeberg Tel: 520-206-6832 PCC is now an accredited school and is getting ready to graduate another class of registry eligible histotechs. Many of those who graduated are now working in the Tucson area but there are some who might be interested in relocating. If interested, you could contact the school with your job opportunities. Andi Grantham ..................................................................... : 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 _______________________________________________ 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 Joseph's Health System, Inc. From novanity <@t> nc.rr.com Wed Nov 16 08:29:06 2005 From: novanity <@t> nc.rr.com (novanity@nc.rr.com) Date: Wed Nov 16 08:29:19 2005 Subject: [Histonet] RE: histology schools/Malcolm Message-ID: <6b7d356b44dd.6b44dd6b7d35@southeast.rr.com> malcolm a school would need to do a number of things. From NAACLS comes "Basic Eligibility Criteria for Becoming An Accredited Program NAACLS applies the following basic eligibility criteria when it considers an applicant program for initial accreditation: 1. The sponsoring institution and affiliates, clinical and/or academic, if any, must be accredited by recongnized regional and or natinal agencies. 2. Academic institutions sponsoring clinical laboratory science education programs must be empowered by a state authority to grant the appropriate degree. 3. The institution must be legally authorized under applicable state law to provide postsecondary education." You can find the rest in pdf format at: http://www.naacls.org/accreditation/ht/ The guide to accreditation goes through a process of self study of the program, paper self review of the program, site visit of program, site review of program and then board approval. In the past this was handled by CAHEA for a fee but now it is handled as far as I can tell by NAACLS for HT and HTL programs among the other allied health programs out there looking for accreditation to qualify its graduates to take the ASCP registrys'. Hope this helps you find the direction for what your school may have in mind. It would be great to see more schools opening than closing. orginal message said From: "Malcolm McCallum" Now this is a new thing to me. What would a school need to do to offer a histotech certificate or degree (bs) program??? Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html From Sandra.Harrison3 <@t> va.gov Wed Nov 16 08:39:22 2005 From: Sandra.Harrison3 <@t> va.gov (Harrison, Sandra C.) Date: Wed Nov 16 08:39:32 2005 Subject: [Histonet] best formalin Message-ID: <736E8889E98B8F4FBBD29FDFEC8074BA49BF3A@VHAV23MSGA2.v23.med.va.gov> Is there a brand of formalin that is optimal for a routine hospital histology lab? We use an automated tissue processor and have approx. 150 blocks per day. From TMcNemar <@t> lmhealth.org Wed Nov 16 09:19:11 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Wed Nov 16 09:23:45 2005 Subject: [Histonet] Biohazard Labeling Message-ID: <6CD94D97ED7D924BA5C2B588FA95282139681D@nt_exchange.lmhealth.org> I would certainly consider fresh tissue to be biohazard but ours are not labeled as such. We only get fresh tissue from surgery and they are always hand delivered to us immediately after collection so they are only handled by trained personnel. We are JCAHO and they have never expressed any concern. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Tuesday, November 15, 2005 3:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Biohazard Labeling Can anyone give me information on the labeling of fresh specimens? In the past, I was told by our infection control coordinator that fresh specimens did not need to be labeled as "Biohazard." Now, I'm being told they do need to be labeled. The new coordinator is getting me more info, but can anyone guide me to info from OSHA, CAP, JCAHO, etc. that refers to this topic? Laurie Colbert Huntington Memorial Hospital _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Nov 16 09:26:46 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 16 09:26:58 2005 Subject: [Histonet] best formalin In-Reply-To: <736E8889E98B8F4FBBD29FDFEC8074BA49BF3A@VHAV23MSGA2.v23.med.va.gov> Message-ID: <20051116152646.41026.qmail@web61219.mail.yahoo.com> Sandra: Formalin, as a neutral buffered solution (NBF), is pretty much the same from any supplier. You should go with the one that provides you with the most advantageous offer from a reputable supplier. Also there is something else to consider: are you going to supply vials with NBF to others or are you just going to use the NBF for your tissue processor? If you are not going to supply the NBF to others, you should know how they are fixing the tissues you are going to process to see if whatever they use is convenient for you. Rene J. "Harrison, Sandra C." wrote: Is there a brand of formalin that is optimal for a routine hospital histology lab? We use an automated tissue processor and have approx. 150 blocks per day. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From jwatson <@t> gnf.org Wed Nov 16 09:28:20 2005 From: jwatson <@t> gnf.org (James Watson) Date: Wed Nov 16 09:28:37 2005 Subject: [Histonet] RE: QIHC change Message-ID: This is my point. With the requirements listed below someone with 25 years of experience doing immuno (single, double, triple antibody staining, making own antibodies, and in situ Hybridization: all with and without using kits, all with and without using an automated stainer) is not qualified for this certification if they work in a research facility where immunophenotyping is not done. There is no system of doing it on your own to prove that you have the capability to do immunophenotyping in order to fullfil this requirement. I guess it is time to start harrassing ASCP about the unfairness of this system. From almost always sunny San Diego Jamie -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of novanity@nc.rr.com Sent: Wed 11/16/2005 5:55 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] RE: QIHC change James, to qualify for the qualification you take the 50 question test and submit an employer reference form + of course satisfy one of the three routes. There is no practical to submit anymore. I think that is what you are asking. It seems to me that it wouldn't matter about specificity antigens/markers or what diseases or human cells. There is no requirement other than what is requested on the employer reference form which you can't see the details until you order and receive your packet. Is it possible for anyone to post a copy of the employer reference form. From the ASCP website this is what it says " Qualification in Immunohistochemistry Experience requirements Applicants must have experience in the following areas * Immunohistochemical and Immunofluorescence Preparation All of the following should have been performed by the applicant o staining technique o selection of proper control material o titration of immunologic reagents * Immunophenotyping in at least one of the following applications o immunodeficiencies o immunoproliferative disorders (neoplastic and non-neoplastic disorders) o transplantation biopsies o other immunophenotyping applications please specify: ______________________ * Quality Assurance The applicant should have participated in Quality Assurance related to all of the following o specimen fixation, processing, microtomy o reagent selection, preparation, storage, disposal o method selection, validation, documentation o quality control o safety " This is the experience which I am assuming is only documented for the ASCP through the employer reference form, hence if you only do A and C and not B you can't qualify unless your employer is dishonest on the form. Because even if you crosstrain into what I assume is flow cytometry but don't actually work it day to day as part of your job you do not qualify because you have not had experience doing it for a minimun of 12 months. As for research, same thing if you do all of it every day then your good to go. If not it is a grey or is it gray area that I'm looking more information/details on. In the past you qualified your work with different immuno stains as a practical , I don't remember there being a flow requirement. Maybe I'm wrong but anyone have this info I'm looking for. G Hurlburt HT(ASCP) sunny and warm NC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MVaughan4 <@t> ucok.edu Wed Nov 16 09:33:54 2005 From: MVaughan4 <@t> ucok.edu (MVaughan4@ucok.edu) Date: Wed Nov 16 09:34:42 2005 Subject: [Histonet] Histology schools; Techniques course suggestions In-Reply-To: Message-ID: Dr. Amy Aulthouse directs a Histotechniques course at Ohio Northern University in Ada, Ohio. I think she teaches it occasionally and has a limited number of students participating. "BIOL 343 Histological Techniques - Principles and procedures used in the preparation of biological specimens for microscopic study. Students learn both routine stains (H&E) and special histochemical techniques." We (University of Central Oklahoma) have a Microtechniques course where the students learn how to fix, section and stain paraffin sections. I also train one or two students a semester doing independent research using these techniques as well as IHC. I haven't yet taught the Microtech course because I am teaching most of the biomedical courses in the department. It sounds like I need to make a case for teaching the course next semester. The course is only capable of carrying 10 students, so it is comparable to other such classes. I don't think a lot of people realize there is a problem finding histotechs. I am currently in the process of updating the course material. I plan to include immunohistochemistry and metaphase spreads/chromosome staining into the curriculum. This discussion group is full of many professionals in the field, and I would welcome any suggestions as to what other kinds of techniques might be useful to train students so they would be competitive for a histologist position. Mel Melville B. Vaughan, Ph. D. Assistant Professor Department of Biology University of Central Oklahoma Edmond, OK 73034 From int09018 <@t> alphahunt.com Wed Nov 16 09:57:17 2005 From: int09018 <@t> alphahunt.com (HCS) Date: Wed Nov 16 09:57:34 2005 Subject: [Histonet] looking for methods for triphenyltetrazolium chloride (TTC) stains Message-ID: <000e01c5eac6$6c2f23a0$6601a8c0@hp> Does anyone have a protocol for triphenyltetrazolium chloride (TTC) stained slides? Thanks. LeRoy LeRoy Brown HT(ASCP) HTL Histology Consultation Services PO Box 770 207 N Harkness St Everson, WA 98247 www.histocs.com 360-966-7300 fax : 360-543-5626 From mistressjsofla <@t> yahoo.com Wed Nov 16 10:16:26 2005 From: mistressjsofla <@t> yahoo.com (MJ) Date: Wed Nov 16 10:16:36 2005 Subject: [Histonet] Help Need with Tissue for Practical Exam 2006 Message-ID: <20051116161626.95288.qmail@web30801.mail.mud.yahoo.com> I need help getting the following tissue for my Practical exam for 2006 Bone must be at least 1.0cm long Esophagus must be at least 1.5cm I would appreciate any help. Anyone who is a Histologist in the South East Florida area. I would be willing to pick up the tissue in person. --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From jcox90 <@t> yahoo.com Wed Nov 16 10:34:49 2005 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Wed Nov 16 10:34:59 2005 Subject: [Histonet] Looking for Histotechs for new lab in Phoenix Message-ID: <20051116163449.65792.qmail@web52109.mail.yahoo.com> We are opening a new Lab here in sunny Phoenix and looking for Histotechs. Lab will be fully automated with state of the art equipment, new building and a wonderful place to work. We also are looking for Lab Assistants, Couriers and Transcriptionists. Please email me if you are interested or know of anyone. We will be probably be opening by March but need to bring people on about a month prior. Have a great day, Jill Jill Cox HT (ASCP) From tpmorken <@t> labvision.com Wed Nov 16 11:07:46 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Wed Nov 16 11:07:59 2005 Subject: [Histonet] RE: QIHC change Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D357@usca0082k08.labvision.apogent.com> Jamie, It seems from what you say that you are working in a research lab. Is that correct? My understanding about the ASCP certification is that it is aimed at providing a modicum of proof that a person is qualified to work in a medical diagnostic lab. Research labs are not considered diagnostic labs. As you imply, a person in a research lab will often work on only a limited sample set. Therefore, it is meaningless to apply the the ASCP standard to research people. If you are planning to move into the diagnostic field, then I'll bet you could easily find a job in a diagnostic lab, get the experience, and qualify to take the test. It may be that some diagnostic labs have a suggested requirement to be ASCP certified as a QIHC, but the vast majority would be happy to find someone with the experience you outline, even if they had not previously worked in a diagnositc lab. 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: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James Watson Sent: Wednesday, November 16, 2005 7:28 AM To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: QIHC change This is my point. With the requirements listed below someone with 25 years of experience doing immuno (single, double, triple antibody staining, making own antibodies, and in situ Hybridization: all with and without using kits, all with and without using an automated stainer) is not qualified for this certification if they work in a research facility where immunophenotyping is not done. There is no system of doing it on your own to prove that you have the capability to do immunophenotyping in order to fullfil this requirement. I guess it is time to start harrassing ASCP about the unfairness of this system. >From almost always sunny San Diego Jamie -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of novanity@nc.rr.com Sent: Wed 11/16/2005 5:55 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] RE: QIHC change James, to qualify for the qualification you take the 50 question test and submit an employer reference form + of course satisfy one of the three routes. There is no practical to submit anymore. I think that is what you are asking. It seems to me that it wouldn't matter about specificity antigens/markers or what diseases or human cells. There is no requirement other than what is requested on the employer reference form which you can't see the details until you order and receive your packet. Is it possible for anyone to post a copy of the employer reference form. From the ASCP website this is what it says " Qualification in Immunohistochemistry Experience requirements Applicants must have experience in the following areas * Immunohistochemical and Immunofluorescence Preparation All of the following should have been performed by the applicant o staining technique o selection of proper control material o titration of immunologic reagents * Immunophenotyping in at least one of the following applications o immunodeficiencies o immunoproliferative disorders (neoplastic and non-neoplastic disorders) o transplantation biopsies o other immunophenotyping applications please specify: ______________________ * Quality Assurance The applicant should have participated in Quality Assurance related to all of the following o specimen fixation, processing, microtomy o reagent selection, preparation, storage, disposal o method selection, validation, documentation o quality control o safety " This is the experience which I am assuming is only documented for the ASCP through the employer reference form, hence if you only do A and C and not B you can't qualify unless your employer is dishonest on the form. Because even if you crosstrain into what I assume is flow cytometry but don't actually work it day to day as part of your job you do not qualify because you have not had experience doing it for a minimun of 12 months. As for research, same thing if you do all of it every day then your good to go. If not it is a grey or is it gray area that I'm looking more information/details on. In the past you qualified your work with different immuno stains as a practical , I don't remember there being a flow requirement. Maybe I'm wrong but anyone have this info I'm looking for. G Hurlburt HT(ASCP) sunny and warm NC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Wed Nov 16 11:08:48 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Wed Nov 16 11:09:02 2005 Subject: [Histonet] best formalin In-Reply-To: <20051116152646.41026.qmail@web61219.mail.yahoo.com> References: <736E8889E98B8F4FBBD29FDFEC8074BA49BF3A@VHAV23MSGA2.v23.med.va.gov> <20051116152646.41026.qmail@web61219.mail.yahoo.com> Message-ID: <6.1.1.1.2.20051116120254.019b4518@mail.vet.upenn.edu> At 10:26 AM 11/16/2005, Rene J Buesa wrote: >Sandra: > Formalin, as a neutral buffered solution (NBF), is pretty much the same > from >any supplier. You should go with the one that provides you with the most > advantageous offer from a reputable supplier. > Also there is something else to consider: are you going to supply vials > with > NBF to others or are you just going to use the NBF for your tissue > processor? > If you are not going to supply the NBF to others, you should know how > they are > fixing the tissues you are going to process to see if whatever they use > is convenient > for you. > Rene J. >"Harrison, Sandra C." wrote: > Is there a brand of formalin that is optimal for a routine hospital >histology lab? We use an automated tissue processor and have approx. >150 blocks per day. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > >--------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet I am sorry I must disagree as there are several different formulas now available in pre-made 10% NBF with different buffer components. It is best to decide which buffer base you require and stay with that formula. The most popular and common is the sodium phosphate base with a combination of mono and dibasic salts. The formalin mixtures with citrate or single salts may not hold buffering capacity as well and can precipitate if temperature ranges vary too much even short time frames. These may also have changes in pH due to the salt concentration change. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From PMonfils <@t> Lifespan.org Wed Nov 16 11:40:06 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Wed Nov 16 11:40:18 2005 Subject: [Histonet] aqueous coverslipping medium Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717617@lsexch.lsmaster.lifespan.org> What brand do you prefer for aqueous mounting medium to be used with a coverslip? I have several good hydrocarbon-based coverslipping media, and also some good aqueous media to be used without coverslips (so-called liquid coverslip), but I don't have an aqueous coverslipping medium I am completely satisfied with. Applications would include immunofluorescence and a few enzyme stains, as well as lipid stains and other techniques that are not compatible with hydrocarbon-based media. Are there any such media that harden after application, to produce a permanent slide? From pmarcum <@t> vet.upenn.edu Wed Nov 16 11:56:02 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Wed Nov 16 11:56:25 2005 Subject: [Histonet] RE: QIHC change In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328D357@usca0082k08.labvi sion.apogent.com> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328D357@usca0082k08.labvision.apogent.com> Message-ID: <6.1.1.1.2.20051116123818.019c43e0@mail.vet.upenn.edu> Hi Tim and Jamie, I know Jamie and have for many years and you are right anyone would be happy to hire him for his experience in clinical or his current research position. However, it is also true that now research positions are often asking for at least an HT or even HTL (ASCP) to fill position with histology as the main focus. Yet we are given a set of criteria for tissue that often excludes animal research applicants from completing the practical easily. I took my HT many years ago and I was told that even in a research position (and I had a BS at the time) it would improve my salary and increase me to higher level in the university if I got my HT. I was told not to use animal tissue (1976) as no one reading the exam could properly read them. Now we have veterinary person there and tissue requirements can still eliminate some people or make it almost impossible to complete the practical with out help in procurement. Why should that happen to some one attempting to improve their position within the histology community? My real problem with what you said about the QIHC is that I would also like to take it and can not qualify either. Yet those of us in research are often finding the very antibodies and test methods companies and diagnostics later fight to get or learn. We are exempt in your mind and ASCP's even though research is what you depend on often for advances. I have never and will never understand this logic and exempt status for those of us who chose not to be clinical. We are still often required to have or get ASCP status as a way to advance and prove we know our field. ASCP needs to get up to date on the fields it is registering or make new categories for those of still contribute to clinical advances every year. Sorry if sounds like I am picking on you Tim. I just don't see how we are required to be registered on one hand for acceptance (even NSH likes to see it) and discounted on the other. Pam Marcum UPENN Vet School New Bolton Center Kennett Square, PA 19384 610-925-6278 At 12:07 PM 11/16/2005, Morken, Tim - Labvision wrote: >Jamie, It seems from what you say that you are working in a research lab. Is >that correct? My understanding about the ASCP certification is that it is >aimed at providing a modicum of proof that a person is qualified to work in >a medical diagnostic lab. Research labs are not considered diagnostic labs. >As you imply, a person in a research lab will often work on only a limited >sample set. Therefore, it is meaningless to apply the the ASCP standard to >research people. > > If you are planning to move into the diagnostic field, then I'll bet you >could easily find a job in a diagnostic lab, get the experience, and qualify >to take the test. It may be that some diagnostic labs have a suggested >requirement to be ASCP certified as a QIHC, but the vast majority would be >happy to find someone with the experience you outline, even if they had not >previously worked in a diagnositc lab. > >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: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James Watson >Sent: Wednesday, November 16, 2005 7:28 AM >To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: QIHC change > > >This is my point. With the requirements listed below someone with 25 years >of experience doing immuno (single, double, triple antibody staining, making >own antibodies, and in situ Hybridization: all with and without using kits, >all with and without using an automated stainer) is not qualified for this >certification if they work in a research facility where immunophenotyping is >not done. There is no system of doing it on your own to prove that you have >the capability to do immunophenotyping in order to fullfil this requirement. >I guess it is time to start harrassing ASCP about the unfairness of this >system. > > >From almost always sunny San Diego >Jamie > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of >novanity@nc.rr.com > Sent: Wed 11/16/2005 5:55 AM > To: histonet@lists.utsouthwestern.edu > Cc: > Subject: [Histonet] RE: QIHC change > > > > James, to qualify for the qualification you take the 50 question >test > and submit an employer reference form + of course satisfy one of the > three routes. There is no practical to submit anymore. I think >that is > what you are asking. It seems to me that it wouldn't matter about > specificity antigens/markers or what diseases or human cells. There >is > no requirement other than what is requested on the employer >reference > form which you can't see the details until you order and receive >your > packet. Is it possible for anyone to post a copy of the employer > reference form. From the ASCP website this is what it says " > > Qualification in Immunohistochemistry > Experience requirements > > Applicants must have experience in the following areas > > * Immunohistochemical and Immunofluorescence Preparation > All of the following should have been performed by the >applicant > o staining technique > o selection of proper control material > o titration of immunologic reagents > > * Immunophenotyping > in at least one of the following applications > o immunodeficiencies > o immunoproliferative disorders (neoplastic and >non-neoplastic > disorders) > o transplantation biopsies > o other immunophenotyping applications > please specify: ______________________ > > * Quality Assurance > The applicant should have participated in Quality Assurance > related to all of the following > o specimen fixation, processing, microtomy > o reagent selection, preparation, storage, disposal > o method selection, validation, documentation > o quality control > o safety > > " This is the experience which I am assuming is only documented for >the > ASCP through the employer reference form, hence if you only do A and >C > and not B you can't qualify unless your employer is dishonest on the > form. Because even if you crosstrain into what I assume is flow > cytometry but don't actually work it day to day as part of your job >you > do not qualify because you have not had experience doing it for a > minimun of 12 months. As for research, same thing if you do all of >it > every day then your good to go. If not it is a grey or is it gray >area > that I'm looking more information/details on. In the past you > qualified your work with different immuno stains as a practical , I > don't remember there being a flow requirement. Maybe I'm wrong but > anyone have this info I'm looking for. > > G Hurlburt HT(ASCP) > sunny and warm NC > > _______________________________________________ > 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 Wed Nov 16 12:09:33 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 16 12:09:46 2005 Subject: [Histonet] best formalin In-Reply-To: <6.1.1.1.2.20051116120254.019b4518@mail.vet.upenn.edu> References: <736E8889E98B8F4FBBD29FDFEC8074BA49BF3A@VHAV23MSGA2.v23.med.va.gov> <20051116152646.41026.qmail@web61219.mail.yahoo.com> <6.1.1.1.2.20051116120254.019b4518@mail.vet.upenn.edu> Message-ID: <6.0.0.22.1.20051116104226.01b6ef38@gemini.msu.montana.edu> We like a neutral buffered formalin (phosphate salts) and purchase it either ready to use or in concentrated form (to make a final working solution) We find NBF concentrates convenient and economical. Several of our labs prefer ready to use phophate buffered formalin i.e. NBF(Fisherbrand) in order to avoid formaldehyde fumes during NBF preparation without an available fume hood. . Richard Allan, Surgipath and others have formalin concentrates available. Surgipath has Carsons Millonigs Buffered Formalin with slightly different buffering formulation using sodium hydroxide and a phosphate salt, and MBF also comes in concentrated form. The joy of this one is - it was developed for use with both FFPE and EM per Freida Carson's publication in JOH many years ago. All of the above provided excellent fixation. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From cforster <@t> umn.edu Wed Nov 16 12:27:10 2005 From: cforster <@t> umn.edu (Colleen Forster) Date: Wed Nov 16 12:27:23 2005 Subject: [Histonet] RE: QIHC change In-Reply-To: <6.1.1.1.2.20051116123818.019c43e0@mail.vet.upenn.edu> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328D357@usca0082k08.labvision.apogent.com> <6.1.1.1.2.20051116123818.019c43e0@mail.vet.upenn.edu> Message-ID: <437B79FE.7090208@umn.edu> I agree with Pam ...completely. I too do research and I am HT certified because I started in the clinical route. While in the clinical lab I tested for the QIHC. It has been a GREAT benefit for me in my research positions and I encourage all people pursuning histology work weather research or clinicla to get registered. One never knows what turns life might take and the added skills/knoweledge will be a benefit!! ASCP definitiely needs to widen their range! Colleen Forster U of MN Pamela Marcum wrote: > Hi Tim and Jamie, > > I know Jamie and have for many years and you are right anyone would be > happy to hire him for his experience in clinical or his current > research position. However, it is also true that now research > positions are often asking for at least an HT or even HTL (ASCP) to > fill position with histology as the main focus. Yet we are given a > set of criteria for tissue that often excludes animal research > applicants from completing the practical easily. I took my HT many > years ago and I was told that even in a research position (and I had a > BS at the time) it would improve my salary and increase me to higher > level in the university if I got my HT. I was told not to use animal > tissue (1976) as no one reading the exam could properly read them. > Now we have veterinary person there and tissue requirements can still > eliminate some people or make it almost impossible to complete the > practical with out help in procurement. Why should that happen to > some one attempting to improve their position within the histology > community? > > My real problem with what you said about the QIHC is that I would also > like to take it and can not qualify either. Yet those of us in > research are often finding the very antibodies and test methods > companies and diagnostics later fight to get or learn. We are exempt > in your mind and ASCP's even though research is what you depend on > often for advances. I have never and will never understand this logic > and exempt status for those of us who chose not to be clinical. We > are still often required to have or get ASCP status as a way to > advance and prove we know our field. ASCP needs to get up to date on > the fields it is registering or make new categories for those of still > contribute to clinical advances every year. > > Sorry if sounds like I am picking on you Tim. I just don't see how > we are required to be registered on one hand for acceptance (even NSH > likes to see it) and discounted on the other. > > Pam Marcum > UPENN Vet School > New Bolton Center > Kennett Square, PA 19384 > 610-925-6278 > > > At 12:07 PM 11/16/2005, Morken, Tim - Labvision wrote: > >> Jamie, It seems from what you say that you are working in a research >> lab. Is >> that correct? My understanding about the ASCP certification is that >> it is >> aimed at providing a modicum of proof that a person is qualified to >> work in >> a medical diagnostic lab. Research labs are not considered >> diagnostic labs. >> As you imply, a person in a research lab will often work on only a >> limited >> sample set. Therefore, it is meaningless to apply the the ASCP >> standard to >> research people. >> >> If you are planning to move into the diagnostic field, then I'll bet >> you >> could easily find a job in a diagnostic lab, get the experience, and >> qualify >> to take the test. It may be that some diagnostic labs have a suggested >> requirement to be ASCP certified as a QIHC, but the vast majority >> would be >> happy to find someone with the experience you outline, even if they >> had not >> previously worked in a diagnositc lab. >> >> 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: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James >> Watson >> Sent: Wednesday, November 16, 2005 7:28 AM >> To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu >> Subject: RE: [Histonet] RE: QIHC change >> >> >> This is my point. With the requirements listed below someone with 25 >> years >> of experience doing immuno (single, double, triple antibody staining, >> making >> own antibodies, and in situ Hybridization: all with and without using >> kits, >> all with and without using an automated stainer) is not qualified for >> this >> certification if they work in a research facility where >> immunophenotyping is >> not done. There is no system of doing it on your own to prove that >> you have >> the capability to do immunophenotyping in order to fullfil this >> requirement. >> I guess it is time to start harrassing ASCP about the unfairness of this >> system. >> >> >From almost always sunny San Diego >> Jamie >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of >> novanity@nc.rr.com >> Sent: Wed 11/16/2005 5:55 AM >> To: histonet@lists.utsouthwestern.edu >> Cc: >> Subject: [Histonet] RE: QIHC change >> >> >> >> James, to qualify for the qualification you take the 50 question >> test >> and submit an employer reference form + of course satisfy one >> of the >> three routes. There is no practical to submit anymore. I think >> that is >> what you are asking. It seems to me that it wouldn't matter >> about >> specificity antigens/markers or what diseases or human >> cells. There >> is >> no requirement other than what is requested on the employer >> reference >> form which you can't see the details until you order and receive >> your >> packet. Is it possible for anyone to post a copy of the employer >> reference form. From the ASCP website this is what it says " >> >> Qualification in Immunohistochemistry >> Experience requirements >> >> Applicants must have experience in the following areas >> >> * Immunohistochemical and Immunofluorescence Preparation >> All of the following should have been performed by the >> applicant >> o staining technique >> o selection of proper control material >> o titration of immunologic reagents >> >> * Immunophenotyping >> in at least one of the following applications >> o immunodeficiencies >> o immunoproliferative disorders (neoplastic and >> non-neoplastic >> disorders) >> o transplantation biopsies >> o other immunophenotyping applications >> please specify: ______________________ >> >> * Quality Assurance >> The applicant should have participated in Quality >> Assurance >> related to all of the following >> o specimen fixation, processing, microtomy >> o reagent selection, preparation, storage, disposal >> o method selection, validation, documentation >> o quality control >> o safety >> >> " This is the experience which I am assuming is only >> documented for >> the >> ASCP through the employer reference form, hence if you only >> do A and >> C >> and not B you can't qualify unless your employer is dishonest >> on the >> form. Because even if you crosstrain into what I assume is flow >> cytometry but don't actually work it day to day as part of >> your job >> you >> do not qualify because you have not had experience doing it >> for a >> minimun of 12 months. As for research, same thing if you do >> all of >> it >> every day then your good to go. If not it is a grey or is it >> gray >> area >> that I'm looking more information/details on. In the past you >> qualified your work with different immuno stains as a >> practical , I >> don't remember there being a flow requirement. Maybe I'm >> wrong but >> anyone have this info I'm looking for. >> >> G Hurlburt HT(ASCP) >> sunny and warm NC >> >> _______________________________________________ >> 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 DeBrosse_Beatrice <@t> Allergan.com Wed Nov 16 12:26:56 2005 From: DeBrosse_Beatrice <@t> Allergan.com (DeBrosse_Beatrice) Date: Wed Nov 16 12:27:46 2005 Subject: [Histonet] RE: QIHC change Message-ID: I agree with Pam that the ASCP needs to update on several things. I was qualified to take the QIHC, which I did and passed, but I'm sure Jamie can run circles around me since he has overall a whole lot more experience. I ran into a similar problem; being educated in Switzerland, where I got a degree as a laboratory technician in biology, according to the ASCP I cannot take the HTL just because my degree is not equivalent to a BS from the US. I went to a technical school, which doesn't qualify, but I've been trained specifically for being a laboratory technician in histology and microbiology. I can fully understand Jamie's frustration! Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pamela Marcum Sent: Wednesday, November 16, 2005 9:56 AM To: Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: QIHC change Hi Tim and Jamie, I know Jamie and have for many years and you are right anyone would be happy to hire him for his experience in clinical or his current research position. However, it is also true that now research positions are often asking for at least an HT or even HTL (ASCP) to fill position with histology as the main focus. Yet we are given a set of criteria for tissue that often excludes animal research applicants from completing the practical easily. I took my HT many years ago and I was told that even in a research position (and I had a BS at the time) it would improve my salary and increase me to higher level in the university if I got my HT. I was told not to use animal tissue (1976) as no one reading the exam could properly read them. Now we have veterinary person there and tissue requirements can still eliminate some people or make it almost impossible to complete the practical with out help in procurement. Why should that happen to some one attempting to improve their position within the histology community? My real problem with what you said about the QIHC is that I would also like to take it and can not qualify either. Yet those of us in research are often finding the very antibodies and test methods companies and diagnostics later fight to get or learn. We are exempt in your mind and ASCP's even though research is what you depend on often for advances. I have never and will never understand this logic and exempt status for those of us who chose not to be clinical. We are still often required to have or get ASCP status as a way to advance and prove we know our field. ASCP needs to get up to date on the fields it is registering or make new categories for those of still contribute to clinical advances every year. Sorry if sounds like I am picking on you Tim. I just don't see how we are required to be registered on one hand for acceptance (even NSH likes to see it) and discounted on the other. Pam Marcum UPENN Vet School New Bolton Center Kennett Square, PA 19384 610-925-6278 At 12:07 PM 11/16/2005, Morken, Tim - Labvision wrote: >Jamie, It seems from what you say that you are working in a research lab. Is >that correct? My understanding about the ASCP certification is that it is >aimed at providing a modicum of proof that a person is qualified to work in >a medical diagnostic lab. Research labs are not considered diagnostic labs. >As you imply, a person in a research lab will often work on only a limited >sample set. Therefore, it is meaningless to apply the the ASCP standard to >research people. > > If you are planning to move into the diagnostic field, then I'll bet you >could easily find a job in a diagnostic lab, get the experience, and qualify >to take the test. It may be that some diagnostic labs have a suggested >requirement to be ASCP certified as a QIHC, but the vast majority would be >happy to find someone with the experience you outline, even if they had not >previously worked in a diagnositc lab. > >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: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James Watson >Sent: Wednesday, November 16, 2005 7:28 AM >To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: QIHC change > > >This is my point. With the requirements listed below someone with 25 years >of experience doing immuno (single, double, triple antibody staining, making >own antibodies, and in situ Hybridization: all with and without using kits, >all with and without using an automated stainer) is not qualified for this >certification if they work in a research facility where immunophenotyping is >not done. There is no system of doing it on your own to prove that you have >the capability to do immunophenotyping in order to fullfil this requirement. >I guess it is time to start harrassing ASCP about the unfairness of this >system. > > >From almost always sunny San Diego >Jamie > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of >novanity@nc.rr.com > Sent: Wed 11/16/2005 5:55 AM > To: histonet@lists.utsouthwestern.edu > Cc: > Subject: [Histonet] RE: QIHC change > > > > James, to qualify for the qualification you take the 50 question >test > and submit an employer reference form + of course satisfy one of the > three routes. There is no practical to submit anymore. I think >that is > what you are asking. It seems to me that it wouldn't matter about > specificity antigens/markers or what diseases or human cells. There >is > no requirement other than what is requested on the employer >reference > form which you can't see the details until you order and receive >your > packet. Is it possible for anyone to post a copy of the employer > reference form. From the ASCP website this is what it says " > > Qualification in Immunohistochemistry > Experience requirements > > Applicants must have experience in the following areas > > * Immunohistochemical and Immunofluorescence Preparation > All of the following should have been performed by the >applicant > o staining technique > o selection of proper control material > o titration of immunologic reagents > > * Immunophenotyping > in at least one of the following applications > o immunodeficiencies > o immunoproliferative disorders (neoplastic and >non-neoplastic > disorders) > o transplantation biopsies > o other immunophenotyping applications > please specify: ______________________ > > * Quality Assurance > The applicant should have participated in Quality Assurance > related to all of the following > o specimen fixation, processing, microtomy > o reagent selection, preparation, storage, disposal > o method selection, validation, documentation > o quality control > o safety > > " This is the experience which I am assuming is only documented for >the > ASCP through the employer reference form, hence if you only do A and >C > and not B you can't qualify unless your employer is dishonest on the > form. Because even if you crosstrain into what I assume is flow > cytometry but don't actually work it day to day as part of your job >you > do not qualify because you have not had experience doing it for a > minimun of 12 months. As for research, same thing if you do all of >it > every day then your good to go. If not it is a grey or is it gray >area > that I'm looking more information/details on. In the past you > qualified your work with different immuno stains as a practical , I > don't remember there being a flow requirement. Maybe I'm wrong but > anyone have this info I'm looking for. > > G Hurlburt HT(ASCP) > sunny and warm NC > > _______________________________________________ > 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 judi.ford <@t> jax.org Wed Nov 16 12:46:52 2005 From: judi.ford <@t> jax.org (judi.ford@jax.org) Date: Wed Nov 16 12:48:47 2005 Subject: [Histonet] Luxol fast blue/cresyl violet stains Message-ID: <714463.1132166812935.JavaMail.ocsadmin@jcs-mid-prod.jax.org> Hi, I have a question concerning the lfb/cv stain. This is the situation we have in our lab. The tissue we are working on is Bouin's fixed mouse brain and spinal cord (cord is left in the vertebrae). The spinal cord may be left in Bouin's for an extended period for decalcification (two-three weeks). The trimmed tissue is then sent to our lab where we rinse it for a day before processing on our VIP processors. The next day we embed the tissue and then it is cut by one of our technicians and stained. This is our staining technique: deparaffinize to 95% alcohol and place in alcoholic luxol fast blue overnight at 37 degrees. The next morning it is rinsed twice in 95% alcohol and twice in distilled water. The slides are separated into racks containing either brain or spinal cord for differentiation. Slides are dipped in 70% alcohol for 20 seconds then lithium carbonate for 20 seconds. This is followed by two rinses in distilled water. Then checked microscopically before determining if differentiation is complete or not. If not, then the slides go another round through 70% and lithium for a few more seconds. Once this part is done the slides are put into a warmed cresyl violet solution (we add 10% glacial acetic acid, 10 drops for every 100mls of stain) for 4-6 minutes then rinsed in 95% alcohol three times. Finally they are dehydrated, cleared and coverslipped. The problem comes when we have 5 slides from the same animal, all processsed and stained together, where in one slide the cresyl violet works but in the other 4 it doesnt' work. Every slide is treated exactly the same, yet we have this difference. Any ideas on what may be happening? We've discussed decalcification and possible lithium effects on the cresyl violet. We're at a loss and our pathologist is very determined to discover the solution for this problem. Thank you advance for any ideas you may send our way..........We greatly appreciate your help! Judi Ford HIstotechnologist HIstopathology & Microscopy The Jackson Laboratory 600 Main St. Bar Harbor, Me 04609 207-288-6193 From tpmorken <@t> labvision.com Wed Nov 16 13:04:00 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Wed Nov 16 13:04:11 2005 Subject: [Histonet] RE: QIHC change Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D35A@usca0082k08.labvision.apogent.com> Pam, I'm not discounting the knowledge researchers have. I just don't think the ASCP certification route fits research. I pointed out that the ASCP, by definition, is intended soley for clinical diagnostic labs. It is not the ASCP's fault that the research institutions are mis-using the ASCP certification for their own in-house qualification standards. Certainly it would help researchers if there was a more general certification, but is it the ASCP's place to do that? I don't know. The ASCP is an organization of Clinical Pathology based in human diagnostics. Because of that the HT, HTL and QIHC are all based on human clinical pathology diagnostic work. I'm not sure the ASCP would be interested in setting up a whole new testing procedure for research. What if a person only does work in ferrets, or fish, or snakes? Who looks at tests based on that work? There are many similarities in procedures, and I'm confident a generalized test could be designed. It simply a question of whether the ASCP is the organization to do that. I guess the research techs have to explore this with ASCP. But I don' think you should blame ASCP for not accomodating research institutions when that is not what they are concerned with. Remember, there are no regulatory requirements at all covering the work of research lab techs but there are regulations covering who can do certain work in the diagnostic lab. ASCP, again by definition, is only concerned with the diagnostic side. It has nothing to do with whether one is "better" than another. There is no "better" in this work, only differences in the work done. I also suggest that you lobby your institution and show them how the ASCP certification may not be fully applicable to what you do. Tim Morken -----Original Message----- From: Pamela Marcum [mailto:pmarcum@vet.upenn.edu] Sent: Wednesday, November 16, 2005 9:56 AM To: Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: QIHC change Hi Tim and Jamie, I know Jamie and have for many years and you are right anyone would be happy to hire him for his experience in clinical or his current research position. However, it is also true that now research positions are often asking for at least an HT or even HTL (ASCP) to fill position with histology as the main focus. Yet we are given a set of criteria for tissue that often excludes animal research applicants from completing the practical easily. I took my HT many years ago and I was told that even in a research position (and I had a BS at the time) it would improve my salary and increase me to higher level in the university if I got my HT. I was told not to use animal tissue (1976) as no one reading the exam could properly read them. Now we have veterinary person there and tissue requirements can still eliminate some people or make it almost impossible to complete the practical with out help in procurement. Why should that happen to some one attempting to improve their position within the histology community? My real problem with what you said about the QIHC is that I would also like to take it and can not qualify either. Yet those of us in research are often finding the very antibodies and test methods companies and diagnostics later fight to get or learn. We are exempt in your mind and ASCP's even though research is what you depend on often for advances. I have never and will never understand this logic and exempt status for those of us who chose not to be clinical. We are still often required to have or get ASCP status as a way to advance and prove we know our field. ASCP needs to get up to date on the fields it is registering or make new categories for those of still contribute to clinical advances every year. Sorry if sounds like I am picking on you Tim. I just don't see how we are required to be registered on one hand for acceptance (even NSH likes to see it) and discounted on the other. Pam Marcum UPENN Vet School New Bolton Center Kennett Square, PA 19384 610-925-6278 At 12:07 PM 11/16/2005, Morken, Tim - Labvision wrote: >Jamie, It seems from what you say that you are working in a research >lab. Is that correct? My understanding about the ASCP certification is >that it is aimed at providing a modicum of proof that a person is >qualified to work in a medical diagnostic lab. Research labs are not >considered diagnostic labs. As you imply, a person in a research lab >will often work on only a limited sample set. Therefore, it is >meaningless to apply the the ASCP standard to research people. > > If you are planning to move into the diagnostic field, then I'll bet >you could easily find a job in a diagnostic lab, get the experience, >and qualify to take the test. It may be that some diagnostic labs have >a suggested requirement to be ASCP certified as a QIHC, but the vast >majority would be happy to find someone with the experience you >outline, even if they had not previously worked in a diagnositc lab. > >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: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James >Watson >Sent: Wednesday, November 16, 2005 7:28 AM >To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: QIHC change > > >This is my point. With the requirements listed below someone with 25 >years of experience doing immuno (single, double, triple antibody >staining, making own antibodies, and in situ Hybridization: all with >and without using kits, all with and without using an automated >stainer) is not qualified for this certification if they work in a >research facility where immunophenotyping is not done. There is no >system of doing it on your own to prove that you have the capability to >do immunophenotyping in order to fullfil this requirement. I guess it >is time to start harrassing ASCP about the unfairness of this system. > > >From almost always sunny San Diego >Jamie > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of >novanity@nc.rr.com > Sent: Wed 11/16/2005 5:55 AM > To: histonet@lists.utsouthwestern.edu > Cc: > Subject: [Histonet] RE: QIHC change > > > > James, to qualify for the qualification you take the 50 >question test > and submit an employer reference form + of course satisfy one of the > three routes. There is no practical to submit anymore. I >think that is > what you are asking. It seems to me that it wouldn't matter about > specificity antigens/markers or what diseases or human cells. >There is > no requirement other than what is requested on the employer >reference > form which you can't see the details until you order and >receive your > packet. Is it possible for anyone to post a copy of the employer > reference form. From the ASCP website this is what it says " > > Qualification in Immunohistochemistry > Experience requirements > > Applicants must have experience in the following areas > > * Immunohistochemical and Immunofluorescence Preparation > All of the following should have been performed by the >applicant > o staining technique > o selection of proper control material > o titration of immunologic reagents > > * Immunophenotyping > in at least one of the following applications > o immunodeficiencies > o immunoproliferative disorders (neoplastic and >non-neoplastic > disorders) > o transplantation biopsies > o other immunophenotyping applications > please specify: ______________________ > > * Quality Assurance > The applicant should have participated in Quality Assurance > related to all of the following > o specimen fixation, processing, microtomy > o reagent selection, preparation, storage, disposal > o method selection, validation, documentation > o quality control > o safety > > " This is the experience which I am assuming is only >documented for the > ASCP through the employer reference form, hence if you only do >A and C > and not B you can't qualify unless your employer is dishonest on the > form. Because even if you crosstrain into what I assume is flow > cytometry but don't actually work it day to day as part of >your job you > do not qualify because you have not had experience doing it for a > minimun of 12 months. As for research, same thing if you do >all of it > every day then your good to go. If not it is a grey or is it >gray area > that I'm looking more information/details on. In the past you > qualified your work with different immuno stains as a practical , I > don't remember there being a flow requirement. Maybe I'm wrong but > anyone have this info I'm looking for. > > G Hurlburt HT(ASCP) > sunny and warm NC > > _______________________________________________ > 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 Barry.R.Rittman <@t> uth.tmc.edu Wed Nov 16 13:19:05 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Wed Nov 16 13:19:11 2005 Subject: [Histonet] RE: QIHC change Message-ID: In the states the major problem seems to me to be that individual organizations here are somewhat too rigid in their requirements and in many instances are unable or unwilling to equate qualifications from outside the USA with USA qualifications. This is not just in Histotechnology but in many other professions. While in some areas the education here may be better, in others it is not of such a high quality as may be found for example in the UK. In those cases it would behoove those organizations to be less introverted and a lot more flexible in their administration of rules and regulations. In Texas many of us have the attitude that rules are guidelines and not rigidly set in stone. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DeBrosse_Beatrice Sent: Wednesday, November 16, 2005 12:27 PM To: Pamela Marcum; Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: QIHC change I agree with Pam that the ASCP needs to update on several things. I was qualified to take the QIHC, which I did and passed, but I'm sure Jamie can run circles around me since he has overall a whole lot more experience. I ran into a similar problem; being educated in Switzerland, where I got a degree as a laboratory technician in biology, according to the ASCP I cannot take the HTL just because my degree is not equivalent to a BS from the US. I went to a technical school, which doesn't qualify, but I've been trained specifically for being a laboratory technician in histology and microbiology. I can fully understand Jamie's frustration! Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pamela Marcum Sent: Wednesday, November 16, 2005 9:56 AM To: Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: QIHC change Hi Tim and Jamie, I know Jamie and have for many years and you are right anyone would be happy to hire him for his experience in clinical or his current research position. However, it is also true that now research positions are often asking for at least an HT or even HTL (ASCP) to fill position with histology as the main focus. Yet we are given a set of criteria for tissue that often excludes animal research applicants from completing the practical easily. I took my HT many years ago and I was told that even in a research position (and I had a BS at the time) it would improve my salary and increase me to higher level in the university if I got my HT. I was told not to use animal tissue (1976) as no one reading the exam could properly read them. Now we have veterinary person there and tissue requirements can still eliminate some people or make it almost impossible to complete the practical with out help in procurement. Why should that happen to some one attempting to improve their position within the histology community? My real problem with what you said about the QIHC is that I would also like to take it and can not qualify either. Yet those of us in research are often finding the very antibodies and test methods companies and diagnostics later fight to get or learn. We are exempt in your mind and ASCP's even though research is what you depend on often for advances. I have never and will never understand this logic and exempt status for those of us who chose not to be clinical. We are still often required to have or get ASCP status as a way to advance and prove we know our field. ASCP needs to get up to date on the fields it is registering or make new categories for those of still contribute to clinical advances every year. Sorry if sounds like I am picking on you Tim. I just don't see how we are required to be registered on one hand for acceptance (even NSH likes to see it) and discounted on the other. Pam Marcum UPENN Vet School New Bolton Center Kennett Square, PA 19384 610-925-6278 At 12:07 PM 11/16/2005, Morken, Tim - Labvision wrote: >Jamie, It seems from what you say that you are working in a research lab. Is >that correct? My understanding about the ASCP certification is that it is >aimed at providing a modicum of proof that a person is qualified to work in >a medical diagnostic lab. Research labs are not considered diagnostic labs. >As you imply, a person in a research lab will often work on only a limited >sample set. Therefore, it is meaningless to apply the the ASCP standard to >research people. > > If you are planning to move into the diagnostic field, then I'll bet you >could easily find a job in a diagnostic lab, get the experience, and qualify >to take the test. It may be that some diagnostic labs have a suggested >requirement to be ASCP certified as a QIHC, but the vast majority would be >happy to find someone with the experience you outline, even if they had not >previously worked in a diagnositc lab. > >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: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James Watson >Sent: Wednesday, November 16, 2005 7:28 AM >To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: QIHC change > > >This is my point. With the requirements listed below someone with 25 years >of experience doing immuno (single, double, triple antibody staining, making >own antibodies, and in situ Hybridization: all with and without using kits, >all with and without using an automated stainer) is not qualified for this >certification if they work in a research facility where immunophenotyping is >not done. There is no system of doing it on your own to prove that you have >the capability to do immunophenotyping in order to fullfil this requirement. >I guess it is time to start harrassing ASCP about the unfairness of this >system. > > >From almost always sunny San Diego >Jamie > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of >novanity@nc.rr.com > Sent: Wed 11/16/2005 5:55 AM > To: histonet@lists.utsouthwestern.edu > Cc: > Subject: [Histonet] RE: QIHC change > > > > James, to qualify for the qualification you take the 50 question >test > and submit an employer reference form + of course satisfy one of the > three routes. There is no practical to submit anymore. I think >that is > what you are asking. It seems to me that it wouldn't matter about > specificity antigens/markers or what diseases or human cells. There >is > no requirement other than what is requested on the employer >reference > form which you can't see the details until you order and receive >your > packet. Is it possible for anyone to post a copy of the employer > reference form. From the ASCP website this is what it says " > > Qualification in Immunohistochemistry > Experience requirements > > Applicants must have experience in the following areas > > * Immunohistochemical and Immunofluorescence Preparation > All of the following should have been performed by the >applicant > o staining technique > o selection of proper control material > o titration of immunologic reagents > > * Immunophenotyping > in at least one of the following applications > o immunodeficiencies > o immunoproliferative disorders (neoplastic and >non-neoplastic > disorders) > o transplantation biopsies > o other immunophenotyping applications > please specify: ______________________ > > * Quality Assurance > The applicant should have participated in Quality Assurance > related to all of the following > o specimen fixation, processing, microtomy > o reagent selection, preparation, storage, disposal > o method selection, validation, documentation > o quality control > o safety > > " This is the experience which I am assuming is only documented for >the > ASCP through the employer reference form, hence if you only do A and >C > and not B you can't qualify unless your employer is dishonest on the > form. Because even if you crosstrain into what I assume is flow > cytometry but don't actually work it day to day as part of your job >you > do not qualify because you have not had experience doing it for a > minimun of 12 months. As for research, same thing if you do all of >it > every day then your good to go. If not it is a grey or is it gray >area > that I'm looking more information/details on. In the past you > qualified your work with different immuno stains as a practical , I > don't remember there being a flow requirement. Maybe I'm wrong but > anyone have this info I'm looking for. > > G Hurlburt HT(ASCP) > sunny and warm NC > > _______________________________________________ > 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 tpmorken <@t> labvision.com Wed Nov 16 13:22:46 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Wed Nov 16 13:22:59 2005 Subject: [Histonet] aqueous coverslipping medium Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D35C@usca0082k08.labvision.apogent.com> Paul, for fluorescence work we use the PVA-DABCO aqueous mounting media from sigma-aldrich, Cat# 10981. It is actully made by Fluka. "Poly vinyl alcohol with DABCO anti-fading agent." one caveat - it has only a 3-month shelf life before it polymerizes in the bottle. And it's one of the few mounting mediums that works with Qdots, if you plan on using those for fluorescence work. I've also found the DAKO Faramount medium to be great for enzyme IHC. It hardens well and is storable in standard slide file drawers. Cat# S3025 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: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Wednesday, November 16, 2005 9:40 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] aqueous coverslipping medium What brand do you prefer for aqueous mounting medium to be used with a coverslip? I have several good hydrocarbon-based coverslipping media, and also some good aqueous media to be used without coverslips (so-called liquid coverslip), but I don't have an aqueous coverslipping medium I am completely satisfied with. Applications would include immunofluorescence and a few enzyme stains, as well as lipid stains and other techniques that are not compatible with hydrocarbon-based media. Are there any such media that harden after application, to produce a permanent slide? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Wed Nov 16 13:36:10 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Wed Nov 16 13:36:30 2005 Subject: [Histonet] RE: QIHC change In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328D35A@usca0082k08.labvi sion.apogent.com> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328D35A@usca0082k08.labvision.apogent.com> Message-ID: <6.1.1.1.2.20051116142731.019fe628@mail.vet.upenn.edu> Tim, I understand the point exactly however, ASCP has also never failed in over thirty years to take my money for registration yearly. Many of the pathologists I have worked with over the years in research, clinical and industry have insisted on ASCP registration for research and clinical as part of the qualifications for application to a job. They were and are clinical pathologist and members of ASCP who should have then been aware in research and industry it should not matter whether I or anyone else was certified by the group. I have worked for several universities since the '70s all wanted the registration in HT and later some wanted HTL if you had no undergraduate or graduate degree. I have both so that was not my issue. ASCP does not discourage anyone from taking the test and if it is only for diagnostics they are the ones who are not adhering to the code not those of us who wish to be as well qualified as possible for any area of histology. By the way I agree with Barry wholeheartedly. Pam At 02:04 PM 11/16/2005, Morken, Tim - Labvision wrote: >Pam, > >I'm not discounting the knowledge researchers have. I just don't think the >ASCP certification route fits research. I pointed out that the ASCP, by >definition, is intended soley for clinical diagnostic labs. It is not the >ASCP's fault that the research institutions are mis-using the ASCP >certification for their own in-house qualification standards. Certainly it >would help researchers if there was a more general certification, but is it >the ASCP's place to do that? I don't know. The ASCP is an organization of >Clinical Pathology based in human diagnostics. Because of that the HT, HTL >and QIHC are all based on human clinical pathology diagnostic work. I'm not >sure the ASCP would be interested in setting up a whole new testing >procedure for research. What if a person only does work in ferrets, or fish, >or snakes? Who looks at tests based on that work? There are many >similarities in procedures, and I'm confident a generalized test could be >designed. It simply a question of whether the ASCP is the organization to do >that. I guess the research techs have to explore this with ASCP. But I don' >think you should blame ASCP for not accomodating research institutions when >that is not what they are concerned with. Remember, there are no regulatory >requirements at all covering the work of research lab techs but there are >regulations covering who can do certain work in the diagnostic lab. ASCP, >again by definition, is only concerned with the diagnostic side. It has >nothing to do with whether one is "better" than another. There is no >"better" in this work, only differences in the work done. I also suggest >that you lobby your institution and show them how the ASCP certification may >not be fully applicable to what you do. > > >Tim Morken > > > >-----Original Message----- >From: Pamela Marcum [mailto:pmarcum@vet.upenn.edu] >Sent: Wednesday, November 16, 2005 9:56 AM >To: Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: QIHC change > > >Hi Tim and Jamie, > >I know Jamie and have for many years and you are right anyone would be >happy to hire him for his experience in clinical or his current research >position. However, it is also true that now research positions are often >asking for at least an HT or even HTL (ASCP) to fill position with >histology as the main focus. Yet we are given a set of criteria for tissue >that often excludes animal research applicants from completing the >practical easily. I took my HT many years ago and I was told that even in >a research position (and I had a BS at the time) it would improve my salary >and increase me to higher level in the university if I got my HT. I was >told not to use animal tissue (1976) as no one reading the exam could >properly read them. Now we have veterinary person there and tissue >requirements can still eliminate some people or make it almost impossible >to complete the practical with out help in procurement. Why should that >happen to some one attempting to improve their position within the >histology community? > >My real problem with what you said about the QIHC is that I would also like >to take it and can not qualify either. Yet those of us in research are >often finding the very antibodies and test methods companies and >diagnostics later fight to get or learn. We are exempt in your mind and >ASCP's even though research is what you depend on often for advances. I >have never and will never understand this logic and exempt status for those >of us who chose not to be clinical. We are still often required to have >or get ASCP status as a way to advance and prove we know our field. ASCP >needs to get up to date on the fields it is registering or make new >categories for those of still contribute to clinical advances every year. > >Sorry if sounds like I am picking on you Tim. I just don't see how we are >required to be registered on one hand for acceptance (even NSH likes to see >it) and discounted on the other. > >Pam Marcum >UPENN Vet School >New Bolton Center >Kennett Square, PA 19384 >610-925-6278 > > >At 12:07 PM 11/16/2005, Morken, Tim - Labvision wrote: > >Jamie, It seems from what you say that you are working in a research > >lab. Is that correct? My understanding about the ASCP certification is > >that it is aimed at providing a modicum of proof that a person is > >qualified to work in a medical diagnostic lab. Research labs are not > >considered diagnostic labs. As you imply, a person in a research lab > >will often work on only a limited sample set. Therefore, it is > >meaningless to apply the the ASCP standard to research people. > > > > If you are planning to move into the diagnostic field, then I'll bet > >you could easily find a job in a diagnostic lab, get the experience, > >and qualify to take the test. It may be that some diagnostic labs have > >a suggested requirement to be ASCP certified as a QIHC, but the vast > >majority would be happy to find someone with the experience you > >outline, even if they had not previously worked in a diagnositc lab. > > > >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: histonet-bounces@lists.utsouthwestern.edu > >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James > >Watson > >Sent: Wednesday, November 16, 2005 7:28 AM > >To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu > >Subject: RE: [Histonet] RE: QIHC change > > > > > >This is my point. With the requirements listed below someone with 25 > >years of experience doing immuno (single, double, triple antibody > >staining, making own antibodies, and in situ Hybridization: all with > >and without using kits, all with and without using an automated > >stainer) is not qualified for this certification if they work in a > >research facility where immunophenotyping is not done. There is no > >system of doing it on your own to prove that you have the capability to > >do immunophenotyping in order to fullfil this requirement. I guess it > >is time to start harrassing ASCP about the unfairness of this system. > > > > >From almost always sunny San Diego > >Jamie > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > >novanity@nc.rr.com > > Sent: Wed 11/16/2005 5:55 AM > > To: histonet@lists.utsouthwestern.edu > > Cc: > > Subject: [Histonet] RE: QIHC change > > > > > > > > James, to qualify for the qualification you take the 50 > >question test > > and submit an employer reference form + of course satisfy one of >the > > three routes. There is no practical to submit anymore. I > >think that is > > what you are asking. It seems to me that it wouldn't matter about > > specificity antigens/markers or what diseases or human cells. > >There is > > no requirement other than what is requested on the employer > >reference > > form which you can't see the details until you order and > >receive your > > packet. Is it possible for anyone to post a copy of the employer > > reference form. From the ASCP website this is what it says " > > > > Qualification in Immunohistochemistry > > Experience requirements > > > > Applicants must have experience in the following areas > > > > * Immunohistochemical and Immunofluorescence Preparation > > All of the following should have been performed by the > >applicant > > o staining technique > > o selection of proper control material > > o titration of immunologic reagents > > > > * Immunophenotyping > > in at least one of the following applications > > o immunodeficiencies > > o immunoproliferative disorders (neoplastic and > >non-neoplastic > > disorders) > > o transplantation biopsies > > o other immunophenotyping applications > > please specify: ______________________ > > > > * Quality Assurance > > The applicant should have participated in Quality Assurance > > related to all of the following > > o specimen fixation, processing, microtomy > > o reagent selection, preparation, storage, disposal > > o method selection, validation, documentation > > o quality control > > o safety > > > > " This is the experience which I am assuming is only > >documented for the > > ASCP through the employer reference form, hence if you only do > >A and C > > and not B you can't qualify unless your employer is dishonest on >the > > form. Because even if you crosstrain into what I assume is flow > > cytometry but don't actually work it day to day as part of > >your job you > > do not qualify because you have not had experience doing it for a > > minimun of 12 months. As for research, same thing if you do > >all of it > > every day then your good to go. If not it is a grey or is it > >gray area > > that I'm looking more information/details on. In the past you > > qualified your work with different immuno stains as a practical , >I > > don't remember there being a flow requirement. Maybe I'm wrong >but > > anyone have this info I'm looking for. > > > > G Hurlburt HT(ASCP) > > sunny and warm NC > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > From pmarcum <@t> vet.upenn.edu Wed Nov 16 13:49:07 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Wed Nov 16 13:49:25 2005 Subject: [Histonet] RE: QIHC change In-Reply-To: <437B79FE.7090208@umn.edu> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328D357@usca0082k08.labvision.apogent.com> <6.1.1.1.2.20051116123818.019c43e0@mail.vet.upenn.edu> <437B79FE.7090208@umn.edu> Message-ID: <6.1.1.1.2.20051116144321.01a017e8@mail.vet.upenn.edu> THANKS, to all who answered and understand why both Jamie and I - as well as many others are so out of sorts with ASCP at times. We are all histologists with something to offer the field and it is something I have always been proud of, just not happy with the way ASCP treats research people. Pam At 01:27 PM 11/16/2005, Colleen Forster wrote: >I agree with Pam ...completely. I too do research and I am HT certified >because I started in the clinical route. While in the clinical lab I >tested for the QIHC. It has been a GREAT benefit for me in my research >positions and I encourage all people pursuning histology work weather >research or clinicla to get registered. One never knows what turns life >might take and the added skills/knoweledge will be a benefit!! > >ASCP definitiely needs to widen their range! > >Colleen Forster >U of MN > >Pamela Marcum wrote: > >>Hi Tim and Jamie, >> >>I know Jamie and have for many years and you are right anyone would be >>happy to hire him for his experience in clinical or his current research >>position. However, it is also true that now research positions are often >>asking for at least an HT or even HTL (ASCP) to fill position with >>histology as the main focus. Yet we are given a set of criteria for >>tissue that often excludes animal research applicants from completing the >>practical easily. I took my HT many years ago and I was told that even >>in a research position (and I had a BS at the time) it would improve my >>salary and increase me to higher level in the university if I got my >>HT. I was told not to use animal tissue (1976) as no one reading the >>exam could properly read them. >>Now we have veterinary person there and tissue requirements can still >>eliminate some people or make it almost impossible to complete the >>practical with out help in procurement. Why should that happen to some >>one attempting to improve their position within the histology community? >> >>My real problem with what you said about the QIHC is that I would also >>like to take it and can not qualify either. Yet those of us in research >>are often finding the very antibodies and test methods companies and >>diagnostics later fight to get or learn. We are exempt in your mind and >>ASCP's even though research is what you depend on often for advances. I >>have never and will never understand this logic and exempt status for >>those of us who chose not to be clinical. We are still often required >>to have or get ASCP status as a way to advance and prove we know our >>field. ASCP needs to get up to date on the fields it is registering or >>make new categories for those of still contribute to clinical advances >>every year. >> >>Sorry if sounds like I am picking on you Tim. I just don't see how we >>are required to be registered on one hand for acceptance (even NSH likes >>to see it) and discounted on the other. >> >>Pam Marcum >>UPENN Vet School >>New Bolton Center >>Kennett Square, PA 19384 >>610-925-6278 >> >> >>At 12:07 PM 11/16/2005, Morken, Tim - Labvision wrote: >> >>>Jamie, It seems from what you say that you are working in a research lab. Is >>>that correct? My understanding about the ASCP certification is that it is >>>aimed at providing a modicum of proof that a person is qualified to work in >>>a medical diagnostic lab. Research labs are not considered diagnostic labs. >>>As you imply, a person in a research lab will often work on only a limited >>>sample set. Therefore, it is meaningless to apply the the ASCP standard to >>>research people. >>> >>> If you are planning to move into the diagnostic field, then I'll bet you >>>could easily find a job in a diagnostic lab, get the experience, and qualify >>>to take the test. It may be that some diagnostic labs have a suggested >>>requirement to be ASCP certified as a QIHC, but the vast majority would be >>>happy to find someone with the experience you outline, even if they had not >>>previously worked in a diagnositc lab. >>> >>>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: histonet-bounces@lists.utsouthwestern.edu >>>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James Watson >>>Sent: Wednesday, November 16, 2005 7:28 AM >>>To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu >>>Subject: RE: [Histonet] RE: QIHC change >>> >>> >>>This is my point. With the requirements listed below someone with 25 years >>>of experience doing immuno (single, double, triple antibody staining, making >>>own antibodies, and in situ Hybridization: all with and without using kits, >>>all with and without using an automated stainer) is not qualified for this >>>certification if they work in a research facility where immunophenotyping is >>>not done. There is no system of doing it on your own to prove that you have >>>the capability to do immunophenotyping in order to fullfil this requirement. >>>I guess it is time to start harrassing ASCP about the unfairness of this >>>system. >>> >>> >From almost always sunny San Diego >>>Jamie >>> >>> -----Original Message----- >>> From: histonet-bounces@lists.utsouthwestern.edu on behalf of >>>novanity@nc.rr.com >>> Sent: Wed 11/16/2005 5:55 AM >>> To: histonet@lists.utsouthwestern.edu >>> Cc: >>> Subject: [Histonet] RE: QIHC change >>> >>> >>> >>> James, to qualify for the qualification you take the 50 question >>>test >>> and submit an employer reference form + of course satisfy one >>> of the >>> three routes. There is no practical to submit anymore. I think >>>that is >>> what you are asking. It seems to me that it wouldn't matter about >>> specificity antigens/markers or what diseases or human >>> cells. There >>>is >>> no requirement other than what is requested on the employer >>>reference >>> form which you can't see the details until you order and receive >>>your >>> packet. Is it possible for anyone to post a copy of the employer >>> reference form. From the ASCP website this is what it says " >>> >>> Qualification in Immunohistochemistry >>> Experience requirements >>> >>> Applicants must have experience in the following areas >>> >>> * Immunohistochemical and Immunofluorescence Preparation >>> All of the following should have been performed by the >>>applicant >>> o staining technique >>> o selection of proper control material >>> o titration of immunologic reagents >>> >>> * Immunophenotyping >>> in at least one of the following applications >>> o immunodeficiencies >>> o immunoproliferative disorders (neoplastic and >>>non-neoplastic >>> disorders) >>> o transplantation biopsies >>> o other immunophenotyping applications >>> please specify: ______________________ >>> >>> * Quality Assurance >>> The applicant should have participated in Quality Assurance >>> related to all of the following >>> o specimen fixation, processing, microtomy >>> o reagent selection, preparation, storage, disposal >>> o method selection, validation, documentation >>> o quality control >>> o safety >>> >>> " This is the experience which I am assuming is only >>> documented for >>>the >>> ASCP through the employer reference form, hence if you only do >>> A and >>>C >>> and not B you can't qualify unless your employer is dishonest >>> on the >>> form. Because even if you crosstrain into what I assume is flow >>> cytometry but don't actually work it day to day as part of your job >>>you >>> do not qualify because you have not had experience doing it for a >>> minimun of 12 months. As for research, same thing if you do all of >>>it >>> every day then your good to go. If not it is a grey or is it gray >>>area >>> that I'm looking more information/details on. In the past you >>> qualified your work with different immuno stains as a practical , I >>> don't remember there being a flow requirement. Maybe I'm wrong but >>> anyone have this info I'm looking for. >>> >>> G Hurlburt HT(ASCP) >>> sunny and warm NC >>> >>> _______________________________________________ >>> 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 BMolinari <@t> heart.thi.tmc.edu Wed Nov 16 14:00:23 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Wed Nov 16 14:03:23 2005 Subject: [Histonet] recycled xylenes Message-ID: Hi, I am interested in knowing if you have preferences for your recycled xylenes. Do you use it just for staining, or processing alone. Or use it for both? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) From jwatson <@t> gnf.org Wed Nov 16 14:08:01 2005 From: jwatson <@t> gnf.org (James Watson) Date: Wed Nov 16 14:08:17 2005 Subject: [Histonet] RE: QIHC change Message-ID: Histonetters interested in QIHC exam, After several phone calls I was finally put into contact with someone at ASCP that handles the QIHC questions. Her question to me was how recent my experience with immunophenotyping on humans was and what I might be doing now that would qualify as immunophenotyping. She was not able to tell me what would specifically be considered as immunophenotyping and did not say that some types of work on animals would or would not be considered immunophenotyping such as B and T cell markers. She said that they try to be flexible about this. She told me that this weekend the committee is meeting about the QIHC exam, so if you have concerns about the exam or qualifications you may want to e-mail Pam Frommelt from ASCP at pamelaf@ascp.org . James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pamela Marcum Sent: Wednesday, November 16, 2005 11:36 AM To: Morken, Tim - Labvision; Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: QIHC change Tim, I understand the point exactly however, ASCP has also never failed in over thirty years to take my money for registration yearly. Many of the pathologists I have worked with over the years in research, clinical and industry have insisted on ASCP registration for research and clinical as part of the qualifications for application to a job. They were and are clinical pathologist and members of ASCP who should have then been aware in research and industry it should not matter whether I or anyone else was certified by the group. I have worked for several universities since the '70s all wanted the registration in HT and later some wanted HTL if you had no undergraduate or graduate degree. I have both so that was not my issue. ASCP does not discourage anyone from taking the test and if it is only for diagnostics they are the ones who are not adhering to the code not those of us who wish to be as well qualified as possible for any area of histology. By the way I agree with Barry wholeheartedly. Pam At 02:04 PM 11/16/2005, Morken, Tim - Labvision wrote: >Pam, > >I'm not discounting the knowledge researchers have. I just don't think >the ASCP certification route fits research. I pointed out that the >ASCP, by definition, is intended soley for clinical diagnostic labs. >It is not the ASCP's fault that the research institutions are mis-using >the ASCP certification for their own in-house qualification standards. >Certainly it would help researchers if there was a more general >certification, but is it the ASCP's place to do that? I don't know. The >ASCP is an organization of Clinical Pathology based in human >diagnostics. Because of that the HT, HTL and QIHC are all based on >human clinical pathology diagnostic work. I'm not sure the ASCP would >be interested in setting up a whole new testing procedure for research. >What if a person only does work in ferrets, or fish, or snakes? Who >looks at tests based on that work? There are many similarities in >procedures, and I'm confident a generalized test could be designed. It >simply a question of whether the ASCP is the organization to do that. >I guess the research techs have to explore this with ASCP. But I don' >think you should blame ASCP for not accomodating research institutions >when that is not what they are concerned with. Remember, there are no >regulatory requirements at all covering the work of research lab techs >but there are regulations covering who can do certain work in the >diagnostic lab. ASCP, again by definition, is only concerned with the >diagnostic side. It has nothing to do with whether one is "better" than >another. There is no "better" in this work, only differences in the >work done. I also suggest that you lobby your institution and show them >how the ASCP certification may not be fully applicable to what you do. > > >Tim Morken > > > >-----Original Message----- >From: Pamela Marcum [mailto:pmarcum@vet.upenn.edu] >Sent: Wednesday, November 16, 2005 9:56 AM >To: Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: QIHC change > > >Hi Tim and Jamie, > >I know Jamie and have for many years and you are right anyone would be >happy to hire him for his experience in clinical or his current >research position. However, it is also true that now research >positions are often asking for at least an HT or even HTL (ASCP) to >fill position with histology as the main focus. Yet we are given a set >of criteria for tissue that often excludes animal research applicants >from completing the practical easily. I took my HT many years ago and >I was told that even in a research position (and I had a BS at the >time) it would improve my salary and increase me to higher level in the >university if I got my HT. I was told not to use animal tissue (1976) >as no one reading the exam could properly read them. Now we have >veterinary person there and tissue requirements can still eliminate >some people or make it almost impossible to complete the practical with >out help in procurement. Why should that happen to some one attempting >to improve their position within the histology community? > >My real problem with what you said about the QIHC is that I would also >like to take it and can not qualify either. Yet those of us in >research are often finding the very antibodies and test methods >companies and diagnostics later fight to get or learn. We are exempt >in your mind and ASCP's even though research is what you depend on >often for advances. I have never and will never understand this logic >and exempt status for those of us who chose not to be clinical. We >are still often required to have or get ASCP status as a way to advance >and prove we know our field. ASCP needs to get up to date on the >fields it is registering or make new categories for those of still >contribute to clinical advances every year. > >Sorry if sounds like I am picking on you Tim. I just don't see how we are >required to be registered on one hand for acceptance (even NSH likes to >see >it) and discounted on the other. > >Pam Marcum >UPENN Vet School >New Bolton Center >Kennett Square, PA 19384 >610-925-6278 > > >At 12:07 PM 11/16/2005, Morken, Tim - Labvision wrote: > >Jamie, It seems from what you say that you are working in a research > >lab. Is that correct? My understanding about the ASCP certification > >is that it is aimed at providing a modicum of proof that a person is > >qualified to work in a medical diagnostic lab. Research labs are not > >considered diagnostic labs. As you imply, a person in a research lab > >will often work on only a limited sample set. Therefore, it is > >meaningless to apply the the ASCP standard to research people. > > > > If you are planning to move into the diagnostic field, then I'll > >bet you could easily find a job in a diagnostic lab, get the > >experience, and qualify to take the test. It may be that some > >diagnostic labs have a suggested requirement to be ASCP certified as > >a QIHC, but the vast majority would be happy to find someone with the > >experience you outline, even if they had not previously worked in a > >diagnositc lab. > > > >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: histonet-bounces@lists.utsouthwestern.edu > >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James > >Watson > >Sent: Wednesday, November 16, 2005 7:28 AM > >To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu > >Subject: RE: [Histonet] RE: QIHC change > > > > > >This is my point. With the requirements listed below someone with 25 > >years of experience doing immuno (single, double, triple antibody > >staining, making own antibodies, and in situ Hybridization: all with > >and without using kits, all with and without using an automated > >stainer) is not qualified for this certification if they work in a > >research facility where immunophenotyping is not done. There is no > >system of doing it on your own to prove that you have the capability > >to do immunophenotyping in order to fullfil this requirement. I guess > >it is time to start harrassing ASCP about the unfairness of this > >system. > > > > >From almost always sunny San Diego > >Jamie > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > >novanity@nc.rr.com > > Sent: Wed 11/16/2005 5:55 AM > > To: histonet@lists.utsouthwestern.edu > > Cc: > > Subject: [Histonet] RE: QIHC change > > > > > > > > James, to qualify for the qualification you take the 50 > >question test > > and submit an employer reference form + of course satisfy > >one of >the > > three routes. There is no practical to submit anymore. I > >think that is > > what you are asking. It seems to me that it wouldn't matter about > > specificity antigens/markers or what diseases or human > >cells. There is > > no requirement other than what is requested on the employer > >reference > > form which you can't see the details until you order and > >receive your > > packet. Is it possible for anyone to post a copy of the employer > > reference form. From the ASCP website this is what it says > >" > > > > Qualification in Immunohistochemistry > > Experience requirements > > > > Applicants must have experience in the following areas > > > > * Immunohistochemical and Immunofluorescence Preparation > > All of the following should have been performed by the > >applicant > > o staining technique > > o selection of proper control material > > o titration of immunologic reagents > > > > * Immunophenotyping > > in at least one of the following applications > > o immunodeficiencies > > o immunoproliferative disorders (neoplastic and > >non-neoplastic > > disorders) > > o transplantation biopsies > > o other immunophenotyping applications > > please specify: ______________________ > > > > * Quality Assurance > > The applicant should have participated in Quality Assurance > > related to all of the following > > o specimen fixation, processing, microtomy > > o reagent selection, preparation, storage, disposal > > o method selection, validation, documentation > > o quality control > > o safety > > > > " This is the experience which I am assuming is only > >documented for the > > ASCP through the employer reference form, hence if you only > >do A and C > > and not B you can't qualify unless your employer is > >dishonest on >the > > form. Because even if you crosstrain into what I assume is flow > > cytometry but don't actually work it day to day as part of > >your job you > > do not qualify because you have not had experience doing it for a > > minimun of 12 months. As for research, same thing if you do > >all of it > > every day then your good to go. If not it is a grey or is > >it gray area > > that I'm looking more information/details on. In the past you > > qualified your work with different immuno stains as a > >practical , >I > > don't remember there being a flow requirement. Maybe I'm > > wrong >but > > anyone have this info I'm looking for. > > > > G Hurlburt HT(ASCP) > > sunny and warm NC > > > > _______________________________________________ > > 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 liz <@t> premierlab.com Wed Nov 16 14:08:53 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Wed Nov 16 14:09:14 2005 Subject: [Histonet] QIHC Message-ID: <000001c5eae9$91e76df0$a7d48a80@AMY> I'm going to have to chime in here. One thing I need to say upfront is that I served on the BOR when they were developing the QIHC. The BOR for the histotech exam committee consists of histology technicians, histotechnologists, MD pathologists and veterinary pathologists, plus the ASCP staff, so it encompassed a wide variety of individuals and skill sets, both in research and clinical. These are the individuals who created the exam and the requirements for the exam. I went to the BOR guidelines online for the QIHC and as far as I can see that have not changed. Immunophenotyping has always been a pre-requisite for sitting for this qualification. This is what BOR requires: Immunophenotyping in at least one of the following applications * immunodeficiencies * immunoproliferative disorders (neoplastic and non-neoplastic disorders) * transplantation biopsies * other immunophenotyping applications please specify: ______________________ After looking online for the definition of immunophenotyping it is the following: " IMMUNOPHENOTYPING refers to the technique of identifying molecules that are associated with lymphoid cells and help to characterize them". Such as cell surface markers, which I would characterize broadly as CD markers. I work in research and sat for the QIHC last year. In research we perform many Immunohistochemical stains utilizing CD markers to determine what specific cell types are present in certain disease states. When I filled out my application last year I selected the forth choice which is "immunophenotyping applications other" and listed what types of CD markers I had worked with and the type of projects and applications associated with them. I was able to sit for the exam and so was my employee. In my opinion many histotechs in research have the necessary experience since they are performing Immunohistochemical staining utilizing a wide variety of CD markers. I just don't see how in working in research you would not have the pre-requisite to sit for this exam. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From Bauer.Karen <@t> mayo.edu Wed Nov 16 14:14:06 2005 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Wed Nov 16 14:16:02 2005 Subject: [Histonet] recycled xylenes References: Message-ID: Hi Betsy, We use are recycled xylene for both processing and staining and have had no problems. Karen L. Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Molinari, Betsy Sent: Wed 11/16/2005 2:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] recycled xylenes Hi, I am interested in knowing if you have preferences for your recycled xylenes. Do you use it just for staining, or processing alone. Or use it for both? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************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 jwatson <@t> gnf.org Wed Nov 16 14:20:09 2005 From: jwatson <@t> gnf.org (James Watson) Date: Wed Nov 16 14:20:19 2005 Subject: [Histonet] QIHC Message-ID: Elizabeth, I have been trying to find out what the other category entails without success, this is why I became involved with these discussions to see if someone on the histo net knew. But I have finally gotten someone at ASCP to talk to as listed on my last posting. She was not very clear as to what qualifies and if work on animal tissue is acceptable. So she wants me to send her a list of what I do that may qualify as immunophenotyping to determine if I qualify. I really appreciate your response because it gives me some guidelines as what I can list as other immunophenotyping applications. Thank you very much. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Wednesday, November 16, 2005 12:09 PM To: 'Histonet' Subject: [Histonet] QIHC I'm going to have to chime in here. One thing I need to say upfront is that I served on the BOR when they were developing the QIHC. The BOR for the histotech exam committee consists of histology technicians, histotechnologists, MD pathologists and veterinary pathologists, plus the ASCP staff, so it encompassed a wide variety of individuals and skill sets, both in research and clinical. These are the individuals who created the exam and the requirements for the exam. I went to the BOR guidelines online for the QIHC and as far as I can see that have not changed. Immunophenotyping has always been a pre-requisite for sitting for this qualification. This is what BOR requires: Immunophenotyping in at least one of the following applications * immunodeficiencies * immunoproliferative disorders (neoplastic and non-neoplastic disorders) * transplantation biopsies * other immunophenotyping applications please specify: ______________________ After looking online for the definition of immunophenotyping it is the following: " IMMUNOPHENOTYPING refers to the technique of identifying molecules that are associated with lymphoid cells and help to characterize them". Such as cell surface markers, which I would characterize broadly as CD markers. I work in research and sat for the QIHC last year. In research we perform many Immunohistochemical stains utilizing CD markers to determine what specific cell types are present in certain disease states. When I filled out my application last year I selected the forth choice which is "immunophenotyping applications other" and listed what types of CD markers I had worked with and the type of projects and applications associated with them. I was able to sit for the exam and so was my employee. In my opinion many histotechs in research have the necessary experience since they are performing Immunohistochemical staining utilizing a wide variety of CD markers. I just don't see how in working in research you would not have the pre-requisite to sit for this exam. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jeffery-Smith <@t> ouhsc.edu Wed Nov 16 14:18:09 2005 From: Jeffery-Smith <@t> ouhsc.edu (Smith, Jeffery D. (HSC)) Date: Wed Nov 16 14:22:00 2005 Subject: [Histonet] RE: QIHC change Message-ID: Tim, I would like to know what your certification is and what qualifies you to decide what the focus of the ASCP certifications entale? Who put you in charge of deciding what the sole purpose of the ASCP is? ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Morken, Tim - Labvision Sent: Wed 11/16/2005 1:04 PM To: 'Pamela Marcum'; Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: QIHC change Pam, I'm not discounting the knowledge researchers have. I just don't think the ASCP certification route fits research. I pointed out that the ASCP, by definition, is intended soley for clinical diagnostic labs. It is not the ASCP's fault that the research institutions are mis-using the ASCP certification for their own in-house qualification standards. Certainly it would help researchers if there was a more general certification, but is it the ASCP's place to do that? I don't know. The ASCP is an organization of Clinical Pathology based in human diagnostics. Because of that the HT, HTL and QIHC are all based on human clinical pathology diagnostic work. I'm not sure the ASCP would be interested in setting up a whole new testing procedure for research. What if a person only does work in ferrets, or fish, or snakes? Who looks at tests based on that work? There are many similarities in procedures, and I'm confident a generalized test could be designed. It simply a question of whether the ASCP is the organization to do that. I guess the research techs have to explore this with ASCP. But I don' think you should blame ASCP for not accomodating research institutions when that is not what they are concerned with. Remember, there are no regulatory requirements at all covering the work of research lab techs but there are regulations covering who can do certain work in the diagnostic lab. ASCP, again by definition, is only concerned with the diagnostic side. It has nothing to do with whether one is "better" than another. There is no "better" in this work, only differences in the work done. I also suggest that you lobby your institution and show them how the ASCP certification may not be fully applicable to what you do. Tim Morken -----Original Message----- From: Pamela Marcum [mailto:pmarcum@vet.upenn.edu] Sent: Wednesday, November 16, 2005 9:56 AM To: Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: QIHC change Hi Tim and Jamie, I know Jamie and have for many years and you are right anyone would be happy to hire him for his experience in clinical or his current research position. However, it is also true that now research positions are often asking for at least an HT or even HTL (ASCP) to fill position with histology as the main focus. Yet we are given a set of criteria for tissue that often excludes animal research applicants from completing the practical easily. I took my HT many years ago and I was told that even in a research position (and I had a BS at the time) it would improve my salary and increase me to higher level in the university if I got my HT. I was told not to use animal tissue (1976) as no one reading the exam could properly read them. Now we have veterinary person there and tissue requirements can still eliminate some people or make it almost impossible to complete the practical with out help in procurement. Why should that happen to some one attempting to improve their position within the histology community? My real problem with what you said about the QIHC is that I would also like to take it and can not qualify either. Yet those of us in research are often finding the very antibodies and test methods companies and diagnostics later fight to get or learn. We are exempt in your mind and ASCP's even though research is what you depend on often for advances. I have never and will never understand this logic and exempt status for those of us who chose not to be clinical. We are still often required to have or get ASCP status as a way to advance and prove we know our field. ASCP needs to get up to date on the fields it is registering or make new categories for those of still contribute to clinical advances every year. Sorry if sounds like I am picking on you Tim. I just don't see how we are required to be registered on one hand for acceptance (even NSH likes to see it) and discounted on the other. Pam Marcum UPENN Vet School New Bolton Center Kennett Square, PA 19384 610-925-6278 At 12:07 PM 11/16/2005, Morken, Tim - Labvision wrote: >Jamie, It seems from what you say that you are working in a research >lab. Is that correct? My understanding about the ASCP certification is >that it is aimed at providing a modicum of proof that a person is >qualified to work in a medical diagnostic lab. Research labs are not >considered diagnostic labs. As you imply, a person in a research lab >will often work on only a limited sample set. Therefore, it is >meaningless to apply the the ASCP standard to research people. > > If you are planning to move into the diagnostic field, then I'll bet >you could easily find a job in a diagnostic lab, get the experience, >and qualify to take the test. It may be that some diagnostic labs have >a suggested requirement to be ASCP certified as a QIHC, but the vast >majority would be happy to find someone with the experience you >outline, even if they had not previously worked in a diagnositc lab. > >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: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James >Watson >Sent: Wednesday, November 16, 2005 7:28 AM >To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: QIHC change > > >This is my point. With the requirements listed below someone with 25 >years of experience doing immuno (single, double, triple antibody >staining, making own antibodies, and in situ Hybridization: all with >and without using kits, all with and without using an automated >stainer) is not qualified for this certification if they work in a >research facility where immunophenotyping is not done. There is no >system of doing it on your own to prove that you have the capability to >do immunophenotyping in order to fullfil this requirement. I guess it >is time to start harrassing ASCP about the unfairness of this system. > > >From almost always sunny San Diego >Jamie > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of >novanity@nc.rr.com > Sent: Wed 11/16/2005 5:55 AM > To: histonet@lists.utsouthwestern.edu > Cc: > Subject: [Histonet] RE: QIHC change > > > > James, to qualify for the qualification you take the 50 >question test > and submit an employer reference form + of course satisfy one of the > three routes. There is no practical to submit anymore. I >think that is > what you are asking. It seems to me that it wouldn't matter about > specificity antigens/markers or what diseases or human cells. >There is > no requirement other than what is requested on the employer >reference > form which you can't see the details until you order and >receive your > packet. Is it possible for anyone to post a copy of the employer > reference form. From the ASCP website this is what it says " > > Qualification in Immunohistochemistry > Experience requirements > > Applicants must have experience in the following areas > > * Immunohistochemical and Immunofluorescence Preparation > All of the following should have been performed by the >applicant > o staining technique > o selection of proper control material > o titration of immunologic reagents > > * Immunophenotyping > in at least one of the following applications > o immunodeficiencies > o immunoproliferative disorders (neoplastic and >non-neoplastic > disorders) > o transplantation biopsies > o other immunophenotyping applications > please specify: ______________________ > > * Quality Assurance > The applicant should have participated in Quality Assurance > related to all of the following > o specimen fixation, processing, microtomy > o reagent selection, preparation, storage, disposal > o method selection, validation, documentation > o quality control > o safety > > " This is the experience which I am assuming is only >documented for the > ASCP through the employer reference form, hence if you only do >A and C > and not B you can't qualify unless your employer is dishonest on the > form. Because even if you crosstrain into what I assume is flow > cytometry but don't actually work it day to day as part of >your job you > do not qualify because you have not had experience doing it for a > minimun of 12 months. As for research, same thing if you do >all of it > every day then your good to go. If not it is a grey or is it >gray area > that I'm looking more information/details on. In the past you > qualified your work with different immuno stains as a practical , I > don't remember there being a flow requirement. Maybe I'm wrong but > anyone have this info I'm looking for. > > G Hurlburt HT(ASCP) > sunny and warm NC > > _______________________________________________ > 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 sjchtascp <@t> yahoo.com Wed Nov 16 14:26:44 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Nov 16 14:26:52 2005 Subject: [Histonet] cryoprotection Message-ID: <20051116202644.45169.qmail@web90201.mail.scd.yahoo.com> How are techs freezing their fixed, 30% infiltrated tissue. Right now we freeze the tissue (liver) using OCT placed in a small freezing cup and float on liquid nitrogen. They section great but the cellular detail is only fair. Does anyone have really good result cryoprotecting tissue. Steve --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From JJacobs <@t> student.uchc.edu Wed Nov 16 14:31:07 2005 From: JJacobs <@t> student.uchc.edu (Jacobs,Jennifer (stu)) Date: Wed Nov 16 14:31:18 2005 Subject: [Histonet] am I snap-freezing correctly? Message-ID: <9F866D418736DD408F43059FF7FC697203B34973@itnsa.uchc.edu> Hi, I am snap freezing brain tissue for sectioning (8uM) on cryostat (-25C). When I snap freeze, I put the brain hemisphere on a piece of aluminum foil (boat) and let it float on the LN2. Most times, some LN2 comes in direct contact with the brain tissue as the boat is not leak-free. Is this bad? Should I make sure the LN2 never comes in contact with the brain? The problem I've been noticing is that, right off the cryostat, my tissue looks "hole-y". This just gets worse as I proceed with the IHC stain (CD-31 Ab to see vessels) and dehydration + xylene steps. By the xylene steps the holes are so stretched out that I can't even recognize vessels in the tissue. I am not sure whether the holes arise from the brain extraction procedure (ie: snap freezing) or from the sectioning. But the person who sections has been doing it for years and has great experience with the cryostat. Any ideas/suggestions would be great! Thanks a lot! Jenn UConn Health Center Dept Pharmacology Farmington, CT From cfavara <@t> niaid.nih.gov Wed Nov 16 14:47:08 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Wed Nov 16 14:47:19 2005 Subject: [Histonet] am I snap-freezing correctly? Message-ID: Look in the archives for the protocol for freezing over isopentane by Gayle Callis. This is what I find this better than LN for morphology - ditto for the cryoprotected specimens. c 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: Jacobs,Jennifer (stu) [mailto:JJacobs@student.uchc.edu] Sent: Wednesday, November 16, 2005 1:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] am I snap-freezing correctly? Hi, I am snap freezing brain tissue for sectioning (8uM) on cryostat (-25C). When I snap freeze, I put the brain hemisphere on a piece of aluminum foil (boat) and let it float on the LN2. Most times, some LN2 comes in direct contact with the brain tissue as the boat is not leak-free. Is this bad? Should I make sure the LN2 never comes in contact with the brain? The problem I've been noticing is that, right off the cryostat, my tissue looks "hole-y". This just gets worse as I proceed with the IHC stain (CD-31 Ab to see vessels) and dehydration + xylene steps. By the xylene steps the holes are so stretched out that I can't even recognize vessels in the tissue. I am not sure whether the holes arise from the brain extraction procedure (ie: snap freezing) or from the sectioning. But the person who sections has been doing it for years and has great experience with the cryostat. Any ideas/suggestions would be great! Thanks a lot! Jenn UConn Health Center Dept Pharmacology Farmington, CT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Nov 16 14:56:17 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 16 14:56:25 2005 Subject: [Histonet] Luxol fast blue/cresyl violet stains In-Reply-To: <714463.1132166812935.JavaMail.ocsadmin@jcs-mid-prod.jax.org> Message-ID: <20051116205617.42144.qmail@web61218.mail.yahoo.com> Judi: When you say "5 slides from the same animal" do you mean slides from different blocks from the same animal or slides from the same block. If the slides are from different blocks then the problem could reside in the blocks and how they were treated. If all the slides are from the same block then the difference could reside in the configuration the slides were placed in the staining jar (although this explanation could be far fetched). I don't think that the differences could be due to the differentiation step because you say that after differentiation the slides are placed in distilled water, had they been "waiting" for the result of one slide to stop the differentiation on the others that could be the cause. Now, if the 37Celsius incubation is done in an oven and the oven does not have a good air circulation it could be that one area in the staining jar could have a difference in temperature at least during some time, although that could be compensated during the long incubation period. So I really don't know what to atribute the problem to. Not exactly these, but the delay itself, prompted us to eliminate this protocol and go to one using microwaves (MW) as follows: 1- dewax and take sections to absolute ethanol (EthOL) 2-place slides in plastic Coplin jar and add 95% EThOL 3- heat slides in the 95% EthOL for 15 secs. at 0.95 kcal ("Level 5" of our 480 W MW oven which was equivalent to 266 Watts) 4- dump the heated 95% EthOL (used only to heat the slides) and substitute it with 40 mL of Luxol Fast Blue (LFB). Heat at the same level (266 Watts) for 20 secs (=1.27 kcal). 5- transfer the Coplin jar with the slides to a water bath at 60Celsius during 5 minutes. 6- Wash slides in distilled water. 7- Differentiate the slides , one at a time, with 0.5%aq. lithium carbonate solution until no more blue color runs off the slides (not necessary under the microscope) 8- Transfer the slides to distilled water. 9- Dump the dist. water and add the Cresyl Echt Violet (CEV) and heat in the MW at full power (480 Watts) for 20 secs.(=2.3 kcal). 10-Transfer the slides to the water bath at 60Celsius for 5 minutes. 11- Dehydrate QUICKLY, clear and mount as usual. With this procedure we did not have any problems and it could be completed in about 30 minutes. Perhaps you could care to try this protocol to find out if the problem persists. This is how we did it in our lab. Hope this will help! Rene J. judi.ford@jax.org wrote: Hi, I have a question concerning the lfb/cv stain. This is the situation we have in our lab. The tissue we are working on is Bouin's fixed mouse brain and spinal cord (cord is left in the vertebrae). The spinal cord may be left in Bouin's for an extended period for decalcification (two-three weeks). The trimmed tissue is then sent to our lab where we rinse it for a day before processing on our VIP processors. The next day we embed the tissue and then it is cut by one of our technicians and stained. This is our staining technique: deparaffinize to 95% alcohol and place in alcoholic luxol fast blue overnight at 37 degrees. The next morning it is rinsed twice in 95% alcohol and twice in distilled water. The slides are separated into racks containing either brain or spinal cord for differentiation. Slides are dipped in 70% alcohol for 20 seconds then lithium carbonate for 20 seconds. This is followed by two rinses in distilled water. Then checked microscopically before determining if differentiation is complete or not. If not, then the slides go another round through 70% and lithium for a few more seconds. Once this part is done the slides are put into a warmed cresyl violet solution (we add 10% glacial acetic acid, 10 drops for every 100mls of stain) for 4-6 minutes then rinsed in 95% alcohol three times. Finally they are dehydrated, cleared and coverslipped. The problem comes when we have 5 slides from the same animal, all processsed and stained together, where in one slide the cresyl violet works but in the other 4 it doesnt' work. Every slide is treated exactly the same, yet we have this difference. Any ideas on what may be happening? We've discussed decalcification and possible lithium effects on the cresyl violet. We're at a loss and our pathologist is very determined to discover the solution for this problem. Thank you advance for any ideas you may send our way..........We greatly appreciate your help! Judi Ford HIstotechnologist HIstopathology & Microscopy The Jackson Laboratory 600 Main St. Bar Harbor, Me 04609 207-288-6193 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From rjbuesa <@t> yahoo.com Wed Nov 16 15:01:46 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 16 15:01:55 2005 Subject: [Histonet] recycled xylenes In-Reply-To: Message-ID: <20051116210146.28437.qmail@web61224.mail.yahoo.com> Betsy: We used to recycle xylene and used for everything. Since the recovery is not 100% when we had to buy new we preferred to use it in the staining and coverslippin instruments. Rene J. "Molinari, Betsy" wrote: Hi, I am interested in knowing if you have preferences for your recycled xylenes. Do you use it just for staining, or processing alone. Or use it for both? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From jkiernan <@t> uwo.ca Wed Nov 16 15:07:06 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Wed Nov 16 15:07:17 2005 Subject: [Histonet] am I snap-freezing correctly? References: <9F866D418736DD408F43059FF7FC697203B34973@itnsa.uchc.edu> Message-ID: <437B9F7A.7E99AF0F@uwo.ca> The more holey than righteous appearance that you describe is typical of slowly frozen tissue - especially if it hasn't been cryoprotected. When liquid nitrogen comes into contact with an object it boils, and the resulting layer of nitrogen gas has an insulating effect that slows down the cooling process. For that reason it is preferable to freeze specimens quickly by immersing in a liquid that has been pre-cooled to a low temperature. The one most often used is isopentane. Put some in a small metal can, and stand the can in liquid nitrogen. Stir the isopentane with a lollipop stick. When it starts to thicken the temperature is approaching its freezing point (-160C) and the specimen (with or without aluminium foil) can be dropped in. Another method is to bring a fairly massive block of metal to liquid nitrogen temperature (-196C) and slap the specimen onto the surface of the block. You will probably get plenty of replies. There has been much Histonetting about quick freezing methods over the years! -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Jacobs,Jennifer (stu)" wrote: > > Hi, > > I am snap freezing brain tissue for sectioning (8uM) on cryostat (-25C). > When I snap freeze, I put the brain hemisphere on a piece of aluminum foil (boat) and let it float on the LN2. Most times, some LN2 comes in direct contact with the brain tissue as the boat is not leak-free. Is this bad? Should I make sure the LN2 never comes in contact with the brain? > > The problem I've been noticing is that, right off the cryostat, my tissue looks "hole-y". This just gets worse as I proceed with the IHC stain (CD-31 Ab to see vessels) and dehydration + xylene steps. By the xylene steps the holes are so stretched out that I can't even recognize vessels in the tissue. > > I am not sure whether the holes arise from the brain extraction procedure (ie: snap freezing) or from the sectioning. But the person who sections has been doing it for years and has great experience with the cryostat. > > Any ideas/suggestions would be great! Thanks a lot! > > Jenn > > UConn Health Center > Dept Pharmacology > Farmington, CT > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Wed Nov 16 15:36:26 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Nov 16 15:36:52 2005 Subject: [Histonet] RE: QIHC change Message-ID: James, et al, Thank you for bringing lively discussion in an area I believe needs it. I have worked for many years in clinical histology labs and now work in the research environment. I have to admit, once I heard they had moved this exam to a 50-question test, I signed up. Previously, I knew it would be nearly impossible for me to do the practical exam with human tissue samples since we don't have any here. I took the exam last week and have to say that while most of the questions were based on general IHC knowledge, a couple of questions regarded clinical samples (cytology specimens) not usually done in research, or were geared toward estrogen receptor-type immunostaining. Most of my issues with the exam related to the wording of the question, or the choices given (I didn't always agree with them, and one was ambiguous). I'm glad I now have a contact to whom I can give my opinion of the exam, thanks James! My conclusion was, the exam can be taken successfully by research techs with a strong background in IHC, and the clinical-only questions they will miss will still give them enough margin to pass. (Provided, of course, they meet the requirements.) I agree as well that the immunophenotyping requirement be phased out or reworded to show knowledge of typical staining patterns for typical markers and not just the leukemia/lymphoma ones. Usually only the large hospital or private IHC labs do these types of panels, so even in the clinical arena this type of experience isn't easy to get. The IHC techs aren' t the ones interpreting these results, nor are they ordering the panels. At best they run patient samples with known positive controls (mostly tonsil), and order and work up new antibodies to run in these panels--which are then evaluated by a pathologist for applicability. How is that different from any other marker run in an IHC lab? Regarding the need for ASCP-certified techs in the research environment, I agree with Tim that it should not necessarily be a requirement. I know that an ASCP-certified tech has general knowledge of histotechnology and has demonstrated skill with a microtome and staining techiques, and has the certification to prove it. But that alone does not make them any more qualified to work in the lab than someone with a BS or MS and histotechnology experience but no certification. Please don't think I'm making the case for random hiring of non-certified techs. I believe firmly in certifications for personal and professional growth. I just don't think the research environment should base their job requirements on it. My employment ad usually indicates the person we are looking for be ASCP certified, or certification eligible. In the absence of the certification, meeting minimum educational requirements is usually acceptable. Finally, I have not paid my dues to ASCP for a very long time and have no plans to start doing so any time soon. I will eagerly hand over my money to the NSH because they offer workshops and a publication that are geared towards the profession. Even the Advance magazine runs more articles on Histology than the ASCP journal ever did. Perhaps that has changed, but somehow I doubt it. I too would be steamed if I had paid all this time and got no value for my $$$s. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From int09018 <@t> alphahunt.com Wed Nov 16 19:50:15 2005 From: int09018 <@t> alphahunt.com (HCS) Date: Wed Nov 16 19:50:46 2005 Subject: [Histonet] Help- triphenyltetrazolium chloride instructions for staining needed Message-ID: <000601c5eb19$4d4b7df0$6601a8c0@hp> Hi, I am still looking for a reference or procedure for doing the TTC stain (triphenyltetrazolium chloride). If you might have a procedure I would be able to use that would be very helpful. Thanks LeRoy Brown HCS . From JUDIEWHIT <@t> aol.com Wed Nov 16 20:51:35 2005 From: JUDIEWHIT <@t> aol.com (JUDIEWHIT@aol.com) Date: Wed Nov 16 20:51:50 2005 Subject: [Histonet] Re: Histonet Digest, Vol 24, Issue 23 Message-ID: <217.e6894da.30ad4a37@aol.com> Hi Jill, Looks like you are getting ready to open your new lab. Congrats girlfriend I knew you could do it! I know some of the techs in Phoenix if you need references let me know. I am getting ready to come out of retirement and go traveling...looks like I will be working in a lab in new Mexico for 13 weeks soon, I think the travel thing will be fun and I can move on to another Histo job in different parts of the country. The very best to you my friend... Judie W. From pathrm35 <@t> adelphia.net Wed Nov 16 22:12:28 2005 From: pathrm35 <@t> adelphia.net (Ron Martin) Date: Wed Nov 16 22:12:55 2005 Subject: [Histonet] biohazard waste removal companies Message-ID: <001001c5eb2d$2100bf80$27233418@Pathrm35> Does any one know of any companies that service southeast Florida for biohazard waste and chemical waste removal? I am currently using Environmental Enterprises out of Orlando but lately this company has been unable to service our needs. Thanks in advance. Ron Martin From c.m.vanderloos <@t> amc.uva.nl Thu Nov 17 02:05:55 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Thu Nov 17 02:06:07 2005 Subject: [Histonet] RE: aqueous coverslipping medium Message-ID: <1b34f811b341d2.1b341d21b34f81@amc.uva.nl> Paul, The media you call the "liquid coverslip" is in fact a kind of plastic polymer that can be mounted up organically after polymerisation. I experienced that without this additional organic mounting the plastic polymer will fall off after some years. If this happens you can remove the plastic completely by soaking the slide in luke warm water. Then "liquid-coverslip" again. Although most of those products state in their data sheet that it should be used without a coverslip, it also works fine WITH a coverslip. The only thing you should not do is harden at 60-70C as you normally do (it causes air bubbles!). Just leave the slides at room temp. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Wed, 16 Nov 2005 12:40:06 -0500 From: "Monfils, Paul" Subject: [Histonet] aqueous coverslipping medium To: "'histonet@lists.utsouthwestern.edu'" What brand do you prefer for aqueous mounting medium to be used with a coverslip? I have several good hydrocarbon-based coverslipping media, and also some good aqueous media to be used without coverslips (so-called liquid coverslip), but I don't have an aqueous coverslipping medium I am completely satisfied with. Applications would include immunofluorescence and a few enzyme stains, as well as lipid stains and other techniques that are not compatible with hydrocarbon-based media. Are there any such media that harden after application, to produce a permanent slide? From louise.renton <@t> gmail.com Thu Nov 17 03:31:40 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Thu Nov 17 03:31:49 2005 Subject: [Histonet] hard tissue distortion Message-ID: Helo all, I've been asked this question by a postgrad and I wasn't able to answer it to my satisfaction. The question was - how much distortion occurs in tissue (in this case teeth) that have been fixed in 70% alcohol, processed by hand and embedded in MMA?, Sections were cut at 6um, stained free floating and mounted in Entellan on glass slides. Any thoughts/references? I would be most grateful for anything The post grad is doing histometry on healing furcation defects, and is wondering if there are any variables that he needs to mentionor take into consideration. Best regards -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....onwards through the fog!" From sluhisto <@t> yahoo.com Thu Nov 17 08:10:19 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Thu Nov 17 08:10:29 2005 Subject: [Histonet] CD31 staining histiocytes??? Message-ID: <20051117141019.29359.qmail@web51003.mail.yahoo.com> Hello all: Is anyone out there using Dako CD31 and getting histiocyte staining in human skin??? Our dermpath is seeing this and is questioning it. We are using it at 1:100 with the biocare pressure cooker with Biocare Diva decloaker solution. Our detection system is Envision +. Any thoughts??? Susan Histopathology Supervisor St. Louis University Medical School --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From angela.mcnabola.b <@t> bayer.com Thu Nov 17 08:52:03 2005 From: angela.mcnabola.b <@t> bayer.com (Angela McNabola) Date: Thu Nov 17 08:51:37 2005 Subject: [Histonet] CD31 Santa Cruz update Message-ID: Hello all, I received a new lot of CD31 yesterday, provided to me by Santa Cruz. I have not yet had time to try it out (FFPE-mouse xenografts), but I have been told that it is working. I know that several of you have been waiting on this, so now might be a good time to give them a call. Angela Angela McNabola MS, HT(ASCP)SLS, QIHC Scientist Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 203-812-5001 fax# 203-812-5820 angela.mcnabola.b@bayer.com From ree3 <@t> leicester.ac.uk Thu Nov 17 08:56:02 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Thu Nov 17 08:56:13 2005 Subject: [Histonet] E: beta-2- microglobulin Message-ID: Looking for an antibody to the above that works, as ever, on paraffin processed mouse tissues... Many thanks Richard Edwards MRC TOX UNIT LEICESTER...U.K.... From david.kinsley <@t> spcorp.com Thu Nov 17 09:36:34 2005 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Thu Nov 17 09:37:21 2005 Subject: [Histonet] fixed jaw clamps/ large size specimens Message-ID: <79A75F7AD4BFAE4CA27C8AABE3F75D9A0E6C42@KENMSG22.us.schp.com> I'm looking to section some larger size specimens, particularly rat hind paw samples and am looking for cassettes that are slightly larger than the mega cassette but not as large as the super mega cassette. (Approximately 2" long by 1.5" wide by 1/2 to 3/4" deep) I have called some vendors and have had little luck. The only suggestion was to purchase a fixed jaw clamp for my microtome (Microm HM355) and try to use that with the super mega cassette. Does anyone have any experience with these types of specimens? Any suggestions on cassettes or microtome adapters? thanks David Kinsley Schering-Plough Research Institute Kenilworth, NJ 07033 908-740-4669 ********************************************************************* 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 Bauer.Karen <@t> mayo.edu Thu Nov 17 10:09:09 2005 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Thu Nov 17 10:11:36 2005 Subject: [Histonet] Dissecting Microscopes Message-ID: Good morning! I need to purchase a new dissecting microscope for our pathologists. Ours is getting old and needs to be updated. We mainly use it for finding glomeruli in kidney biopsies to make sure there is adequate tissue for certain types of testing. I've looked online and have requested some quotes, but I'm curious to know what others are using. Any suggestions? As always, thank you. Karen L. Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************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 rjbuesa <@t> yahoo.com Thu Nov 17 11:31:15 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 17 11:31:25 2005 Subject: [Histonet] fixed jaw clamps/ large size specimens In-Reply-To: <79A75F7AD4BFAE4CA27C8AABE3F75D9A0E6C42@KENMSG22.us.schp.com> Message-ID: <20051117173115.47213.qmail@web61215.mail.yahoo.com> David: In 1988 we began cutting whole prostate slices and we could not find neither megacassettes nor molds, so we made ours. For cassettes I used small round plastic containers, cut to an appropriate height with drilled holes and stappled 2 together, with the specimen in between. They work perfectly. As molds I cut pieces of wood of the required size and used aluminum sheets bought in a hardware to "wrap" around the wood piece and made them "melted paraffin leak proff". They also worked beautifully and, even when at last we bought the available cassettes and molds, for really large specimens (up to 3" x 4" ) we still use them. For those large paraffin blocks I used the very very old technique of adhering them to small 1"x1"x 2" blocks of wood, using a hot spatula, to be clamped to the microtome. Hope this "barrage" of "oldies" will help! Rene J. "Kinsley, David" wrote: I'm looking to section some larger size specimens, particularly rat hind paw samples and am looking for cassettes that are slightly larger than the mega cassette but not as large as the super mega cassette. (Approximately 2" long by 1.5" wide by 1/2 to 3/4" deep) I have called some vendors and have had little luck. The only suggestion was to purchase a fixed jaw clamp for my microtome (Microm HM355) and try to use that with the super mega cassette. Does anyone have any experience with these types of specimens? Any suggestions on cassettes or microtome adapters? thanks David Kinsley Schering-Plough Research Institute Kenilworth, NJ 07033 908-740-4669 ********************************************************************* 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 --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From immrstambo <@t> hotmail.com Thu Nov 17 12:15:19 2005 From: immrstambo <@t> hotmail.com (Christine Tambasco) Date: Thu Nov 17 12:15:28 2005 Subject: [Histonet] Slide Adhesion;toenail for fungus stain In-Reply-To: <9B614E04-8475-4F11-ABEF-EA21961E61A5@infosight.com> Message-ID: Dear John, I have a lot of trouble getting toenails to stay on the slide, especially doing the GMS stain for fungus. I have used Poly-L-Lysine coated slides, probe slides and even Elmers glue. I would be very interested in a slide that could keep the toenail attached. Thanks for R&R. Christine Tambasco, HT (ASCP) St. Mary's Hospital Laboratory Amsterdam, New York 12010 ph-518-841-7287 ______________________________________________________________ From: John Robertson To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide Adhesion Date: Wed, 16 Nov 2005 08:42:57 -0500 >We are interested in seeing if a new slide coating method has >advantages for the adhesion of "difficult" sections. It should work >well in all applications and ultimately be less costly than existing > charged slides. >If you have an especially difficult adhesion problem , I would like >to hear from you and offer you a few prototype samples for testing. >Thanks > >John > > > > >John Robertson P.E. Ph.D. jr@infosight.com >CEO InfoSight Corporation http://www.infosight.com " We > Barcode Difficult Stuff" tm >P.O. Box 5000 >20700 Rt. 23 Chillicothe , OH 45601 >Phone (740) 642 3600 (Eastern time zone) >Fax (740) 642 5001 >Private Fax. (740) 642 3106 >PGP Key 0x8EDD65A2 > >Never Confuse Motion with Action > Ben Franklin > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From beckyjo <@t> email.unc.edu Thu Nov 17 12:15:28 2005 From: beckyjo <@t> email.unc.edu (Rebecca Jo) Date: Thu Nov 17 12:16:08 2005 Subject: [Histonet] H&E Staining of Frozen Sections Message-ID: <20051117131528.t3ng9d1cu8wgw0kw@webmail1.isis.unc.edu> Hi Histoneters- I was wondering if someone could give me a protocol for H&E staining on frozen sections. I assume I can just skip over the deparaffinizing steps and go straight to the staining, but I wanted to know how long I should keep the sections in hematoxylin. Just enough to stain the sections? Any help would be greatly appreciated. -Cheers -- Rebecca E. Jo UNC-School of Medicine Department of Cell and Developmental Biology (919)843-9648 CB# 7090 From rjbuesa <@t> yahoo.com Thu Nov 17 12:29:31 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 17 12:29:40 2005 Subject: [Histonet] H&E Staining of Frozen Sections In-Reply-To: <20051117131528.t3ng9d1cu8wgw0kw@webmail1.isis.unc.edu> Message-ID: <20051117182931.35394.qmail@web61214.mail.yahoo.com> We used to follow the following protocol: 1- fix the frozen section in acetone X 10 secs. 2- distilled water x 10 secs 3- hematoxylin x 30 secs 4- tap water 5- quick dip in acid alcohol 6- eosin 5-10 secs. 7- 95% ethanol > 100% ethanol x2 > xylene X2 > coverslip. The pathologists had the slide ready in about 2-3 minutes. Rene J. Rebecca Jo wrote: Hi Histoneters- I was wondering if someone could give me a protocol for H&E staining on frozen sections. I assume I can just skip over the deparaffinizing steps and go straight to the staining, but I wanted to know how long I should keep the sections in hematoxylin. Just enough to stain the sections? Any help would be greatly appreciated. -Cheers -- Rebecca E. Jo UNC-School of Medicine Department of Cell and Developmental Biology (919)843-9648 CB# 7090 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From rjbuesa <@t> yahoo.com Thu Nov 17 12:35:43 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 17 12:35:53 2005 Subject: [Histonet] Slide Adhesion;toenail for fungus stain In-Reply-To: Message-ID: <20051117183543.88485.qmail@web61211.mail.yahoo.com> If the toenails are well processed (which in its turn will require an alkaline "digestion" before processing until the toenail is pliable) they will usually stay in the positive slide. Have you tried adding carpenter's yellow glue mixed in the water bath (about 0.5% sol.)? Another thing, we used PAS to detect fungi (they stain fantastic) GMS is too "violent" for this type of so "capriceous" specimen. Would you give it a try to PAS for fungi in trying to circunvent this problem? Just a thought! Rene J. Christine Tambasco wrote: Dear John, I have a lot of trouble getting toenails to stay on the slide, especially doing the GMS stain for fungus. I have used Poly-L-Lysine coated slides, probe slides and even Elmers glue. I would be very interested in a slide that could keep the toenail attached. Thanks for R&R. Christine Tambasco, HT (ASCP) St. Mary's Hospital Laboratory Amsterdam, New York 12010 ph-518-841-7287 ______________________________________________________________ From: John Robertson To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide Adhesion Date: Wed, 16 Nov 2005 08:42:57 -0500 >We are interested in seeing if a new slide coating method has >advantages for the adhesion of "difficult" sections. It should work >well in all applications and ultimately be less costly than existing > charged slides. >If you have an especially difficult adhesion problem , I would like >to hear from you and offer you a few prototype samples for testing. >Thanks > >John > > > > >John Robertson P.E. Ph.D. jr@infosight.com >CEO InfoSight Corporation http://www.infosight.com " We > Barcode Difficult Stuff" tm >P.O. Box 5000 >20700 Rt. 23 Chillicothe , OH 45601 >Phone (740) 642 3600 (Eastern time zone) >Fax (740) 642 5001 >Private Fax. (740) 642 3106 >PGP Key 0x8EDD65A2 > >Never Confuse Motion with Action > Ben Franklin > > > >_______________________________________________ >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 --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From sharon.osborn <@t> dnax.org Thu Nov 17 12:40:15 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Thu Nov 17 12:40:54 2005 Subject: [Histonet] Recycled xylenes Message-ID: <29B25753F6B1D51196110002A589D444032C48CF@PALMSG30.us.schp.com> Betsy, The recycled xylenes become more pure over time than that you purchase commercially. Xylenes is by definition, plural and a mixture of ortho, meta and para isomers of xylenes with some ethyl benzene. The recycled works well in all functions of the histology laboratory except one. If you are using the Sakura tape coverslipper, you will need to use some newly purchased xylenes to run it. The recycled, ie., "purified" xylenes do not work as well with the tape coverslipper's adhesive causing it to not activate to "stick tight". I found this out through experience; the tape coverslipper needs the impurities in the xylenes to do its job(as best as it can). We solved this problem by having the last two stations of xylene on the autostainer to be the commercial xylenes rather than the recycled xylenes; or have two stations that you hand dip through before loading the carrier on the tape slipper. There seems to be no problem with the mounting solutions used in glass coverslippers by using the recycled xylenes. Enjoy your savings of money and the environment with your recyclers! sharon osborn DNAX, Schering-Plough BioPharma Palo Alto, Ca "Molinari, Betsy" wrote: Hi, I am interested in knowing if you have preferences for your recycled xylenes. Do you use it just for staining, or processing alone. Or use it for both? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) ********************************************************************* 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 pruegg <@t> ihctech.net Thu Nov 17 12:50:22 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Nov 17 12:50:34 2005 Subject: [Histonet] CD31 staining histiocytes??? In-Reply-To: <20051117141019.29359.qmail@web51003.mail.yahoo.com> Message-ID: <200511171850.jAHIoMxT014147@chip.viawest.net> You may be over heating your samples for cd31, I had to go to a waterbath at 80dc for 3 hours using the low ph hier buffer. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: Thursday, November 17, 2005 7:10 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD31 staining histiocytes??? Hello all: Is anyone out there using Dako CD31 and getting histiocyte staining in human skin??? Our dermpath is seeing this and is questioning it. We are using it at 1:100 with the biocare pressure cooker with Biocare Diva decloaker solution. Our detection system is Envision +. Any thoughts??? Susan Histopathology Supervisor St. Louis University Medical School --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Nov 17 12:55:45 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Nov 17 12:55:59 2005 Subject: [Histonet] Slide Adhesion;toenail for fungus stain In-Reply-To: Message-ID: <200511171855.jAHItjxT015745@chip.viawest.net> It is really tuff to get toe nails and cartilage to stay on the slide, especially if you use heat when staining. My very best method includes pre-coating slides with a 5% elmers glue solution, let the slides airdry very well after coating, I usually airdry overnight. Use these slides to pick up your sample and then airdry them with the section on them again overnight. To melt the paraffin I place the airdryed sections on a flat slide warming plate at 55dc for 10 min., followed again by complete airdrying. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Christine Tambasco Sent: Thursday, November 17, 2005 11:15 AM To: jr@infosight.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Slide Adhesion;toenail for fungus stain Dear John, I have a lot of trouble getting toenails to stay on the slide, especially doing the GMS stain for fungus. I have used Poly-L-Lysine coated slides, probe slides and even Elmers glue. I would be very interested in a slide that could keep the toenail attached. Thanks for R&R. Christine Tambasco, HT (ASCP) St. Mary's Hospital Laboratory Amsterdam, New York 12010 ph-518-841-7287 ______________________________________________________________ From: John Robertson To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide Adhesion Date: Wed, 16 Nov 2005 08:42:57 -0500 >We are interested in seeing if a new slide coating method has >advantages for the adhesion of "difficult" sections. It should work >well in all applications and ultimately be less costly than existing > charged slides. >If you have an especially difficult adhesion problem , I would like >to hear from you and offer you a few prototype samples for testing. >Thanks > >John > > > > >John Robertson P.E. Ph.D. jr@infosight.com >CEO InfoSight Corporation http://www.infosight.com " We > Barcode Difficult Stuff" tm >P.O. Box 5000 >20700 Rt. 23 Chillicothe , OH 45601 >Phone (740) 642 3600 (Eastern time zone) >Fax (740) 642 5001 >Private Fax. (740) 642 3106 >PGP Key 0x8EDD65A2 > >Never Confuse Motion with Action > Ben Franklin > > > >_______________________________________________ >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 jamie.erickson <@t> abbott.com Thu Nov 17 13:46:01 2005 From: jamie.erickson <@t> abbott.com (Jamie E Erickson) Date: Thu Nov 17 13:46:09 2005 Subject: [Histonet] Question about De-calcifying mouse paws Message-ID: HI All, Here is my problem, We are a research histology lab which supports groups doing Mouse/ Rat Collagen Induced Arthritis (CIA). The researchers collected the paws and knees for routine processing (Paraffin) and we take it from there. We are trying to give a quick turn around time (2 weeks) on these studies so we are fixing the paws for 24 hours in 10% neutral buffered formalin then switching it to CAL RITE (a formaldehyde/Methanol/ Formic acid mixture from Richard Allan) for decalcification. The paws sit in Cal-Rite for 2 days on a shaker and are then trimmed by cutting the skin and one toe on each side off the paws or trimming 1/3 off the knee Sagittal section (knee and some of the long bones), this helps in decaling the paws and knee quicker. Then the samples are put into fresh De-cal again for 2 more days before washing for 1 hour in tap water and processing. Problem: Knees are great , section great look great but some but not all of the paws are chalky, white deposits in toes and ankles. This only happens to about 1/3 of the samples. I guess they are not de-calcified long enough? Is there another way people are De-caling quickly with better results? Our Pathologist is happy with the slides for the most part but the bad ones are more difficult to section as you can imagine and sometimes the ankle joints don't section well at all. Any Ideas would be greatly appreciated. Thanks _______________________________ Jamie Erickson Sr. Research Associate Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com From gcallis <@t> montana.edu Thu Nov 17 14:32:17 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Nov 17 14:32:38 2005 Subject: [Histonet] Question about De-calcifying mouse paws In-Reply-To: References: Message-ID: <6.0.0.22.1.20051117130329.01b77b50@gemini.msu.montana.edu> Jamie, Well you are NOT alone in this, you have described a common problem we experienced also. Liz Chilpala should also have input on this. Mouse paws with ankles are really dense (tightly packed tiny bones and connective tissues). We fix paws much longer than 24 hours due to their density, often up to a week. Have you thought of perfusion to help with speeding up fixation? Decalcification should be done in the time frame you gave. Did you look to see what the concentration of formic is in your CalRite MSDS? I think maybe as high as 10% but if concentration of methanol is high, then decalcification slows down. We have used 70% alcohol to stop decalcification since calcium does not ionize well in presence of bipolar alcohols. Use of alcohol, historically, slows down decalcification rate, hence damaging effects of mineral acids on tissues (called overdecalcification, although overexposure may be a better term) , etc i.e nitric formulations. Perenyi's decalcifying solution an example If bone is totally fixed, we prefer 10 to 15 % formic acid from 90% formic acid stock (V/V) and paws can still take a couple of days. I do a weight gain/weight loss endpoint check and since most samples are the same size, I work with several from both the control and OA induced group for endpoint testing. Rather than weigh each and every sample in a group of 50 mice!! You did not say how long you process, but the chalky, opaque look can be caused by 1. Underdecalcification (endpoint test helps unless you have an xray machine or microCT scanner, even better) 2. Poor dehydration/clearing (increase times in all stations - I do 2 hours per station in a VIP and use Tissue Prep 2 (harder) for infiltration and embedding) but infiltrate at 58 to 60C with this, 3 to 4 changes always using vacuum/pressure/ambient temps for processing. 3. OR both of the above. When that happens, heavy sigh and see what has been done, then correct accordingly. You can try and melt down the blocks, go back into vacuum/paraffin for a few hours - 2 or so, reembed and see it that improves things. If tissue swells away from trimmed blocks after soak, back off - warm water followed by ice cold water often helps. Oversoaking only exacerbates the problem!! I also know of another decalcifier that works faster and with cartilage. It may be more damaging to IHC though but works with cartilage stains and H&E's - I will be happy to send recipe to you. Good luck Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From craig.case <@t> thermo.com Thu Nov 17 15:17:01 2005 From: craig.case <@t> thermo.com (Case, Craig T.) Date: Thu Nov 17 15:17:17 2005 Subject: [Histonet] biohazard waste removal companies Message-ID: Ron: I saw your posting on histonet. You may want to look into Stericyle as they process red bag waste and certain chemical waste. They can be found on the web. I hope this helps. Craig Craig T. Case Midwest Sales Representative Clinical Diagnostics Anatomical Pathology Thermo Electron Corporation 800-245-6212 ext. 4721 563-663-0428 (cell) 563-583-9244 (fax) craig.case@thermo.com From liz <@t> premierlab.com Thu Nov 17 15:20:53 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Thu Nov 17 15:21:12 2005 Subject: [Histonet] Question about De-calcifying mouse paws (long) In-Reply-To: Message-ID: <000001c5ebbc$ca9aaa80$a7d48a80@AMY> Jamie Gayle is correct especially about fixation. We like to fix for a minimum of 48 hours, placement of specimen in decal prior to adequate fixation can cause extremely poor results. Depending upon how you section your mouse joints (we normally have 8 joints per animal, 2 knees, 2 ankles, 2 front paws and 2 rear paws) once we receive the joints, prior to decalcification we trim the knees to remove most of the muscle and trim the ankle from the rear paw and trim the front paws to the size we need. If this can be done by the individuals at necropsy it will aid in fixation, if not then we do as soon as we get the specimens, if we feel that they have been adequately fixed then we place the tissues in decal. We process two joints, sometimes four depending upon the client per block. If I had my way it would be no more than two like tissues (ie two ankles, etc) but even with the mouse model since the disease is not symmetrical you might have one ankle that's swollen and the other normal, ultimately on those blocks you are cutting two levels so you can demonstrate what needs to be demonstrated. We decal in the cassette with 5 to 10% formic acid depending upon the time frame. I would use at least 10% formic acid, change frequently if you want to decrease the time in decal. Knees will take 48 hours but ankles and paws take longer. We never cut any of the tissue or skin. Use of a shaker is going to help decrease the decalcification time. If you are getting white spots in your samples then your decal solution is saturated with calcium and is depositing back into the tissue, when you stain them do those white spots appear purple? Regardless of the time in decal the day before we are going to process these specimens we always change the decal one more time, rinse well before processing and process on a cycle that is about 16 hours long, three changes in absolute, and xylene and 4 paraffin baths. We like to decal for about a week ideally, but you can speed that up if necessary. For the rat samples the same applies, if you can get the necropsy techs to cut the toes off at necropsy you will get better fixation. Rat joints take a while to decal, knees in 48 to 72 hours and ankles 5 to 7 days, but you can speed it up with more frequent changes of decal. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jamie E Erickson Sent: Thursday, November 17, 2005 12:46 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Question about De-calcifying mouse paws HI All, Here is my problem, We are a research histology lab which supports groups doing Mouse/ Rat Collagen Induced Arthritis (CIA). The researchers collected the paws and knees for routine processing (Paraffin) and we take it from there. We are trying to give a quick turn around time (2 weeks) on these studies so we are fixing the paws for 24 hours in 10% neutral buffered formalin then switching it to CAL RITE (a formaldehyde/Methanol/ Formic acid mixture from Richard Allan) for decalcification. The paws sit in Cal-Rite for 2 days on a shaker and are then trimmed by cutting the skin and one toe on each side off the paws or trimming 1/3 off the knee Sagittal section (knee and some of the long bones), this helps in decaling the paws and knee quicker. Then the samples are put into fresh De-cal again for 2 more days before washing for 1 hour in tap water and processing. Problem: Knees are great , section great look great but some but not all of the paws are chalky, white deposits in toes and ankles. This only happens to about 1/3 of the samples. I guess they are not de-calcified long enough? Is there another way people are De-caling quickly with better results? Our Pathologist is happy with the slides for the most part but the bad ones are more difficult to section as you can imagine and sometimes the ankle joints don't section well at all. Any Ideas would be greatly appreciated. Thanks _______________________________ Jamie Erickson Sr. Research Associate Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From caron_fournier <@t> yahoo.ca Thu Nov 17 15:23:08 2005 From: caron_fournier <@t> yahoo.ca (caron fournier) Date: Thu Nov 17 15:23:18 2005 Subject: [Histonet] re: stereo microscopes Message-ID: <20051117212308.85259.qmail@web36307.mail.mud.yahoo.com> When I was doing renal biopsies and needed to check tissue amount we just used a basic microscope when we got back to the lab. At ultrasound when the biopsy is being done we only used a 10x eyepiece to check the sample. You would only need a basic stereo microscope like the student grade one's from Leica or Olympus (these are probably the cheapest), even a Motic would probably do because you just have to see the glomeruli which are in most cases easy to see. Remember that most vendors have a lot of margin on the microscopes so try to wheel and deal and if they want to keep followup business then they will accomadate you. Hope this helps --------------------------------- Find your next car at Yahoo! Canada Autos From sturaganiwai <@t> health.gov.fj Thu Nov 17 16:44:06 2005 From: sturaganiwai <@t> health.gov.fj (Simione R. Turaganiwai) Date: Thu Nov 17 16:49:00 2005 Subject: [Histonet] Haematoxylin absorption Message-ID: 1.A haematoxylin and eosin slide was submitted to our Pathologist and she asked as to why did some portion of stained tissue absorbed more haematoxylin and some portion were light meaning absorbed less haematoxylin? 2.How can this be improved? Regard, Simione From al.floyd <@t> juno.com Thu Nov 17 18:47:33 2005 From: al.floyd <@t> juno.com (al.floyd@juno.com) Date: Thu Nov 17 19:04:19 2005 Subject: [Histonet] Haematoxylin absorption Message-ID: <20051117.200228.192.0.al.floyd@juno.com> Hello Simione, Your problem sounds like incomplete deparaffinization. Many modern paraffins have plastisizers that may become difficult to remove if the sections are dried at a high temperature. Patchy staining is a good indicator that some of the embeddment is still present, and interfering with staining. Al Floyd From tahseen <@t> brain.net.pk Thu Nov 17 11:30:09 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Thu Nov 17 19:33:32 2005 Subject: [Histonet] Haematoxylin absorption References: Message-ID: <002d01c5eb9c$907d8d60$972bfea9@m7c0y4> Hello Simione, Your problem sounds like uneven section cutting on microtomy or section was not complete dried after microtome. Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Lahore Pakistan. ----- Original Message ----- From: Simione R. Turaganiwai To: Sent: Friday, November 18, 2005 11:44 AM Subject: [Histonet] Haematoxylin absorption 1.A haematoxylin and eosin slide was submitted to our Pathologist and she asked as to why did some portion of stained tissue absorbed more haematoxylin and some portion were light meaning absorbed less haematoxylin? 2.How can this be improved? Regard, Simione _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sasa <@t> jovanovic.co.za Thu Nov 17 23:07:52 2005 From: sasa <@t> jovanovic.co.za (Sasa Jovanovic) Date: Thu Nov 17 23:08:08 2005 Subject: [Histonet] FW: mimicry Message-ID: <000201c5ebfe$073b1420$640aa8c0@SinisaHome> -----Original Message----- From: Sasa Jovanovic [mailto:sasa@jovanovic.co.za] Sent: 18 November 2005 07:05 AM To: 'hstonet@lists.utsouthwestern.edu' Subject: FW: mimicry -----Original Message----- From: Sasa Jovanovic [mailto:sasa@jovanovic.co.za] Sent: 17 November 2005 11:16 AM To: 'Johnson, Teri' Subject: mimicry Dear Teri Johnson, I am referring to your previous HIstonet correspondence of Teri Johnson where they mentioned the following :....we also tried using an anti-IgG2a anti mouse/HRP labelled secondary antibody, and in the negative control there was no staining..... I have red few of the Alon R papers regarding the mimicry between streptavidin and fibronectin since I presume I might have the same problem In my project I am trying to determine presence/absence of alpha4beta 1 integrin in the rat uterus by using immunocytochemistry on the frozen tissue....At the moment I have DAKO ARK kit and applying the following procedure: Cut 6um thick sections (use positive charged slides and silane pre-treated ) dry sections overnight at room temperature fix them the next day in cold acetone for 10 minutes proceed with staining: Biotinylate primary antibody (monoclonal -mouse anti human CD49d (integrin alpha 4 beta 1)P4G9. Step 1: Mix diluent, concentrated primary Ab (positive or negative) and biotinylation reagent, incubate for 15 min Step 2: add blocking reagent incubate for 5 min Apply peroxidase block for 15 min Rinse Apply prepared biotinylated primary Ab (positive or negative), incubate for 1 hour Rinse Apply streptavidin HRP incubate for 15 min Rinse Apply prepared DAB incubate for 5 min Rinse Counterstain Mount coverslips I am using Tris-HCL pH9 (with added Tween 20) as a washing buffer I am using human tonsil as a positive control where I did not see any specific immunolocalization on the sections where I had omit primary Ab. But, I am experiencing "staining" of my negative control, actually positive and negative uterus look the same.....they "stain" on the same place. Could this be the sterptavidin mimicry and would you have any suggestions how to incorporate anti IgG2a antibody in my kit or maybe do you have any other solution to my problem? You advice is greatly appreciated!!! Thank you in advance Sasha From cytologer <@t> gmail.com Fri Nov 18 00:26:13 2005 From: cytologer <@t> gmail.com (Ul Soo Choi) Date: Fri Nov 18 00:26:44 2005 Subject: [Histonet] preparation of chromic acid solution for CAS.... Message-ID: <001101c5ec08$fdc05120$593a2e93@ksvcpuo8c26n3k> Hi, histonetters! I am interested in CAS chromic acid Schiff stain, instead of PAS. The following article prompted me to change it with CAS. http://www.sigmaaldrich.com/Area_of_Interest/Laboratory_Essentials/Hematology_and_Histology/Key_Resources/Article_Highlights/Staining_for_Fungi.html But, I am not aware of the protocol how to prepare chromic acid solution. Also I am still just wondering if CAS produces more distinct and good contrast than PAS in the tissues, and cytology samples. I usually use cytological specimens. I would like to hear your experiences on CAS. Let me hear..... Thank you always. Regards, Ul Soo Choi DVM., PhD. Dept of clinical pathology, CVM, SNU Seoul, Korea email. cytologer@gmail.com tel 82-02-880-8688 fax 82-02-880-8662 From gillian.2.brown <@t> gsk.com Fri Nov 18 02:36:22 2005 From: gillian.2.brown <@t> gsk.com (gillian.2.brown@gsk.com) Date: Fri Nov 18 02:36:41 2005 Subject: [Histonet] H&E Staining of Frozen Sections In-Reply-To: <20051117182931.35394.qmail@web61214.mail.yahoo.com> Message-ID: Any bets on the next question being 'what kind of haematoxylin'? Gill Brown T GlaxoSmithKline Medicines Research Centre, STEVENAGE, Hertfordshire. UK SG1 2NY "Rene J Buesa" Sent by: histonet-bounces@lists.utsouthwestern.edu 17-Nov-2005 18:29 To "Rebecca Jo" , "Histonet Listserv" cc Subject Re: [Histonet] H&E Staining of Frozen Sections We used to follow the following protocol: 1- fix the frozen section in acetone X 10 secs. 2- distilled water x 10 secs 3- hematoxylin x 30 secs 4- tap water 5- quick dip in acid alcohol 6- eosin 5-10 secs. 7- 95% ethanol > 100% ethanol x2 > xylene X2 > coverslip. The pathologists had the slide ready in about 2-3 minutes. Rene J. Rebecca Jo wrote: Hi Histoneters- I was wondering if someone could give me a protocol for H&E staining on frozen sections. I assume I can just skip over the deparaffinizing steps and go straight to the staining, but I wanted to know how long I should keep the sections in hematoxylin. Just enough to stain the sections? Any help would be greatly appreciated. -Cheers -- Rebecca E. Jo UNC-School of Medicine Department of Cell and Developmental Biology (919)843-9648 CB# 7090 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gillian.2.brown <@t> gsk.com Fri Nov 18 02:54:13 2005 From: gillian.2.brown <@t> gsk.com (gillian.2.brown@gsk.com) Date: Fri Nov 18 02:54:37 2005 Subject: [Histonet] Question about De-calcifying mouse paws In-Reply-To: Message-ID: Jamie, presumably you are facilitating fixative penetration by opening the skin? You could try trimming the fixed sample to size by using a small bone saw, we got one from this UK site, if you navigate you will come to some great images of nasal turbinates in which the soft tissue is very well preserved. We have used the same saw for LS sections of mouse femurs. www.materials-science-nw.co.uk I also use a decalcifying agent which is formic acid based (plus ingredients XXX) which does my whole fixed mouse heads or bone sawed head slices of rat and guinea-pig in 24hrs. I also use it as a surface decal of blocks if the teeth remain a bit solid. All of the immuno we have done to date has worked as well as any tissues we have decalcified in EDTA for much longer periods. Price 139.90 Euros Cat #1414 1 gallon Quartett Immunodiagnostika - Biotechnologie GMBH Schichauweg 16 - Berlin - Germany 12307 Phone: 49(030)765-925-0 Fax:49 (030) 765-925-55 Website: http://www.quartett.com email: quartett@t-online.de Contact Name J?rgen Gorczyza Regards Gill Brown GlaxoSmithKline Medicines Research Centre, STEVENAGE, Hertfordshire. UK "Jamie E Erickson" Sent by: histonet-bounces@lists.utsouthwestern.edu 17-Nov-2005 19:46 To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Question about De-calcifying mouse paws HI All, Here is my problem, We are a research histology lab which supports groups doing Mouse/ Rat Collagen Induced Arthritis (CIA). The researchers collected the paws and knees for routine processing (Paraffin) and we take it from there. We are trying to give a quick turn around time (2 weeks) on these studies so we are fixing the paws for 24 hours in 10% neutral buffered formalin then switching it to CAL RITE (a formaldehyde/Methanol/ Formic acid mixture from Richard Allan) for decalcification. The paws sit in Cal-Rite for 2 days on a shaker and are then trimmed by cutting the skin and one toe on each side off the paws or trimming 1/3 off the knee Sagittal section (knee and some of the long bones), this helps in decaling the paws and knee quicker. Then the samples are put into fresh De-cal again for 2 more days before washing for 1 hour in tap water and processing. Problem: Knees are great , section great look great but some but not all of the paws are chalky, white deposits in toes and ankles. This only happens to about 1/3 of the samples. I guess they are not de-calcified long enough? Is there another way people are De-caling quickly with better results? Our Pathologist is happy with the slides for the most part but the bad ones are more difficult to section as you can imagine and sometimes the ankle joints don't section well at all. Any Ideas would be greatly appreciated. Thanks _______________________________ Jamie Erickson Sr. Research Associate Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AFeatherstone <@t> KaleidaHealth.Org Fri Nov 18 05:22:11 2005 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Nov 18 05:20:34 2005 Subject: [Histonet] RE: Histonet Digest, Vol 24, Issue 25 Message-ID: <139141F8BAF4A642A945ECC528511AF0012A7ABD@kalmb02.kaleidahealth.org> We use recycled xylene in processing and staining. We have sent samples of our recycled xylene to be tested for purity and it is actually better than new. Annette Featherstone -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Thursday, November 17, 2005 13:02 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 24, Issue 25 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. QIHC (Elizabeth Chlipala) 2. RE: recycled xylenes (Bauer, Karen) 3. RE: QIHC (James Watson) 4. RE: RE: QIHC change (Smith, Jeffery D. (HSC)) 5. cryoprotection (Steven Coakley) 6. am I snap-freezing correctly? (Jacobs,Jennifer (stu)) 7. RE: am I snap-freezing correctly? (Favara, Cynthia (NIH/NIAID)) 8. Re: Luxol fast blue/cresyl violet stains (Rene J Buesa) 9. Re: recycled xylenes (Rene J Buesa) 10. Re: am I snap-freezing correctly? (John A. Kiernan) 11. RE: QIHC change (Johnson, Teri) 12. Help- triphenyltetrazolium chloride instructions for staining needed (HCS) 13. Re: Histonet Digest, Vol 24, Issue 23 (JUDIEWHIT@aol.com) 14. biohazard waste removal companies (Ron Martin) 15. RE: aqueous coverslipping medium (C.M. van der Loos) 16. hard tissue distortion (louise renton) 17. CD31 staining histiocytes??? (Histology SLU) 18. CD31 Santa Cruz update (Angela McNabola) 19. RE: E: beta-2- microglobulin (Edwards, R.E.) 20. fixed jaw clamps/ large size specimens (Kinsley, David) 21. Dissecting Microscopes (Bauer, Karen) 22. Re: fixed jaw clamps/ large size specimens (Rene J Buesa) ---------------------------------------------------------------------- Message: 1 Date: Wed, 16 Nov 2005 13:08:53 -0700 From: "Elizabeth Chlipala" Subject: [Histonet] QIHC To: "'Histonet'" Message-ID: <000001c5eae9$91e76df0$a7d48a80@AMY> Content-Type: text/plain; charset="us-ascii" I'm going to have to chime in here. One thing I need to say upfront is that I served on the BOR when they were developing the QIHC. The BOR for the histotech exam committee consists of histology technicians, histotechnologists, MD pathologists and veterinary pathologists, plus the ASCP staff, so it encompassed a wide variety of individuals and skill sets, both in research and clinical. These are the individuals who created the exam and the requirements for the exam. I went to the BOR guidelines online for the QIHC and as far as I can see that have not changed. Immunophenotyping has always been a pre-requisite for sitting for this qualification. This is what BOR requires: Immunophenotyping in at least one of the following applications * immunodeficiencies * immunoproliferative disorders (neoplastic and non-neoplastic disorders) * transplantation biopsies * other immunophenotyping applications please specify: ______________________ After looking online for the definition of immunophenotyping it is the following: " IMMUNOPHENOTYPING refers to the technique of identifying molecules that are associated with lymphoid cells and help to characterize them". Such as cell surface markers, which I would characterize broadly as CD markers. I work in research and sat for the QIHC last year. In research we perform many Immunohistochemical stains utilizing CD markers to determine what specific cell types are present in certain disease states. When I filled out my application last year I selected the forth choice which is "immunophenotyping applications other" and listed what types of CD markers I had worked with and the type of projects and applications associated with them. I was able to sit for the exam and so was my employee. In my opinion many histotechs in research have the necessary experience since they are performing Immunohistochemical staining utilizing a wide variety of CD markers. I just don't see how in working in research you would not have the pre-requisite to sit for this exam. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 ------------------------------ Message: 2 Date: Wed, 16 Nov 2005 14:14:06 -0600 From: "Bauer, Karen" Subject: RE: [Histonet] recycled xylenes To: "Molinari, Betsy" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi Betsy, We use are recycled xylene for both processing and staining and have had no problems. Karen L. Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Molinari, Betsy Sent: Wed 11/16/2005 2:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] recycled xylenes Hi, I am interested in knowing if you have preferences for your recycled xylenes. Do you use it just for staining, or processing alone. Or use it for both? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************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. ------------------------------ Message: 3 Date: Wed, 16 Nov 2005 12:20:09 -0800 From: "James Watson" Subject: RE: [Histonet] QIHC To: "Elizabeth Chlipala" , Message-ID: Content-Type: text/plain; charset="us-ascii" Elizabeth, I have been trying to find out what the other category entails without success, this is why I became involved with these discussions to see if someone on the histo net knew. But I have finally gotten someone at ASCP to talk to as listed on my last posting. She was not very clear as to what qualifies and if work on animal tissue is acceptable. So she wants me to send her a list of what I do that may qualify as immunophenotyping to determine if I qualify. I really appreciate your response because it gives me some guidelines as what I can list as other immunophenotyping applications. Thank you very much. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Wednesday, November 16, 2005 12:09 PM To: 'Histonet' Subject: [Histonet] QIHC I'm going to have to chime in here. One thing I need to say upfront is that I served on the BOR when they were developing the QIHC. The BOR for the histotech exam committee consists of histology technicians, histotechnologists, MD pathologists and veterinary pathologists, plus the ASCP staff, so it encompassed a wide variety of individuals and skill sets, both in research and clinical. These are the individuals who created the exam and the requirements for the exam. I went to the BOR guidelines online for the QIHC and as far as I can see that have not changed. Immunophenotyping has always been a pre-requisite for sitting for this qualification. This is what BOR requires: Immunophenotyping in at least one of the following applications * immunodeficiencies * immunoproliferative disorders (neoplastic and non-neoplastic disorders) * transplantation biopsies * other immunophenotyping applications please specify: ______________________ After looking online for the definition of immunophenotyping it is the following: " IMMUNOPHENOTYPING refers to the technique of identifying molecules that are associated with lymphoid cells and help to characterize them". Such as cell surface markers, which I would characterize broadly as CD markers. I work in research and sat for the QIHC last year. In research we perform many Immunohistochemical stains utilizing CD markers to determine what specific cell types are present in certain disease states. When I filled out my application last year I selected the forth choice which is "immunophenotyping applications other" and listed what types of CD markers I had worked with and the type of projects and applications associated with them. I was able to sit for the exam and so was my employee. In my opinion many histotechs in research have the necessary experience since they are performing Immunohistochemical staining utilizing a wide variety of CD markers. I just don't see how in working in research you would not have the pre-requisite to sit for this exam. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Wed, 16 Nov 2005 14:18:09 -0600 From: "Smith, Jeffery D. \(HSC\)" Subject: RE: [Histonet] RE: QIHC change To: "Morken, Tim - Labvision" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Tim, I would like to know what your certification is and what qualifies you to decide what the focus of the ASCP certifications entale? Who put you in charge of deciding what the sole purpose of the ASCP is? ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Morken, Tim - Labvision Sent: Wed 11/16/2005 1:04 PM To: 'Pamela Marcum'; Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: QIHC change Pam, I'm not discounting the knowledge researchers have. I just don't think the ASCP certification route fits research. I pointed out that the ASCP, by definition, is intended soley for clinical diagnostic labs. It is not the ASCP's fault that the research institutions are mis-using the ASCP certification for their own in-house qualification standards. Certainly it would help researchers if there was a more general certification, but is it the ASCP's place to do that? I don't know. The ASCP is an organization of Clinical Pathology based in human diagnostics. Because of that the HT, HTL and QIHC are all based on human clinical pathology diagnostic work. I'm not sure the ASCP would be interested in setting up a whole new testing procedure for research. What if a person only does work in ferrets, or fish, or snakes? Who looks at tests based on that work? There are many similarities in procedures, and I'm confident a generalized test could be designed. It simply a question of whether the ASCP is the organization to do that. I guess the research techs have to explore this with ASCP. But I don' think you should blame ASCP for not accomodating research institutions when that is not what they are concerned with. Remember, there are no regulatory requirements at all covering the work of research lab techs but there are regulations covering who can do certain work in the diagnostic lab. ASCP, again by definition, is only concerned with the diagnostic side. It has nothing to do with whether one is "better" than another. There is no "better" in this work, only differences in the work done. I also suggest that you lobby your institution and show them how the ASCP certification may not be fully applicable to what you do. Tim Morken -----Original Message----- From: Pamela Marcum [mailto:pmarcum@vet.upenn.edu] Sent: Wednesday, November 16, 2005 9:56 AM To: Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: QIHC change Hi Tim and Jamie, I know Jamie and have for many years and you are right anyone would be happy to hire him for his experience in clinical or his current research position. However, it is also true that now research positions are often asking for at least an HT or even HTL (ASCP) to fill position with histology as the main focus. Yet we are given a set of criteria for tissue that often excludes animal research applicants from completing the practical easily. I took my HT many years ago and I was told that even in a research position (and I had a BS at the time) it would improve my salary and increase me to higher level in the university if I got my HT. I was told not to use animal tissue (1976) as no one reading the exam could properly read them. Now we have veterinary person there and tissue requirements can still eliminate some people or make it almost impossible to complete the practical with out help in procurement. Why should that happen to some one attempting to improve their position within the histology community? My real problem with what you said about the QIHC is that I would also like to take it and can not qualify either. Yet those of us in research are often finding the very antibodies and test methods companies and diagnostics later fight to get or learn. We are exempt in your mind and ASCP's even though research is what you depend on often for advances. I have never and will never understand this logic and exempt status for those of us who chose not to be clinical. We are still often required to have or get ASCP status as a way to advance and prove we know our field. ASCP needs to get up to date on the fields it is registering or make new categories for those of still contribute to clinical advances every year. Sorry if sounds like I am picking on you Tim. I just don't see how we are required to be registered on one hand for acceptance (even NSH likes to see it) and discounted on the other. Pam Marcum UPENN Vet School New Bolton Center Kennett Square, PA 19384 610-925-6278 At 12:07 PM 11/16/2005, Morken, Tim - Labvision wrote: >Jamie, It seems from what you say that you are working in a research >lab. Is that correct? My understanding about the ASCP certification is >that it is aimed at providing a modicum of proof that a person is >qualified to work in a medical diagnostic lab. Research labs are not >considered diagnostic labs. As you imply, a person in a research lab >will often work on only a limited sample set. Therefore, it is >meaningless to apply the the ASCP standard to research people. > > If you are planning to move into the diagnostic field, then I'll bet >you could easily find a job in a diagnostic lab, get the experience, >and qualify to take the test. It may be that some diagnostic labs have >a suggested requirement to be ASCP certified as a QIHC, but the vast >majority would be happy to find someone with the experience you >outline, even if they had not previously worked in a diagnositc lab. > >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: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James >Watson >Sent: Wednesday, November 16, 2005 7:28 AM >To: novanity@nc.rr.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: QIHC change > > >This is my point. With the requirements listed below someone with 25 >years of experience doing immuno (single, double, triple antibody >staining, making own antibodies, and in situ Hybridization: all with >and without using kits, all with and without using an automated >stainer) is not qualified for this certification if they work in a >research facility where immunophenotyping is not done. There is no >system of doing it on your own to prove that you have the capability to >do immunophenotyping in order to fullfil this requirement. I guess it >is time to start harrassing ASCP about the unfairness of this system. > > >From almost always sunny San Diego >Jamie > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of >novanity@nc.rr.com > Sent: Wed 11/16/2005 5:55 AM > To: histonet@lists.utsouthwestern.edu > Cc: > Subject: [Histonet] RE: QIHC change > > > > James, to qualify for the qualification you take the 50 >question test > and submit an employer reference form + of course satisfy one of the > three routes. There is no practical to submit anymore. I >think that is > what you are asking. It seems to me that it wouldn't matter about > specificity antigens/markers or what diseases or human cells. >There is > no requirement other than what is requested on the employer >reference > form which you can't see the details until you order and >receive your > packet. Is it possible for anyone to post a copy of the employer > reference form. From the ASCP website this is what it says " > > Qualification in Immunohistochemistry > Experience requirements > > Applicants must have experience in the following areas > > * Immunohistochemical and Immunofluorescence Preparation > All of the following should have been performed by the >applicant > o staining technique > o selection of proper control material > o titration of immunologic reagents > > * Immunophenotyping > in at least one of the following applications > o immunodeficiencies > o immunoproliferative disorders (neoplastic and >non-neoplastic > disorders) > o transplantation biopsies > o other immunophenotyping applications > please specify: ______________________ > > * Quality Assurance > The applicant should have participated in Quality Assurance > related to all of the following > o specimen fixation, processing, microtomy > o reagent selection, preparation, storage, disposal > o method selection, validation, documentation > o quality control > o safety > > " This is the experience which I am assuming is only >documented for the > ASCP through the employer reference form, hence if you only do >A and C > and not B you can't qualify unless your employer is dishonest on the > form. Because even if you crosstrain into what I assume is flow > cytometry but don't actually work it day to day as part of >your job you > do not qualify because you have not had experience doing it for a > minimun of 12 months. As for research, same thing if you do >all of it > every day then your good to go. If not it is a grey or is it >gray area > that I'm looking more information/details on. In the past you > qualified your work with different immuno stains as a practical , I > don't remember there being a flow requirement. Maybe I'm wrong but > anyone have this info I'm looking for. > > G Hurlburt HT(ASCP) > sunny and warm NC > > _______________________________________________ > 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 ------------------------------ Message: 5 Date: Wed, 16 Nov 2005 12:26:44 -0800 (PST) From: Steven Coakley Subject: [Histonet] cryoprotection To: Histonet@lists.utsouthwestern.edu Message-ID: <20051116202644.45169.qmail@web90201.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 How are techs freezing their fixed, 30% infiltrated tissue. Right now we freeze the tissue (liver) using OCT placed in a small freezing cup and float on liquid nitrogen. They section great but the cellular detail is only fair. Does anyone have really good result cryoprotecting tissue. Steve --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 6 Date: Wed, 16 Nov 2005 15:31:07 -0500 From: "Jacobs,Jennifer \(stu\)" Subject: [Histonet] am I snap-freezing correctly? To: Message-ID: <9F866D418736DD408F43059FF7FC697203B34973@itnsa.uchc.edu> Content-Type: text/plain; charset="Windows-1252" Hi, I am snap freezing brain tissue for sectioning (8uM) on cryostat (-25C). When I snap freeze, I put the brain hemisphere on a piece of aluminum foil (boat) and let it float on the LN2. Most times, some LN2 comes in direct contact with the brain tissue as the boat is not leak-free. Is this bad? Should I make sure the LN2 never comes in contact with the brain? The problem I've been noticing is that, right off the cryostat, my tissue looks "hole-y". This just gets worse as I proceed with the IHC stain (CD-31 Ab to see vessels) and dehydration + xylene steps. By the xylene steps the holes are so stretched out that I can't even recognize vessels in the tissue. I am not sure whether the holes arise from the brain extraction procedure (ie: snap freezing) or from the sectioning. But the person who sections has been doing it for years and has great experience with the cryostat. Any ideas/suggestions would be great! Thanks a lot! Jenn UConn Health Center Dept Pharmacology Farmington, CT ------------------------------ Message: 7 Date: Wed, 16 Nov 2005 15:47:08 -0500 From: "Favara, Cynthia \(NIH/NIAID\)" Subject: RE: [Histonet] am I snap-freezing correctly? To: "Jacobs,Jennifer \(stu\)" , Message-ID: Content-Type: text/plain; charset="us-ascii" Look in the archives for the protocol for freezing over isopentane by Gayle Callis. This is what I find this better than LN for morphology - ditto for the cryoprotected specimens. c 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: Jacobs,Jennifer (stu) [mailto:JJacobs@student.uchc.edu] Sent: Wednesday, November 16, 2005 1:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] am I snap-freezing correctly? Hi, I am snap freezing brain tissue for sectioning (8uM) on cryostat (-25C). When I snap freeze, I put the brain hemisphere on a piece of aluminum foil (boat) and let it float on the LN2. Most times, some LN2 comes in direct contact with the brain tissue as the boat is not leak-free. Is this bad? Should I make sure the LN2 never comes in contact with the brain? The problem I've been noticing is that, right off the cryostat, my tissue looks "hole-y". This just gets worse as I proceed with the IHC stain (CD-31 Ab to see vessels) and dehydration + xylene steps. By the xylene steps the holes are so stretched out that I can't even recognize vessels in the tissue. I am not sure whether the holes arise from the brain extraction procedure (ie: snap freezing) or from the sectioning. But the person who sections has been doing it for years and has great experience with the cryostat. Any ideas/suggestions would be great! Thanks a lot! Jenn UConn Health Center Dept Pharmacology Farmington, CT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Wed, 16 Nov 2005 12:56:17 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Luxol fast blue/cresyl violet stains To: judi.ford@jax.org, histonet@lists.utsouthwestern.edu Message-ID: <20051116205617.42144.qmail@web61218.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Judi: When you say "5 slides from the same animal" do you mean slides from different blocks from the same animal or slides from the same block. If the slides are from different blocks then the problem could reside in the blocks and how they were treated. If all the slides are from the same block then the difference could reside in the configuration the slides were placed in the staining jar (although this explanation could be far fetched). I don't think that the differences could be due to the differentiation step because you say that after differentiation the slides are placed in distilled water, had they been "waiting" for the result of one slide to stop the differentiation on the others that could be the cause. Now, if the 37Celsius incubation is done in an oven and the oven does not have a good air circulation it could be that one area in the staining jar could have a difference in temperature at least during some time, although that could be compensated during the long incubation period. So I really don't know what to atribute the problem to. Not exactly these, but the delay itself, prompted us to eliminate this protocol and go to one using microwaves (MW) as follows: 1- dewax and take sections to absolute ethanol (EthOL) 2-place slides in plastic Coplin jar and add 95% EThOL 3- heat slides in the 95% EthOL for 15 secs. at 0.95 kcal ("Level 5" of our 480 W MW oven which was equivalent to 266 Watts) 4- dump the heated 95% EthOL (used only to heat the slides) and substitute it with 40 mL of Luxol Fast Blue (LFB). Heat at the same level (266 Watts) for 20 secs (=1.27 kcal). 5- transfer the Coplin jar with the slides to a water bath at 60Celsius during 5 minutes. 6- Wash slides in distilled water. 7- Differentiate the slides , one at a time, with 0.5%aq. lithium carbonate solution until no more blue color runs off the slides (not necessary under the microscope) 8- Transfer the slides to distilled water. 9- Dump the dist. water and add the Cresyl Echt Violet (CEV) and heat in the MW at full power (480 Watts) for 20 secs.(=2.3 kcal). 10-Transfer the slides to the water bath at 60Celsius for 5 minutes. 11- Dehydrate QUICKLY, clear and mount as usual. With this procedure we did not have any problems and it could be completed in about 30 minutes. Perhaps you could care to try this protocol to find out if the problem persists. This is how we did it in our lab. Hope this will help! Rene J. judi.ford@jax.org wrote: Hi, I have a question concerning the lfb/cv stain. This is the situation we have in our lab. The tissue we are working on is Bouin's fixed mouse brain and spinal cord (cord is left in the vertebrae). The spinal cord may be left in Bouin's for an extended period for decalcification (two-three weeks). The trimmed tissue is then sent to our lab where we rinse it for a day before processing on our VIP processors. The next day we embed the tissue and then it is cut by one of our technicians and stained. This is our staining technique: deparaffinize to 95% alcohol and place in alcoholic luxol fast blue overnight at 37 degrees. The next morning it is rinsed twice in 95% alcohol and twice in distilled water. The slides are separated into racks containing either brain or spinal cord for differentiation. Slides are dipped in 70% alcohol for 20 seconds then lithium carbonate for 20 seconds. This is followed by two rinses in distilled water. Then checked microscopically before determining if differentiation is complete or not. If not, then the slides go another round through 70% and lithium for a few more seconds. Once this part is done the slides are put into a warmed cresyl violet solution (we add 10% glacial acetic acid, 10 drops for every 100mls of stain) for 4-6 minutes then rinsed in 95% alcohol three times. Finally they are dehydrated, cleared and coverslipped. The problem comes when we have 5 slides from the same animal, all processsed and stained together, where in one slide the cresyl violet works but in the other 4 it doesnt' work. Every slide is treated exactly the same, yet we have this difference. Any ideas on what may be happening? We've discussed decalcification and possible lithium effects on the cresyl violet. We're at a loss and our pathologist is very determined to discover the solution for this problem. Thank you advance for any ideas you may send our way..........We greatly appreciate your help! Judi Ford HIstotechnologist HIstopathology & Microscopy The Jackson Laboratory 600 Main St. Bar Harbor, Me 04609 207-288-6193 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 9 Date: Wed, 16 Nov 2005 13:01:46 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] recycled xylenes To: "Molinari, Betsy" , histonet@lists.utsouthwestern.edu Message-ID: <20051116210146.28437.qmail@web61224.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Betsy: We used to recycle xylene and used for everything. Since the recovery is not 100% when we had to buy new we preferred to use it in the staining and coverslippin instruments. Rene J. "Molinari, Betsy" wrote: Hi, I am interested in knowing if you have preferences for your recycled xylenes. Do you use it just for staining, or processing alone. Or use it for both? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 10 Date: Wed, 16 Nov 2005 16:07:06 -0500 From: "John A. Kiernan" Subject: Re: [Histonet] am I snap-freezing correctly? To: "Jacobs,Jennifer (stu)" Cc: histonet@lists.utsouthwestern.edu Message-ID: <437B9F7A.7E99AF0F@uwo.ca> Content-Type: text/plain; charset=us-ascii The more holey than righteous appearance that you describe is typical of slowly frozen tissue - especially if it hasn't been cryoprotected. When liquid nitrogen comes into contact with an object it boils, and the resulting layer of nitrogen gas has an insulating effect that slows down the cooling process. For that reason it is preferable to freeze specimens quickly by immersing in a liquid that has been pre-cooled to a low temperature. The one most often used is isopentane. Put some in a small metal can, and stand the can in liquid nitrogen. Stir the isopentane with a lollipop stick. When it starts to thicken the temperature is approaching its freezing point (-160C) and the specimen (with or without aluminium foil) can be dropped in. Another method is to bring a fairly massive block of metal to liquid nitrogen temperature (-196C) and slap the specimen onto the surface of the block. You will probably get plenty of replies. There has been much Histonetting about quick freezing methods over the years! -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Jacobs,Jennifer (stu)" wrote: > > Hi, > > I am snap freezing brain tissue for sectioning (8uM) on cryostat (-25C). > When I snap freeze, I put the brain hemisphere on a piece of aluminum foil (boat) and let it float on the LN2. Most times, some LN2 comes in direct contact with the brain tissue as the boat is not leak-free. Is this bad? Should I make sure the LN2 never comes in contact with the brain? > > The problem I've been noticing is that, right off the cryostat, my tissue looks "hole-y". This just gets worse as I proceed with the IHC stain (CD-31 Ab to see vessels) and dehydration + xylene steps. By the xylene steps the holes are so stretched out that I can't even recognize vessels in the tissue. > > I am not sure whether the holes arise from the brain extraction procedure (ie: snap freezing) or from the sectioning. But the person who sections has been doing it for years and has great experience with the cryostat. > > Any ideas/suggestions would be great! Thanks a lot! > > Jenn > > UConn Health Center > Dept Pharmacology > Farmington, CT > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Wed, 16 Nov 2005 15:36:26 -0600 From: "Johnson, Teri" Subject: [Histonet] RE: QIHC change To: Message-ID: Content-Type: text/plain; charset="us-ascii" James, et al, Thank you for bringing lively discussion in an area I believe needs it. I have worked for many years in clinical histology labs and now work in the research environment. I have to admit, once I heard they had moved this exam to a 50-question test, I signed up. Previously, I knew it would be nearly impossible for me to do the practical exam with human tissue samples since we don't have any here. I took the exam last week and have to say that while most of the questions were based on general IHC knowledge, a couple of questions regarded clinical samples (cytology specimens) not usually done in research, or were geared toward estrogen receptor-type immunostaining. Most of my issues with the exam related to the wording of the question, or the choices given (I didn't always agree with them, and one was ambiguous). I'm glad I now have a contact to whom I can give my opinion of the exam, thanks James! My conclusion was, the exam can be taken successfully by research techs with a strong background in IHC, and the clinical-only questions they will miss will still give them enough margin to pass. (Provided, of course, they meet the requirements.) I agree as well that the immunophenotyping requirement be phased out or reworded to show knowledge of typical staining patterns for typical markers and not just the leukemia/lymphoma ones. Usually only the large hospital or private IHC labs do these types of panels, so even in the clinical arena this type of experience isn't easy to get. The IHC techs aren' t the ones interpreting these results, nor are they ordering the panels. At best they run patient samples with known positive controls (mostly tonsil), and order and work up new antibodies to run in these panels--which are then evaluated by a pathologist for applicability. How is that different from any other marker run in an IHC lab? Regarding the need for ASCP-certified techs in the research environment, I agree with Tim that it should not necessarily be a requirement. I know that an ASCP-certified tech has general knowledge of histotechnology and has demonstrated skill with a microtome and staining techiques, and has the certification to prove it. But that alone does not make them any more qualified to work in the lab than someone with a BS or MS and histotechnology experience but no certification. Please don't think I'm making the case for random hiring of non-certified techs. I believe firmly in certifications for personal and professional growth. I just don't think the research environment should base their job requirements on it. My employment ad usually indicates the person we are looking for be ASCP certified, or certification eligible. In the absence of the certification, meeting minimum educational requirements is usually acceptable. Finally, I have not paid my dues to ASCP for a very long time and have no plans to start doing so any time soon. I will eagerly hand over my money to the NSH because they offer workshops and a publication that are geared towards the profession. Even the Advance magazine runs more articles on Histology than the ASCP journal ever did. Perhaps that has changed, but somehow I doubt it. I too would be steamed if I had paid all this time and got no value for my $$$s. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 ------------------------------ Message: 12 Date: Wed, 16 Nov 2005 17:50:15 -0800 From: "HCS" Subject: [Histonet] Help- triphenyltetrazolium chloride instructions for staining needed To: "Histonet" Message-ID: <000601c5eb19$4d4b7df0$6601a8c0@hp> Content-Type: text/plain; charset="iso-8859-1" Hi, I am still looking for a reference or procedure for doing the TTC stain (triphenyltetrazolium chloride). If you might have a procedure I would be able to use that would be very helpful. Thanks LeRoy Brown HCS . ------------------------------ Message: 13 Date: Wed, 16 Nov 2005 21:51:35 EST From: JUDIEWHIT@aol.com Subject: [Histonet] Re: Histonet Digest, Vol 24, Issue 23 To: histonet@lists.utsouthwestern.edu Message-ID: <217.e6894da.30ad4a37@aol.com> Content-Type: text/plain; charset="US-ASCII" Hi Jill, Looks like you are getting ready to open your new lab. Congrats girlfriend I knew you could do it! I know some of the techs in Phoenix if you need references let me know. I am getting ready to come out of retirement and go traveling...looks like I will be working in a lab in new Mexico for 13 weeks soon, I think the travel thing will be fun and I can move on to another Histo job in different parts of the country. The very best to you my friend... Judie W. ------------------------------ Message: 14 Date: Wed, 16 Nov 2005 23:12:28 -0500 From: "Ron Martin" Subject: [Histonet] biohazard waste removal companies To: Message-ID: <001001c5eb2d$2100bf80$27233418@Pathrm35> Content-Type: text/plain; charset="iso-8859-1" Does any one know of any companies that service southeast Florida for biohazard waste and chemical waste removal? I am currently using Environmental Enterprises out of Orlando but lately this company has been unable to service our needs. Thanks in advance. Ron Martin ------------------------------ Message: 15 Date: Thu, 17 Nov 2005 09:05:55 +0100 From: "C.M. van der Loos" Subject: [Histonet] RE: aqueous coverslipping medium To: histonet@lists.utsouthwestern.edu Cc: PMonfils@Lifespan.org Message-ID: <1b34f811b341d2.1b341d21b34f81@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Paul, The media you call the "liquid coverslip" is in fact a kind of plastic polymer that can be mounted up organically after polymerisation. I experienced that without this additional organic mounting the plastic polymer will fall off after some years. If this happens you can remove the plastic completely by soaking the slide in luke warm water. Then "liquid-coverslip" again. Although most of those products state in their data sheet that it should be used without a coverslip, it also works fine WITH a coverslip. The only thing you should not do is harden at 60-70C as you normally do (it causes air bubbles!). Just leave the slides at room temp. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Wed, 16 Nov 2005 12:40:06 -0500 From: "Monfils, Paul" Subject: [Histonet] aqueous coverslipping medium To: "'histonet@lists.utsouthwestern.edu'" What brand do you prefer for aqueous mounting medium to be used with a coverslip? I have several good hydrocarbon-based coverslipping media, and also some good aqueous media to be used without coverslips (so-called liquid coverslip), but I don't have an aqueous coverslipping medium I am completely satisfied with. Applications would include immunofluorescence and a few enzyme stains, as well as lipid stains and other techniques that are not compatible with hydrocarbon-based media. Are there any such media that harden after application, to produce a permanent slide? ------------------------------ Message: 16 Date: Thu, 17 Nov 2005 11:31:40 +0200 From: louise renton Subject: [Histonet] hard tissue distortion To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Helo all, I've been asked this question by a postgrad and I wasn't able to answer it to my satisfaction. The question was - how much distortion occurs in tissue (in this case teeth) that have been fixed in 70% alcohol, processed by hand and embedded in MMA?, Sections were cut at 6um, stained free floating and mounted in Entellan on glass slides. Any thoughts/references? I would be most grateful for anything The post grad is doing histometry on healing furcation defects, and is wondering if there are any variables that he needs to mentionor take into consideration. Best regards -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....onwards through the fog!" ------------------------------ Message: 17 Date: Thu, 17 Nov 2005 06:10:19 -0800 (PST) From: Histology SLU Subject: [Histonet] CD31 staining histiocytes??? To: "histonet@lists.utsouthwestern.edu" Message-ID: <20051117141019.29359.qmail@web51003.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all: Is anyone out there using Dako CD31 and getting histiocyte staining in human skin??? Our dermpath is seeing this and is questioning it. We are using it at 1:100 with the biocare pressure cooker with Biocare Diva decloaker solution. Our detection system is Envision +. Any thoughts??? Susan Histopathology Supervisor St. Louis University Medical School --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ Message: 18 Date: Thu, 17 Nov 2005 09:52:03 -0500 From: Angela McNabola Subject: [Histonet] CD31 Santa Cruz update To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hello all, I received a new lot of CD31 yesterday, provided to me by Santa Cruz. I have not yet had time to try it out (FFPE-mouse xenografts), but I have been told that it is working. I know that several of you have been waiting on this, so now might be a good time to give them a call. Angela Angela McNabola MS, HT(ASCP)SLS, QIHC Scientist Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 203-812-5001 fax# 203-812-5820 angela.mcnabola.b@bayer.com ------------------------------ Message: 19 Date: Thu, 17 Nov 2005 14:56:02 -0000 From: "Edwards, R.E." Subject: RE: [Histonet] E: beta-2- microglobulin To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Looking for an antibody to the above that works, as ever, on paraffin processed mouse tissues... Many thanks Richard Edwards MRC TOX UNIT LEICESTER...U.K.... ------------------------------ Message: 20 Date: Thu, 17 Nov 2005 10:36:34 -0500 From: "Kinsley, David" Subject: [Histonet] fixed jaw clamps/ large size specimens To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <79A75F7AD4BFAE4CA27C8AABE3F75D9A0E6C42@KENMSG22.us.schp.com> Content-Type: text/plain I'm looking to section some larger size specimens, particularly rat hind paw samples and am looking for cassettes that are slightly larger than the mega cassette but not as large as the super mega cassette. (Approximately 2" long by 1.5" wide by 1/2 to 3/4" deep) I have called some vendors and have had little luck. The only suggestion was to purchase a fixed jaw clamp for my microtome (Microm HM355) and try to use that with the super mega cassette. Does anyone have any experience with these types of specimens? Any suggestions on cassettes or microtome adapters? thanks David Kinsley Schering-Plough Research Institute Kenilworth, NJ 07033 908-740-4669 ********************************************************************* 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. ------------------------------ Message: 21 Date: Thu, 17 Nov 2005 10:09:09 -0600 From: "Bauer, Karen" Subject: [Histonet] Dissecting Microscopes To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Good morning! I need to purchase a new dissecting microscope for our pathologists. Ours is getting old and needs to be updated. We mainly use it for finding glomeruli in kidney biopsies to make sure there is adequate tissue for certain types of testing. I've looked online and have requested some quotes, but I'm curious to know what others are using. Any suggestions? As always, thank you. Karen L. Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************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. ------------------------------ Message: 22 Date: Thu, 17 Nov 2005 09:31:15 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] fixed jaw clamps/ large size specimens To: "Kinsley, David" , "'histonet@lists.utsouthwestern.edu'" Message-ID: <20051117173115.47213.qmail@web61215.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 David: In 1988 we began cutting whole prostate slices and we could not find neither megacassettes nor molds, so we made ours. For cassettes I used small round plastic containers, cut to an appropriate height with drilled holes and stappled 2 together, with the specimen in between. They work perfectly. As molds I cut pieces of wood of the required size and used aluminum sheets bought in a hardware to "wrap" around the wood piece and made them "melted paraffin leak proff". They also worked beautifully and, even when at last we bought the available cassettes and molds, for really large specimens (up to 3" x 4" ) we still use them. For those large paraffin blocks I used the very very old technique of adhering them to small 1"x1"x 2" blocks of wood, using a hot spatula, to be clamped to the microtome. Hope this "barrage" of "oldies" will help! Rene J. "Kinsley, David" wrote: I'm looking to section some larger size specimens, particularly rat hind paw samples and am looking for cassettes that are slightly larger than the mega cassette but not as large as the super mega cassette. (Approximately 2" long by 1.5" wide by 1/2 to 3/4" deep) I have called some vendors and have had little luck. The only suggestion was to purchase a fixed jaw clamp for my microtome (Microm HM355) and try to use that with the super mega cassette. Does anyone have any experience with these types of specimens? Any suggestions on cassettes or microtome adapters? thanks David Kinsley Schering-Plough Research Institute Kenilworth, NJ 07033 908-740-4669 ********************************************************************* 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 --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 24, Issue 25 **************************************** 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 NHeath <@t> Lifespan.org Fri Nov 18 06:22:56 2005 From: NHeath <@t> Lifespan.org (Heath, Nancy L.) Date: Fri Nov 18 06:23:08 2005 Subject: [Histonet] help getting on list Message-ID: <09C945920A6B654199F7A58A1D7D1FDE65ED3C@lsexch.lsmaster.lifespan.org> Hi, Can someone refresh my memory on how to sign up to be included on Histonet?? Tx :) From Barry.R.Rittman <@t> uth.tmc.edu Fri Nov 18 06:39:39 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Nov 18 06:39:52 2005 Subject: [Histonet] Haematoxylin absorption Message-ID: Simone Difficult to diagnose without seeing the sections. If your sections contain bone or cartilage , it is possible that these have lifted from the slide. Soft tissue may also on occasions lift because it is not adhering to the slide. This often allows stain to reach the section from both sides and stain the section more strongly in those areas. If this is the case then, as suggested by Muhammad careful drying of sections may be the answer. Also the use of plus slides may help. The most common cause, as noted earlier is due to in complete deparaffinization. Good luck. Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Muhammad Tahseen Sent: Thu 11/17/2005 11:30 AM To: Simione R. Turaganiwai; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Haematoxylin absorption Hello Simione, Your problem sounds like uneven section cutting on microtomy or section was not complete dried after microtome. Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Lahore Pakistan. ----- Original Message ----- From: Simione R. Turaganiwai To: Sent: Friday, November 18, 2005 11:44 AM Subject: [Histonet] Haematoxylin absorption 1.A haematoxylin and eosin slide was submitted to our Pathologist and she asked as to why did some portion of stained tissue absorbed more haematoxylin and some portion were light meaning absorbed less haematoxylin? 2.How can this be improved? Regard, Simione _______________________________________________ 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 Sjohnson <@t> sarapath.com Fri Nov 18 06:40:56 2005 From: Sjohnson <@t> sarapath.com (Saundra Johnson) Date: Fri Nov 18 06:40:57 2005 Subject: [Histonet] formalin precipitate/Iron artifact Message-ID: <211AEAE1E7C4974BB8B62A7BF8E4FF801BA70D@webber_nt.sarapath.com> I am having a problem with needle core liver Bx. The iron stains has a funny blue artifact staining around the peripheral. Bone marrows and other tissues with Iron stains do no exhibit this. We recently started recycling our zink formalin with Creative Waste Managment's filitering system but make the correct adjustments as instructed. Additionally, there is a slight precipate left in the retort chamber at the end of processing,even after the clean cycle. We have a 50% alcohol after the formalin and then go to 95%. Any suggestions with these two problems and are they related to recycling of formalin Saundra Johnson Histology-IHC Supervisor Sarasota Pathology **************************************************************************** SARASOTA PATHOLOGY CONFIDENTIALITY NOTICE: The information contained in this E-Mail message may be privileged, confidential, and protected under applicable law and is intended solely for the use of the individual or en- tity to whom it is addressed. If you are not the intended recipient or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited. It is strictly prohi- bited from using the internet email system to send messages with patient identifiable information per HIPAA Privacy regulations and Sarasota Patho- logy policy. If you have received this communication in error, please notify the sender immediately and delete this message. **************************************************************************** From Kemlo.Rogerson <@t> elht.nhs.uk Fri Nov 18 06:56:41 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Nov 18 06:56:52 2005 Subject: [Histonet] formalin precipitate/Iron artifact Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9EB236EE@elht-exch1.xelht.nhs.uk> Could it be from the needle? Are needles made of iron? Am I talking rubbish? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Saundra Johnson Sent: 18 November 2005 12:41 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] formalin precipitate/Iron artifact I am having a problem with needle core liver Bx. The iron stains has a funny blue artifact staining around the peripheral. Bone marrows and other tissues with Iron stains do no exhibit this. We recently started recycling our zink formalin with Creative Waste Managment's filitering system but make the correct adjustments as instructed. Additionally, there is a slight precipate left in the retort chamber at the end of processing,even after the clean cycle. We have a 50% alcohol after the formalin and then go to 95%. Any suggestions with these two problems and are they related to recycling of formalin Saundra Johnson Histology-IHC Supervisor Sarasota Pathology ************************************************************************ **** SARASOTA PATHOLOGY CONFIDENTIALITY NOTICE: The information contained in this E-Mail message may be privileged, confidential, and protected under applicable law and is intended solely for the use of the individual or en- tity to whom it is addressed. If you are not the intended recipient or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited. It is strictly prohi- bited from using the internet email system to send messages with patient identifiable information per HIPAA Privacy regulations and Sarasota Patho- logy policy. If you have received this communication in error, please notify the sender immediately and delete this message. ************************************************************************ **** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Fri Nov 18 08:13:05 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Nov 18 08:16:33 2005 Subject: [Histonet] RE: Slide adhesion - toe nail for fungi Message-ID: We embed the slide of toe nail so it will be cut vertically; then soften the block surface after trimming in Baker's fluid (10% glycerol in 70% IMS) for about 20 mins, then coat a slide with a film of gel-chrome alum (0.1% chrome alum in 1% gelatine), let it dry, mount the section on the slide and dry in 60C oven for a couple of hours then stain. I would avoid a silver stain though and use PAS or Gomori's aldehyde fuchsin. Jacqui Malam Lancaster 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 rjbuesa <@t> yahoo.com Fri Nov 18 08:24:35 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 18 08:24:44 2005 Subject: [Histonet] formalin precipitate/Iron artifact In-Reply-To: <53A22BBB01D6184EBF7A4DC18A021E9EB236EE@elht-exch1.xelht.nhs.uk> Message-ID: <20051118142435.72711.qmail@web61216.mail.yahoo.com> Saundra: Let me see if I can be of some help. Emmel (1964) quoting Mallory wrote that formalin solutions can produce a brown-black finely crystaline precipitate from laked hematoxylin that can be eliminated by treating sections for 30 min. in a 0.5% ammonia water sol. in 75% ethanol. Liver is more compact than bone marrow and perhaps the action of the fixative will be different and you will not find the ppdo. in bone marrow. I don't think that it would "harm" trying to see if the precipitate you found can be eliminated with the weak ammonia in ethanol which will point to the source of the problem. Also, did you begin to have the problem only after you started recycling your fixative? If that is the case, there you will have a lead to finding the cause. By the way, I would never ever recycle formalin fixatives, too toxic a procedure! "Disclaimer": perhaps I am also in the "talking rubbish" realm, as Kemlo pointed before me! Rene J. "Rogerson Kemlo (ELHT) Pathology" wrote: Could it be from the needle? Are needles made of iron? Am I talking rubbish? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Saundra Johnson Sent: 18 November 2005 12:41 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] formalin precipitate/Iron artifact I am having a problem with needle core liver Bx. The iron stains has a funny blue artifact staining around the peripheral. Bone marrows and other tissues with Iron stains do no exhibit this. We recently started recycling our zink formalin with Creative Waste Managment's filitering system but make the correct adjustments as instructed. Additionally, there is a slight precipate left in the retort chamber at the end of processing,even after the clean cycle. We have a 50% alcohol after the formalin and then go to 95%. Any suggestions with these two problems and are they related to recycling of formalin Saundra Johnson Histology-IHC Supervisor Sarasota Pathology ************************************************************************ **** SARASOTA PATHOLOGY CONFIDENTIALITY NOTICE: The information contained in this E-Mail message may be privileged, confidential, and protected under applicable law and is intended solely for the use of the individual or en- tity to whom it is addressed. If you are not the intended recipient or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited. It is strictly prohi- bited from using the internet email system to send messages with patient identifiable information per HIPAA Privacy regulations and Sarasota Patho- logy policy. If you have received this communication in error, please notify the sender immediately and delete this message. ************************************************************************ **** _______________________________________________ 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 --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From KevinMcGovern <@t> catholichealth.net Fri Nov 18 08:38:17 2005 From: KevinMcGovern <@t> catholichealth.net (McGovern, Kevin) Date: Fri Nov 18 08:38:32 2005 Subject: [Histonet] Dissolving paraffin Message-ID: Happy Friday, Histonetians! It wasn't known to the folks in our Histo lab, so I thought I'd pass this tip along to all of you: the home cleaner "Goo-Gone" does a right nice job of dissolving paraffin, without fuss. A great way to clean up all that stuff that seems to accumulate here and there! Kevin Kevin S. McGovern, BMETII Good Samaritan Hospital Clinical Engineering Dept. 10 East 31st Street Kearney, NE 68836 Phone: (308) 865-7051 Fax: (308) 865-2914 e-Mail: kevinmcgovern@catholichealth.net Confidentiality Notice: This email, including any attachments, contains CONFIDENTIAL information. The information is intended only for the use of the individual(s) or entity name above. If you are not the intended recipient, you are notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this email is not permissible. If you have received this email in error, please notify the sender immediately by reply mail, and destroy all copies of the original message. From LewisS <@t> pediatrics.ohio-state.edu Wed Nov 16 10:10:13 2005 From: LewisS <@t> pediatrics.ohio-state.edu (Lewis, Sarah) Date: Fri Nov 18 09:11:06 2005 Subject: [Histonet] Auto stainer IHC/SS Message-ID: <714B9F12B4E18C4C843B66E8E190F2AD4475F7@res2k3ms01.CRII.ORG> Just a quick question in regards to the Autostainer Plus from Dako. I am currently looking to purchase an Automated Immuno Stainer. I have demoed The i6000, Mozaic, and the Nemesis. I will soon demo the Autostainer Plus from Dako. They claim to be able to do special stains and IHC. Are any of you running specials on your Autostainer plus? Any thoughts on this? Also, If you have any experience with any of the units any feedback would be appreciated!!! The i6000 also claims to do specials as well, I only ran a few, I never tried to run IHC and specials together on the same run.(They have disposable pipette tips which eliminates the cross contamination issue.) Any thoughts would be great!!! Thanks in advance!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net From gcallis <@t> montana.edu Fri Nov 18 10:12:28 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Nov 18 10:12:46 2005 Subject: [Histonet] Strepavidin blocking kit with your mimicry problem In-Reply-To: <000201c5ebfe$073b1420$640aa8c0@SinisaHome> References: <000201c5ebfe$073b1420$640aa8c0@SinisaHome> Message-ID: <6.0.0.22.1.20051118090208.01b4f9d8@gemini.msu.montana.edu> Sasa, Please read the following private communcation I had with Vector Technical services concerning the issue you just mentioned. You may want to change your avidin/biotin blocking method to a Strepavidin/biotin blocking method (Vector kit) and use the DAKO kit with Strepavidin-HRP as it is intended with no secondary antibody involved. I did not see any avidin nor Strepavidin/biotin blocking in your method. Also, was your negative control the antibody isotype, which should be ARK'd the same as the primary and at the same concentration? ***************************************************************************************************************************** Craig Pow (Vector) wrote: Streptavidin does have some subtle binding differences than avidin. Streptavidin has been shown to bind to fibronectin receptors that are expressed on cell types. These integrins (extracellular matrix receptors) mediate cell adhesion, migration etc and streptavidin binds to these too. So the bottomline this that streptavidin based detection systems may give background/non-specific staining due to this inetgrin binding that is not blocked by a standard avidin/biotin blocking step. Hence the idea behind the streptavidin/biotin kit. See the following reference: Alon, R. et al (1991) Cell-adhesive properties of streptavidin are mediated by the exposure of an RGD-like RYD site. Eur. J. Cell Biol. 58:271-279. The nomenclature in this article is dated. The GpIIbIIIa is now known as the alpha 5 beta 1 dimer that specifically binds fibronectin via the RGD amino acid sequence. There are antibodies available against the alpha 5 subunit itself. Migratory cells, such as embryonic cells, some tumor cells etc would be high expressors of this integrin. Obviously you could either block with the streptavidin kit or use an avidin based detection system. ********************************************************************************************************* At 10:07 PM 11/17/2005, you wrote: >I am referring to your previous HIstonet correspondence of Teri Johnson >where they mentioned the following :....we also tried using an anti-IgG2a >anti mouse/HRP labelled secondary antibody, and in the negative control >there was no staining..... >I have red few of the Alon R papers regarding the mimicry between >streptavidin and fibronectin since I presume I might have the same problem >In my project I am trying to determine presence/absence of alpha4beta 1 >integrin in the rat uterus by using immunocytochemistry on the frozen >tissue....At the moment I have DAKO ARK kit and applying the following >procedure: >Cut 6um thick sections (use positive charged slides and silane pre-treated ) >dry sections overnight at room temperature >fix them the next day in cold acetone for 10 minutes >proceed with staining: >Biotinylate primary antibody (monoclonal -mouse anti human CD49d (integrin >alpha 4 beta 1)P4G9. >Step 1: Mix diluent, concentrated primary Ab (positive or negative) and >biotinylation reagent, incubate for 15 min >Step 2: add blocking reagent incubate for 5 min >Apply peroxidase block for 15 min >Rinse >Apply prepared biotinylated primary Ab (positive or negative), incubate for >1 hour >Rinse >Apply streptavidin HRP incubate for 15 min >Rinse >Apply prepared DAB incubate for 5 min >Rinse >Counterstain >Mount coverslips > >I am using Tris-HCL pH9 (with added Tween 20) as a washing buffer >I am using human tonsil as a positive control where I did not see any >specific immunolocalization on the sections where I had omit primary Ab. >But, I am experiencing "staining" of my negative control, actually >positive and >negative uterus look the same.....they "stain" on the same place. >Could this be the sterptavidin mimicry and would you have any suggestions >how to incorporate anti IgG2a antibody in my kit or maybe do you have any >other solution to my problem? Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Fri Nov 18 10:17:20 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Nov 18 10:17:36 2005 Subject: [Histonet] Re: preparation of chromic acid solution for CAS.... In-Reply-To: <001101c5ec08$fdc05120$593a2e93@ksvcpuo8c26n3k> References: <001101c5ec08$fdc05120$593a2e93@ksvcpuo8c26n3k> Message-ID: <6.0.0.22.1.20051118091248.01b5b9d0@gemini.msu.montana.edu> The chromic acid concentration used with a Grocotts methenamine silver stain for fungus will work - 4% to 5% Chromium Trioxide aka chromic acid. It should work nicely as periodic acid may not completely oxidize some fungal walls and you can get a false negative staining result. Carson and Fredenburgh have a very nice publication on this in Journal of Histotechnology a few years ago. This was excellent discussion on CAS versus PAS not too long ago on Histonet, so be sure to check Histonet Archives for further information. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From TMcNemar <@t> lmhealth.org Fri Nov 18 10:34:23 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Nov 18 10:38:58 2005 Subject: [Histonet] Auto stainer IHC/SS Message-ID: <6CD94D97ED7D924BA5C2B588FA95282139681F@nt_exchange.lmhealth.org> The special stains thatI have run have all been very nice except for masson's trichrome. We just don't like it. It is an open system though and I can use my own reagents and tweak it, just haven't had time. You cannot do IHC and special stains at the same time... they have to be separate runs. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Lewis, Sarah [mailto:LewisS@pediatrics.ohio-state.edu] Sent: Wednesday, November 16, 2005 11:10 AM To: HISTONET (E-mail) Subject: [Histonet] Auto stainer IHC/SS Just a quick question in regards to the Autostainer Plus from Dako. I am currently looking to purchase an Automated Immuno Stainer. I have demoed The i6000, Mozaic, and the Nemesis. I will soon demo the Autostainer Plus from Dako. They claim to be able to do special stains and IHC. Are any of you running specials on your Autostainer plus? Any thoughts on this? Also, If you have any experience with any of the units any feedback would be appreciated!!! The i6000 also claims to do specials as well, I only ran a few, I never tried to run IHC and specials together on the same run.(They have disposable pipette tips which eliminates the cross contamination issue.) Any thoughts would be great!!! Thanks in advance!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Nov 18 10:49:26 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Nov 18 10:49:36 2005 Subject: [Histonet] formalin precipitate/Iron artifact In-Reply-To: <53A22BBB01D6184EBF7A4DC18A021E9EB236EE@elht-exch1.xelht.nh s.uk> References: <53A22BBB01D6184EBF7A4DC18A021E9EB236EE@elht-exch1.xelht.nhs.uk> Message-ID: <6.0.0.22.1.20051118093335.01b76730@gemini.msu.montana.edu> You are not talking rubbish. We occasionally "pin" tissues to cork for fixation but use (aluminum?) hypodermic,disposable needles - they don't rust. However, you bring up an excellent point as some pins, particularly those we thought useful and purchased from a sewing store did rust - tossed those out!! A possible source of iron contamination is the specimen container with the lid as the culprit. Years ago, before the advent of good specimen containers, people recycled baby food jars whose lids rusted excessively and exfoliated rust/iron contamination onto the samples during fixation. If one has metal lids on containers, check them for rust and don't use them. Or possibly, are they returning any reagent in some type of metal can? We had metal cans whose liners failed, and rusted. We used these for refilling/bulk alcohol storage and rusty gunk poured out! Check the storage containers also. At 05:56 AM 11/18/2005, you wrote: >Could it be from the needle? Are needles made of iron? Am I talking >rubbish? Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From p.rastogi <@t> biogenex.com Fri Nov 18 11:02:39 2005 From: p.rastogi <@t> biogenex.com (Promila Rastogi) Date: Fri Nov 18 11:03:08 2005 Subject: [Histonet] Auto stainer IHC/SS Message-ID: <37DC9F93CF7F864182D0463EF93D571B0C1227@ISLETON2.california.biogenex.com> FYI, one CAN do IHC, ISH and special stains in the same run on the i6000 Automated Stainer from BioGenex. Thanks, ~Promila~ Promila A. Rastogi Product Marketing Manager p.rastogi@biogenex.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Friday, November 18, 2005 8:34 AM To: HISTONET (E-mail) Subject: RE: [Histonet] Auto stainer IHC/SS The special stains thatI have run have all been very nice except for masson's trichrome. We just don't like it. It is an open system though and I can use my own reagents and tweak it, just haven't had time. You cannot do IHC and special stains at the same time... they have to be separate runs. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Lewis, Sarah [mailto:LewisS@pediatrics.ohio-state.edu] Sent: Wednesday, November 16, 2005 11:10 AM To: HISTONET (E-mail) Subject: [Histonet] Auto stainer IHC/SS Just a quick question in regards to the Autostainer Plus from Dako. I am currently looking to purchase an Automated Immuno Stainer. I have demoed The i6000, Mozaic, and the Nemesis. I will soon demo the Autostainer Plus from Dako. They claim to be able to do special stains and IHC. Are any of you running specials on your Autostainer plus? Any thoughts on this? Also, If you have any experience with any of the units any feedback would be appreciated!!! The i6000 also claims to do specials as well, I only ran a few, I never tried to run IHC and specials together on the same run.(They have disposable pipette tips which eliminates the cross contamination issue.) Any thoughts would be great!!! Thanks in advance!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.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 sarah <@t> kidneybiopsy.com Fri Nov 18 11:17:03 2005 From: sarah <@t> kidneybiopsy.com (Sarah Holmes) Date: Fri Nov 18 11:06:05 2005 Subject: [Histonet] Dissecting Microscopes References: Message-ID: <003701c5ec63$e5738dc0$780a010a@wp.comcast.net> We are very satisfied with Van Guard brand stereomicroscopes model 1254SH for the purpose of allocating renal tissue. We have purchased several from different suppliers so I don't think they're too hard to find. All of these have been reliable and durable, having been carted around quite a bit. Email, or call me if more info needed. Sarah Holmes Laboratory Manager Laboratory for Kidney Pathology, Inc. Nashville, TN 615-321-5729 ----- Original Message ----- From: "Bauer, Karen" To: Sent: Thursday, November 17, 2005 10:09 AM Subject: [Histonet] Dissecting Microscopes Good morning! I need to purchase a new dissecting microscope for our pathologists. Ours is getting old and needs to be updated. We mainly use it for finding glomeruli in kidney biopsies to make sure there is adequate tissue for certain types of testing. I've looked online and have requested some quotes, but I'm curious to know what others are using. Any suggestions? As always, thank you. Karen L. Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Fri Nov 18 11:15:43 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Nov 18 11:15:55 2005 Subject: [Histonet] Fixing question References: <20051115200445.64231.qmail@web61219.mail.yahoo.com> Message-ID: <437E0C3F.C8DC263B@uwo.ca> A little clarification is called for here. 1. The diffusion of a fixative into a specimen follows the law of diffusion: d = K sqrt(t) where d is the distance penetrated (mm) and t is time (hours). [sqrt(t) means the square root of the time.] For formaldehyde entering a gelatin-albumin gel the value of the constant K is given by JR Baker as 3.6 (Principles of Biol. Microtechnique, p.40). 2. Although formaldehyde penetrates rapidly its chemical reactions with proteins occur rather slowly. Studies with radioactively labelled formaldehyde indicate that the amount bound reaches a maximum at 24 hours. Most of this can be removed by prolonged washing in water and probably is not involved in cross-links. Cross linking, which increases the resilience of the tissue in the face of osmotic gradients, alcohols etc, takes about 2 weeks, but is often considered to be sufficient after 24 hours. With shorter times in formaldehyde, tissues are still osmotically responsive, and subsequent immersion in alcohol causes coagulation of proteins that have not been cross-linked. This is not necessarily a bad thing. Cell nuclei stain more strongly after coagulant fixation than after thorough fixation with formadehyde. For a short account of how aldehyde fixatives work, see http://publish.uwo.ca/~jkiernan/formglut.htm For more details, Pearse's Histochemistry 4th ed Vol 1, or Hopwood's chapter in the 2002 edition of Bancroft & Gamble's Theory and Practice of hisatol. Techniques. John Kiernan Anatomy, UWO London, Canada ________________________________ Rene J Buesa wrote: > > Christian: > There are several issues to consider including the type of fixative. I you are referring > to neutral buffered formalin (NBF) overfixation occurs when tissues are left more time > than necessary and this will depend on the thickness of the tissue slice to be fixed. > NBF has a diffusability coefficient (K) of 0.78 mm/hour so a slice 4 mm thick will require > 2.5 hours to fix (remember that the NBF will be "entering" the tissues from both sides!). > Any time after that will be "theoretically" and overfixation, i.e. not needed time in NBF > and this can translate into more crosslinkage of the proteins and more difficulties for > immunohistochemistry (IHC) procedures. > Ethanol has a K of 1.0 mm/h so the same type of slice will require only 2 hours, will > fix by quagulation and theoretically will not interfere with IHC but, again, after 2 hours > it is not necessary to keep the tissue in ethanol. > Having said all that, in tissue processors like the Sakura VIP all stations can have > vacuum/pressure only depending in how long the station is programmed to last. > With thin enough slices of tissue I don't think you have to worry about overfixation > as long as you keep the tissues in the fixative the time required for its penetration, > whichever the fixative is. > Hope this will help (although I think I gave a somewhat "convoluted" answer!). > Rene J. > > Christian Franci wrote: > dear folks, pardon my ignorance but.... > > It seems that pressure is applied to the processing of tissues to > ensure evenness and to force out any residual alcohol and/or xylene > that might over-dry one's delicate samples. > I gather that depending on what machine you have you can apply pressure > at each step or just at the final paraffin stage ( as in my case). > Here is my question... would applying pressure during the fixation of > the tissues also be beneficial? > Would it speed up the process therefore minimizing the possibility of > over-fixation? > Do any of you fix tissue under pressure and, if so, how do you go about > it? Just curious.... > > Cheers > > Chris > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > --------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Fri Nov 18 11:17:26 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Nov 18 11:16:39 2005 Subject: [Histonet] formalin precipitate/Iron artifact Message-ID: Am I in a parallel universe? Iron from a few seconds in a stainless steel needle? Is this some soluble iron salt? If so how does it survive watery fixation and processing? Is it insoluble iron? If so what is it doing on the surface of stainless steel? Saundra Johnson did not describe it as particulate. Why is everyone assuming it is so? Surely what is being described is simply an edge artefact which is seen *very* commonly on the edges of liver biopsies stained with Perl's. It is a faint blush - no more, and the clue to its being an artefact is that it is non-particulate and at the edge. PAS sometimes shows a similar phenomenon. Beam me up Scottie. 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: Gayle Callis [mailto:gcallis@montana.edu] Sent: 18 November 2005 16:49 To: Rogerson Kemlo (ELHT) Pathology; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] formalin precipitate/Iron artifact You are not talking rubbish. We occasionally "pin" tissues to cork for fixation but use (aluminum?) hypodermic,disposable needles - they don't rust. However, you bring up an excellent point as some pins, particularly those we thought useful and purchased from a sewing store did rust - tossed those out!! A possible source of iron contamination is the specimen container with the lid as the culprit. Years ago, before the advent of good specimen containers, people recycled baby food jars whose lids rusted excessively and exfoliated rust/iron contamination onto the samples during fixation. If one has metal lids on containers, check them for rust and don't use them. Or possibly, are they returning any reagent in some type of metal can? We had metal cans whose liners failed, and rusted. We used these for refilling/bulk alcohol storage and rusty gunk poured out! Check the storage containers also. At 05:56 AM 11/18/2005, you wrote: >Could it be from the needle? Are needles made of iron? Am I talking >rubbish? Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From doninn <@t> mail.nih.gov Fri Nov 18 12:14:30 2005 From: doninn <@t> mail.nih.gov (Donin, Nick (NIH/NCI)) Date: Fri Nov 18 12:14:46 2005 Subject: [Histonet] HRP conjugation Message-ID: <16A0583FB1644E4DB8C0A0265028B6FD02B4E83F@nihexchange13.nih.gov> Hello Everyone, This isn't a Histology question, but it relates to antibodies, so I thought someone could help me. I'm doing a western blot, and I'm running the protein from mice tumors. I'm using a mouse primary antibody to stain for a particular antigen, but the problem is that when I apply anti-mouse secondary, it stains everything on the blot (presumably because the tumor tissue is rich in mouse IgG antibodies). I thought I could get around this by conjugating my primary antibody directly to H.R.Peroxidase, thus eliminating the need for a secondary. My first question is: 1. Does this strategy even make sense? 2. Does anyone have a technique or product for carrying out this conjugation? Thanks for all your help, any thoughts or suggestions are greatly appreciated. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 From sluhisto <@t> yahoo.com Fri Nov 18 12:30:37 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Nov 18 12:30:47 2005 Subject: [Histonet] charged slides for IHC Message-ID: <20051118183037.24025.qmail@web51010.mail.yahoo.com> Hello All: What kind of adhesion slides are you using and who is your vendor? Thanks Susan Hay-Antoff Histopathology Supervisor St. Louis University --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From JurciseJ <@t> pediatrics.ohio-state.edu Fri Nov 18 12:42:56 2005 From: JurciseJ <@t> pediatrics.ohio-state.edu (Jurcisek, Joseph) Date: Fri Nov 18 12:43:42 2005 Subject: [Histonet] Dissecting Microscopes Message-ID: <714B9F12B4E18C4C843B66E8E190F2ADB1EC70@res2k3ms01.CRII.ORG> I have always been very pleased with the Zeiss line of microscopes. I am currently using the SV-11 stereomicroscope. It is very versatile and dependable. Also, the optics can't be beat. Joseph A. Jurcisek Senior Research Associate Center for Microbial Pathogenesis Columbus Children's Research Institute 700 Children's Dr. Columbus, OH 43205-2696 (614) 355-3521 fax (614) 722-2818 jurcisej@pediatrics.ohio-state.edu We are very satisfied with Van Guard brand stereomicroscopes model 1254SH for the purpose of allocating renal tissue. We have purchased several from different suppliers so I don't think they're too hard to find. All of these have been reliable and durable, having been carted around quite a bit. Email, or call me if more info needed. Sarah Holmes Laboratory Manager Laboratory for Kidney Pathology, Inc. Nashville, TN 615-321-5729 ----- Original Message ----- From: "Bauer, Karen" To: Sent: Thursday, November 17, 2005 10:09 AM Subject: [Histonet] Dissecting Microscopes Good morning! I need to purchase a new dissecting microscope for our pathologists. Ours is getting old and needs to be updated. We mainly use it for finding glomeruli in kidney biopsies to make sure there is adequate tissue for certain types of testing. I've looked online and have requested some quotes, but I'm curious to know what others are using. Any suggestions? As always, thank you. Karen L. Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tahseen <@t> brain.net.pk Fri Nov 18 04:45:25 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Nov 18 12:48:52 2005 Subject: [Histonet] Auto stainer IHC/SS References: <714B9F12B4E18C4C843B66E8E190F2AD4475F7@res2k3ms01.CRII.ORG> Message-ID: <005601c5ec2d$329d9a00$972bfea9@m7c0y4> Dear Sarah E. Lewis, The special stains that we have run, all been very nice.You cannot do IHC and special stains at the same time... they have to be separate runs, separate software. I use DakoAutostainer in my lab and have absolutely no problems. We have beautiful reproducible staining with every antibody. Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Lahore Pakistan. ----- Original Message ----- From: Lewis, Sarah To: HISTONET (E-mail) Sent: Thursday, November 17, 2005 5:10 AM Subject: [Histonet] Auto stainer IHC/SS Just a quick question in regards to the Autostainer Plus from Dako. I am currently looking to purchase an Automated Immuno Stainer. I have demoed The i6000, Mozaic, and the Nemesis. I will soon demo the Autostainer Plus from Dako. They claim to be able to do special stains and IHC. Are any of you running specials on your Autostainer plus? Any thoughts on this? Also, If you have any experience with any of the units any feedback would be appreciated!!! The i6000 also claims to do specials as well, I only ran a few, I never tried to run IHC and specials together on the same run.(They have disposable pipette tips which eliminates the cross contamination issue.) Any thoughts would be great!!! Thanks in advance!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JOliver <@t> asterand.com Fri Nov 18 12:59:17 2005 From: JOliver <@t> asterand.com (Jeff Oliver) Date: Fri Nov 18 13:00:03 2005 Subject: [Histonet] HRP conjugation Message-ID: <1B0B60D56CA6A440A240E866BA3458D810101A@ATL1VEXC017.usdom004.tco.tc> Unfortunately you will need to buy a new primary. A mouse primary also may have cross-reactivity problems with mouse samples, so I would stay away from any preconjugated mouse antibodies too. Preconjugating your own antibodies can be a timesaver, but I found it introduced to many random variables. The most concerning one in your case is you can't guarantee all of the secondary is bound to the primary. I would expect the same result if you try to preconjugate with an anti-mouse secondary. -Jeff -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Donin, Nick (NIH/NCI) Sent: Friday, November 18, 2005 1:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HRP conjugation Hello Everyone, This isn't a Histology question, but it relates to antibodies, so I thought someone could help me. I'm doing a western blot, and I'm running the protein from mice tumors. I'm using a mouse primary antibody to stain for a particular antigen, but the problem is that when I apply anti-mouse secondary, it stains everything on the blot (presumably because the tumor tissue is rich in mouse IgG antibodies). I thought I could get around this by conjugating my primary antibody directly to H.R.Peroxidase, thus eliminating the need for a secondary. My first question is: 1. Does this strategy even make sense? 2. Does anyone have a technique or product for carrying out this conjugation? Thanks for all your help, any thoughts or suggestions are greatly appreciated. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GDawson <@t> dynacaremilwaukee.com Fri Nov 18 14:15:49 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Nov 18 14:17:14 2005 Subject: [Histonet] pH meter Message-ID: All, My prehistoric pH meter is in the process of biting the dust so I'm in the market for a new one. I'm interested in a pH meter that is very easy to use and affordable. Please sound off on your favorites histo-land. Vendors are more than welcome to respond. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI From pathrm35 <@t> adelphia.net Fri Nov 18 15:16:56 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Nov 18 15:17:04 2005 Subject: [Histonet] Auto stainer IHC/SS Message-ID: <21066377.1132348616862.JavaMail.root@web7.mail.adelphia.net> Sarah, We purchased the Biogenex i6000 about 5 months ago and have had good results with it. We run special stains (alcian blue/pas and PAS for fungus) along with our immunos (mostly Melan A and S-100's). We are using about 17 of their antibodies. The antigen retrieval system is easy to use and consistant. The Biogenex rep., Alan Younger is very good and is a pleasure to work with. We wanted to demo the Dako stainer but the Dako rep. was going to charge us $1,700.00 shipping fee for the demo. My Dr. said "No Way". Ron Martin, BS, HT (ASCP) HTL ---- "Lewis wrote: > Just a quick question in regards to the Autostainer Plus from Dako. I am currently looking to purchase an Automated Immuno Stainer. I have demoed The i6000, Mozaic, and the Nemesis. I will soon demo the Autostainer Plus from Dako. They claim to be able to do special stains and IHC. Are any of you running specials on your Autostainer plus? Any thoughts on this? Also, If you have any experience with any of the units any feedback would be appreciated!!! The i6000 also claims to do specials as well, I only ran a few, I never tried to run IHC and specials together on the same run.(They have disposable pipette tips which eliminates the cross contamination issue.) Any thoughts would be great!!! Thanks in advance!!! > > Sarah E. Lewis > Histotechnician > Childrens Research > Center for Gene Therapy > 700 Childrens Dr Rm WA3112 > Columbus OH 43205 > (614)-722-2204 > LewisS@ccri.net > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Nov 18 16:47:18 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Nov 18 16:47:34 2005 Subject: [Histonet] Auto stainer IHC/SS In-Reply-To: <21066377.1132348616862.JavaMail.root@web7.mail.adelphia.net> Message-ID: <200511182247.jAIMlGxT015646@chip.viawest.net> I use the reg. DakoAutostainer and have done some special stains on it, I mostly use it for IHC and I have not ever had a cross contamination problem even though it does not have disposable tips it uses one probe for everything and cleans it with a sonicator between each reagent. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net 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. 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. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pathrm35@adelphia.net Sent: Friday, November 18, 2005 2:17 PM To: LewisS@pediatrics.ohio-state.edu Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Auto stainer IHC/SS Sarah, We purchased the Biogenex i6000 about 5 months ago and have had good results with it. We run special stains (alcian blue/pas and PAS for fungus) along with our immunos (mostly Melan A and S-100's). We are using about 17 of their antibodies. The antigen retrieval system is easy to use and consistant. The Biogenex rep., Alan Younger is very good and is a pleasure to work with. We wanted to demo the Dako stainer but the Dako rep. was going to charge us $1,700.00 shipping fee for the demo. My Dr. said "No Way". Ron Martin, BS, HT (ASCP) HTL ---- "Lewis wrote: > Just a quick question in regards to the Autostainer Plus from Dako. I am currently looking to purchase an Automated Immuno Stainer. I have demoed The i6000, Mozaic, and the Nemesis. I will soon demo the Autostainer Plus from Dako. They claim to be able to do special stains and IHC. Are any of you running specials on your Autostainer plus? Any thoughts on this? Also, If you have any experience with any of the units any feedback would be appreciated!!! The i6000 also claims to do specials as well, I only ran a few, I never tried to run IHC and specials together on the same run.(They have disposable pipette tips which eliminates the cross contamination issue.) Any thoughts would be great!!! Thanks in advance!!! > > Sarah E. Lewis > Histotechnician > Childrens Research > Center for Gene Therapy > 700 Childrens Dr Rm WA3112 > Columbus OH 43205 > (614)-722-2204 > LewisS@ccri.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 shive003 <@t> umn.edu Fri Nov 18 17:24:20 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Nov 18 17:24:50 2005 Subject: [Histonet] Rotavirus antibody source Message-ID: <00be01c5ec97$34396280$41065486@auxs.umn.edu> My past supplier (DAKO) of Rotavirus A antiserum has discontinued that item, so does anyone else know of a new vendor I could try? I use it on both bovine and porcine tissue, so it'd be ideal to find another polyclonal that binds to both species, rather than to buy 2 species-specific monoclonal antibodies. Thanks in advance for your suggestions. Jan Shivers U of MN Vet Diag Lab From vanann702 <@t> skmc.gov.ae Fri Nov 18 22:10:00 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Fri Nov 18 22:07:47 2005 Subject: [Histonet] best formalin Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02F6D6BD@SKMCEMAIL.skmc.gov.ae> We use Millonigs here too - we make it up in the lab. Also used it for many years in my previous lab, always with excellent results Annieinarabia -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Wednesday, November 16, 2005 10:10 PM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] best formalin We like a neutral buffered formalin (phosphate salts) and purchase it either ready to use or in concentrated form (to make a final working solution) We find NBF concentrates convenient and economical. Several of our labs prefer ready to use phophate buffered formalin i.e. NBF(Fisherbrand) in order to avoid formaldehyde fumes during NBF preparation without an available fume hood. . Richard Allan, Surgipath and others have formalin concentrates available. Surgipath has Carsons Millonigs Buffered Formalin with slightly different buffering formulation using sodium hydroxide and a phosphate salt, and MBF also comes in concentrated form. The joy of this one is - it was developed for use with both FFPE and EM per Freida Carson's publication in JOH many years ago. All of the above provided excellent fixation. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PKamalavenkatesh <@t> wockhardtin.com Fri Nov 18 23:43:08 2005 From: PKamalavenkatesh <@t> wockhardtin.com (PKamalavenkatesh@wockhardtin.com) Date: Fri Nov 18 23:40:46 2005 Subject: [Histonet] cytocentrifuge Message-ID: Dear Histonetters, We are willing to procure one cytocentrifuge for our experimental studies. Any of the histonetters who is having one which they want to recommend for us. Regards Dr.P. Kamalavenkatesh DRUG DISCOVERY RESEARCH- BIOLOGY Wockhardt Research Center D-4, MIDC, Chikalthana Aurangabad Mobile : 09822629964 E.mail : Pkamalavenkatesh@wockhardtin.com Please Visit our New Corporate Web Site www.wockhardt.com --------------------- Disclaimer ------------------------------------------------------------------------------ Information transmitted by this E-MAIL is proprietary to Wockhardt Ltd. and/ or its Group Companies and/or,its Customers and is intended for use only by the individual or entity to which it is addressed, and may contain information that is privileged, confidential or exempt from disclosure under applicable law. If you are not the intended recipient or it appears that this mail has been forwarded to you without proper authority, you are notified that any use or dissemination of this information in any manner is strictly prohibited. In such cases, please delete this mail from your records. --------------------------------------------------------------------------------------------------------------- From jkiernan <@t> uwo.ca Sat Nov 19 01:09:48 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Sat Nov 19 01:09:57 2005 Subject: [Histonet] pH meter References: Message-ID: <437ECFBC.A7201C90@uwo.ca> Dear Glen, Are you sure it's the meter that's moribund, not the electrode? There's not a lot that can go wrong with an old fashioned pH meter; it's basically just a simple amplifier connected to a microammeter. The vacuum tube in the amplifier - it's called an electrometer tube - ought to be replaceable, though they last for years if switched on all the time. The d'Arsonval movement of the meter is very rugged despite its being able to move the needle in response to tiny currents. The only way to kill such a meter is by electrocution - burning out the wire of the moving coil. That doesn't happen with even the amplified output of a pH electrode recording pH 14. Glass and similar pH electrodes, on the other hand, are very sensitive to physical and chemical insults. They can even deteriorate badly from not being used for a few weeks. Solutions that can deposit thin layers of a metal are bad news for pH electrodes. That includes some formulations of silver and gold compounds. Some cationic dyes stain glass, as we have all seen. A dyed pH electrode may not be entirely trustworthy. The wick on the side of a pH electrode, above the delicate glass bulb, or on the side of the protective plastic sleeve, can become encrusted with crystals from the solution inside the electrode (KCl with a tiny amount of AgCl) or it can be stained by dyes or other chemicals into which it gets dipped. Contamination is as inevitable as taxation, and it changes the way the pH electrode (which is really a cell) varies its electrical output in response to the hydrogen ion concentration of the surrounding liquid. If a pH meter is behaving strangely, try to revive the electrode. (The manufacturer's bumph should have instructions; it usually means various washes, replacing the internal solution, and then soaking for a few days in saturated aqueous KCl. MUCH cheaper than buying a new electrode.) If that fails, buy a new electrode. Modern pH meters use solid-state amplifiers and provide "digital" display. If precision to 0.1 pH units is all you need, a little hand-held meter with an electrode costs ?$200. A new electrode for any kind of pH meter costs the same or more. Expensive modern pH meters make calibration against standard solutions easy. The on-board computer replaces turning a screw and twiddling a knob, and it can also take care of some temperature effects. Checking pH has always been a horrid chore, because standard solutions must be checked every time the meter is used. That's the law even with 21st century smart pH meters, because errors come from the dipstick (pH electrode), not from the meter. End of sermon. John Kiernan Anatomy, UWO London, Canada ___________________________________________________________________ "Dawson, Glen" wrote: > > All, > > My prehistoric pH meter is in the process of biting the dust so I'm in the > market for a new one. I'm interested in a pH meter that is very easy to use > and affordable. Please sound off on your favorites histo-land. Vendors are > more than welcome to respond. > > Thank-you In Advance, > > Glen Dawson BS, HT & QIHC (ASCP) > IHC Manager > Milwaukee, WI > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MajorFocus <@t> aol.com Sat Nov 19 01:44:17 2005 From: MajorFocus <@t> aol.com (MajorFocus@aol.com) Date: Sat Nov 19 01:44:35 2005 Subject: [Histonet] Olympus CUT 4060E motorized rotary microtome available Message-ID: <21f.3495e50.30b031d1@aol.com> I recently acquired an Olympus motorized CUT 4060E microtome from a pathology laboratory that has merged with their sister hospital. This microtome is in like new condition and can be used in automatic or manual mode. You can view this item on Ebay (Item number: 7564375954) or contact me directly. Greg Good, HT(ASCP) Major Focus (800) 888-1152 From rmeyers <@t> ctcsurge.com Sat Nov 19 12:02:06 2005 From: rmeyers <@t> ctcsurge.com (Roger Meyers) Date: Sat Nov 19 12:05:42 2005 Subject: [Histonet] automated response Message-ID: <10511191302.AA02926@mail1.ctcsurge.com> I will be out of the office from Monday, November 21, 2005 through Tuesday, November 22, 2005. If the matter is urgent, please call Computer Trust Corporation at 617-557-9264 for immediate assistance. From kaleid11 <@t> yahoo.com Sun Nov 20 09:39:10 2005 From: kaleid11 <@t> yahoo.com (Adam Perry) Date: Sun Nov 20 09:39:19 2005 Subject: [Histonet] HRP conjugation In-Reply-To: <16A0583FB1644E4DB8C0A0265028B6FD02B4E83F@nihexchange13.nih.gov> Message-ID: <20051120153910.20993.qmail@web30404.mail.mud.yahoo.com> What do you mean by "It stains everything on the blot"? Are you detecting bands across the entire MW spectrum? If so, that sounds more like your antibody concentrations are too high. I would think if it really were cross-reaction with endogenous mouse IgG that you would detect bands corresponding to heavy and light chain IgG, etc. and not pick up everything in the blot? I am fairly new to western blotting, but maybe other folks could weigh in with their thoughts... Adam Department of Physiology and Biophysics University of Illinois at Chicago Chicago, IL 60612 "Donin, Nick (NIH/NCI)" wrote: Hello Everyone, This isn't a Histology question, but it relates to antibodies, so I thought someone could help me. I'm doing a western blot, and I'm running the protein from mice tumors. I'm using a mouse primary antibody to stain for a particular antigen, but the problem is that when I apply anti-mouse secondary, it stains everything on the blot (presumably because the tumor tissue is rich in mouse IgG antibodies). I thought I could get around this by conjugating my primary antibody directly to H.R.Peroxidase, thus eliminating the need for a secondary. My first question is: 1. Does this strategy even make sense? 2. Does anyone have a technique or product for carrying out this conjugation? Thanks for all your help, any thoughts or suggestions are greatly appreciated. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From cbass <@t> bidmc.harvard.edu Sun Nov 20 12:06:50 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Sun Nov 20 12:07:01 2005 Subject: [Histonet] HA tag antibody/protocol Message-ID: Hi Everyone, I am looking for a good HA-tag antibody for use in mouse livers. Does anyone have a recommendation? I would be particularly interested in any protocols that you have as well. I am still new to immunostaining, so I would appreciate any advice you can give me. thanks, Caroline Bass From cwscouten <@t> myneurolab.com Sun Nov 20 16:57:08 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Sun Nov 20 16:57:53 2005 Subject: [Histonet] am I snap-freezing correctly? Message-ID: <5784D843593D874C93E9BADCB87342AB9168CB@tpiserver03.Coretech-holdings.com> See the following link, which will appear in Microscopy Today soon. http://www.myneurolab.com/global/Manuals/Tips%20and%20Techniques%20Freez ing%20Artifact.pdf Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 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 Jacobs,Jennifer (stu) Sent: Wednesday, November 16, 2005 2:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] am I snap-freezing correctly? Hi, I am snap freezing brain tissue for sectioning (8uM) on cryostat (-25C). When I snap freeze, I put the brain hemisphere on a piece of aluminum foil (boat) and let it float on the LN2. Most times, some LN2 comes in direct contact with the brain tissue as the boat is not leak-free. Is this bad? Should I make sure the LN2 never comes in contact with the brain? The problem I've been noticing is that, right off the cryostat, my tissue looks "hole-y". This just gets worse as I proceed with the IHC stain (CD-31 Ab to see vessels) and dehydration + xylene steps. By the xylene steps the holes are so stretched out that I can't even recognize vessels in the tissue. I am not sure whether the holes arise from the brain extraction procedure (ie: snap freezing) or from the sectioning. But the person who sections has been doing it for years and has great experience with the cryostat. Any ideas/suggestions would be great! Thanks a lot! Jenn UConn Health Center Dept Pharmacology Farmington, CT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From yang5yc <@t> yahoo.com Sun Nov 20 20:36:55 2005 From: yang5yc <@t> yahoo.com (Kelly Yang) Date: Sun Nov 20 20:37:04 2005 Subject: [Histonet] questions about immunofluorescence on frozen tissue Message-ID: <20051121023655.46193.qmail@web33908.mail.mud.yahoo.com> Hi Histolanders, I have the following questions: I?m trying to do immunofluorescence on frozen tissue. I had tried both 4% formaldehyde and AA fixations. It seems that 4% formaldehyde give the better morphology but weaker signal than AA fixation. I wonder whether it is necessary to apply the permeabilization step in a frozen tissue section. I had saw two different opinions about this. Some people said we don?t need to permeabilztion, since the cell interior is fully exposed to antibody in the cutting sections. Some said there is need to add this step to get better results. In previous, Chris van der Loos mentioned that ?At the end of the IHC staining procedure, after the chromogen step wash your slides with tap water. NEVER use distilled water as this will ruin the tissue section completely?. We also conterstain DNA with DAPI. My question is that should we wash slides with tap water after 2nd antibody-conjugated with chromogen or after conterstaining with DAPI? Also Chris mentioned about avoiding using tween-20 in AA fixed slides. If elimiating tween-20 from the washing buffer, I encountered a bubble problem while putting the coverglass. There is no or weaker signal from those regions cover by the bubbles. Can anybody teach me how to avoid creating bubble while applying the coverglass? The mounting medium was bought from Vector. Any help will be highly appreciated. Kelly Yang Graduate student Department of Epidemiology School of Public Heath University of California, Los Angeles 310-409-9179 ext. 57795 yangyc@ucla.edu --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From prouty27 <@t> msn.com Sun Nov 20 22:53:16 2005 From: prouty27 <@t> msn.com (Sally Prouty) Date: Sun Nov 20 22:53:25 2005 Subject: [Histonet] oil red o Message-ID: I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From vanann702 <@t> skmc.gov.ae Sun Nov 20 23:07:49 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Sun Nov 20 23:05:56 2005 Subject: [Histonet] oil red o Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02F6D6C8@SKMCEMAIL.skmc.gov.ae> I did ORO on musce bx's for years - successfully - try the method in Bancroft and when you coverslip, be sure to use AQUEOUS mountant Annieinarabia -----Original Message----- From: Sally Prouty [mailto:prouty27@msn.com] Sent: Monday, November 21, 2005 8:53 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Nov 21 03:17:59 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Mon Nov 21 03:27:36 2005 Subject: [Histonet] formalin precipitate/Iron artifact Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9EB236F6@elht-exch1.xelht.nhs.uk> Hello Captain, how was the 'away team'? Did you see alien life? If Gayle says it's so, then it's so; she's always right. I did issue a warning with my flight of fancy. Meg sends her love............... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Friday, November 18, 2005 5:17 PM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] formalin precipitate/Iron artifact Am I in a parallel universe? Iron from a few seconds in a stainless steel needle? Is this some soluble iron salt? If so how does it survive watery fixation and processing? Is it insoluble iron? If so what is it doing on the surface of stainless steel? Saundra Johnson did not describe it as particulate. Why is everyone assuming it is so? Surely what is being described is simply an edge artefact which is seen *very* commonly on the edges of liver biopsies stained with Perl's. It is a faint blush - no more, and the clue to its being an artefact is that it is non-particulate and at the edge. PAS sometimes shows a similar phenomenon. Beam me up Scottie. 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: Gayle Callis [mailto:gcallis@montana.edu] Sent: 18 November 2005 16:49 To: Rogerson Kemlo (ELHT) Pathology; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] formalin precipitate/Iron artifact You are not talking rubbish. We occasionally "pin" tissues to cork for fixation but use (aluminum?) hypodermic,disposable needles - they don't rust. However, you bring up an excellent point as some pins, particularly those we thought useful and purchased from a sewing store did rust - tossed those out!! A possible source of iron contamination is the specimen container with the lid as the culprit. Years ago, before the advent of good specimen containers, people recycled baby food jars whose lids rusted excessively and exfoliated rust/iron contamination onto the samples during fixation. If one has metal lids on containers, check them for rust and don't use them. Or possibly, are they returning any reagent in some type of metal can? We had metal cans whose liners failed, and rusted. We used these for refilling/bulk alcohol storage and rusty gunk poured out! Check the storage containers also. At 05:56 AM 11/18/2005, you wrote: >Could it be from the needle? Are needles made of iron? Am I talking >rubbish? Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ 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 Mon Nov 21 04:37:37 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Mon Nov 21 04:37:47 2005 Subject: [Histonet] RE: HRP conjugation Message-ID: <135e5321360929.1360929135e532@amc.uva.nl> Hello Nick, I am wondering if you could use a Mouse-on-mouse detection system for this purpose. Try Vector MOM kit or Dako ARKit. These kit are definitely much more simpler than "true" HRP-conjugation. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Fri, 18 Nov 2005 13:14:30 -0500 From: "Donin, Nick \(NIH/NCI\)" Subject: [Histonet] HRP conjugation To: Hello Everyone, This isn't a Histology question, but it relates to antibodies, so I thought someone could help me. I'm doing a western blot, and I'm running the protein from mice tumors. I'm using a mouse primary antibody to stain for a particular antigen, but the problem is that when I apply anti-mouse secondary, it stains everything on the blot (presumably because the tumor tissue is rich in mouse IgG antibodies). I thought I could get around this by conjugating my primary antibody directly to H.R.Peroxidase, thus eliminating the need for a secondary. My first question is: 1. Does this strategy even make sense? 2. Does anyone have a technique or product for carrying out this conjugation? Thanks for all yo! ur help, From BMolinari <@t> heart.thi.tmc.edu Mon Nov 21 06:26:58 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Mon Nov 21 06:30:11 2005 Subject: [Histonet] oil red o Message-ID: Hi Sally, I use the ORO stain found in Freida Carson's Histotechnology text book with consistent results. You do have to use a gentle hand when applying the coverslip and use an aqueous mounting media. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 10:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kbroomal <@t> NEMOURS.ORG Mon Nov 21 10:19:15 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Mon Nov 21 10:19:39 2005 Subject: [Histonet] oil red o Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824494@wlmmsx02.nemours.org> Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Nov 21 10:28:45 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Mon Nov 21 10:28:55 2005 Subject: [Histonet] oil red o Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9EB236FE@elht-exch1.xelht.nhs.uk> "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ 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 llewllew <@t> shaw.ca Mon Nov 21 11:06:34 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Mon Nov 21 11:07:04 2005 Subject: [Histonet] oil red o References: <53A22BBB01D6184EBF7A4DC18A021E9EB236FE@elht-exch1.xelht.nhs.uk> Message-ID: <000801c5eebd$ed618cb0$690a4246@yourlk4rlmsu> The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound) would be someone (or something) who irritates someone else. Is there another meaning? Bryan ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Kristen Broomall" ; "Sally Prouty" ; Sent: Monday, November 21, 2005 8:28 AM Subject: RE: [Histonet] oil red o "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ 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 rjbuesa <@t> yahoo.com Mon Nov 21 11:54:45 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 21 11:54:54 2005 Subject: [Histonet] oil red o In-Reply-To: Message-ID: <20051121175445.63758.qmail@web61221.mail.yahoo.com> Hi Sally: The following is the Oil red O (ORO) stain I used for more than 20 years, with consistent results always. 1- Stock solution: ORO dye---1.0 g + isopropyl alcohol ---500 mL. Let stand for at least 24 hours to dissolve. This solution is stable if capped well. 2- Working solution: ORO stock solution---24 mL + distilled water ---16 mL; mix, cover and let it stand for 10 minutes. Filter through 2 thicknesses of paper because the dye will precipitate on exposure to air (be patient, it will take sometime to filter). The staining procedure: 1- cut the frozen sections. 2- place the slide with the frozen sections in a Coplin jar with 5 mL of "pure" formaldehyde in a way that the sections are not in contact with the 36% formalin (5 mL will be enought). Cover the jar and place it in a water bath at 45-50 Celsius for 10-15 minutes. The idea is that the sections are fixed by the formalin fumes, not by the liquid itself. After placing the Coplin jar in the water bath, start preparing the ORO working solution. 3- Take the slides and wash them with distilled water to eliminate the OCT used to embed the tissue for the frozen sections. 4- Stain the section with the hematoxylin ("Harris" type) at this moment for 30 secs. If you try to stain the nuclei after the ORO it will be weaken, so nuclear staining has to be done before the fat stain. 5- Place the slides in running tap water for 5 minute. 6- Wash slides with distilled water. 7- Place the slides in the working ORO solution for 10-15 minutes. 8- Wash the slides with distilled water, quickly, and cover with a water soluble medium that will harden after a few hours. Do NOT press the coverslipp trying to eliminate bubbles, rather use a "generous" amount of the mounting medium to assure there are no bubbles, and clean around the coverslip afterwards. There are many water soluble media in the market but I used one prepared "in house": A-Gelatin (same as used for the water bath)---40 g + distilled water--210 mL. Let the gelatin soak in water for 2 hours. B- Glycerine ---250 mL + phenol in crystals ---1.0 g. Mix A and B and heat gently until all the gelatin is dissolved and the solution is clear. Store in refrigerator. This protocol served me well for years, I hope it will be useful for you also! Rene J. Sally Prouty wrote: I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don?t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From sluhisto <@t> yahoo.com Mon Nov 21 11:55:16 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Mon Nov 21 11:55:24 2005 Subject: [Histonet] Cardinal information Message-ID: <20051121175516.44401.qmail@web51011.mail.yahoo.com> Hello All Does anyone have contact information for Cardinal the medical supply place? Thanks in advance. Susan SLU Histopathology --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From kbroomal <@t> NEMOURS.ORG Mon Nov 21 13:25:16 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Mon Nov 21 13:25:44 2005 Subject: [Histonet] oil red o Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824496@wlmmsx02.nemours.org> You guys are PITA's. (Do I need to spell that one out?) Bryan got the meaning exactly. (Good Job!) As in, I hate coverslipping ORO's because you can't press on the slip to get air bubbles out because the ORO will move. It's irritating. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bryan Llewellyn Sent: Monday, November 21, 2005 12:07 PM To: Histonet Subject: Re: [Histonet] oil red o The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound) would be someone (or something) who irritates someone else. Is there another meaning? Bryan ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Kristen Broomall" ; "Sally Prouty" ; Sent: Monday, November 21, 2005 8:28 AM Subject: RE: [Histonet] oil red o "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ 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 gentras <@t> vetmed.auburn.edu Mon Nov 21 14:30:35 2005 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Mon Nov 21 14:31:00 2005 Subject: Fwd: [Histonet] Cardinal information Message-ID: <6.0.1.1.0.20051121142935.01aad8e0@mailhost.vetmed.auburn.edu> The phone number that I have for Cardinal Health is : 1800-326-6457. Best wishes, Atoska >DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; s=s1024; d=yahoo.com; > >h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; > >b=SsMLnISHc50aE/mYL3xVf1xyfrICP3+dZEpSft1/xp+fBWgFThU7WjcxFca14Uxn8kKXA9FX8ESeEv+7JNCRAqpQQM13k6w94XER65WoaMkkPVEu7rRvo/T2YqI5bMyHzLZJgidJwk3T1i8Nb+rx9yuus+ulFmbTyrgKWMLvRIo= > ; >Date: Mon, 21 Nov 2005 09:55:16 -0800 (PST) >From: Histology SLU >To: "histonet@lists.utsouthwestern.edu" >X-Scan-Signature: 05628b3a735271e1629bc35bb8abda07 >X-Content-Filtered-By: Mailman/MimeDel 2.1.5 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >List-Id: For the exchange of information pertaining to histotechnology and > related fields >List-Unsubscribe: >, > > >List-Archive: >List-Post: >List-Help: >List-Subscribe: , > >Sender: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: ef1ad7719a1154f9450b1c67eafa0a9b >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] Cardinal information >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=5.5 tests=TO_ADDRESS_EQ_REAL > autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >X-spam: ESP<-82>=RBL:<-139> RDNS:<0> SHA:<57> UHA:<0> SLS:<0> BAYES:<0> > SenderID:<0> URL Substring Dictionary (TRU8):<0> Spam > Dictionary (TRU8):<0> NigeriaScam Dictionary (TRU8):<0> HTML > Dictionary (TRU8):<0> Porn Dictionary (TRU8):<0> Embed HTML > Dictionary (TRU8):<0> Obscenities Dictionary (TRU8):<0> URL > Dictionary (TRU8):<0> CAN-SPAM Compliance Dictionary (TRU8):<0> > >Hello All > > Does anyone have contact information for Cardinal the medical supply > place? Thanks in advance. > > Susan > SLU Histopathology > > >--------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Mon Nov 21 16:00:08 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Mon Nov 21 16:00:29 2005 Subject: [Histonet] Cardinal information Message-ID: 800-964-5227 or 800-326-6457 From nige.hammond <@t> blueyonder.co.uk Mon Nov 21 16:07:08 2005 From: nige.hammond <@t> blueyonder.co.uk (Nige) Date: Mon Nov 21 16:07:09 2005 Subject: [Histonet] Adult Mouse Skin Cryosections Message-ID: <001d01c5eee7$eb3f08c0$af82c050@cheekynige> Hi, I'm having trouble sectioning adult mouse skin prepared for cryosections. The dorsal skin has been taken freshly and put into plastic dispomoulds with Tissue Tek O.C.T, orientated and then slowly frozen by placing on a metal block which is nearly submerged in liquid nitrogen. The sample freezes nicely with no cracks, but when it comes to sectioning on a cryostat the tissue tends to scrunch up and come away from the tissue tek on the fatty side. Shaving the skin had no effect on the outcome. I've tried lots of different temperatures/section thickness but no luck. I'm assuming the problem has something to do with the fatty layer that is present with the skin, but it is very difficult (if not impossible) to separate this from the skin. Can anyone suggest how to get nice adult skin cryosections, any other processing needed to be done before mounting in O.C.T? Many thanks, nige From Barry.R.Rittman <@t> uth.tmc.edu Mon Nov 21 16:37:10 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Mon Nov 21 16:37:25 2005 Subject: [Histonet] Adult Mouse Skin Cryosections Message-ID: Nige It is not easy to consistently cut frozen sections of adult mouse skin. How you orient the block and whether you have already shaved most of the hair from the skin are critical factors. One reason may be that you do not have the OCT binding sufficiently to the tissue; this can be remedied by soaking in OCT for 5 - 10 minutes before freezing. Theoretically you should be cutting the connective tissue side first and the external surface with hair shafts last if possible. However if you cut along the entire connective tissue length at one time you will usually get some separation from the OCT. I prefer to orient the block so that if the block is rectangular in shape, the first part to meet the knife edge is a connective tissue corner of the block. This allows for a gentler introduction of the tissue block and more uniformity in sections. Hairs will also often cause the block to cut unevenly and may result in separation from the OCT. As far as shaving I prefer an electric razor for my beard; however we had a lot of problems training the mice to use this. We have found that the best way to remove the hair from mouse skin is to use hard backed industrial blades after the skin has been removed from the animal. These blades are less likely to dig into the skin and cause damage than the regular sharper razor blades. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Nige Sent: Monday, November 21, 2005 4:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Adult Mouse Skin Cryosections Hi, I'm having trouble sectioning adult mouse skin prepared for cryosections. The dorsal skin has been taken freshly and put into plastic dispomoulds with Tissue Tek O.C.T, orientated and then slowly frozen by placing on a metal block which is nearly submerged in liquid nitrogen. The sample freezes nicely with no cracks, but when it comes to sectioning on a cryostat the tissue tends to scrunch up and come away from the tissue tek on the fatty side. Shaving the skin had no effect on the outcome. I've tried lots of different temperatures/section thickness but no luck. I'm assuming the problem has something to do with the fatty layer that is present with the skin, but it is very difficult (if not impossible) to separate this from the skin. Can anyone suggest how to get nice adult skin cryosections, any other processing needed to be done before mounting in O.C.T? Many thanks, nige _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Nov 21 17:26:50 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Nov 21 17:27:08 2005 Subject: [Histonet] Adult Mouse Skin Cryosections In-Reply-To: <001d01c5eee7$eb3f08c0$af82c050@cheekynige> References: <001d01c5eee7$eb3f08c0$af82c050@cheekynige> Message-ID: <6.0.0.22.1.20051121161526.01b46c18@gemini.msu.montana.edu> Nige How slowly are you freezing? The freezing method you describe is not really a slow freeze and should work perfectly as a snap freezing method. If it IS slow, then make sure the liquid nitrogen is up almost to top of metal block but NOT over the top of the metal block. We remove mouse skin (shaved or without much hair) and folded it into a v shape so the hair side is on the inside of the V, and then embedded the V on edge. The skin orientation is such - that the knife (very sharp, disposable blade - changed frequently for skin) passes through the soft layers first, and hair side last. If you do it the opposite direction, the layers separate worse as the outer layer tends to get scrunched up or pushed into the softer layers. Sectioning at -20C produced excellent 5 um thick sections especially if the skin is inflamed. By temperatures, which ones? And thicknesses? Good luck At 03:07 PM 11/21/2005, you wrote: >Hi, > >I'm having trouble sectioning adult mouse skin prepared for cryosections. >The dorsal skin has been taken freshly and put into plastic dispomoulds with >Tissue Tek O.C.T, orientated and then slowly frozen by placing on a metal >block which is nearly submerged in liquid nitrogen. The sample freezes >nicely with no cracks, but when it comes to sectioning on a cryostat the >tissue tends to scrunch up and come away from the tissue tek on the fatty >side. Shaving the skin had no effect on the outcome. I've tried lots of >different temperatures/section thickness but no luck. > >I'm assuming the problem has something to do with the fatty layer that is >present with the skin, but it is very difficult (if not impossible) to >separate this from the skin. Can anyone suggest how to get nice adult skin >cryosections, any other processing needed to be done before mounting in >O.C.T? > >Many thanks, > > >nige > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From heyitsra <@t> gmail.com Mon Nov 21 19:59:45 2005 From: heyitsra <@t> gmail.com (Ra) Date: Mon Nov 21 19:59:54 2005 Subject: [Histonet] Seeking histology art Message-ID: <358e3b6c0511211759k69f4dc4ej28dbd3aa7e500bb4@mail.gmail.com> Does anyone out there in HistoLand know where I could purchase any kind of artwork relating to Histology? I'm not picky - printed photographs that I could have framed, posters, or any type of artwork related to the field would suffice. I would appreciate any input. Thanks, Rhonda ******************************* Make a fast friend, ADOPT A GREYHOUND ******************************* From ohenry <@t> dfw.net Mon Nov 21 21:12:39 2005 From: ohenry <@t> dfw.net (Susan Owens) Date: Mon Nov 21 21:13:04 2005 Subject: [Histonet] equipment(used) Message-ID: <000501c5ef12$9d3f0550$79dd3040@your4f1261a8e5> I was on eBAY and saw 5 used, working they say, "FISHER SCIENTIFIC MAGNETIC STIRRER". They look in great shape. There are two models. 3 units - Model #11-500-7S, ceramic top, 120 vac,18W, 50/60 Hz, 0.3 amps(Red color) 2 units - Model # 11-500-16S,ceramic top, 120 vac, 60 Hz, 0.4 amps(Light gray or white color) You can find them in.... eBay Stores/Reliable Tool Store, then type in fisher scientific. Auction ends Wed.Nov.23 at 19:50PST Right now they are at $11.50 each....How high can they go? Susan From nige.hammond <@t> blueyonder.co.uk Tue Nov 22 02:03:26 2005 From: nige.hammond <@t> blueyonder.co.uk (Nige) Date: Tue Nov 22 02:03:25 2005 Subject: [Histonet] Adult Mouse Skin Cryosections In-Reply-To: <6.0.0.22.1.20051121161526.01b46c18@gemini.msu.montana.edu> Message-ID: <000201c5ef3b$37ef3660$af82c050@cheekynige> Hi Gayle, Thanks for your reply. Yeah it is more a snap freezing method. Is is better for sectioning to freeze it quicker or slower? I have been orientating dorsal skin side on for longitudinal hair follicle sections but have not being folding it. With the folding technique, do you press the hairy sides together or is it better to leave a gap between the two sides of the tissue? The cryostat settings have been: chamber temp -30, operating temp -20. I've been trying 7um sections. Many thanks, nige -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 21 November 2005 23:27 To: Nige; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Adult Mouse Skin Cryosections Nige How slowly are you freezing? The freezing method you describe is not really a slow freeze and should work perfectly as a snap freezing method. If it IS slow, then make sure the liquid nitrogen is up almost to top of metal block but NOT over the top of the metal block. We remove mouse skin (shaved or without much hair) and folded it into a v shape so the hair side is on the inside of the V, and then embedded the V on edge. The skin orientation is such - that the knife (very sharp, disposable blade - changed frequently for skin) passes through the soft layers first, and hair side last. If you do it the opposite direction, the layers separate worse as the outer layer tends to get scrunched up or pushed into the softer layers. Sectioning at -20C produced excellent 5 um thick sections especially if the skin is inflamed. By temperatures, which ones? And thicknesses? Good luck At 03:07 PM 11/21/2005, you wrote: >Hi, > >I'm having trouble sectioning adult mouse skin prepared for >cryosections. The dorsal skin has been taken freshly and put into >plastic dispomoulds with Tissue Tek O.C.T, orientated and then slowly >frozen by placing on a metal block which is nearly submerged in liquid >nitrogen. The sample freezes nicely with no cracks, but when it comes >to sectioning on a cryostat the tissue tends to scrunch up and come >away from the tissue tek on the fatty side. Shaving the skin had no >effect on the outcome. I've tried lots of different >temperatures/section thickness but no luck. > >I'm assuming the problem has something to do with the fatty layer that >is present with the skin, but it is very difficult (if not impossible) >to separate this from the skin. Can anyone suggest how to get nice >adult skin cryosections, any other processing needed to be done before >mounting in O.C.T? > >Many thanks, > > >nige > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From Kemlo.Rogerson <@t> elht.nhs.uk Tue Nov 22 03:12:04 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Tue Nov 22 03:12:19 2005 Subject: [Histonet] oil red o Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9EB236FF@elht-exch1.xelht.nhs.uk> Um nope, not even near. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: 21 November 2005 17:07 To: Histonet Subject: Re: [Histonet] oil red o The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound) would be someone (or something) who irritates someone else. Is there another meaning? Bryan ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Kristen Broomall" ; "Sally Prouty" ; Sent: Monday, November 21, 2005 8:28 AM Subject: RE: [Histonet] oil red o "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ 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 Kemlo.Rogerson <@t> elht.nhs.uk Tue Nov 22 03:21:11 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Tue Nov 22 03:21:21 2005 Subject: [Histonet] oil red o Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9EB23701@elht-exch1.xelht.nhs.uk> PITA's? Professional International Technical Advisors? Well thank you so much, it's nice for our talents to be recognised. But I'm still confused over who is buggering whom (note correct use of grammar). -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: 21 November 2005 19:25 To: 'Bryan Llewellyn'; Histonet Subject: RE: [Histonet] oil red o You guys are PITA's. (Do I need to spell that one out?) Bryan got the meaning exactly. (Good Job!) As in, I hate coverslipping ORO's because you can't press on the slip to get air bubbles out because the ORO will move. It's irritating. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bryan Llewellyn Sent: Monday, November 21, 2005 12:07 PM To: Histonet Subject: Re: [Histonet] oil red o The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound) would be someone (or something) who irritates someone else. Is there another meaning? Bryan ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Kristen Broomall" ; "Sally Prouty" ; Sent: Monday, November 21, 2005 8:28 AM Subject: RE: [Histonet] oil red o "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ 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 oshel1pe <@t> cmich.edu Tue Nov 22 07:06:41 2005 From: oshel1pe <@t> cmich.edu (Oshel, Philip Eugene) Date: Tue Nov 22 07:07:18 2005 Subject: [Histonet] oil red o & phrases Message-ID: <07BD45BB3EE8A0468C94776E7A0C660E3A3067@cmail4.central.cmich.local> Nope, spot on. The difference is British vs. the purer form of English we know over on this side of the puddle. This difference possibly comes from days spent as a youth in British Public Schools. Phil Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rogerson Kemlo (ELHT) Pathology Sent: Tue 05/11/22 04:12 To: Bryan Llewellyn; Histonet Subject: RE: [Histonet] oil red o Um nope, not even near. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: 21 November 2005 17:07 To: Histonet Subject: Re: [Histonet] oil red o The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound) would be someone (or something) who irritates someone else. Is there another meaning? Bryan ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Kristen Broomall" ; "Sally Prouty" ; Sent: Monday, November 21, 2005 8:28 AM Subject: RE: [Histonet] oil red o "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Tue Nov 22 07:27:40 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Tue Nov 22 07:28:04 2005 Subject: [Histonet] oil red o & phrases Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9EBBC196@elht-exch1.xelht.nhs.uk> Um no, still wrong. You need to explore the UK accepted meaning of the term rather than it being a derivative of 'bug'; I concede bunging a 'ger; may lead you to think that it's someone who bugs you but in the UK it has a more anal meaning. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Oshel, Philip Eugene Sent: 22 November 2005 13:07 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o & phrases Nope, spot on. The difference is British vs. the purer form of English we know over on this side of the puddle. This difference possibly comes from days spent as a youth in British Public Schools. Phil Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rogerson Kemlo (ELHT) Pathology Sent: Tue 05/11/22 04:12 To: Bryan Llewellyn; Histonet Subject: RE: [Histonet] oil red o Um nope, not even near. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: 21 November 2005 17:07 To: Histonet Subject: Re: [Histonet] oil red o The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound) would be someone (or something) who irritates someone else. Is there another meaning? Bryan ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Kristen Broomall" ; "Sally Prouty" ; Sent: Monday, November 21, 2005 8:28 AM Subject: RE: [Histonet] oil red o "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ 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 oshel1pe <@t> cmich.edu Tue Nov 22 07:38:59 2005 From: oshel1pe <@t> cmich.edu (Oshel, Philip Eugene) Date: Tue Nov 22 07:39:35 2005 Subject: [Histonet] oil red o & phrases Message-ID: <07BD45BB3EE8A0468C94776E7A0C660E3A306A@cmail4.central.cmich.local> I meant what you mean ... why else refer to Public Schools? "We have really everything in common with America nowadays except, of course, language. Oscar Wilde (1854 - 1900), The Canterville Ghost, 1882" -----Original Message----- From: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: Tue 05/11/22 08:27 To: Oshel, Philip Eugene; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o & phrases Um no, still wrong. You need to explore the UK accepted meaning of the term rather than it being a derivative of 'bug'; I concede bunging a 'ger; may lead you to think that it's someone who bugs you but in the UK it has a more anal meaning. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Oshel, Philip Eugene Sent: 22 November 2005 13:07 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o & phrases Nope, spot on. The difference is British vs. the purer form of English we know over on this side of the puddle. This difference possibly comes from days spent as a youth in British Public Schools. Phil Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rogerson Kemlo (ELHT) Pathology Sent: Tue 05/11/22 04:12 To: Bryan Llewellyn; Histonet Subject: RE: [Histonet] oil red o Um nope, not even near. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: 21 November 2005 17:07 To: Histonet Subject: Re: [Histonet] oil red o The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound) would be someone (or something) who irritates someone else. Is there another meaning? Bryan ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Kristen Broomall" ; "Sally Prouty" ; Sent: Monday, November 21, 2005 8:28 AM Subject: RE: [Histonet] oil red o "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ 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 DDittus787 <@t> aol.com Tue Nov 22 08:04:51 2005 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Tue Nov 22 08:05:33 2005 Subject: [Histonet] Seeking histology art Message-ID: <201.e4b3aeb.30b47f83@aol.com> sturkey has some beautiful histology art work dana From BennettW <@t> pac.dfo-mpo.gc.ca Tue Nov 22 10:06:06 2005 From: BennettW <@t> pac.dfo-mpo.gc.ca (BennettW@pac.dfo-mpo.gc.ca) Date: Tue Nov 22 10:03:54 2005 Subject: [Histonet] Quality Management Systems Message-ID: <7CBBD627E4E688499349A5D11D07831603130923@msgpacpbs.pac.dfo-mpo.ca> Hi to all in histoland, Are there any histo labs out there that are ISO 17025 accredited? We are starting to go through the process here and we were wondering if anyone with this accreditation could provide some insight into the process. Thanks for your time. Cheers Bill Bennett Fisheries and Oceans Canada From brett_connolly <@t> merck.com Tue Nov 22 10:18:09 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Tue Nov 22 10:18:54 2005 Subject: [Histonet] anti- Golgi antibodies for FFPE sections Message-ID: <355C35514FEAC9458F75947F5270974D07971C@usctmx1103.merck.com> Any recommendations for IHC ? Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ 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 MICHAEL.OWEN <@t> fda.gov Tue Nov 22 10:22:52 2005 From: MICHAEL.OWEN <@t> fda.gov (Owen, Michael P) Date: Tue Nov 22 10:24:05 2005 Subject: [Histonet] Quality Management Systems Message-ID: Dear Dr. Bennett, My laboratory obtained ISO 17025 status this spring. The facility is not a histology laboratory but some of the work does require histochemical techniques. Feel free to e-mail me so we can compare notes on preparation and maintaining the system once a facility is accredited. Wishing you a successful qualification audit, The FDA Lab Rat Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From liz <@t> premierlab.com Tue Nov 22 10:26:29 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Tue Nov 22 10:26:44 2005 Subject: [Histonet] Seeking histology art In-Reply-To: <358e3b6c0511211759k69f4dc4ej28dbd3aa7e500bb4@mail.gmail.com> Message-ID: <001001c5ef81$7e4d9340$a7d48a80@AMY> Rhonda Damian Laudier has beautiful images of histology sections that he has printed on cards, his e-mail is dmlaud@hotmail.com I saw some of his cards at NSH this year, they were stunning. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ra Sent: Monday, November 21, 2005 7:00 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Seeking histology art Does anyone out there in HistoLand know where I could purchase any kind of artwork relating to Histology? I'm not picky - printed photographs that I could have framed, posters, or any type of artwork related to the field would suffice. I would appreciate any input. Thanks, Rhonda ******************************* Make a fast friend, ADOPT A GREYHOUND ******************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TillRenee <@t> uams.edu Tue Nov 22 10:44:27 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Tue Nov 22 10:44:52 2005 Subject: [Histonet] weak antibody signal Message-ID: We have a BTEB1 goat polyclonal antibody that is staining very weakly using a Vectastain kit and DAB. I personally like to order my secondaries and streptavidin separate, that way I can get F(ab')2 fragements or what have you, if I want. I was going to suggest trying this, as it is has worked better for me on some stains than the Vector kits. Is there anything else that could be done to boost the signal? I had thought of CSA kits and the like, but from what I can tell the Dako kit at least isn't for goat. Would it work better perhaps using fluorescence instead of DAB? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ From llewllew <@t> shaw.ca Tue Nov 22 10:48:34 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Tue Nov 22 10:49:14 2005 Subject: [Histonet] oil red o Message-ID: <000901c5ef84$94499920$690a4246@yourlk4rlmsu> I think an American would use an accepted American meaning of the term rather than a British one, but even in Britain "Bugger" doesn't always mean someone who engages in anal intercourse. It also has the meaning of "fool" or "pitiable person", as a synonym for "poor sod" for example, or something like that. The original user was from the US, so presumably she used one of that dialect's definitions. I originally derived it from "bug" with tongue in cheek, but since then I have looked it up in dictionaries and got the following. It seems that the use to mean "something annoying" is actually derived from "bug" referring to an insect, so I was (quite unintentially) spot on. There is also a third meaning. Do a Google search on "dictionary" and check the meaning in some of the ones that come up. Note Merriam-Webster's definition 1-3. Merrriam-Webster online dictionary Main Entry: 1 bug?ger Pronunciation: 'b&-g&r, 'bu-g&r Function: noun Etymology: Middle English bougre heretic, from Middle French, from Medieval Latin Bulgarus, literally, Bulgarian; from the association of Bulgaria with the Bogomils, who were accused of sodomy 1 : SODOMITE 2 a : a worthless person : RASCAL b : FELLOW, CHAP 3 : a small or annoying thing Main Entry: 2 bug?ger Pronunciation: 'b&-g&r Function: transitive verb 1 usually vulgar : to commit sodomy with 2 : DAMN Main Entry: 3 bugger Function: noun : a person who plants electronic bugs Bryan Llewellyn ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Oshel, Philip Eugene" ; Sent: Tuesday, November 22, 2005 5:27 AM Subject: RE: [Histonet] oil red o & phrases Um no, still wrong. You need to explore the UK accepted meaning of the term rather than it being a derivative of 'bug'; I concede bunging a 'ger; may lead you to think that it's someone who bugs you but in the UK it has a more anal meaning. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Oshel, Philip Eugene Sent: 22 November 2005 13:07 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o & phrases Nope, spot on. The difference is British vs. the purer form of English we know over on this side of the puddle. This difference possibly comes from days spent as a youth in British Public Schools. Phil Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rogerson Kemlo (ELHT) Pathology Sent: Tue 05/11/22 04:12 To: Bryan Llewellyn; Histonet Subject: RE: [Histonet] oil red o Um nope, not even near. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: 21 November 2005 17:07 To: Histonet Subject: Re: [Histonet] oil red o The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound) would be someone (or something) who irritates someone else. Is there another meaning? Bryan ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Kristen Broomall" ; "Sally Prouty" ; Sent: Monday, November 21, 2005 8:28 AM Subject: RE: [Histonet] oil red o "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ 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 Tue Nov 22 10:37:16 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Tue Nov 22 10:49:32 2005 Subject: [Histonet] Seeking histology art Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D3A1@usca0082k08.labvision.apogent.com> Fisher scientific has hundreds of science posters, many relating to biology. Also, Ebay has a huge listing. Many of the companies have posters or calenders of various aspect of histology. My favorite was a print series from Fisher of old-world paintings of different biology and chemistry fields - The Alchemist, The Anatomist, etc. I don't know if they still carry those. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ra Sent: Monday, November 21, 2005 6:00 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Seeking histology art Does anyone out there in HistoLand know where I could purchase any kind of artwork relating to Histology? I'm not picky - printed photographs that I could have framed, posters, or any type of artwork related to the field would suffice. I would appreciate any input. Thanks, Rhonda ******************************* Make a fast friend, ADOPT A GREYHOUND ******************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kbroomal <@t> NEMOURS.ORG Tue Nov 22 12:30:41 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Tue Nov 22 12:31:06 2005 Subject: [Histonet] Bugger - originally ORO Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824499@wlmmsx02.nemours.org> Are all of you really that bored? Do I need to clarify this AGAIN? (I already said that Bryan's original response was correct.) "The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound)would be someone (or something) who irritates someone else." When you bug someone (about anything, like asking similar questions over & over again), you are being annoying or irritating. Like a bug flying around your head & buzzing in your ear would be annoying. So if something is annoying, it's a bugger. Is this really that difficult? Kristen aka. "the original user" -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bryan Llewellyn Sent: Tuesday, November 22, 2005 11:49 AM To: Histonet Subject: Re: [Histonet] oil red o I think an American would use an accepted American meaning of the term rather than a British one, but even in Britain "Bugger" doesn't always mean someone who engages in anal intercourse. It also has the meaning of "fool" or "pitiable person", as a synonym for "poor sod" for example, or something like that. The original user was from the US, so presumably she used one of that dialect's definitions. I originally derived it from "bug" with tongue in cheek, but since then I have looked it up in dictionaries and got the following. It seems that the use to mean "something annoying" is actually derived from "bug" referring to an insect, so I was (quite unintentially) spot on. There is also a third meaning. Do a Google search on "dictionary" and check the meaning in some of the ones that come up. Note Merriam-Webster's definition 1-3. Merrriam-Webster online dictionary Main Entry: 1 bug?ger Pronunciation: 'b&-g&r, 'bu-g&r Function: noun Etymology: Middle English bougre heretic, from Middle French, from Medieval Latin Bulgarus, literally, Bulgarian; from the association of Bulgaria with the Bogomils, who were accused of sodomy 1 : SODOMITE 2 a : a worthless person : RASCAL b : FELLOW, CHAP 3 : a small or annoying thing Main Entry: 2 bug?ger Pronunciation: 'b&-g&r Function: transitive verb 1 usually vulgar : to commit sodomy with 2 : DAMN Main Entry: 3 bugger Function: noun : a person who plants electronic bugs Bryan Llewellyn ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Oshel, Philip Eugene" ; Sent: Tuesday, November 22, 2005 5:27 AM Subject: RE: [Histonet] oil red o & phrases Um no, still wrong. You need to explore the UK accepted meaning of the term rather than it being a derivative of 'bug'; I concede bunging a 'ger; may lead you to think that it's someone who bugs you but in the UK it has a more anal meaning. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Oshel, Philip Eugene Sent: 22 November 2005 13:07 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o & phrases Nope, spot on. The difference is British vs. the purer form of English we know over on this side of the puddle. This difference possibly comes from days spent as a youth in British Public Schools. Phil Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rogerson Kemlo (ELHT) Pathology Sent: Tue 05/11/22 04:12 To: Bryan Llewellyn; Histonet Subject: RE: [Histonet] oil red o Um nope, not even near. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: 21 November 2005 17:07 To: Histonet Subject: Re: [Histonet] oil red o The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound) would be someone (or something) who irritates someone else. Is there another meaning? Bryan ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Kristen Broomall" ; "Sally Prouty" ; Sent: Monday, November 21, 2005 8:28 AM Subject: RE: [Histonet] oil red o "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ 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 Matthew_Frank <@t> URMC.Rochester.edu Tue Nov 22 12:41:36 2005 From: Matthew_Frank <@t> URMC.Rochester.edu (Frank, Matthew) Date: Tue Nov 22 12:41:50 2005 Subject: [Histonet] Bugger - originally ORO Message-ID: UNSUBSCRIBE PLEASE -----Original Message----- From: Kristen Broomall [mailto:kbroomal@NEMOURS.ORG] Sent: Tuesday, November 22, 2005 1:31 PM To: Histonet Subject: [Histonet] Bugger - originally ORO Are all of you really that bored? Do I need to clarify this AGAIN? (I already said that Bryan's original response was correct.) "The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound)would be someone (or something) who irritates someone else." When you bug someone (about anything, like asking similar questions over & over again), you are being annoying or irritating. Like a bug flying around your head & buzzing in your ear would be annoying. So if something is annoying, it's a bugger. Is this really that difficult? Kristen aka. "the original user" -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bryan Llewellyn Sent: Tuesday, November 22, 2005 11:49 AM To: Histonet Subject: Re: [Histonet] oil red o I think an American would use an accepted American meaning of the term rather than a British one, but even in Britain "Bugger" doesn't always mean someone who engages in anal intercourse. It also has the meaning of "fool" or "pitiable person", as a synonym for "poor sod" for example, or something like that. The original user was from the US, so presumably she used one of that dialect's definitions. I originally derived it from "bug" with tongue in cheek, but since then I have looked it up in dictionaries and got the following. It seems that the use to mean "something annoying" is actually derived from "bug" referring to an insect, so I was (quite unintentially) spot on. There is also a third meaning. Do a Google search on "dictionary" and check the meaning in some of the ones that come up. Note Merriam-Webster's definition 1-3. Merrriam-Webster online dictionary Main Entry: 1 bug?ger Pronunciation: 'b&-g&r, 'bu-g&r Function: noun Etymology: Middle English bougre heretic, from Middle French, from Medieval Latin Bulgarus, literally, Bulgarian; from the association of Bulgaria with the Bogomils, who were accused of sodomy 1 : SODOMITE 2 a : a worthless person : RASCAL b : FELLOW, CHAP 3 : a small or annoying thing Main Entry: 2 bug?ger Pronunciation: 'b&-g&r Function: transitive verb 1 usually vulgar : to commit sodomy with 2 : DAMN Main Entry: 3 bugger Function: noun : a person who plants electronic bugs Bryan Llewellyn ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Oshel, Philip Eugene" ; Sent: Tuesday, November 22, 2005 5:27 AM Subject: RE: [Histonet] oil red o & phrases Um no, still wrong. You need to explore the UK accepted meaning of the term rather than it being a derivative of 'bug'; I concede bunging a 'ger; may lead you to think that it's someone who bugs you but in the UK it has a more anal meaning. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Oshel, Philip Eugene Sent: 22 November 2005 13:07 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o & phrases Nope, spot on. The difference is British vs. the purer form of English we know over on this side of the puddle. This difference possibly comes from days spent as a youth in British Public Schools. Phil Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rogerson Kemlo (ELHT) Pathology Sent: Tue 05/11/22 04:12 To: Bryan Llewellyn; Histonet Subject: RE: [Histonet] oil red o Um nope, not even near. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: 21 November 2005 17:07 To: Histonet Subject: Re: [Histonet] oil red o The ending "er" in English denotes a person (or thing) who/which does an action. To "bug" is a colloquial expression meaning to irritate, so a "bugger" (the letter g is doubled to preserve the short vowel sound) would be someone (or something) who irritates someone else. Is there another meaning? Bryan ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Kristen Broomall" ; "Sally Prouty" ; Sent: Monday, November 21, 2005 8:28 AM Subject: RE: [Histonet] oil red o "Quite a bugger" I quote. Do you know what one is? It doesn't seem to fit the context in which you are using it. Are you using it as a noun or a verb? I'm sure you aren't talking about Sally. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Monday, November 21, 2005 4:19 PM To: 'Sally Prouty'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red o Make sure that you are not pressing (at all!) on your coverslip, even to get out air bubbles. The ORO will come right out of the fat cells. Quite a bugger. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sally Prouty Sent: Sunday, November 20, 2005 11:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] oil red o I am trying to locate a protocol for a consistant Oil red O stain. I don't know if it is in coverslipping where the stain is moving off the fat cells or in my differentiation step. I am staining on frozen skeletal muscle sections. Any suggestions would be helpful. Thank you, Sally Sally J. Prouty HHMI/University of Iowa Kevin Campbell Laboratory 400 EMRB Iowa City, Iowa 52246 319-335-6944 _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ 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 shive003 <@t> umn.edu Tue Nov 22 12:44:38 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Tue Nov 22 12:44:48 2005 Subject: [Histonet] weak antibody signal References: Message-ID: <006b01c5ef94$cb102b80$41065486@auxs.umn.edu> DAKO sells an LSAB+ kit (labeled streptavidin-biotin-HRP) that is against mouse, rabbit, AND goat IgGs. I use it for my goat primary antibodies and it works well. Jan Shivers U of MN Vet Diag Lab shive003@umn.edu ----- Original Message ----- From: "Till, Renee" To: Sent: Tuesday, November 22, 2005 10:44 AM Subject: [Histonet] weak antibody signal We have a BTEB1 goat polyclonal antibody that is staining very weakly using a Vectastain kit and DAB. I personally like to order my secondaries and streptavidin separate, that way I can get F(ab')2 fragements or what have you, if I want. I was going to suggest trying this, as it is has worked better for me on some stains than the Vector kits. Is there anything else that could be done to boost the signal? I had thought of CSA kits and the like, but from what I can tell the Dako kit at least isn't for goat. Would it work better perhaps using fluorescence instead of DAB? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 ============================================================================ ==================== Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ============================================================================ ==================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From doninn <@t> mail.nih.gov Tue Nov 22 14:25:48 2005 From: doninn <@t> mail.nih.gov (Donin, Nick (NIH/NCI)) Date: Tue Nov 22 14:26:05 2005 Subject: [Histonet] mouse nestin Message-ID: <16A0583FB1644E4DB8C0A0265028B6FD02B4E84D@nihexchange13.nih.gov> Hey all, Does anyone know of a good anti-mouse-nestin antibody for frozen sections. (i.e. an antibody that will detect the nestin protein produced in mouse neuro-stem-cells). I'm trying the Chemicon MAB353 and not really having much luck. Thanks, hope all is well, happy holidays. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 From steven.p.postl <@t> abbott.com Tue Nov 22 15:40:02 2005 From: steven.p.postl <@t> abbott.com (Steven P Postl) Date: Tue Nov 22 15:40:31 2005 Subject: [Histonet] Calcium control slides Message-ID: I am looking for a company that sells calcium control slides. Any recommendations? From Djemge <@t> aol.com Wed Nov 23 01:56:55 2005 From: Djemge <@t> aol.com (Djemge@aol.com) Date: Wed Nov 23 01:57:07 2005 Subject: [Histonet] IEC Minotome & Reichart 855 Cryostat Manuals Needed Help! Message-ID: <288.37976b.30b57ac7@aol.com> Hello all. Does anyone have a manual for a Damon / IEC Minotome model Mino Analog cryostat (IM-175 rev 1 I believe is the manual I need for the IEC) and a Reichart 855 cryostat. A pdf file attachment of these full manuals to my e-mail address will be fine. I am willing to pay for a copy, pdf, or original. I need them for our labs CLIA inspection ASAP. Thank you for your help. Donna Emge, HT-ASCP 4111 Clinton Ave Stickney, IL 60402 _djemge@aol.com_ (mailto:djemge@aol.com) 708-788-1694 W 312-503-2036 (9 to 5) From pruegg <@t> ihctech.net Wed Nov 23 09:36:30 2005 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Wed Nov 23 09:36:51 2005 Subject: [Histonet] weak antibody signal In-Reply-To: <006b01c5ef94$cb102b80$41065486@auxs.umn.edu> Message-ID: <200511231536.jANFaTGV036438@pro12.abac.com> What I do for goat primaries is to use a rab. Anti-goat secondary and then apply the rabbit labeled polymer (I use DAko Envision), it works a charm, with a very strong signal. Invitrogen sent me a goat labeled polymer to try but it did not do well in my hands. Dako has been promising me goat labeled polymer for years but so far I haven't seen it, most of these companies seem to still be more interested in clinical than research IHC. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Tuesday, November 22, 2005 11:45 AM To: Till, Renee Cc: histonet Subject: Re: [Histonet] weak antibody signal DAKO sells an LSAB+ kit (labeled streptavidin-biotin-HRP) that is against mouse, rabbit, AND goat IgGs. I use it for my goat primary antibodies and it works well. Jan Shivers U of MN Vet Diag Lab shive003@umn.edu ----- Original Message ----- From: "Till, Renee" To: Sent: Tuesday, November 22, 2005 10:44 AM Subject: [Histonet] weak antibody signal We have a BTEB1 goat polyclonal antibody that is staining very weakly using a Vectastain kit and DAB. I personally like to order my secondaries and streptavidin separate, that way I can get F(ab')2 fragements or what have you, if I want. I was going to suggest trying this, as it is has worked better for me on some stains than the Vector kits. Is there anything else that could be done to boost the signal? I had thought of CSA kits and the like, but from what I can tell the Dako kit at least isn't for goat. Would it work better perhaps using fluorescence instead of DAB? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 ============================================================================ ==================== Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ============================================================================ ==================== _______________________________________________ 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 Wed Nov 23 10:29:36 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 23 10:29:56 2005 Subject: Well said on RE: [Histonet] weak antibody signal In-Reply-To: <200511231536.jANFaTGV036438@pro12.abac.com> References: <006b01c5ef94$cb102b80$41065486@auxs.umn.edu> <200511231536.jANFaTGV036438@pro12.abac.com> Message-ID: <6.0.0.22.1.20051123092441.01b6bcc0@gemini.msu.montana.edu> Patsy, You couldn't have said it better!!!! A rat kit i.e. for rat antimouse antibody IHC would be nice too and still on my hopechest/wish list. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 At 08:36 AM 11/23/2005, you wrote: >What I do for goat primaries is to use a rab. Anti-goat secondary and then >apply the rabbit labeled polymer (I use DAko Envision), it works a charm, >with a very strong signal. Invitrogen sent me a goat labeled polymer to try >but it did not do well in my hands. Dako has been promising me goat labeled >polymer for years but so far I haven't seen it, most of these companies seem >to still be more interested in clinical than research IHC. >Patsy > >Patsy Ruegg, HT(ASCP)QIHC >IHCtech >12635 Montview Blvd. #216 >Aurora, CO 80010 >720-859-4060 >fax 720-859-4110 >pruegg@ihctech.net >www.ihctech.net > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers >Sent: Tuesday, November 22, 2005 11:45 AM >To: Till, Renee >Cc: histonet >Subject: Re: [Histonet] weak antibody signal > >DAKO sells an LSAB+ kit (labeled streptavidin-biotin-HRP) that is against >mouse, rabbit, AND goat IgGs. I use it for my goat primary antibodies and >it works well. > >Jan Shivers >U of MN Vet Diag Lab >shive003@umn.edu > >----- Original Message ----- >From: "Till, Renee" >To: >Sent: Tuesday, November 22, 2005 10:44 AM >Subject: [Histonet] weak antibody signal > > >We have a BTEB1 goat polyclonal antibody that is staining very weakly >using a Vectastain kit and DAB. I personally like to order my >secondaries and streptavidin separate, that way I can get F(ab')2 >fragements or what have you, if I want. I was going to suggest trying >this, as it is has worked better for me on some stains than the Vector >kits. Is there anything else that could be done to boost the signal? I >had thought of CSA kits and the like, but from what I can tell the Dako >kit at least isn't for goat. Would it work better perhaps using >fluorescence instead of DAB? > > > >Renee' Till, HT > > > >Arkansas Childrens Nutrition Center > >1120 Marshall > >Little Rock, AR > >72202 > > > >(501) 364-2774 > > > > >============================================================================ >==================== > >Confidentiality Notice: This e-mail message, including any attachments, is >for the sole use of the intended recipient(s) and may contain confidential >and privileged information. Any unauthorized review, use, disclosure or >distribution is prohibited. If you are not the intended recipient, please >contact the sender by reply e-mail and destroy all copies of the original >message. >============================================================================ >==================== >_______________________________________________ >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 SAllen <@t> exchange.hsc.mb.ca Wed Nov 23 10:57:36 2005 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Wed Nov 23 10:58:43 2005 Subject: [Histonet] Gallyas method Message-ID: <36FE435D6D2F1D489174B22362A961B66B82F5@hscxntmx0006.hsc.mb.ca> Hi, Does anyone have a good, reliable method for Gallyas Stain? I would appreciate a copy of it. The two methods I have tried haven't been great & I'm really short on time to troubleshoot the method. Thanks for any help you can give me. Sharon sallen@hsc.mb.ca 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 cbass <@t> bidmc.harvard.edu Wed Nov 23 11:02:14 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Wed Nov 23 11:04:25 2005 Subject: [Histonet] best perfusion solution for spinal cord Message-ID: <00F3EA73-9172-4EE6-B0D3-C6CAA04DFDEE@bidmc.harvard.edu> Hello, I was hoping you guys could help me with something. I have a friend who is using rat spinal cord injury models. He needs a good method for fixing the spinal cord to examine both morphology of the spinal cord tissue and various proteins by IHC. What is the best method of fixation for this? I assume that what is best for the brain is usually best for the spinal cord. I recently read that some use a sucrose solution in their perfusion protocol. I have always just dropped the brain into 25% sucrose overnight to cryoprotect. I never considered using it in the perfusion. Any help would be appreciated. I am particularly interested in which formulations of fixatives are best and why. For example, could NBF be used? I mostly see paraformaldehyde in use, usually 2 or 4%. What are the advantages of this over NBF? Thanks, Caroline Bass From jfish <@t> gladstone.ucsf.edu Wed Nov 23 11:45:50 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Wed Nov 23 11:46:08 2005 Subject: [Histonet] Gallyas method In-Reply-To: <36FE435D6D2F1D489174B22362A961B66B82F5@hscxntmx0006.hsc.mb.ca> Message-ID: <000001c5f055$be12b2b0$8903010a@JFISH> Good morning, Sharon, I use the method from Bancroft and Stevens, if you can't get your hands on the book, let me know and I will send the protocol. The most important factor, I stress "important", is cleaning your glassware before doing the stain, acid wash with concentrated nitric acid. If you don't your slides will stain unevenly, if at all! Bancroft, JD, et al. Theory and Practice of Histological Techniques. New York:Churchill Livingstone; 1996:365-366. Good luck, Jo Dee Fish -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Allen Sent: Wednesday, November 23, 2005 8:58 AM To: Histonet (E-mail) Subject: [Histonet] Gallyas method Hi, Does anyone have a good, reliable method for Gallyas Stain? I would appreciate a copy of it. The two methods I have tried haven't been great & I'm really short on time to troubleshoot the method. Thanks for any help you can give me. Sharon sallen@hsc.mb.ca 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 jengirl1014 <@t> yahoo.com Wed Nov 23 12:18:30 2005 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Wed Nov 23 12:18:40 2005 Subject: [Histonet] Happy Thanksgiving Message-ID: <20051123181830.17042.qmail@web60615.mail.yahoo.com> To all who celebrate and to those who don't, May you all have a very Happy Thanksgiving and eat lots of turkey......don't worry about dissecting it! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 410-955-9688 e-mail: jengirl1014@yahoo.com --------------------------------- Yahoo! FareChase - Search multiple travel sites in one click. From lsunhwa <@t> gmail.com Wed Nov 23 12:37:16 2005 From: lsunhwa <@t> gmail.com (Sunhwa Lee) Date: Wed Nov 23 12:37:25 2005 Subject: [Histonet] zebrafish brain fixative Message-ID: <177172430511231037r2754e0f9nf6b1eddb8f519fc9@mail.gmail.com> Hello, I am asking you guys could help me with fixative for zebra fish brain. I am working on zebra fish brain, I used to use 4% PFA and AFA (alcohol formalin Acetic acid). So far, AFA works better than PFA. And there is a woman who is using PFA for her zebra fish arbor in my lab, and PFA seems to work very well for her zebra fish arbor. But I cannot understand why 4% PFA does not work with zebra fish brain. Actually AFA smells stinky and very toxic so sometimes, it makes nausea. Does anyone who has experience with zebra fish brain? I never work with zebra fish arbor so I cannot tell the difference, but she said it works well. Any help would be appreciated. I am particularly interested in which formulations of fixative are the best for zebra fish brain and why. In advance, thank you so much. Sunny From billingconsultants <@t> yahoo.com Wed Nov 23 12:42:41 2005 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Wed Nov 23 12:42:50 2005 Subject: [Histonet] Burnt tissue Message-ID: <20051123184241.13213.qmail@web54213.mail.yahoo.com> Hi everyone, Does anyone have any suggestions as to how to fix tissue that has a burned appearance after processing? Any suggestions you may provide would be greatly appreciated. Happy Thanksgiving. Louri __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From dharclerode <@t> cytoritx.com Wed Nov 23 14:20:13 2005 From: dharclerode <@t> cytoritx.com (Donna Harclerode) Date: Wed Nov 23 14:20:31 2005 Subject: [Histonet] Nestin and perfusion for spinal cord Message-ID: <3DE0F644E093DF4BAE80C254176696A503CF89@mp-mailserver.macropore.com> We have had no success with Nestin from Chemicon (Neu N either) and would appreciate anyone having an antibody that works to share the info. We are currently trying on chamber slides so they should, but do not stain anything on the slides. We used 4%PFA (from concentrate for 30 minutes to fix human cells. Other antibodies work great. For perfusion I always used freshly prepared filtered 4%PFA in PBS unless there was a reason not to. I have also used the zinc formalin solutions with good results. I would not use NBF as it has methanol in it to stabilize the solution and the pH is not always controled. There is a concentrated solution of PFA (up to 32%) that you can get from Electron Microscopy Sciences that you can order through VWR that probably would be ok. I never let PFA solutions heat over 65oC (breakdown of aldehydes) and it is a time consuming and not so nice solution to make. Be sure to use prill (Electron Microscopy Sciences- very inexpensive and high quality) or granular if you are making the solution from scratch as the powdered PFA gives a lot of dust and gives you more exposure to the paraformaldehyde. Some antibodies preferred 2%PFA and a couple we used gluteraldehyde and PFA. I perfused with RT saline and then with cold PFA until the brain was white and the animal (rat, pigeon or chicken) was very stiff using the left ventricle with the right atria cut to allow the solution to leave the body. We left the brain and or spinal cord in the same fixative as the perfusion overnight and transferred to 30% sucrose in PBS and left it in the fridge for a couple more days. This worked great for the majority of antibodies on free floating sections. Lower concentrations of PFA gave me soft tissue that was more difficult to handle for staining and mounting. Thanks and good luck Donna Harclerode, HT, (ASCP), HTL, QIHC Immunohistochemist Cytori Therapeutics 3020 Callan Rd. San Diego, CA 92121 858-458-0900 ext 322 dharclerode@cytoritx.com From MKing <@t> PeaceHealth.org Wed Nov 23 14:49:21 2005 From: MKing <@t> PeaceHealth.org (King, Mary Ann) Date: Wed Nov 23 14:49:35 2005 Subject: [Histonet] Job Opportunity Message-ID: St. Joseph Hospital in Bellingham Washington has an immediate opening for a full time registered histology technician or histotechnologist. This person must be focused, able to conduct themselves in a professional manner and organize themselves and their work in such a way as to promote accuracy and efficiency. This person must be self motivated and able to assume a leadership role if necessary. Training and Licensure: AA/AS and HT/HTL Job Description includes: Embedding, microtomy, routine and special staining, assisting the pathologist in frozen sections, fine needle aspirations, helping cover path assisting duties including gross assisting, order entry, and reference lab send outs. Must be willing to help in assist in autopsies if necessary. The Pacific Northwest is a beautiful area to call home. Bellingham is located between the North Cascade Mountains and the shores of Puget Sound. We enjoy four distinct seasons with an overall mild climate, and an abundance of varied recreational opportunities. To access the hospital website: peacehealth.org To submit an application: jmcauley@peacehealth.org ------------------------------------------------------------___________________________________________________________ This message is intended solely for the 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 state and federal laws. If you are not the addressee, or are not authorized to receive for the intended addressee, you are hereby notified that you may not use, copy, distribute, or disclose to anyone this message or the information contained herein. If you have received this message in error, immediately advise the sender by reply email and destroy this message. From tahseen <@t> brain.net.pk Wed Nov 23 07:45:40 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Wed Nov 23 15:49:13 2005 Subject: [Histonet] Biopsy for Leishmania Message-ID: <00f501c5f034$321aa5e0$972bfea9@m7c0y4> Has anyone had any experience in performing stain for Leishmania (I have received fresh biopsy specimen)? Any opinions would be much appreciated. Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Lahore Pakistan. From c.m.vanderloos <@t> amc.uva.nl Thu Nov 24 02:08:24 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Thu Nov 24 02:08:40 2005 Subject: [Histonet] RE: weak antibody signal Message-ID: <14dd8be14e18b0.14e18b014dd8be@amc.uva.nl> Renee, I have three detection/visualization procedures for you to consider for testing (no. 3 is based on the assumption that you are intended to stain FFPE sections; no. 1/2 can be used for cryo's too): 1. Most simple is just to repeat what you did so far but ending with Vector NovaRed kit instead of DAB. This very sensitive kit has helped us in similar situations a couple of times here. 2. We have been pretty successful lately with the detection of goat antibodies using a 2-step method ending with a polymer. The second step was a rabbit anti-goat Ig (Dako) 1:5000, 30 min RT and the final step EnVision+ anti-rabbit. DAB+ was used for chromogen. 3. As you said the CSA kit might be a good solution too. In case of a goat antibody your second step should be rabbit anti-goat Ig/HRP (Dako) 1:100, 15 min, RT. Then continue with Amplification reagent, etc. Please realize that the reaction product after tyramide amplification is somewhat more diffusely localized than with other IHC detection procedures. In all three cases you should perform new dilution series of your primary. Good luck! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 Date: Tue, 22 Nov 2005 10:44:27 -0600 From: "Till, Renee" Subject: [Histonet] weak antibody signal To: histonet@lists.utsouthwestern.edu We have a BTEB1 goat polyclonal antibody that is staining very weakly using a Vectastain kit and DAB. I personally like to order my secondaries and streptavidin separate, that way I can get F(ab')2 fragements or what have you, if I want. I was going to suggest trying this, as it is has worked better for me on some stains than the Vector kits. Is there anything else that could be done to boost the signal? I had thought of CSA kits and the like, but from what I can tell the Dako kit at least isn't for goat. Would it work better perhaps using fluorescence instead of DAB? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR From carl.hobbs <@t> kcl.ac.uk Thu Nov 24 02:34:48 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Thu Nov 24 02:35:43 2005 Subject: [Histonet] re Nestin Message-ID: <000701c5f0d1$ee25c150$112b5c9f@Carlos> I use rat 401 anti Nestin AB for SVR progenitor cells in rat pwax sections, after HIER. It's very good. It also works well in mouse pwax. I have also used it in perfused fixed- frozen rat and mouse brains. Not in fresh-frozen tissue tho. Buy it from DSHB....$25/ml TCS. Excellent value Hope that helps I'll post a pic here: http://www.immunoportal.com/modules.php?set_albumName=album01&op=modload&name=Gallery&file=index&include=view_album.php From Kemlo.Rogerson <@t> elht.nhs.uk Thu Nov 24 02:37:37 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Thu Nov 24 02:37:47 2005 Subject: [Histonet] Burnt tissue Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9EB23714@elht-exch1.xelht.nhs.uk> Shouldn't you have fixed it before? Maybe that's why it appears 'burnt'. I think a little more detail could help. Type of tissue, type of primary fixation, etc, etc. Best wishes -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Billing Consultants, LLC Sent: 23 November 2005 18:43 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Burnt tissue Hi everyone, Does anyone have any suggestions as to how to fix tissue that has a burned appearance after processing? Any suggestions you may provide would be greatly appreciated. Happy Thanksgiving. Louri __________________________________________________ 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 lbeaudet <@t> ctcsurge.com Thu Nov 24 12:03:49 2005 From: lbeaudet <@t> ctcsurge.com (Laura Beaudet) Date: Thu Nov 24 12:07:36 2005 Subject: [Histonet] automated response Message-ID: <10511241303.AA10078@mail1.ctcsurge.com> Computer Trust Corporation is closed Thu-Sun Nov 24-27 2005 for Thanksgiving. From Robert.Fauck <@t> ccdhb.org.nz Thu Nov 24 18:17:19 2005 From: Robert.Fauck <@t> ccdhb.org.nz (Robert Fauck) Date: Thu Nov 24 18:18:05 2005 Subject: [Histonet] p16 Message-ID: Hi there! Is there anybody that has a good protocol to do the marker p16?! Any recommendation on the source for this p16 antibody? Thanking you all in advance for your advice. Cheers, Robert Fauck NZ, Wellington Hospital _______________________________________________________________________ This email or attachment(s) may contain confidential or legally privileged information intended for the sole use of the addressee(s). Any use, redistribution, disclosure, or reproduction of this message, except as intended, is prohibited. If you received this email in error, please notify the sender and remove all copies of the message, including any attachments. Any views or opinions expressed in this email (unless otherwise stated) may not represent those of Capital and Coast District Health Board. http://www.ccdhb.org.nz (AC_S001) No Viruses were detected in this message. _______________________________________________________________________ HealthIntelligence eMail Filter Service. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Fri Nov 25 04:11:34 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Nov 25 04:15:05 2005 Subject: [Histonet] RE: Histology art Message-ID: Just come across what looks to be a useful and fun website: histology-world.com which has histology art amongst other things Jacqui 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 ROrr <@t> enh.org Fri Nov 25 12:16:34 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Nov 25 12:16:48 2005 Subject: [Histonet] p16 antibody Message-ID: Hi Robert, I use the p16 antibody from Biocare Medical. I have used it 1:100 on the Benchmark with the Iview kit. I am currently using it 1:200 on the Nemesis stainer with Mach4 detection. Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Friday, November 25, 2005 11:57 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 24, 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. automated response (Laura Beaudet) 2. p16 (Robert Fauck) 3. RE: Histology art (Malam Jacqueline) ---------------------------------------------------------------------- Message: 1 Date: Thu, 24 Nov 2005 13:03:49 -0500 From: "Laura Beaudet" Subject: [Histonet] automated response To: histonet@lists.utsouthwestern.edu Message-ID: <10511241303.AA10078@mail1.ctcsurge.com> Content-Type: text/plain; charset=us-ascii Computer Trust Corporation is closed Thu-Sun Nov 24-27 2005 for Thanksgiving. ------------------------------ Message: 2 Date: Fri, 25 Nov 2005 13:17:19 +1300 From: "Robert Fauck" Subject: [Histonet] p16 To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Hi there! Is there anybody that has a good protocol to do the marker p16?! Any recommendation on the source for this p16 antibody? Thanking you all in advance for your advice. Cheers, Robert Fauck NZ, Wellington Hospital _______________________________________________________________________ This email or attachment(s) may contain confidential or legally privileged information intended for the sole use of the addressee(s). Any use, redistribution, disclosure, or reproduction of this message, except as intended, is prohibited. If you received this email in error, please notify the sender and remove all copies of the message, including any attachments. Any views or opinions expressed in this email (unless otherwise stated) may not represent those of Capital and Coast District Health Board. http://www.ccdhb.org.nz (AC_S001) No Viruses were detected in this message. _______________________________________________________________________ HealthIntelligence eMail Filter Service. ------------------------------ Message: 3 Date: Fri, 25 Nov 2005 10:11:34 -0000 From: Malam Jacqueline Subject: [Histonet] RE: Histology art To: "Histonet submissions (histonet@lists.utsouthwestern.edu)" Message-ID: Content-Type: text/plain Just come across what looks to be a useful and fun website: histology-world.com which has histology art amongst other things Jacqui 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. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 24, Issue 34 **************************************** From Rcartun <@t> harthosp.org Fri Nov 25 13:15:20 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Nov 25 13:15:59 2005 Subject: [Histonet] c4d Message-ID: We use a rabbit polyclonal antibody from American Research Products, Inc. located in Belmont, MA. The catalog number is 12-5000. 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 >>> 11/02/05 02:30PM >>> Could anyone recommend a monoclonal C4d antibody for FFPE human tissue? Thanks Tracey CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify the sender immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From lpwenk <@t> sbcglobal.net Sat Nov 26 14:30:30 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Sat Nov 26 14:31:58 2005 Subject: [Histonet] Biopsy for Leishmania In-Reply-To: <00f501c5f034$321aa5e0$972bfea9@m7c0y4> Message-ID: According to the AFIP (Armed Force Institute of Pathology) website http://www.afip.org/Departments/infectious/lm/06.html Either H&E, or Brown & Hopps (modified Gram) are the best. Photos included in the website Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Muhammad Tahseen Sent: Wednesday, November 23, 2005 8:46 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Biopsy for Leishmania Has anyone had any experience in performing stain for Leishmania (I have received fresh biopsy specimen)? Any opinions would be much appreciated. Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Lahore Pakistan. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Sat Nov 26 14:34:25 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Sat Nov 26 14:35:55 2005 Subject: [Histonet] Gallyas method In-Reply-To: <36FE435D6D2F1D489174B22362A961B66B82F5@hscxntmx0006.hsc.mb.ca> Message-ID: <1079085529-233819592@pathology.swmed.edu> Just curious. Is the positive control you are using normal brain, or brain with Alzheimer's plaques? The Gallyas doesn't stain the normal brain, only the unphosphorylated tau proteins in the Alzheimers, if memory serves me correctly. If no one has sent you the procedure, email me at work pwenk@beaumont.edu on Monday, and I'll send you our version. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Allen Sent: Wednesday, November 23, 2005 11:58 AM To: Histonet (E-mail) Subject: [Histonet] Gallyas method Hi, Does anyone have a good, reliable method for Gallyas Stain? I would appreciate a copy of it. The two methods I have tried haven't been great & I'm really short on time to troubleshoot the method. Thanks for any help you can give me. Sharon sallen@hsc.mb.ca 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 lr_hsvlle2 <@t> yahoo.com.au Sun Nov 27 19:53:04 2005 From: lr_hsvlle2 <@t> yahoo.com.au (Lorraine Rolston) Date: Sun Nov 27 19:53:13 2005 Subject: [Histonet] Help! NBF fixed tissue and subsequent first step for dehydration. Message-ID: <20051128015304.17579.qmail@web30608.mail.mud.yahoo.com> I would appreciate any advice concerning possible detrimental effects that can be caused to the morphology of NBF fixed tissue when placed directly into 70%ethanol from NBF. This tissue (pancreas, spleen, kidney)is for research purposes (not clinical diagnosis) and good preservation of cell morphology is of extreme importance. One of the research groups that uses our processing facility brings the tissues to us immersed in 70% ethanol ready for processing.They are then embedded and cut for H&E staining. Before their tissues are bought to the lab at the 70% ethanol step, they are washed in water after fixation, then immersed in 50%alcohol. At some stage this protocol has been changed and the 50%ethanol step has been omitted. Coinciding with this step being discontinued has come the appearance of bad cracking of the tissue.The head of the research group now feels that the changes to the morphology of these samples is caused by the jump from the NBF fixative to 70%ethanol. The pancreas samples are processed separately using chloroform for clearing. I have yet to view the slides but may be able to submit a picture(if given permission) when I do. Any helpful advice or ideas would be very much appreciated. Thanks in advance, Lorraine. --------------------------------- Do you Yahoo!? Messenger 7.0: Free worldwide PC to PC calls From TMcNemar <@t> lmhealth.org Mon Nov 28 05:48:23 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Mon Nov 28 05:53:11 2005 Subject: [Histonet] Histology assistants Message-ID: <6CD94D97ED7D924BA5C2B588FA952821396820@nt_exchange.lmhealth.org> I would really like some input on this..... How many places use assistants in Histology? What kind of things do they do? Gross? Enter/obtain specimens? File? Would really appreciate any and all thoughts. Thanks. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 From rjbuesa <@t> yahoo.com Mon Nov 28 07:02:20 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 28 07:02:33 2005 Subject: [Histonet] Histology assistants In-Reply-To: <6CD94D97ED7D924BA5C2B588FA952821396820@nt_exchange.lmhealth.org> Message-ID: <20051128130220.97265.qmail@web61224.mail.yahoo.com> Hi Tom: The following is my experience: lab assistants are the best way of increasing laboratory productivity and allowing the histotechs to do the work they know and like to do. In the last place I worked as supervisor, before retiring, a large reference lab, the lag of uncut blocks left from days before amounted to 3-4 days work. I was able to hire 2 lab asistants and at the end of my second month there were no uncut blocks left. The tasks the 2 lab asistants (1 came at 5:30 AM and the other at 1:00 PM, with an overlap of 1 hour) were the following: prepare the water baths for the first shift of HTs, change all reagents in the autostainer and in the tissue processors, operate the slides and cassettes numbering machines, keep the record of the blocks assigned to each HT and file them back, take care of drying the slides, run the H&E autostainer and the automatic coverslipper, match paper/work with the slides, deliver the slides to the pathologists, pull blocks for special requests and some other tasks that would be more costly if done by the HTs (the earn more money than the lab assistants) and that should not be done by them. Sometimes they helped in grossing but just the tasks related to keeping records. The lab assistants were also supposed to start getting an idea about the histology work with the purpose to later on train in-house as a complement to their studies, because since the beginning the idea was for them to study as histotechs in the near future. If you can hire lab assistants and train them well you wil see how things will begin to run more smoothly in your lab. Rene J. Tom McNemar wrote: I would really like some input on this..... How many places use assistants in Histology? What kind of things do they do? Gross? Enter/obtain specimens? File? Would really appreciate any and all thoughts. Thanks. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less From rjbuesa <@t> yahoo.com Mon Nov 28 07:37:23 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 28 07:37:33 2005 Subject: [Histonet] Help! NBF fixed tissue and subsequent first step for dehydration. In-Reply-To: <20051128015304.17579.qmail@web30608.mail.mud.yahoo.com> Message-ID: <20051128133724.17782.qmail@web61224.mail.yahoo.com> Lorraine: Although there are numerous considerations and theories regarding the adverse effects that osmotic and liquid flow currents have on the cellular morphology it is a fact that the water in the cells has to be eliminated and that the space between all its constituents have to be replaced by paraffin. To get to that point, as you well know, the water has to be extracted generally by alcohol, the alcohol has to be exchanged by an antemedium miscible with both ethanol and paraffin and at the end the paraffin has to infiltrate the tissue. There are several "rules of the thumb" (ROT) for the whole process, that will vary with the type of tissue, being some more "resistant" to those changes (like connective tissue) than others (glands, like the pancreas in your case, and whole embryos). The first ROT is that the "gentler" the exchange, the better meaning that the dehydratoin should start with the lowest concentration possible or allowed by your tissue processor and the number of station it has. Many years ago (about 50) I used to work with lizard embryos and I designed a small bottle with a cork with 2 perforations. Through the perforations I inserted 2 glass tubes, the "IN" touched the bottom, and the "OUT" was almost at the surface. After washing the embryos I placed them in the bottle and let alcohol "IN" dropping until after 24-48 hours all the water had been substituted by 100% EthOL and the embryos were dehydrated very gently. The 2nd ROT is that when going from one reagent to the next use one step that contains half and half, if going from 100%EthOL to xylene, 1 step should be 1:2 of each reagent. In the same way at the end of my career I stopped using xylene and changed to mineral oil with the intermediate steps of 1:2 of consecutive reagents. Now about your question on the "cracking" appearence (I gather that it looks as if the cells look like broken crackers with "breaking" lines through the cytoplasm) this can be due either to excessive dehydration or could be due to the antemedium (the one between the alcohol and the paraffin). Delicate tissues require delicate procedures and the best approach is to get several samples and process them with different protocols to find out the best. Also it is not a good practice for you to receive semiprocessed tissues, you will never know how they did it. Try to get the tissues in the fixative and then you will for certain know how you processed it. Escuse me this long answer, I only hope that it will help you. Maybe the head of the research is right but you will never know until you control the whole process. Rene J. Lorraine Rolston wrote: I would appreciate any advice concerning possible detrimental effects that can be caused to the morphology of NBF fixed tissue when placed directly into 70%ethanol from NBF. This tissue (pancreas, spleen, kidney)is for research purposes (not clinical diagnosis) and good preservation of cell morphology is of extreme importance. One of the research groups that uses our processing facility brings the tissues to us immersed in 70% ethanol ready for processing.They are then embedded and cut for H&E staining. Before their tissues are bought to the lab at the 70% ethanol step, they are washed in water after fixation, then immersed in 50%alcohol. At some stage this protocol has been changed and the 50%ethanol step has been omitted. Coinciding with this step being discontinued has come the appearance of bad cracking of the tissue.The head of the research group now feels that the changes to the morphology of these samples is caused by the jump from the NBF fixative to 70%ethanol. The pancreas samples are processed separately using chloroform for clearing. I have yet to view the slides but may be able to submit a picture(if given permission) when I do. Any helpful advice or ideas would be very much appreciated. Thanks in advance, Lorraine. --------------------------------- Do you Yahoo!? Messenger 7.0: Free worldwide PC to PC calls _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From jcline <@t> wchsys.org Mon Nov 28 08:56:09 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Mon Nov 28 08:56:24 2005 Subject: [Histonet] Tom Nemar/Histology Assistant Message-ID: <000d01c5f42b$deff6de0$1d2a14ac@wchsys.org> I have three Histology Assistants that have been trained in the accessioning and cytology prep areas. They log specimens, label cassettes and slides, stand with the pathologists, perform frozen sections, do all the cytology prep and staining for gyn and non gyn specimens, coverslip, run the microwave processor, changing some staining set ups and filing slides. I have two sites, two assistants at one site, one assistant at the other. They perform a wonderful time saving value for the histology lab. The histotech shortage made me find another way of filling vacancies. ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From Dorothy.L.Webb <@t> HealthPartners.Com Mon Nov 28 09:51:17 2005 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Mon Nov 28 09:51:30 2005 Subject: [Histonet] Dark staining Message-ID: <0E394B648E5284478A6CCB78E5AFDA2718C4D1@hpes1.HealthPartners.int> I am wondering what causes some of our tissues to stain with hematoxylin darker than others? The pathologist is noticing it mostly on GI biopsies or colon biopsies, not all of them on the same day either. I can't believe it is due to improper fixation, as all of them are getting practically the same amount of time in formalin and it is very adequate, often more than 24 hours. Any ideas would be appreciated. We use Gills I hematoxylin and Eosin from sigma. ________________________________________ 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 gentras <@t> vetmed.auburn.edu Mon Nov 28 10:17:45 2005 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Mon Nov 28 10:17:56 2005 Subject: [Histonet] Fwd: microtomes Message-ID: <6.0.1.1.0.20051128101632.01996160@mailhost.vetmed.auburn.edu> Hello All, since I do not currently need any of the items listed below I decided to share it with you. Atoska >From: "James" >To: >Subject: microtomes >Date: Tue, 22 Nov 2005 17:30:15 -0700 >X-Mailer: Microsoft Outlook Express 6.00.2800.1106 >X-spam: ESP<42>=RBL:<26> RDNS:<0> SHA:<15> UHA:<0> SLS:<0> BAYES:<0> > SenderID:<0> URL Substring Dictionary (TRU8):<0> Spam > Dictionary (TRU8):<0> NigeriaScam Dictionary (TRU8):<0> HTML > Dictionary (TRU8):<1> Porn Dictionary (TRU8):<0> Embed HTML > Dictionary (TRU8):<0> Obscenities Dictionary (TRU8):<0> URL > Dictionary (TRU8):<0> CAN-SPAM Compliance Dictionary (TRU8):<0> > >Dear Atoska, >If you might be needing a microtome or coverslipper I have some attractive >deals available: > >Microm 335E microtome, fully refurbished, like new condition, 1 year >warranty, $4500. This is an electronic microtome featuring motor driven >coarse feed, trim feed and section advance, manually driven >handwheel. Equivalent in quality to Leica and Sakura equipment, price >offered is below market and lower than what many strictly manual >microtomes sell for. > >Refurbished Reichert/Leica RM2030 for $3,000. > >Refurbished Microm CTM6 glass coverslipper for $10,500 > >Please let me know if you are interested and I will provide more >info. Also have cryostats, embedding centers, tissue processors, >stainers, paraffin dispensers and slide warmers. > >Thanks, >Jim Thompson Jr. >Schlufenhausen Instrument Sales >505-384-5443 From froyer <@t> bitstream.net Mon Nov 28 10:35:18 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Mon Nov 28 10:35:43 2005 Subject: [Histonet] Fwd: microtomes In-Reply-To: <6.0.1.1.0.20051128101632.01996160@mailhost.vetmed.auburn.edu> Message-ID: <004801c5f439$b8502320$0e00a8c0@fords> Also available from: Ford M. Royer, MT(ASCP) Sales Manager, Histology Product Division Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Golden Valley, MN 55427-3601 763-542-8725 phone 888-790-9686 toll free 763-546-4830 fax email web New and Refurbished (with Warranties) Histology Equipment. All Makes and Models. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Atoska S. Gentry Sent: Monday, November 28, 2005 10:18 AM To: Histonet Subject: [Histonet] Fwd: microtomes Hello All, since I do not currently need any of the items listed below I decided to share it with you. Atoska >From: "James" >To: >Subject: microtomes >Date: Tue, 22 Nov 2005 17:30:15 -0700 >X-Mailer: Microsoft Outlook Express 6.00.2800.1106 >X-spam: ESP<42>=RBL:<26> RDNS:<0> SHA:<15> UHA:<0> SLS:<0> BAYES:<0> > SenderID:<0> URL Substring Dictionary (TRU8):<0> Spam > Dictionary (TRU8):<0> NigeriaScam Dictionary (TRU8):<0> HTML > Dictionary (TRU8):<1> Porn Dictionary (TRU8):<0> Embed HTML > Dictionary (TRU8):<0> Obscenities Dictionary (TRU8):<0> URL > Dictionary (TRU8):<0> CAN-SPAM Compliance Dictionary (TRU8):<0> > >Dear Atoska, >If you might be needing a microtome or coverslipper I have some attractive >deals available: > >Microm 335E microtome, fully refurbished, like new condition, 1 year >warranty, $4500. This is an electronic microtome featuring motor driven >coarse feed, trim feed and section advance, manually driven >handwheel. Equivalent in quality to Leica and Sakura equipment, price >offered is below market and lower than what many strictly manual >microtomes sell for. > >Refurbished Reichert/Leica RM2030 for $3,000. > >Refurbished Microm CTM6 glass coverslipper for $10,500 > >Please let me know if you are interested and I will provide more >info. Also have cryostats, embedding centers, tissue processors, >stainers, paraffin dispensers and slide warmers. > >Thanks, >Jim Thompson Jr. >Schlufenhausen Instrument Sales >505-384-5443 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cdemarinis <@t> SARATOGACARE.ORG Mon Nov 28 10:57:34 2005 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis, Carolyn) Date: Mon Nov 28 10:57:47 2005 Subject: [Histonet] moh's surgery Message-ID: Do you have to be a histotech to work in a Moh's surgery center doing frozen sections? If not, what are the qualifications for the technical position? Thanks. From cfavara <@t> niaid.nih.gov Mon Nov 28 11:09:58 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Mon Nov 28 11:10:12 2005 Subject: [Histonet] moh's surgery Message-ID: I know there are some areas of the country where the person doing the frozen sectioning does not need to be a histotech and the person reading the slide does not need to be a pathologist. I am fairly ignorant about these regulations but imagine they are state controlled. c 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: Demarinis, Carolyn [mailto:cdemarinis@SARATOGACARE.ORG] Sent: Monday, November 28, 2005 9:58 AM To: HISTONET (E-mail) Subject: [Histonet] moh's surgery Do you have to be a histotech to work in a Moh's surgery center doing frozen sections? If not, what are the qualifications for the technical position? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Nov 28 11:12:12 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 28 11:12:20 2005 Subject: [Histonet] Dark staining In-Reply-To: <0E394B648E5284478A6CCB78E5AFDA2718C4D1@hpes1.HealthPartners.int> Message-ID: <20051128171212.67842.qmail@web61214.mail.yahoo.com> Are the GI biopsies fixed immediately after being taken? Do some by any chance air-dry before fixing? Are all the sections of the same thickness? I would check on these Rene J. "Webb, Dorothy L" wrote: I am wondering what causes some of our tissues to stain with hematoxylin darker than others? The pathologist is noticing it mostly on GI biopsies or colon biopsies, not all of them on the same day either. I can't believe it is due to improper fixation, as all of them are getting practically the same amount of time in formalin and it is very adequate, often more than 24 hours. Any ideas would be appreciated. We use Gills I hematoxylin and Eosin from sigma. ________________________________________ 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 --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From c.ingles <@t> hosp.wisc.edu Mon Nov 28 11:18:22 2005 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Mon Nov 28 11:18:43 2005 Subject: [Histonet] moh's surgery References: Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE78@uwhis-xchng2.hosp.wisc.edu> Here at the UW Hospitals in Madison we have trained numerous people "off the street" , ok, ok, they at least have a biology degree and a brain in their heads. but no other histology experience. Our current tech is as good as an HT at cuttinggood quality sections at a fast pace. He lets us handle all the troubleshooting, but other than that he is a very valuable member of the team. I wouldn't recommend totally staffing your lab with non-HT's though. Claire Ingles ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Demarinis, Carolyn Sent: Mon 11/28/2005 10:57 AM To: HISTONET (E-mail) Subject: [Histonet] moh's surgery Do you have to be a histotech to work in a Moh's surgery center doing frozen sections? If not, what are the qualifications for the technical position? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Mon Nov 28 11:19:33 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Mon Nov 28 11:19:37 2005 Subject: [Histonet] Dark staining Message-ID: Dorothy Don't know if this is what you are referring to but... Some tissue will always stain more deeply with hematoxylin than others. Tissues that contain large numbers of inflammatory cells such as in the lamina propria of the digestive tract are often more deeply stained. In the lamina propria the cells are often much more crowded together. Cells that carry out a lot of protein synthesis (such as liver cells) are often much more basophilic than many other cells. Barry Those tissues -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Monday, November 28, 2005 11:12 AM To: Webb, Dorothy L; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Dark staining Are the GI biopsies fixed immediately after being taken? Do some by any chance air-dry before fixing? Are all the sections of the same thickness? I would check on these Rene J. "Webb, Dorothy L" wrote: I am wondering what causes some of our tissues to stain with hematoxylin darker than others? The pathologist is noticing it mostly on GI biopsies or colon biopsies, not all of them on the same day either. I can't believe it is due to improper fixation, as all of them are getting practically the same amount of time in formalin and it is very adequate, often more than 24 hours. Any ideas would be appreciated. We use Gills I hematoxylin and Eosin from sigma. ________________________________________ 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 --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dusko.trajkovic <@t> pfizer.com Mon Nov 28 12:07:11 2005 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Mon Nov 28 12:07:23 2005 Subject: [Histonet] Osteopontin and GST-a or GST A1-1 Ab for IHC staining on FFPE Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B20206F0DF@lajamrexm01.amer.pfizer.com> Does anyone have a protocol for these antibodies? I would appreciate any response. Thank you, Dusko Trajkovic ---------------------------------------------------------------------- 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 TJJ <@t> Stowers-Institute.org Mon Nov 28 12:15:31 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Mon Nov 28 12:15:49 2005 Subject: [Histonet] Re: Help! NBF fixed tissue and subsequent first step Message-ID: Lorraine, I would also find out if the length of fixation has changed. Rodent tissue (mouse in particular) tends to be very suseptible to length of fixation time. Extended washing can reverse some of the cross links. Extended fixation can make your tissues overly brittle. We have no problem routinely going from fixative to 70% alcohol, although many of our researchers do a quick PBS wash before placing it into 70% alcohol, again with no detrimental effects. The only problem we have had is trying to process zinc formalin fixed pancreas. It seems no matter the processing schedule, we cannot keep the tissue from cracking and looking awful. Perhaps a lower percentage of alcohol might help this. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From settembr <@t> umdnj.edu Mon Nov 28 12:24:33 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Mon Nov 28 12:25:51 2005 Subject: [Histonet] p16 Message-ID: Hello Robert, I use DakoCytomation's p16 antibody that comes in a kit. I use it at a 1:100 diluton incubating at room temp for 15 minutes. Dana Settembre University Hospital - UMDNJ Newark, NJ USA >>> Robert Fauck 11/24/2005 7:17:19 PM >>> Hi there! Is there anybody that has a good protocol to do the marker p16?! Any recommendation on the source for this p16 antibody? Thanking you all in advance for your advice. Cheers, Robert Fauck NZ, Wellington Hospital _______________________________________________________________________ This email or attachment(s) may contain confidential or legally privileged information intended for the sole use of the addressee(s). Any use, redistribution, disclosure, or reproduction of this message, except as intended, is prohibited. If you received this email in error, please notify the sender and remove all copies of the message, including any attachments. Any views or opinions expressed in this email (unless otherwise stated) may not represent those of Capital and Coast District Health Board. http://www.ccdhb.org.nz (AC_S001) No Viruses were detected in this message. _______________________________________________________________________ HealthIntelligence eMail Filter Service. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MadaryJ <@t> MedImmune.com Mon Nov 28 12:27:10 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Mon Nov 28 12:27:24 2005 Subject: [Histonet] 70% from NBF Message-ID: <746FDB897740814EA52BDDCB5ED1DDBC02EB13CF@medimmune4.medimmune.com> Lorraine, going from NBF to 70 is very common. What you are doing is perhaps overtreating the samples in alcohol. Of all specimens spleen, pancreas are really susceptible to dehydrations negative effects. If you are using chloroform, my fitst question is why? You would want to consider Chlorofrm as a dehydrant as well as a clearant, but get rid of that stuff. Just go fromt NBF inot to 50 or 70, do the washes if you need and cut down on the time or heat. The samples that have been done should just be soaked. You will note in Freidas book she states that 70 is the highest alcohol you can go into from NBF(unwashed), anything higher and you risk the buffer salts not dissolving. I am lazy, I do not wash after formalin since I use it on the processor. I go into about a 60% alcohol, and move on. From settembr <@t> umdnj.edu Mon Nov 28 12:27:48 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Mon Nov 28 12:28:43 2005 Subject: [Histonet] Osteopontin and GST-a or GST A1-1 Ab for IHC staining on FFPE Message-ID: Hello Dusko, I use osteopontin from Novocastra Laboratories in the UK catalog # NCL-O-Pontin I pretreat with a steamer with DakoCytomations Target Retrieval Solution for 40 minutes I use the antibody at a dilution of 1:100 for 15 minutes room temp. Good Luck Dana Settembre University Hospital - UMDNJ Newark, NJ USA >>> "Trajkovic, Dusko" 11/28/2005 1:07:11 PM >>> Does anyone have a protocol for these antibodies? I would appreciate any response. Thank you, Dusko Trajkovic ---------------------------------------------------------------------- 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From billingconsultants <@t> yahoo.com Mon Nov 28 12:28:56 2005 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Mon Nov 28 12:29:05 2005 Subject: [Histonet] Histology Technician Position Open - Georgia Message-ID: <20051128182856.83072.qmail@web54207.mail.yahoo.com> Hi Everyone, There is a histology technician position open at a private laboratory in Athens, GA. If you are interested, please fax your resume to 706-546-4084. Thanks, Louri --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From JMacDonald <@t> mtsac.edu Mon Nov 28 13:37:17 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Mon Nov 28 13:33:40 2005 Subject: [Histonet] Dark staining In-Reply-To: <20051128171212.67842.qmail@web61214.mail.yahoo.com> Message-ID: We noticed this when someone used the freezing spray too much. It gives a burn affect and very dark staining. Rene J Buesa Sent by: histonet-bounces@lists.utsouthwestern.edu 11/28/2005 09:12 AM To "Webb, Dorothy L" , Histonet@lists.utsouthwestern.edu cc Subject Re: [Histonet] Dark staining Are the GI biopsies fixed immediately after being taken? Do some by any chance air-dry before fixing? Are all the sections of the same thickness? I would check on these Rene J. "Webb, Dorothy L" wrote: I am wondering what causes some of our tissues to stain with hematoxylin darker than others? The pathologist is noticing it mostly on GI biopsies or colon biopsies, not all of them on the same day either. I can't believe it is due to improper fixation, as all of them are getting practically the same amount of time in formalin and it is very adequate, often more than 24 hours. Any ideas would be appreciated. We use Gills I hematoxylin and Eosin from sigma. ________________________________________ 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 --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ana.merino-trigo <@t> wanadoo.fr Mon Nov 28 13:45:02 2005 From: ana.merino-trigo <@t> wanadoo.fr (ana.merino-trigo) Date: Mon Nov 28 13:45:15 2005 Subject: [Histonet] Tissue processors Message-ID: <040f01c5f454$39a2fbe0$02fea8c0@BABA> Hi, I have been asked to look for a not expensive tissue processor. I'm working in histology for the last two years and I just know the Leica ASP300. However, I do believe that Leica is not one of the cheapest brands. Could anyone recommend me a nice and no too expensive one? Thanks in advance to everyone, Ana From ftryka <@t> tetonhospital.org Mon Nov 28 14:06:56 2005 From: ftryka <@t> tetonhospital.org (Dr. F. Tryka) Date: Mon Nov 28 14:07:07 2005 Subject: [Histonet] Tissue processors In-Reply-To: <040f01c5f454$39a2fbe0$02fea8c0@BABA> References: <040f01c5f454$39a2fbe0$02fea8c0@BABA> Message-ID: Hello Ana. Whichever brand you decide to purchase, be sure to check on their service record and support. I have a Thermo Citadel, which has been out of service for over a month. We are struggling with getting the manufacturer (Thermo Electron) to get it serviced. In the meantime we are hand processing our cases. Franci Tryka From pathrm35 <@t> adelphia.net Mon Nov 28 15:21:24 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Mon Nov 28 15:21:33 2005 Subject: [Histonet] batch controls Message-ID: <29724345.1133212884603.JavaMail.root@web4.mail.adelphia.net> Fellow techs, Does anyone use only one special stain control slide for multiple cases? We want to run only one control slide for several AB PAS patient slides. How would you cross reference the patient cases to the control? By date or case number or something else? Thanks in Advance. Ron Martin From rjbuesa <@t> yahoo.com Mon Nov 28 15:43:03 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 28 15:43:12 2005 Subject: [Histonet] batch controls In-Reply-To: <29724345.1133212884603.JavaMail.root@web4.mail.adelphia.net> Message-ID: <20051128214303.9587.qmail@web61214.mail.yahoo.com> Hi Ron: I used to batch several cases with 1 + control. For example: when we did the HER2/neu tests with the Dako kit we did all the tests on Thursday and we used 1 Dako + control for all the cases. I prepared a list of all the cases run during the day and sent it to the pathologist with the + control. He signed the list and it was filed in a way that the cases could be referred to that + control. We used the same procedure for H. pyloris staining (we used modefied Steiner) and there was also a list of cases with 1 control every 8 cases that could be stained simultaneously in a Coplin jar, so if we had 20 cases we would distribute them in 3 groups, each with 1 + control. In this way the fundamental thing is the documentation that you have to have in the event that it is necessary to should the + control for a given case. Hope this will help! Rene J. pathrm35@adelphia.net wrote: Fellow techs, Does anyone use only one special stain control slide for multiple cases? We want to run only one control slide for several AB PAS patient slides. How would you cross reference the patient cases to the control? By date or case number or something else? Thanks in Advance. Ron Martin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From SLB <@t> Stowers-Institute.org Mon Nov 28 15:46:19 2005 From: SLB <@t> Stowers-Institute.org (Beckham, Sharon) Date: Mon Nov 28 15:46:55 2005 Subject: [Histonet] batch controls Message-ID: I would label a blank slide for each extra case using the correct patient's label and then write on the slide itself "see case #" and refer to the case that I had put the control slide with. Hope that makes sense. Sharon Beckham -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pathrm35@adelphia.net Sent: Monday, November 28, 2005 3:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] batch controls Fellow techs, Does anyone use only one special stain control slide for multiple cases? We want to run only one control slide for several AB PAS patient slides. How would you cross reference the patient cases to the control? By date or case number or something else? Thanks in Advance. Ron Martin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AGrobe2555 <@t> aol.com Mon Nov 28 16:53:54 2005 From: AGrobe2555 <@t> aol.com (AGrobe2555@aol.com) Date: Mon Nov 28 16:54:04 2005 Subject: [Histonet] Collagen & Elastin IHC on Frozens Message-ID: <1de.490e3dea.30bce482@aol.com> Hello Folks, I would guess that many of you have done IHC for collagen and elastin on frozen sections. Are there any antibodies you would suggest and any tricks for using them. I will be working with sheep tissues, primarily lung and isolated pulmonary vessels. Thanks, Albert From mayhew <@t> vaxxine.com Mon Nov 28 17:01:02 2005 From: mayhew <@t> vaxxine.com (Dave & Lee Mayhew) Date: Mon Nov 28 17:01:16 2005 Subject: [Histonet] batch controls Message-ID: Hi Ron, We use one control slide for several cases for all our special stains. In our LIS (we use Meditech) we create a QC specimen each working day. These QC specimens are numbered sequentially. For all the control slides that are done on a particular day, they all receive and are labelled with that days number. This QC number is entered into each patients record that had a special stain done that day and becomes part of the final report. The control slides are handed in to the pathologist along with the case(s). If more than one pathologist has ordered the same stain for that day, the controls are put in a folder labelled Common Controls and put in a central place for each pathologist to look at when they need to. The control slides are filed numerically in their own file boxes. If anyone needs to see a particular control slide for a case, they look at the patient report, find the QC number, and go to the QC file and pull the appropriate slides. Hope this is clear Lee Mayhew MLT Niagara Health System St. Catharines, Ontario From jkiernan <@t> uwo.ca Mon Nov 28 23:15:38 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Nov 28 23:15:30 2005 Subject: [Histonet] Collagen & Elastin IHC on Frozens References: <1de.490e3dea.30bce482@aol.com> Message-ID: <438BE3FA.153652B7@uwo.ca> Why use expensive immunohistochemical reagents to stain collagen and elastin? There are plenty of less expensive ways to do the job using dyes, which are VERY much cheaper than antibodies. Immunohistochemistry is needed only for specific identification of particular types of collagen. You don't say who and where you are. Are you a graduate student, a technician, a senior professor, a pathology resident, a captain of industry, or a laboratory mouse cloned from some of the DNA in Einstein's formadehyde-fixed brain? You will get more and better replies if you give more information about what you are trying to investigate. John Kiernan Anatomy, UWO London, Canada. ---------------------------------------------- AGrobe2555@aol.com wrote: > > Hello Folks, > I would guess that many of you have done IHC for collagen and elastin on > frozen sections. Are there any antibodies you would suggest and any tricks for > using them. I will be working with sheep tissues, primarily lung and > isolated pulmonary vessels. > Thanks, > Albert > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From moogoolt <@t> sbcglobal.net Mon Nov 28 23:41:15 2005 From: moogoolt <@t> sbcglobal.net (Lance Thomas) Date: Mon Nov 28 23:41:07 2005 Subject: [Histonet] Unscribe to Histonet Message-ID: <068201c5f4a7$84159590$39f78547@lancekt8tt92f5> Please take me off the histonet. Thank you.........Lance Thomas From Anni-Maija.Linden <@t> helsinki.fi Tue Nov 29 00:10:57 2005 From: Anni-Maija.Linden <@t> helsinki.fi (Anni-Maija Linden) Date: Tue Nov 29 00:11:08 2005 Subject: [Histonet] Unsuscribe Message-ID: <1133244657.438bf0f1849c0@www3.helsinki.fi> Please take me off the histonet. Thank you. Best Regards, Anni-Maija From grandclement <@t> lyon.inserm.fr Tue Nov 29 01:03:14 2005 From: grandclement <@t> lyon.inserm.fr (=?iso-8859-1?Q?B=E9atrice_Zabawski?=) Date: Tue Nov 29 01:03:31 2005 Subject: [Histonet] unsubscribe Message-ID: <20051129070318.332044A5E9@dns.lyon.inserm.fr> Please take me off the histonet. Thank you. Best Regards, B?atrice From gmondragon <@t> gsopath.com Tue Nov 29 03:47:47 2005 From: gmondragon <@t> gsopath.com (Gus Mondragon) Date: Tue Nov 29 03:48:00 2005 Subject: [Histonet] Message 4:NBF fixed tissue Message-ID: <03B63DE44B5C014D84F9A9D8A968B5174C4D3C@lithium.corp.gsopath.com> You can avoid deleteriuos artifact in tissue by limiting your fixation to 10% NBF. Alcohol is considered an "additive" fixative which means that alcohol molecules are incorporated into the tissue irreversibly; altough this is OK histochemically, there is a harsh re-configuration of the protein aminoacid in tissue by the alcohol. This reaction is electrostatically and it is permanent. In contradistinction, Neutral buffered formalin is a"Non-additive fixative" that is the linking between the aldehyde molecules and aminoacids is covalent and therefore gentle on the tissue without an irreversible change in the morphology of the proteins. This is why we can do antigen retrieval in IHC, where the aldehyde crosslink is breaking apart with the aid of heat. More on this issue in the next Histologic issue coming out soon. You may want to supply some 10% NBFto your tissue source. Hope this can help you, Lorraine. Gustave Mondragon, Lab Manager gmondragon @gsopath.com From Karen.Heckford <@t> CHW.edu Tue Nov 29 07:11:20 2005 From: Karen.Heckford <@t> CHW.edu (Heckford, Karen - SMMC-SF) Date: Tue Nov 29 07:11:04 2005 Subject: [Histonet] JCAHO and IHC Message-ID: Dear Histonetters, I am trying to find some information on what JCAHO will look for in my IHC Dept. Is there anything in particular they look for??? I am having trouble trying to find information on this subject. Any ideas out there in Histology Land??? Karen Heckford HT (ASCP) CE Lead Histology Technician St. Mary's Medical Center San Francisco, California From rjbuesa <@t> yahoo.com Tue Nov 29 07:25:09 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 29 07:25:18 2005 Subject: [Histonet] JCAHO and IHC In-Reply-To: Message-ID: <20051129132509.6564.qmail@web61219.mail.yahoo.com> Hi Karen: I can tell you what I used to do: treat each inspection as if it were a CAP inspection. In the JCAHO case take a hard look at patients' issues and confidentiality. Develop a general strategy for inspections, it will be simpler than having several strategies. Rene J. "Heckford, Karen - SMMC-SF" wrote: Dear Histonetters, I am trying to find some information on what JCAHO will look for in my IHC Dept. Is there anything in particular they look for??? I am having trouble trying to find information on this subject. Any ideas out there in Histology Land??? Karen Heckford HT (ASCP) CE Lead Histology Technician St. Mary's Medical Center San Francisco, California _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From gu.lang <@t> gmx.at Tue Nov 29 09:17:35 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Tue Nov 29 09:17:44 2005 Subject: AW: [Histonet] c4d In-Reply-To: Message-ID: Dear Richard, We are going to introduce this antibody in our lab. We work with the Benchmark XT. Can you give me some hints about the titer and incubation time you use? Or what tissue is preferd as a positiv control? Does it work constantly? Thank you Gudrun Lang Histotech, Akh Linz Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Richard Cartun Gesendet: Freitag, 25. November 2005 20:15 An: Tracey.Lenek@CLS.ab.ca; histonet@lists.utsouthwestern.edu Betreff: Re: [Histonet] c4d We use a rabbit polyclonal antibody from American Research Products, Inc. located in Belmont, MA. The catalog number is 12-5000. 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 >>> 11/02/05 02:30PM >>> Could anyone recommend a monoclonal C4d antibody for FFPE human tissue? Thanks Tracey CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify the sender immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Tue Nov 29 09:51:14 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Tue Nov 29 09:51:39 2005 Subject: [Histonet] c4d Message-ID: Hi Gudrun, For frozen sections we use Biogenesis C4d at 1:600 on our Benchmark XT for 24" with AB block. We pretreat our acetone fixed sections in VMS Morphosave for 2" but this isn't a critical step and frozens will stain without it. For paraffin sections, we perform HIER in EDTA in a Biocare decloaking chamber for 3" followed by a 32" incubation and VMS amplification as well as AB block. We use rejecting kidney as a control as our patient specimens are 95% renal biopsies being tested for possible rejection. Hope this is of some help, 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: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Tuesday, November 29, 2005 9:18 AM To: Histonetliste (Histonetliste) Subject: AW: [Histonet] c4d Dear Richard, We are going to introduce this antibody in our lab. We work with the Benchmark XT. Can you give me some hints about the titer and incubation time you use? Or what tissue is preferd as a positiv control? Does it work constantly? Thank you Gudrun Lang Histotech, Akh Linz Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Richard Cartun Gesendet: Freitag, 25. November 2005 20:15 An: Tracey.Lenek@CLS.ab.ca; histonet@lists.utsouthwestern.edu Betreff: Re: [Histonet] c4d We use a rabbit polyclonal antibody from American Research Products, Inc. located in Belmont, MA. The catalog number is 12-5000. 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 >>> 11/02/05 02:30PM >>> Could anyone recommend a monoclonal C4d antibody for FFPE human tissue? Thanks Tracey CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify the sender immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ 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 pzeitlow <@t> bbpllab.com Tue Nov 29 10:00:23 2005 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Tue Nov 29 10:02:04 2005 Subject: [Histonet] unsubscribe Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A47D763@bbplsrv1.bbpl> Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From gcallis <@t> montana.edu Tue Nov 29 10:23:44 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Nov 29 10:23:58 2005 Subject: [Histonet] Re: 70% from NBF In-Reply-To: <746FDB897740814EA52BDDCB5ED1DDBC02EB13CF@medimmune4.medimm une.com> References: <746FDB897740814EA52BDDCB5ED1DDBC02EB13CF@medimmune4.medimmune.com> Message-ID: <6.0.0.22.1.20051129083656.01b51a08@gemini.msu.montana.edu> Joseph made some excellent points here Chloroform is an excellent clearing agent (used it back in the 60's in open dip and dunk processors - O.K. so I'm old!) but no one warned us about its carcinogenic nature and there were no safety issue regulations then. Take his advice! Clearite 3 and Propar, two xylene substitutes we have used, do not harden tissues excessively but they are sensitive to water. Careful monitoring of how many tissues pass through substitutes is advisable then either rotate and/or change these more frequently. On our VIP, we have one xylene followed by one Clearite 3 to help reduce hardening while giving better tolerance level/removal of water by xylene in first step. We work with rodents and 70% alcohol storage after NBF fixation and a brief rinse presents no problems. We do store brain and spinal cord in 50% alcohol but all processing starts in 70%. We use custom processing schedules for brain with shorter schedule smaller tissues, sliced brain and spinal cord and/or tissues that tend to be brittle after over exposure (a.k.a. over processing) to alcohols, clearing agents and heat of paraffins. Heat is never added to any tissue processing of animal tissues especially murine. Animal tissues are very lean and any extra heat dries out the tissues even more. Oh wise one, Jerry Fredenburgh (in a panel discussion) said you want to remove free water from the tissue spaces but not the water bound to the proteins as the latter contributes to hard, brittle tissues at microtomy. Acid decalcified bone is not harmed by 70% alcohol storage, in fact, this is a way to stop acid decalcificaton (calcium does not ionize in presence of alcohol) and prevent over exposure to acid (aka over decalcification). Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 Although At 11:27 AM 11/28/2005, you wrote: >Lorraine, going from NBF to 70 is very common. What you are doing is >perhaps overtreating the samples in alcohol. Of all specimens spleen, >pancreas are really susceptible to dehydrations negative effects. If you >are using chloroform, my fitst question is why? You would want to >consider Chlorofrm as a dehydrant as well as a clearant, but get rid of >that stuff. Just go fromt NBF inot to 50 or 70, do the washes if you need >and cut down on the time or heat. The samples that have been done should >just be soaked. You will note in Freidas book she states that 70 is the >highest alcohol you can go into from NBF(unwashed), anything higher and >you risk the buffer salts not dissolving. I am lazy, I do not wash after >formalin since I use it on the processor. I go into about a 60% alcohol, >and move on. From llewllew <@t> shaw.ca Tue Nov 29 11:15:06 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Tue Nov 29 11:15:28 2005 Subject: [Histonet] Re: 70% from NBF References: <746FDB897740814EA52BDDCB5ED1DDBC02EB13CF@medimmune4.medimmune.com> <6.0.0.22.1.20051129083656.01b51a08@gemini.msu.montana.edu> Message-ID: <000501c5f508$7210e320$690a4246@yourlk4rlmsu> Talking about chloroform use in the past. I trained as a lab tech in the early 60s, and the hospital where I did histology specialist training in London used chloroform exclusively. The pathologist apparently thought that xylene made tissues too brittle. I have many times dunked my hand in a litre container of it to get a block out. You wouldn't believe how much it stings, apparently by slightly defatting the skin! I was told this after I had done it a few times, of course. We not only used chloroform without any safety concerns at all, but we also redistilled it to save money. We had an upright, water cooled still with a Liebig type condensor. It was heated with an electrical coil in a sand bath, which surrounded a 2 litre round bottom flask. There was no automatic cut off for the electricity, and we had to keep an eye on it and switch it off when the flask was almost empty. Of course, being a very attentive teenager at the time I missed it more often than not. This was usually announced by a disgusting smell and billows of white smoke. I was never sure whether the smoke was burning fat and stuff, or phosgene, which is produced from chloroform. For those not aware, phosgene is one of the gases used during World War II as a nerve gas. I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue regulations > then. Take his advice! From Barry.R.Rittman <@t> uth.tmc.edu Tue Nov 29 12:41:30 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Tue Nov 29 12:41:35 2005 Subject: [Histonet] Re: 70% from NBF Message-ID: We also used chloroform extensively and found that for some tissues it was greatly superior to xylene. Some tissue such as bone can be left in chloroform for extended periods of time without becoming too hard or brittle. We first noticed this when some rat adrenals were left in chloroform for 2 weeks (in error) and still cut beautifully. The problems we observed were toxicity (we were verrrry careful),the fact that chloroform does not clear and so care must be taken to mix contents occasionally to ensure mixing, and most of all the expense compared to other intermediary agents. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: Tuesday, November 29, 2005 11:15 AM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF Talking about chloroform use in the past. I trained as a lab tech in the early 60s, and the hospital where I did histology specialist training in London used chloroform exclusively. The pathologist apparently thought that xylene made tissues too brittle. I have many times dunked my hand in a litre container of it to get a block out. You wouldn't believe how much it stings, apparently by slightly defatting the skin! I was told this after I had done it a few times, of course. We not only used chloroform without any safety concerns at all, but we also redistilled it to save money. We had an upright, water cooled still with a Liebig type condensor. It was heated with an electrical coil in a sand bath, which surrounded a 2 litre round bottom flask. There was no automatic cut off for the electricity, and we had to keep an eye on it and switch it off when the flask was almost empty. Of course, being a very attentive teenager at the time I missed it more often than not. This was usually announced by a disgusting smell and billows of white smoke. I was never sure whether the smoke was burning fat and stuff, or phosgene, which is produced from chloroform. For those not aware, phosgene is one of the gases used during World War II as a nerve gas. I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue regulations > then. Take his advice! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Tue Nov 29 12:52:27 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Tue Nov 29 12:52:38 2005 Subject: [Histonet] Movat's Pentachrome question Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717621@lsexch.lsmaster.lifespan.org> Does anyone do Movat's Pentachrome stain? I have a researcher who wants to do it. My question involves the "alkaline alcohol" the procedure calls for. The protocol says to add ammonium hydroxide drop by drop to 95% ethanol until the pH reaches 8.0 or higher. Question 1, a theoretical one - How can you accurately measure pH in a nearly water-free solution? Presumably an ionic compound would not ionize very well in such a solution? Question 2, a practical one - When I put the pH probe into 95% ethanol, the meter already reads higher than 9.0, even without adding any ammonium hydroxide. Is this your experience? From drgrant <@t> cmh.edu Tue Nov 29 13:14:06 2005 From: drgrant <@t> cmh.edu (Grant, Debra, R) Date: Tue Nov 29 13:19:08 2005 Subject: [Histonet] Rapid same day processing Message-ID: <6F434E27D5DE944B9E898984CADDD14904397C@exchmail.CMH.Internal> Hi Histonetters, For those of you who process needle core biopsies (liver, kidney, heart) the same day, could you please send me your protocol? i.e.... Manually, on tissue processor, fixative brand, times in alcohol, paraffin. Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 From sbreeden <@t> nmda.nmsu.edu Tue Nov 29 14:13:01 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Tue Nov 29 14:13:09 2005 Subject: [Histonet] Dr. John Kiernan, please? Message-ID: Dr. Kiernan - I've tried to contact you unsuccessfully via email address from Histonet. Could you please contact me at sbreeden@nmda.nmsu.edu? Thank you! Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From mplhisto <@t> aol.com Tue Nov 29 14:14:53 2005 From: mplhisto <@t> aol.com (mplhisto@aol.com) Date: Tue Nov 29 14:15:09 2005 Subject: [Histonet] Cassette slide label systems In-Reply-To: <6F434E27D5DE944B9E898984CADDD14904397C@exchmail.CMH.Internal> References: <6F434E27D5DE944B9E898984CADDD14904397C@exchmail.CMH.Internal> Message-ID: <8C7C338B1B6FE94-15E4-D6FF@FWM-R01.sysops.aol.com> Can anyone give me any information on the Leica cassette writer as well as any others on the market ? Also anyone that is using the system with barcodes? Any help would be appreciated . Thanks, Meredith Hale HT (ASCP) Histology Supervisor Pathology Partners Inc. Irving, Texas From jkiernan <@t> uwo.ca Tue Nov 29 17:25:43 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Tue Nov 29 17:25:44 2005 Subject: [Histonet] Movat's Pentachrome question References: <09C945920A6B654199F7A58A1D7D1FDE01717621@lsexch.lsmaster.lifespan.org> Message-ID: <438CE377.1505CFAD@uwo.ca> Answers below begin with **** John Kiernan London, Canada. ------------------------- "Monfils, Paul" wrote: > > Does anyone do Movat's Pentachrome stain? I have a researcher who wants to > do it. My question involves the "alkaline alcohol" the procedure calls for. > The protocol says to add ammonium hydroxide drop by drop to 95% ethanol > until the pH reaches 8.0 or higher. > > Question 1, a theoretical one - How can you accurately measure pH in a > nearly water-free solution? Presumably an ionic compound would not ionize > very well in such a solution? **** You can't! There is a quantity called pH* (pee-aitch-star) that is can be calculated from a pH meter reading taken from a non-aqueous solution. (see Perrin & Dempsey's book on Buffers). It's complicated stuff, and I don't claim to understand it. The book says you can get pH* standard buffers to calibrate a glass electrode pH meter for solvents that contain "at least a few percent" of water. > > Question 2, a practical one - When I put the pH probe into 95% ethanol, the > meter already reads higher than 9.0, even without adding any ammonium > hydroxide. Is this your experience? **** Never tried! > From moolovescow <@t> yahoo.com.sg Tue Nov 29 21:24:05 2005 From: moolovescow <@t> yahoo.com.sg (Phua Shirley) Date: Tue Nov 29 21:24:14 2005 Subject: [Histonet] FineFix Fixative Message-ID: <20051130032405.3597.qmail@web35204.mail.mud.yahoo.com> Hi All Anyone out there uses FineFix as a routine fixative? Any comments on it? Also, have you ever tried extracting DNA/RNA from those FineFixed processed tissue blocks. Were they successful? Shirley Phua Histopathology Laboratory Centre for Forensic Medicine Health Sciences Authority Singapore --------------------------------- Meet your soulmate! Yahoo! Asia presents Meetic - where millions of singles gather From Tony_Reilly <@t> health.qld.gov.au Tue Nov 29 23:26:09 2005 From: Tony_Reilly <@t> health.qld.gov.au (Anthony Reilly) Date: Tue Nov 29 23:26:44 2005 Subject: [Histonet] Movat's Pentachrome question Message-ID: The alcoholoic ammonia step is the critical stage of Movat's as it is required to convert the Alcian Blue to insoluble Monastral Blue so that it will not be removed during the remainder of the staining method. Just follow the AFIP method which uses 10ml of ammonium hydroxide in 90ml of 95% alcohol. If you want to simplify things then you can do what we do. Just stain using your usual Alcian blue method(pH2.5), treat with alcoholoic ammonia for 2 hours then do a routine VVG. This will stain your elastic fibres, collagen and glycoproteins as the Movat does with routine stains that are already on your shelf for other methods. Good luck regards Tony Reilly Chief Scientist Anatomical Pathology QHPS-Prince Charles Hospital Rode rd Chermside Q 4032 Australia Ph: 07 3350 8543 Fax: 07 33508546 tony_reilly@health.qld.gov.au >>> "Monfils, Paul" 11/30/05 4:52 am >>> Does anyone do Movat's Pentachrome stain? I have a researcher who wants to do it. My question involves the "alkaline alcohol" the procedure calls for. The protocol says to add ammonium hydroxide drop by drop to 95% ethanol until the pH reaches 8.0 or higher. Question 1, a theoretical one - How can you accurately measure pH in a nearly water-free solution? Presumably an ionic compound would not ionize very well in such a solution? Question 2, a practical one - When I put the pH probe into 95% ethanol, the meter already reads higher than 9.0, even without adding any ammonium hydroxide. Is this your experience? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/ received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is strictly prohibited. The information contained in this email, including any attachment sent with it, may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone collect on Australia +61 1800 198 175 or by return email. You should also delete this email, and any copies, from your computer system network and destroy any hard copies produced. If not an intended recipient of this email, you must not copy, distribute or take any action(s) that relies on it; any form of disclosure, modification, distribution and/or publication of this email is also prohibited. Although Queensland Health takes all reasonable steps to ensure this email does not contain malicious software, Queensland Health does not accept responsibility for the consequences if any person's computer inadvertently suffers any disruption to services, loss of information, harm or is infected with a virus, other malicious computer programme or code that may occur as a consequence of receiving this email. Unless stated otherwise, this email represents only the views of the sender and not the views of the Queensland Government. **************************************************************** From louise.renton <@t> gmail.com Wed Nov 30 04:13:17 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Wed Nov 30 04:13:29 2005 Subject: [Histonet] IHC staining intensity. Message-ID: Dear all, I am currently in discussion with someone doing research who is using a semi quantitative method of reporting stainig using INTENSITY of cytoplasmic staining as a means of quantifying results. Staining for a specific antibody is reported as ranging from zero to 3+. What I would like to know is whether IHC intensity is influenced by the thickness of the section. What I mean is, if staining is seen in a whole cell, or one that has been bisected - will there not be a variation in that one will appear darker when compared to the other? Or is this variation minimal? Is my logic fuzzy? I appreciate any input on this matter best regards Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....onwards through the fog!" From brandk <@t> gmail.com Wed Nov 30 05:07:40 2005 From: brandk <@t> gmail.com (Karl) Date: Wed Nov 30 05:07:50 2005 Subject: [Histonet] cryosection bubbles Message-ID: <65146eb50511300307i488de29ep8dbcf8ad2589a4a@mail.gmail.com> Dear Histonetters, When making cryosections of mouse tissues (brain and liver, which is all i section these days) i get bubble formation between the slide and the section. The bubbles are 0.2 -1.0 mm in diameter, and sufficiently large to contact the blade holder when picking up additional sections on the same slide. There are no bubbles visible in the OCT, so it seems clear the bubbles are introduced at the moment the slide is contacting the sections. I get these bubbles regardless of section thickness (10ums - 25ums). I'm very satisfied with the sections, but which i could eliminate these bubbles! Any tips or aspects to pay attention to in this regard are greatly appreciated. many thanks in advance, Karl From BMolinari <@t> heart.thi.tmc.edu Wed Nov 30 05:39:18 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Wed Nov 30 05:42:54 2005 Subject: [Histonet] Movat's Pentachrome question Message-ID: I do many Movats. I make my alkaline alcohol w/ 90ml 95% ETOH and add 10 ml ammonium hydroxide. I do not worry about pH. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Tuesday, November 29, 2005 12:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Movat's Pentachrome question Does anyone do Movat's Pentachrome stain? I have a researcher who wants to do it. My question involves the "alkaline alcohol" the procedure calls for. The protocol says to add ammonium hydroxide drop by drop to 95% ethanol until the pH reaches 8.0 or higher. Question 1, a theoretical one - How can you accurately measure pH in a nearly water-free solution? Presumably an ionic compound would not ionize very well in such a solution? Question 2, a practical one - When I put the pH probe into 95% ethanol, the meter already reads higher than 9.0, even without adding any ammonium hydroxide. Is this your experience? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Nov 30 07:02:41 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 30 07:02:55 2005 Subject: [Histonet] IHC staining intensity. In-Reply-To: Message-ID: <20051130130241.98189.qmail@web61218.mail.yahoo.com> Hi Louise: Quantifying IHC staining intensity as an evaluation of the overall reaction and epitope abundance has been used in several procedures and consistency on several qualitative aspects of the sections have been identified as the limiting parameter in several procedures. There are several papers published in the Journal of Histotechnology that deal with this aspect (15: 123-126, 1992; 18: 119-125,1995; 19:23-25,1996; 22:109-111,1999). This year I have also published a paper in the JOH [Quantifying Quality: a review and scale proposal; 28: 89-97, 2005) that, as the title indicates, deals with this subject. If you don't have access to my paper, you can e-mail me your FAX number and I can send you a copy. You are not "logic fuzzy" because there will be cases of cells that have lost their "integrity" due to sectioning. The approach would be to define which cells or parts of the cells are going to be measured, and only use them. Rene J. louise renton wrote: Dear all, I am currently in discussion with someone doing research who is using a semi quantitative method of reporting stainig using INTENSITY of cytoplasmic staining as a means of quantifying results. Staining for a specific antibody is reported as ranging from zero to 3+. What I would like to know is whether IHC intensity is influenced by the thickness of the section. What I mean is, if staining is seen in a whole cell, or one that has been bisected - will there not be a variation in that one will appear darker when compared to the other? Or is this variation minimal? Is my logic fuzzy? I appreciate any input on this matter best regards Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....onwards through the fog!" _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From Don.Birgerson <@t> leica-microsystems.com Wed Nov 30 08:04:08 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Wed Nov 30 08:04:23 2005 Subject: [Histonet] cryosection bubbles In-Reply-To: <65146eb50511300307i488de29ep8dbcf8ad2589a4a@mail.gmail.com> Message-ID: Hi Karl, It sounds like you are cutting to cold. Try raising the temperature to -10 to -15. The tissues you mention contain a lot of water and the ice crystals that form get harder at the colder temperatures. The softer crystals (warmer) are broken by your cutting edge when it passes through the tissue. If the crystals are to cold, they are scraped out of the tissue surface leaving little craters. Try testing this by cutting at 3 to 4 microns. If this "to cold" condition exists. the tissue will crumble rather than produce a slice. If you have further questions, please call me. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 (7:00 to 4:00 CT) Karl Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2005 05:07 AM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] cryosection bubbles Dear Histonetters, When making cryosections of mouse tissues (brain and liver, which is all i section these days) i get bubble formation between the slide and the section. The bubbles are 0.2 -1.0 mm in diameter, and sufficiently large to contact the blade holder when picking up additional sections on the same slide. There are no bubbles visible in the OCT, so it seems clear the bubbles are introduced at the moment the slide is contacting the sections. I get these bubbles regardless of section thickness (10ums - 25ums). I'm very satisfied with the sections, but which i could eliminate these bubbles! Any tips or aspects to pay attention to in this regard are greatly appreciated. many thanks in advance, Karl _______________________________________________ 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 Barry.R.Rittman <@t> uth.tmc.edu Wed Nov 30 08:10:19 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Wed Nov 30 08:10:24 2005 Subject: [Histonet] Off topic and long Message-ID: As there was so much discussion and so many opinions about the term "bugger". I am listing below an extract from a 1937 book I have as you may find this interesting and that you may all be correct. Remember this was printed before 1939 so GW here refers to the Great War (first world war). Barry A DICTIONARY OF SLANG AND UNCONVENTIONAL ENGLISH. Slang - including the language of the underworld. Colloquialisms and Catch -phrases Solecisms and Catachreses Nicknames Vulgarisms And Such Americanisms as have been naturalized. By Eric Partridge Published by George Rutledge & Sons, Limited London, 1937 buggah. A variant, rare in C. 20, of sense 2 of: bugger. In c., a stealer of breast-pins from C drunks: C. 19. Ex bug, n., 2.-2. A man: fellow; chap: low coil.; 1719, D'Urfey. In S.E. (C. 16- P 20), a sodomite. In low coll. and in dial., as in the U.S., the word has no offensive connotation whatsoever: cf. the remark at pakeha, q.v., and the gradual and complete decolorisation of bastard, q.v., and of Fr. bougre, as in C. 19-20 un bon bougre, a good chap. But also as a pejorative: disagreeable person of either sex; an unpleasant, very difficult, or dangerous thing, project, episode, circumstance, as in G.W. 'It's a bugger making a raid on a wet night.' In 1929, still an actionable word if: printed (Norah James: Sleeveless Errand); in 1934, no longer so (R.Blaker: Night-Shift; GeoffreyDennis: Bloody Mary's). See also bugger, not a. Ex L. Bulgaris, a Bulgarian: the Albigensian heretics were often perverts. O.E.D.; E.D.D.; and the introduction to B. & P. bugger, v. To spoil; ruin; check or change drastically: from ca. 1880; in 1914 +, badly wounded, done for. In the G.W. the Tommy and his Colonial peers were often heard to say, Well, that's buggered it.' Doubtless & development from the S.E. sense, to commit sodomy with. The past PJ ppl. passive, buggered, occurs in expletive phrases, e.g. 'Well, I'm buggered!', damned; . you be buggered ! ' (cf. . 'bugger you !'), go to the devil! - 2. Vi. and t., to cheat at cards: c. or low: late C. 19-20; ob.-3. See bugger about. bugger! A strong expletive: latish C. 19-20 Manchon. bugger, not a. Not at all, as in not care a bugger: low coil.: C.20. Geoffrey Dennis, 1934. bugger about, v. Potter about; fuss; act ineffectually; waste time on a thing, with a person. Hence, bugger about with, to caress intimately; interfere with (person or thing). C. 20 : colI. rather than s.; in Australia more than in Britain. bugger all. A low variant of damn all q.v. bugger up. To spoil, ruin; nullify: low: late 101 C.19-20. Cf. bugger, v., 1. bugger you! A strong expletive: low (- 1887). Baumann. buggered. See bugger, V., 1, latter part. buggerlugs. An offensive term of address: mainly nautical: late C. 19-20. (J. Brophy, Waterfront, 1934.) buggery. (In S.E., sodomy: like bugger and to bugger, it is the correct legal term: see O.E.D. and S.O.D.) In unconventional English, in two phrases: (all) to buggery, completely, destructively, ruinously: C. 20. In G. W., ' Our batteries shelled poor old Jerry to buggery'; Manchon.-2. like buggery: either vigorously, cruelly, vindictively; or, as an expletive, certainly not! From ca. 1890. buggly, v.t. To exchange, to swap: military: C.20. F. & Gibbons. ? ex Hindustani. *bugher; occ. as in Coles, 1676, bughar. .A dog, esp. if a mongrel or given to yelping or barking: ca. 1670-1820: orig. c., then low. Cf. buffer, 1, and see bufe. bugs. A dirty seaman: nautical: late C. 19-20. Bowen. Cf. bug.trap.-2. Bacteria; bacteriology: medical students' (- 1933). Slang, p. 191. From contact <@t> excaliburpathology.com Wed Nov 30 09:55:28 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Wed Nov 30 09:55:38 2005 Subject: [Histonet] Probe-On Slides Message-ID: <20051130155528.85433.qmail@web50103.mail.yahoo.com> Hello everyone, if anyone is still using Probe-On slides, I just saw an auction on Ebay for almost 5000, with a starting bid of $40! The item number is 7567090814 or do a search for blank slides. Paula From gentras <@t> vetmed.auburn.edu Wed Nov 30 10:07:18 2005 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Wed Nov 30 10:07:31 2005 Subject: [Histonet] Acidified PTAH Message-ID: <6.0.1.1.0.20051130100300.019a59d0@mailhost.vetmed.auburn.edu> Hello All, a colleague of mine has asked that I inquire about an acidified version of PTAH and it's formula(s). Therefore, if anyone has one will you be so kind as to share it with me via histonet or direct reply. Your prompt replies will be greatly appreciated. Atoska From judi.ford <@t> jax.org Wed Nov 30 10:07:58 2005 From: judi.ford <@t> jax.org (judi.ford@jax.org) Date: Wed Nov 30 10:08:17 2005 Subject: [Histonet] Bile Staining in Mice Message-ID: <5878570.1133366878176.JavaMail.ocsadmin@jcs-mid-prod.jax.org> I was wondering if any one could offer suggestions as to why our bile stain on mouse liver isn't working. The human positive control works well each time we run the stain, but there is no staining on any of the mouse liver tissue. We've tried different fixes, different strains of mice, etc. Even mice that should be very positive with bile show no positive staining. We're stumped........ Judi Ford HIstotechnologist HIstopathology & Microscopy The Jackson Laboratory 600 Main St. Bar Harbor, Me 04609 207-288-6193 From pmarcum <@t> vet.upenn.edu Wed Nov 30 10:23:49 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Wed Nov 30 10:24:10 2005 Subject: [Histonet] Probe-On Slides In-Reply-To: <20051130155528.85433.qmail@web50103.mail.yahoo.com> References: <20051130155528.85433.qmail@web50103.mail.yahoo.com> Message-ID: <6.1.1.1.2.20051130112059.019c2df8@mail.vet.upenn.edu> At 10:55 AM 11/30/2005, Paula Pierce wrote: >Hello everyone, > > if anyone is still using Probe-On slides, I just saw an auction on Ebay > for almost 5000, with a starting bid of $40! > > The item number is 7567090814 or do a search for blank slides. > > Paula >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Be Careful!! Charged slides can lose the charge over time and the expiration date should be taken seriously. If anyone looks into these slides see if you can get the expiration date and how they have been stored over the time the lab had them. I learned this the hard way with some older stored slides from another lab and got burned when the sections came off. Well kept and stored slides at consistent temperature are safer longer. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From PIXLEYSK <@t> UCMAIL.UC.EDU Wed Nov 30 10:39:26 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Wed Nov 30 10:39:32 2005 Subject: [Histonet] NBF or paraformaldehyde Message-ID: Dear Histonet: I should probably know this, but I don't, so I am asking. What is the difference between using neutral buffered formalin and paraformaldehyde? Does one give better morphology or better recovery of antigens for immunostaining? I am a researcher, not a histotech, and I have always used para. However, with all this talk of NBF, I am wondering why. I believe that I have been told at one time or another that the crosslinking is better with para, but since I am always doing immunostaining, I don't want strong crosslinking. I apologize if this is a totally "you should know" question. If there are websites describing this, I would appreciate getting them, because that will save me some time looking them up. Thanks, this is a great listserve! Sarah Pixley Univ. of Cincinnati From PMonfils <@t> Lifespan.org Wed Nov 30 10:42:49 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Wed Nov 30 10:43:02 2005 Subject: [Histonet] Problem with Vectashield Hard Set Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717622@lsexch.lsmaster.lifespan.org> A couple of weeks ago I inquired about aqueous coverslipping media that harden. Vectashield Hard Set was recommended by a few people. Then I remembered that I actually had some of the product, which I purchased a couple of months ago, in my refrigerator. I tried it, and at first was very pleased with it. It seemed to have a refractive index closer to that of glass than most aqueous products, and the resolution was excellent. However, I am now finding that after being coverslipped for a couple of weeks a large amount of air space develops, first immediately around the tissue section, and eventually extending onto the tissue section. This does not appear to be the typical problem of air being drawn under the edges of the coverslip due to evaporation of solvent, as there is no air space near the edges, only on and immediately around the tissue. Besides, the stuff is supposed to harden overnight, according to the package insert. Has anyone else encountered this problem? What is the nature of the problem? Are the spaces actually air? Or some evaporated component of the product? And any hints on how to deal with it, other than soaking off the coverslips and recoverslipping? From ian.montgomery <@t> bio.gla.ac.uk Wed Nov 30 10:51:27 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Wed Nov 30 10:51:39 2005 Subject: Fwd: [Histonet] Off topic and long Message-ID: <7.0.0.16.2.20051130163746.035f65c0@bio.gla.ac.uk> > >As there was so much discussion and so many opinions about the term >"bugger". I am listing below an extract from a 1937 book I have as you >may find this interesting and that you may all be correct. Remember this >was printed before 1939 so GW here refers to the Great War (first world >war). > >Barry Such is the beauty of the English language that the term 'bugger' can be used several times in the same sentence and have totally different meanings. Then we come to the magnificence of Lallan's Scots, same word, different meanings and different pronunciation. Small island but so much linguistics on offer. Ian. Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From cforster <@t> umn.edu Wed Nov 30 10:51:39 2005 From: cforster <@t> umn.edu (Colleen Forster) Date: Wed Nov 30 10:51:53 2005 Subject: [Histonet] E.Coli and enterococcus IHC Message-ID: <438DD89B.4020502@umn.edu> To all in Histoland, Has anyone ever done IHC staining in FFPE tissue for e.coli or enterococcus? If so, PLEASE share your info. Yhanks, Colleen Forster U of MN From tpmorken <@t> labvision.com Wed Nov 30 10:53:53 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Wed Nov 30 10:54:10 2005 Subject: [Histonet] NBF or paraformaldehyde Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D3DE@usca0082k08.labvision.apogent.com> Sarah, Besides the various buffer components, nd assuming identical concentration, functionally they are exactly the same - paraformaldehyde is just polymerized formaldehyde. Commercial neutral buffered formalin may have methanol added to prevent polyermization over long periods of time, but not enough to make a difference in fixation. The reason to use paraformaldehyde to make your formalin is to control the ingredients so you know what is in it. There may also be other ingredients you wish to add for various experimental reasons. Therefore, it may be better to use formalin you make yourself so you can control your experiments properly. A commercial NBF (Carson's modified Millonig's buffered formalin) is also excellent for electron microscopy. For general pathology, any NBF is usually fine. 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: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pixley, Sarah (pixleysk) Sent: Wednesday, November 30, 2005 8:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] NBF or paraformaldehyde Dear Histonet: I should probably know this, but I don't, so I am asking. What is the difference between using neutral buffered formalin and paraformaldehyde? Does one give better morphology or better recovery of antigens for immunostaining? I am a researcher, not a histotech, and I have always used para. However, with all this talk of NBF, I am wondering why. I believe that I have been told at one time or another that the crosslinking is better with para, but since I am always doing immunostaining, I don't want strong crosslinking. I apologize if this is a totally "you should know" question. If there are websites describing this, I would appreciate getting them, because that will save me some time looking them up. Thanks, this is a great listserve! Sarah Pixley Univ. of Cincinnati _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Wed Nov 30 10:55:25 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Wed Nov 30 10:55:38 2005 Subject: [Histonet] NBF or paraformaldehyde Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717623@lsexch.lsmaster.lifespan.org> The active ingredient in both products is formaldehyde. Commercial formaldehyde solution contains at least 10% methanol as a stabilizer. Paraformaldehyde, which is simply polymerized formaldehyde, is a pure compound - nothing but formaldehyde molecules. When you dissolve it in distilled water you get a formaldehyde solution free of methanol and other compounds that might be present in commercial formaldehyde solution. In my opinion, either "10% formalin" solution (which contains about 4% formaldehyde) and 4% paraformaldehyde solution (which also contains 4% formaldehyde) provide identical results for most histological applications, certainly for purely morphological studies. However, some biochemical studies could be compromised by the presence of methanol, and in such cases the addditional trouble and expense of using paraformaldehyde are justifiable. I do projects for many different researchers, and quite a few of them routinely use para when formalin would work just as well for what they are doing. From c.ingles <@t> hosp.wisc.edu Wed Nov 30 10:55:13 2005 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Wed Nov 30 10:59:16 2005 Subject: [Histonet] Re: 70% from NBF References: <746FDB897740814EA52BDDCB5ED1DDBC02EB13CF@medimmune4.medimmune.com><6.0.0.22.1.20051129083656.01b51a08@gemini.msu.montana.edu> <000501c5f508$7210e320$690a4246@yourlk4rlmsu> Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE79@uwhis-xchng2.hosp.wisc.edu> I have wondered the same thing many times myself. Whether it was naturally me or the addition of the chemicals that made me a bit strange. I think it may be partly both. I usually blame it on the chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn Sent: Tue 11/29/2005 11:15 AM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue regulations > then. Take his advice! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bhewlett <@t> cogeco.ca Wed Nov 30 11:03:46 2005 From: bhewlett <@t> cogeco.ca (bhewlett@cogeco.ca) Date: Wed Nov 30 11:05:05 2005 Subject: [Histonet] NBF or paraformaldehyde Message-ID: <438ddb72.13e.3bdb.10351@cogeco.ca> > Sarah, There is no difference between 4% NBF and 4% paraformaldehyde in the same buffer! That is because paraformaldehyde in solution IS formaldehyde. Bryan > > Dear Histonet: > I should probably know this, but I don't, so I am asking. What is the > difference between using neutral buffered formalin and paraformaldehyde? > Does one give better morphology or better recovery of antigens for > immunostaining? I am a researcher, not a histotech, and I have always > used para. However, with all this talk of NBF, I am wondering why. I > believe that I have been told at one time or another that the > crosslinking is better with para, but since I am always doing > immunostaining, I don't want strong crosslinking. > > I apologize if this is a totally "you should know" question. If there > are websites describing this, I would appreciate getting them, because > that will save me some time looking them up. > Thanks, this is a great listserve! > > Sarah Pixley > Univ. of Cincinnati > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Wed Nov 30 11:10:54 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Wed Nov 30 11:11:13 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D3DF@usca0082k08.labvision.apogent.com> The first time I walked into a histology lab it was the day after the 4th of July and there were 4 blackened fingers sitting on the grossing bench (one guess how they got there - and it's nothing to do with anything Cajun!). My first thought was : 'this is going to be a strange job.' I've seen much stranger things since, so I think histotechs become strange due to exposure to unnatural sights (among other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Wednesday, November 30, 2005 8:55 AM To: Histonet Subject: RE: [Histonet] Re: 70% from NBF I have wondered the same thing many times myself. Whether it was naturally me or the addition of the chemicals that made me a bit strange. I think it may be partly both. I usually blame it on the chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn Sent: Tue 11/29/2005 11:15 AM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue > regulations then. Take his advice! _______________________________________________ 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.ingles <@t> hosp.wisc.edu Wed Nov 30 11:12:11 2005 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Wed Nov 30 11:17:04 2005 Subject: [Histonet] cryosection bubbles References: <65146eb50511300307i488de29ep8dbcf8ad2589a4a@mail.gmail.com> Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE7B@uwhis-xchng2.hosp.wisc.edu> We occasionally get bubbles between the section and the slide. It ususally helps if you warm the slide with your thumb, etc, before putting the section on so it adheres faster. I find the slower the section sticks to the slide, the more prevelant air bubbles and wrinkles are. Softly breathing on the slide while the section is adhering helps too. Claire Ingles Mohs Clinic, UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Karl Sent: Wed 11/30/2005 5:07 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cryosection bubbles Dear Histonetters, When making cryosections of mouse tissues (brain and liver, which is all i section these days) i get bubble formation between the slide and the section. The bubbles are 0.2 -1.0 mm in diameter, and sufficiently large to contact the blade holder when picking up additional sections on the same slide. There are no bubbles visible in the OCT, so it seems clear the bubbles are introduced at the moment the slide is contacting the sections. I get these bubbles regardless of section thickness (10ums - 25ums). I'm very satisfied with the sections, but which i could eliminate these bubbles! Any tips or aspects to pay attention to in this regard are greatly appreciated. many thanks in advance, Karl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Wed Nov 30 11:22:13 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Wed Nov 30 11:22:19 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: Brian Suspect that it is both. This is a strange profession but also satisfying at the same time. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Wednesday, November 30, 2005 11:11 AM To: 'Ingles Claire'; Histonet Subject: [Histonet] Histotechs: born or made? The first time I walked into a histology lab it was the day after the 4th of July and there were 4 blackened fingers sitting on the grossing bench (one guess how they got there - and it's nothing to do with anything Cajun!). My first thought was : 'this is going to be a strange job.' I've seen much stranger things since, so I think histotechs become strange due to exposure to unnatural sights (among other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Wednesday, November 30, 2005 8:55 AM To: Histonet Subject: RE: [Histonet] Re: 70% from NBF I have wondered the same thing many times myself. Whether it was naturally me or the addition of the chemicals that made me a bit strange. I think it may be partly both. I usually blame it on the chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn Sent: Tue 11/29/2005 11:15 AM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue > regulations then. Take his advice! _______________________________________________ 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 froyer <@t> bitstream.net Wed Nov 30 11:28:18 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Wed Nov 30 11:29:12 2005 Subject: [Histonet] Histotechs: born or made? In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328D3DF@usca0082k08.labvision.apogent.com> Message-ID: <20051130172902.D4CF82B174A@smtpout-2.iphouse.net> When I was a practicing laboratory scientist (27 years ago), we would have some of the wildest lab parties and everyone seemed to be on the same page as far as having a weird sense of humor. A work day didn't go by without some form of laughter in our lab. Non-laboratory people often asked me why this was. The only thing that I could come up with is it was how we dealt with the profession that we chose. I won't go into details or give examples. We all know what I am talking about. It does take a special kind of person to this sometimes morbid (some would say hideous) work and I for one am glad that there are these types of persons to take it on. Thank you all for your dedication to your profession and the people that you serve - mankind. ~ Ford Ford M. Royer, MT(ASCP) Minneapolis, MN -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Wednesday, November 30, 2005 11:11 AM To: 'Ingles Claire'; Histonet Subject: [Histonet] Histotechs: born or made? The first time I walked into a histology lab it was the day after the 4th of July and there were 4 blackened fingers sitting on the grossing bench (one guess how they got there - and it's nothing to do with anything Cajun!). My first thought was : 'this is going to be a strange job.' I've seen much stranger things since, so I think histotechs become strange due to exposure to unnatural sights (among other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Wednesday, November 30, 2005 8:55 AM To: Histonet Subject: RE: [Histonet] Re: 70% from NBF I have wondered the same thing many times myself. Whether it was naturally me or the addition of the chemicals that made me a bit strange. I think it may be partly both. I usually blame it on the chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn Sent: Tue 11/29/2005 11:15 AM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue > regulations then. Take his advice! _______________________________________________ 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 cking <@t> rallansci.com Wed Nov 30 11:36:37 2005 From: cking <@t> rallansci.com (King, Curtis - RAS) Date: Wed Nov 30 11:36:47 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <490C1EC04BA10F4891494D0ED756AC9303424254@usmi0012k03.rallansci.apogent.com> I would never make it through a day without craking up over something most people would consider IN BAD TASTE HUMOR. I Love My Job Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ford Royer Sent: Wednesday, November 30, 2005 12:28 PM To: 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? When I was a practicing laboratory scientist (27 years ago), we would have some of the wildest lab parties and everyone seemed to be on the same page as far as having a weird sense of humor. A work day didn't go by without some form of laughter in our lab. Non-laboratory people often asked me why this was. The only thing that I could come up with is it was how we dealt with the profession that we chose. I won't go into details or give examples. We all know what I am talking about. It does take a special kind of person to this sometimes morbid (some would say hideous) work and I for one am glad that there are these types of persons to take it on. Thank you all for your dedication to your profession and the people that you serve - mankind. ~ Ford Ford M. Royer, MT(ASCP) Minneapolis, MN -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Wednesday, November 30, 2005 11:11 AM To: 'Ingles Claire'; Histonet Subject: [Histonet] Histotechs: born or made? The first time I walked into a histology lab it was the day after the 4th of July and there were 4 blackened fingers sitting on the grossing bench (one guess how they got there - and it's nothing to do with anything Cajun!). My first thought was : 'this is going to be a strange job.' I've seen much stranger things since, so I think histotechs become strange due to exposure to unnatural sights (among other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Wednesday, November 30, 2005 8:55 AM To: Histonet Subject: RE: [Histonet] Re: 70% from NBF I have wondered the same thing many times myself. Whether it was naturally me or the addition of the chemicals that made me a bit strange. I think it may be partly both. I usually blame it on the chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn Sent: Tue 11/29/2005 11:15 AM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue > regulations then. Take his advice! _______________________________________________ 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 gcallis <@t> montana.edu Wed Nov 30 11:56:57 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Nov 30 11:57:13 2005 Subject: [Histonet] Problem with Vectashield Hard Set In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE01717622@lsexch.lsmaster.l ifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE01717622@lsexch.lsmaster.lifespan.org> Message-ID: <6.0.0.22.1.20051130105122.01b3dc68@gemini.msu.montana.edu> Paul, What you need to do with Vectashield Hard Set and also with Molecular Probes Prolong Gold antifade, ready to use (hard set) is seal the edges of the coverslip with nail polish (only if you do not have GFP in the tissue section). These hard set mounting medias retract, as you describe, when they dry. Sometimes the inserts do not tell you these little things - I learned of this from both companies tech services. We like to dump the clear nail polish out, rinse the nail polish bottle with acetone, let dry then put toluene or xylene diluted mounting media in the little bottle since we like the tidy little brush for application to coverglass edges. When we have GFP or DsRed present in tissue section, we use the diluted mounting media - since it does NOT contain alcohol. At 09:42 AM 11/30/2005, you wrote: >A couple of weeks ago I inquired about aqueous coverslipping media that >harden. Vectashield Hard Set was recommended by a few people. Then I >remembered that I actually had some of the product, which I purchased a >couple of months ago, in my refrigerator. I tried it, and at first was very >pleased with it. It seemed to have a refractive index closer to that of >glass than most aqueous products, and the resolution was excellent. >However, I am now finding that after being coverslipped for a couple of >weeks a large amount of air space develops, first immediately around the >tissue section, and eventually extending onto the tissue section. This does >not appear to be the typical problem of air being drawn under the edges of >the coverslip due to evaporation of solvent, as there is no air space near >the edges, only on and immediately around the tissue. Besides, the stuff is >supposed to harden overnight, according to the package insert. > >Has anyone else encountered this problem? What is the nature of the >problem? Are the spaces actually air? Or some evaporated component of the >product? And any hints on how to deal with it, other than soaking off the >coverslips and recoverslipping? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From rjbuesa <@t> yahoo.com Wed Nov 30 12:07:54 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 30 12:08:03 2005 Subject: [Histonet] Histotechs: born or made? In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328D3DF@usca0082k08.labvision.apogent.com> Message-ID: <20051130180754.60705.qmail@web61215.mail.yahoo.com> This can be brought to the old and always controversial debate about "nature" or "nurture", I think. Our job requires dexterity, attention to detail, good color appreciation, concentration power, some physical stamina, dedication and s/he who does not like it, will always be a mediocre histotech. Which of these, and other similar qualities, are "born" and which are "made"? You can choose, but for me is a combination where I think that "nature" has a bigger component. At least this is what I think! Rene J. "Morken, Tim - Labvision" wrote: The first time I walked into a histology lab it was the day after the 4th of July and there were 4 blackened fingers sitting on the grossing bench (one guess how they got there - and it's nothing to do with anything Cajun!). My first thought was : 'this is going to be a strange job.' I've seen much stranger things since, so I think histotechs become strange due to exposure to unnatural sights (among other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Wednesday, November 30, 2005 8:55 AM To: Histonet Subject: RE: [Histonet] Re: 70% from NBF I have wondered the same thing many times myself. Whether it was naturally me or the addition of the chemicals that made me a bit strange. I think it may be partly both. I usually blame it on the chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn Sent: Tue 11/29/2005 11:15 AM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue > regulations then. Take his advice! _______________________________________________ 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 --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From mucram11 <@t> comcast.net Wed Nov 30 12:22:20 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Wed Nov 30 12:22:31 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <113020051822.25284.438DEDDC0002D17E000062C42200734748CECE030E9D0C9A03@comcast.net> Ford, I don't think we are allowed to tell the new people in the field how much fun we had in the old days. I have always loved my job however, sometimes I have to watch my sense of humor in non-histologist/medical company as we don't ususally see things the same way they do. Heck, I thought Qunicy and now CSI were sit coms at first as it was so far from what we were doing and really funny for mistakes. All my first boss in histology (and also a city/county coroner) wanted for several years was a Sam like Qunicy had with all the equipment of course. He figured he could rid of at least 5 people in the lab with one Sam. Pam Marcum > When I was a practicing laboratory scientist (27 years ago), we would have > some of the wildest lab parties and everyone seemed to be on the same page > as far as having a weird sense of humor. A work day didn't go by without > some form of laughter in our lab. Non-laboratory people often asked me why > this was. The only thing that I could come up with is it was how we dealt > with the profession that we chose. I won't go into details or give > examples. We all know what I am talking about. It does take a special kind > of person to this sometimes morbid (some would say hideous) work and I for > one am glad that there are these types of persons to take it on. Thank you > all for your dedication to your profession and the people that you serve - > mankind. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minneapolis, MN > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim > - Labvision > Sent: Wednesday, November 30, 2005 11:11 AM > To: 'Ingles Claire'; Histonet > Subject: [Histonet] Histotechs: born or made? > > The first time I walked into a histology lab it was the day after the 4th of > July and there were 4 blackened fingers sitting on the grossing bench (one > guess how they got there - and it's nothing to do with anything Cajun!). My > first thought was : 'this is going to be a strange job.' I've seen much > stranger things since, so I think histotechs become strange due to exposure > to unnatural sights (among other things!). And, of course, the pathology > staff of any hospital is infamous for their "gross" humor. > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > Claire > Sent: Wednesday, November 30, 2005 8:55 AM > To: Histonet > Subject: RE: [Histonet] Re: 70% from NBF > > > I have wondered the same thing many times myself. Whether it was naturally > me or the addition of the chemicals that made me a bit strange. I think it > may be partly both. I usually blame it on the chemical fumes though. :) > Claire Ingles Mohs Clinic, UW Hosp. Madison WI > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn > Sent: Tue 11/29/2005 11:15 AM > To: Histonet > Subject: Re: [Histonet] Re: 70% from NBF > > > > I have often wondered whether I became a histotech because I was born > strange, or whether I became strange because of the time I spent training in > a place like that! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, November 29, 2005 8:23 AM > Subject: [Histonet] Re: 70% from NBF > > > > Joseph made some excellent points here > > > > Chloroform is an excellent clearing agent (used it back in the 60's in > > open dip and dunk processors - O.K. so I'm old!) but no one warned us > > about its carcinogenic nature and there were no safety issue > > regulations then. Take his advice! > > > _______________________________________________ > 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 lrichey <@t> u.washington.edu Wed Nov 30 12:54:52 2005 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Wed Nov 30 12:55:07 2005 Subject: [Histonet] Movat's Pentachrome question In-Reply-To: References: Message-ID: <438DF57C.2020203@u.washington.edu> We do the Movat's stain. The Alkaline alcohol recipe that we use is 45 ml 95% ETOH, 5 ml Concentrated Ammonium Hydroxide. We preheat, the alkaline alcohol sol'n in a 60 degree oven ( I know this is very flammable), and stain in preheated sol'n for 10 min. We don't pH at all. Anthony Reilly wrote: >The alcoholoic ammonia step is the critical stage of Movat's as it is >required to convert the Alcian Blue to insoluble Monastral Blue >so that it will not be removed during the remainder of the > staining method. Just follow the AFIP method which uses 10ml of >ammonium hydroxide in 90ml of 95% alcohol. >If you want to simplify things then you can do what we do. Just >stain using your usual Alcian blue method(pH2.5), treat with alcoholoic >ammonia for 2 hours then do a routine VVG. This will stain your elastic >fibres, collagen and glycoproteins as the Movat does with routine >stains that are already on your shelf for other methods. > >Good luck >regards > >Tony Reilly >Chief Scientist >Anatomical Pathology >QHPS-Prince Charles Hospital >Rode rd Chermside Q 4032 >Australia >Ph: 07 3350 8543 >Fax: 07 33508546 >tony_reilly@health.qld.gov.au > > > > >>>>"Monfils, Paul" 11/30/05 4:52 am >>> >>>> >>>> >Does anyone do Movat's Pentachrome stain? I have a researcher who wants to >do it. My question involves the "alkaline alcohol" the procedure calls for. >The protocol says to add ammonium hydroxide drop by drop to 95% ethanol >until the pH reaches 8.0 or higher. > >Question 1, a theoretical one - How can you accurately measure pH in a >nearly water-free solution? Presumably an ionic compound would not ionize >very well in such a solution? > >Question 2, a practical one - When I put the pH probe into 95% ethanol, the >meter already reads higher than 9.0, even without adding any ammonium >hydroxide. Is this your experience? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >***************************************************************** >This email, including any attachments sent with it, is >confidential and for the sole use of the intended recipient(s). >This confidentiality is not waived or lost, if you receive it and >you are not the intended recipient(s), or if it is transmitted/ >received in error. > >Any unauthorised use, alteration, disclosure, distribution or >review of this email is strictly prohibited. The information >contained in this email, including any attachment sent with >it, may be subject to a statutory duty of confidentiality if it >relates to health service matters. > >If you are not the intended recipient(s), or if you have >received this email in error, you are asked to immediately >notify the sender by telephone collect on Australia >+61 1800 198 175 or by return email. You should also >delete this email, and any copies, from your computer >system network and destroy any hard copies produced. > >If not an intended recipient of this email, you must not copy, >distribute or take any action(s) that relies on it; any form of >disclosure, modification, distribution and/or publication of this >email is also prohibited. > >Although Queensland Health takes all reasonable steps to >ensure this email does not contain malicious software, >Queensland Health does not accept responsibility for the >consequences if any person's computer inadvertently suffers >any disruption to services, loss of information, harm or is >infected with a virus, other malicious computer programme or >code that may occur as a consequence of receiving this >email. > >Unless stated otherwise, this email represents only the views >of the sender and not the views of the Queensland Government. >**************************************************************** > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From jqb7 <@t> cdc.gov Wed Nov 30 12:52:23 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Wed Nov 30 12:59:34 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: And wasn't it amazing how much Sam and Quincy got done in one hour!? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam Marcum Sent: Wednesday, November 30, 2005 1:22 PM To: Ford Royer; 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? Ford, I don't think we are allowed to tell the new people in the field how much fun we had in the old days. I have always loved my job however, sometimes I have to watch my sense of humor in non-histologist/medical company as we don't ususally see things the same way they do. Heck, I thought Qunicy and now CSI were sit coms at first as it was so far from what we were doing and really funny for mistakes. All my first boss in histology (and also a city/county coroner) wanted for several years was a Sam like Qunicy had with all the equipment of course. He figured he could rid of at least 5 people in the lab with one Sam. Pam Marcum > When I was a practicing laboratory scientist (27 years ago), we would > have some of the wildest lab parties and everyone seemed to be on the > same page as far as having a weird sense of humor. A work day didn't > go by without some form of laughter in our lab. Non-laboratory people > often asked me why this was. The only thing that I could come up with > is it was how we dealt with the profession that we chose. I won't go > into details or give examples. We all know what I am talking about. > It does take a special kind of person to this sometimes morbid (some > would say hideous) work and I for one am glad that there are these > types of persons to take it on. Thank you all for your dedication to > your profession and the people that you serve - mankind. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minneapolis, MN > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Morken, Tim > - Labvision > Sent: Wednesday, November 30, 2005 11:11 AM > To: 'Ingles Claire'; Histonet > Subject: [Histonet] Histotechs: born or made? > > The first time I walked into a histology lab it was the day after the > 4th of July and there were 4 blackened fingers sitting on the grossing > bench (one guess how they got there - and it's nothing to do with > anything Cajun!). My first thought was : 'this is going to be a > strange job.' I've seen much stranger things since, so I think > histotechs become strange due to exposure to unnatural sights (among > other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > Claire > Sent: Wednesday, November 30, 2005 8:55 AM > To: Histonet > Subject: RE: [Histonet] Re: 70% from NBF > > > I have wondered the same thing many times myself. Whether it was > naturally me or the addition of the chemicals that made me a bit > strange. I think it may be partly both. I usually blame it on the > chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison > WI > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > Llewellyn > Sent: Tue 11/29/2005 11:15 AM > To: Histonet > Subject: Re: [Histonet] Re: 70% from NBF > > > > I have often wondered whether I became a histotech because I was born > strange, or whether I became strange because of the time I spent > training in a place like that! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, November 29, 2005 8:23 AM > Subject: [Histonet] Re: 70% from NBF > > > > Joseph made some excellent points here > > > > Chloroform is an excellent clearing agent (used it back in the 60's > > in open dip and dunk processors - O.K. so I'm old!) but no one > > warned us about its carcinogenic nature and there were no safety > > issue regulations then. Take his advice! > > > _______________________________________________ > 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 liz <@t> premierlab.com Wed Nov 30 13:02:26 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Wed Nov 30 13:02:44 2005 Subject: [Histonet] murine herpes virus Message-ID: <001201c5f5e0$9a79f350$a7d48a80@AMY> Does anyone out there know of a probe or antibody that will detect murine herpes virus in tissue sections? Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From lpjones <@t> srhs-pa.org Wed Nov 30 13:03:29 2005 From: lpjones <@t> srhs-pa.org (Jones, Laura) Date: Wed Nov 30 13:03:41 2005 Subject: [Histonet] Paraffin Block Storage Message-ID: <8E7AD740937B954F947F0DB4467EFEE056E967@mail.srhs-pa.org> Greetings to everyone in Histoland! We are hoping you all can help us with a question. We store our paraffin blocks in a small closet/room (5' x 10', we're guessing) and have had a Halon fire extinguishing system in there for years. We have apparently been cited by the PA Dept. of Health (?) for this system not being adequate. So, the question is, what type of system does everyone else have? We realize that regulations may differ from state to state, but would appreciate any input, as our safety officer seems to be puzzled as well. Thanks in advance to all the experts. Laura Jones From mucram11 <@t> comcast.net Wed Nov 30 13:03:36 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Wed Nov 30 13:03:49 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <113020051903.4254.438DF787000497B50000109E2207020953CECE030E9D0C9A03@comcast.net> It is almost as good as the 3 minute turn around time for DNA in CSI. Of course Sam did it all. He didn't need all these other technicans running around getting in his way when a crime needed to be solved. Just goes to show we oldies got more done with less (don't I wish). Pam > And wasn't it amazing how much Sam and Quincy got done in one hour!? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam > Marcum > Sent: Wednesday, November 30, 2005 1:22 PM > To: Ford Royer; 'Histonet' > Subject: RE: [Histonet] Histotechs: born or made? > > Ford, > > I don't think we are allowed to tell the new people in the field how > much fun we had in the old days. I have always loved my job however, > sometimes I have to watch my sense of humor in non-histologist/medical > company as we don't ususally see things the same way they do. > > Heck, I thought Qunicy and now CSI were sit coms at first as it was so > far from what we were doing and really funny for mistakes. All my first > boss in histology (and also a city/county coroner) wanted for several > years was a Sam like Qunicy had with all the equipment of course. He > figured he could rid of at least 5 people in the lab with one Sam. > > Pam Marcum > > > > When I was a practicing laboratory scientist (27 years ago), we would > > have some of the wildest lab parties and everyone seemed to be on the > > same page as far as having a weird sense of humor. A work day didn't > > go by without some form of laughter in our lab. Non-laboratory people > > > often asked me why this was. The only thing that I could come up with > > > is it was how we dealt with the profession that we chose. I won't go > > into details or give examples. We all know what I am talking about. > > It does take a special kind of person to this sometimes morbid (some > > would say hideous) work and I for one am glad that there are these > > types of persons to take it on. Thank you all for your dedication to > > your profession and the people that you serve - mankind. > > > > ~ Ford > > > > Ford M. Royer, MT(ASCP) > > Minneapolis, MN > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > > Morken, Tim > > - Labvision > > Sent: Wednesday, November 30, 2005 11:11 AM > > To: 'Ingles Claire'; Histonet > > Subject: [Histonet] Histotechs: born or made? > > > > The first time I walked into a histology lab it was the day after the > > 4th of July and there were 4 blackened fingers sitting on the grossing > > > bench (one guess how they got there - and it's nothing to do with > > anything Cajun!). My first thought was : 'this is going to be a > > strange job.' I've seen much stranger things since, so I think > > histotechs become strange due to exposure to unnatural sights (among > > other things!). And, of course, the pathology staff of any hospital is > infamous for their "gross" humor. > > > > > > Tim Morken > > > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > > > Claire > > Sent: Wednesday, November 30, 2005 8:55 AM > > To: Histonet > > Subject: RE: [Histonet] Re: 70% from NBF > > > > > > I have wondered the same thing many times myself. Whether it was > > naturally me or the addition of the chemicals that made me a bit > > strange. I think it may be partly both. I usually blame it on the > > chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison > > WI > > > > ________________________________ > > > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > > Llewellyn > > Sent: Tue 11/29/2005 11:15 AM > > To: Histonet > > Subject: Re: [Histonet] Re: 70% from NBF > > > > > > > > I have often wondered whether I became a histotech because I was born > > strange, or whether I became strange because of the time I spent > > training in a place like that! > > > > Bryan Llewellyn > > > > > > ----- Original Message ----- > > From: "Gayle Callis" > > To: > > Sent: Tuesday, November 29, 2005 8:23 AM > > Subject: [Histonet] Re: 70% from NBF > > > > > > > Joseph made some excellent points here > > > > > > Chloroform is an excellent clearing agent (used it back in the 60's > > > in open dip and dunk processors - O.K. so I'm old!) but no one > > > warned us about its carcinogenic nature and there were no safety > > > issue regulations then. Take his advice! > > > > > > _______________________________________________ > > 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 tpmorken <@t> labvision.com Wed Nov 30 13:28:52 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Wed Nov 30 13:29:02 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D3E2@usca0082k08.labvision.apogent.com> One of my classmates in electron microscopy was a consultant for the Quincy show. On one show they wanted to look for needle holes in an orange peel using a transmission electron microscope. My friend said it couldn't be done that way, but might be possible in a scanning EM, if the sample wasn't altered by processing. They didn't like the SEM - not impressive enough size-wise (it matters!). So they did it their way - putting an entire orange in a TEM (with Sam on the EM, of course) and gave us some laughs. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine Sent: Wednesday, November 30, 2005 10:52 AM To: Pam Marcum; Ford Royer; Histonet Subject: RE: [Histonet] Histotechs: born or made? And wasn't it amazing how much Sam and Quincy got done in one hour!? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam Marcum Sent: Wednesday, November 30, 2005 1:22 PM To: Ford Royer; 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? Ford, I don't think we are allowed to tell the new people in the field how much fun we had in the old days. I have always loved my job however, sometimes I have to watch my sense of humor in non-histologist/medical company as we don't ususally see things the same way they do. Heck, I thought Qunicy and now CSI were sit coms at first as it was so far from what we were doing and really funny for mistakes. All my first boss in histology (and also a city/county coroner) wanted for several years was a Sam like Qunicy had with all the equipment of course. He figured he could rid of at least 5 people in the lab with one Sam. Pam Marcum > When I was a practicing laboratory scientist (27 years ago), we would > have some of the wildest lab parties and everyone seemed to be on the > same page as far as having a weird sense of humor. A work day didn't > go by without some form of laughter in our lab. Non-laboratory people > often asked me why this was. The only thing that I could come up with > is it was how we dealt with the profession that we chose. I won't go > into details or give examples. We all know what I am talking about. > It does take a special kind of person to this sometimes morbid (some > would say hideous) work and I for one am glad that there are these > types of persons to take it on. Thank you all for your dedication to > your profession and the people that you serve - mankind. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minneapolis, MN > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Morken, Tim > - Labvision > Sent: Wednesday, November 30, 2005 11:11 AM > To: 'Ingles Claire'; Histonet > Subject: [Histonet] Histotechs: born or made? > > The first time I walked into a histology lab it was the day after the > 4th of July and there were 4 blackened fingers sitting on the grossing > bench (one guess how they got there - and it's nothing to do with > anything Cajun!). My first thought was : 'this is going to be a > strange job.' I've seen much stranger things since, so I think > histotechs become strange due to exposure to unnatural sights (among > other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > Claire > Sent: Wednesday, November 30, 2005 8:55 AM > To: Histonet > Subject: RE: [Histonet] Re: 70% from NBF > > > I have wondered the same thing many times myself. Whether it was > naturally me or the addition of the chemicals that made me a bit > strange. I think it may be partly both. I usually blame it on the > chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison > WI > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > Llewellyn > Sent: Tue 11/29/2005 11:15 AM > To: Histonet > Subject: Re: [Histonet] Re: 70% from NBF > > > > I have often wondered whether I became a histotech because I was born > strange, or whether I became strange because of the time I spent > training in a place like that! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, November 29, 2005 8:23 AM > Subject: [Histonet] Re: 70% from NBF > > > > Joseph made some excellent points here > > > > Chloroform is an excellent clearing agent (used it back in the 60's > > in open dip and dunk processors - O.K. so I'm old!) but no one > > warned us about its carcinogenic nature and there were no safety > > issue regulations then. Take his advice! > > > _______________________________________________ > 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From king.laurie <@t> marshfieldclinic.org Wed Nov 30 13:29:46 2005 From: king.laurie <@t> marshfieldclinic.org (king.laurie@marshfieldclinic.org) Date: Wed Nov 30 13:29:59 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <1daa01c5f5e4$6c3d1c20$8e0110ac@mfldclinframe.org> I used to love that show, until the episode where Quincy complained that Sam was demoted to work in histology, who were, after all, only a bunch of "bottle labelers". Them's fightin' words! Laurie King ------Original Message------ From: "Bartlett, Jeanine" Date: Wed Nov 30, 2005 -- 01:15:00 PM To: "Pam Marcum" , "Ford Royer" , "Histonet" Subject: RE: [Histonet] Histotechs: born or made? And wasn't it amazing how much Sam and Quincy got done in one hour!? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam Marcum Sent: Wednesday, November 30, 2005 1:22 PM To: Ford Royer; 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? Ford, I don't think we are allowed to tell the new people in the field how much fun we had in the old days. I have always loved my job however, sometimes I have to watch my sense of humor in non-histologist/medical company as we don't ususally see things the same way they do. Heck, I thought Qunicy and now CSI were sit coms at first as it was so far from what we were doing and really funny for mistakes. All my first boss in histology (and also a city/county coroner) wanted for several years was a Sam like Qunicy had with all the equipment of course. He figured he could rid of at least 5 people in the lab with one Sam. Pam Marcum > When I was a practicing laboratory scientist (27 years ago), we would > have some of the wildest lab parties and everyone seemed to be on the > same page as far as having a weird sense of humor. A work day didn't > go by without some form of laughter in our lab. Non-laboratory people > often asked me why this was. The only thing that I could come up with > is it was how we dealt with the profession that we chose. I won't go > into details or give examples. We all know what I am talking about. > It does take a special kind of person to this sometimes morbid (some > would say hideous) work and I for one am glad that there are these > types of persons to take it on. Thank you all for your dedication to > your profession and the people that you serve - mankind. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minneapolis, MN > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Morken, Tim > - Labvision > Sent: Wednesday, November 30, 2005 11:11 AM > To: 'Ingles Claire'; Histonet > Subject: [Histonet] Histotechs: born or made? > > The first time I walked into a histology lab it was the day after the > 4th of July and there were 4 blackened fingers sitting on the grossing > bench (one guess how they got there - and it's nothing to do with > anything Cajun!). My first thought was : 'this is going to be a > strange job.' I've seen much stranger things since, so I think > histotechs become strange due to exposure to unnatural sights (among > other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > Claire > Sent: Wednesday, November 30, 2005 8:55 AM > To: Histonet > Subject: RE: [Histonet] Re: 70% from NBF > > > I have wondered the same thing many times myself. Whether it was > naturally me or the addition of the chemicals that made me a bit > strange. I think it may be partly both. I usually blame it on the > chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison > WI > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > Llewellyn > Sent: Tue 11/29/2005 11:15 AM > To: Histonet > Subject: Re: [Histonet] Re: 70% from NBF > > > > I have often wondered whether I became a histotech because I was born > strange, or whether I became strange because of the time I spent > training in a place like that! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, November 29, 2005 8:23 AM > Subject: [Histonet] Re: 70% from NBF > > > > Joseph made some excellent points here > > > > Chloroform is an excellent clearing agent (used it back in the 60's > > in open dip and dunk processors - O.K. so I'm old!) but no one > > warned us about its carcinogenic nature and there were no safety > > issue regulations then. Take his advice! > > > _______________________________________________ > 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet >From histonet-bounces@lists.utsouthwestern.edu Wed Nov 30 13:03:55 2005 From POWELL_SA <@t> Mercer.edu Wed Nov 30 13:45:29 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Wed Nov 30 13:46:00 2005 Subject: [Histonet] Histotechs: born or made? In-Reply-To: <1daa01c5f5e4$6c3d1c20$8e0110ac@mfldclinframe.org> Message-ID: <01LW0H9YR58S8WX40X@Macon2.Mercer.edu> Yep, that is when I stopped watching Quincy. Made me want to fight too. I do watch CSI but usually not for accuracy, mostly for the scenery. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of king.laurie@marshfieldclinic.org Sent: Wednesday, November 30, 2005 2:30 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotechs: born or made? I used to love that show, until the episode where Quincy complained that Sam was demoted to work in histology, who were, after all, only a bunch of "bottle labelers". Them's fightin' words! Laurie King ------Original Message------ From: "Bartlett, Jeanine" Date: Wed Nov 30, 2005 -- 01:15:00 PM To: "Pam Marcum" , "Ford Royer" , "Histonet" Subject: RE: [Histonet] Histotechs: born or made? And wasn't it amazing how much Sam and Quincy got done in one hour!? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam Marcum Sent: Wednesday, November 30, 2005 1:22 PM To: Ford Royer; 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? Ford, I don't think we are allowed to tell the new people in the field how much fun we had in the old days. I have always loved my job however, sometimes I have to watch my sense of humor in non-histologist/medical company as we don't ususally see things the same way they do. Heck, I thought Qunicy and now CSI were sit coms at first as it was so far from what we were doing and really funny for mistakes. All my first boss in histology (and also a city/county coroner) wanted for several years was a Sam like Qunicy had with all the equipment of course. He figured he could rid of at least 5 people in the lab with one Sam. Pam Marcum > When I was a practicing laboratory scientist (27 years ago), we would > have some of the wildest lab parties and everyone seemed to be on the > same page as far as having a weird sense of humor. A work day didn't > go by without some form of laughter in our lab. Non-laboratory people > often asked me why this was. The only thing that I could come up with > is it was how we dealt with the profession that we chose. I won't go > into details or give examples. We all know what I am talking about. > It does take a special kind of person to this sometimes morbid (some > would say hideous) work and I for one am glad that there are these > types of persons to take it on. Thank you all for your dedication to > your profession and the people that you serve - mankind. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minneapolis, MN > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Morken, Tim > - Labvision > Sent: Wednesday, November 30, 2005 11:11 AM > To: 'Ingles Claire'; Histonet > Subject: [Histonet] Histotechs: born or made? > > The first time I walked into a histology lab it was the day after the > 4th of July and there were 4 blackened fingers sitting on the grossing > bench (one guess how they got there - and it's nothing to do with > anything Cajun!). My first thought was : 'this is going to be a > strange job.' I've seen much stranger things since, so I think > histotechs become strange due to exposure to unnatural sights (among > other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > Claire > Sent: Wednesday, November 30, 2005 8:55 AM > To: Histonet > Subject: RE: [Histonet] Re: 70% from NBF > > > I have wondered the same thing many times myself. Whether it was > naturally me or the addition of the chemicals that made me a bit > strange. I think it may be partly both. I usually blame it on the > chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison > WI > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > Llewellyn > Sent: Tue 11/29/2005 11:15 AM > To: Histonet > Subject: Re: [Histonet] Re: 70% from NBF > > > > I have often wondered whether I became a histotech because I was born > strange, or whether I became strange because of the time I spent > training in a place like that! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, November 29, 2005 8:23 AM > Subject: [Histonet] Re: 70% from NBF > > > > Joseph made some excellent points here > > > > Chloroform is an excellent clearing agent (used it back in the 60's > > in open dip and dunk processors - O.K. so I'm old!) but no one > > warned us about its carcinogenic nature and there were no safety > > issue regulations then. Take his advice! > > > _______________________________________________ > 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet >From histonet-bounces@lists.utsouthwestern.edu Wed Nov 30 13:03:55 2005 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From t-sherman <@t> comcast.net Wed Nov 30 13:59:13 2005 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Wed Nov 30 14:00:02 2005 Subject: [Histonet] Re: IHC staining intensity Message-ID: <438E0491.1030303@comcast.net> Louise, It's been a while since I've done semi-quant analysis of IHC sections, and you are right to be concerned about such issues. Section thickness will impact staining intensity. We used a similar integer scale of 0 to +4 to indicate chromogen intensity within a given experiment. Generally, it was sufficient though admittedly imprecise. Such are the limitations of quantitative (even if semi-) observations in IHC... or ISH for that matter. Consider also variable levels of intracellular expression and the plane of the section within particular cells. If you have a perinuclear antigen, for example, and one cell with exposed surface (from cutting) with nuclear material is evident, then that cell will "stain" more intensely than a neighboring cell which might have quantitatively more antigen but has not been cleaved near the nucleus, but which may actually appear to be a measurably larger cell due to the nucleus being localized to one pole. Without getting too exact about an inexact observation technique, the checks we implemented were to note the relative counterstain intensity relative to the IHC chromogen within sections of a given experiment, to not extrapolate observations from one species to the next or from one tissue to the next, and to get several observers corroborate any "measurable" observations. To elaborate on the first point, if the counterstain intensity in one section is measurably higher than another, complementary section, then that usually indicated to us that the section was thicker; consequently, that comparison was discarded. Also, apart from having multiple observers quantify the final product, it helps to have terrific and highly experienced staff to create the source palette. And lots of previous experience observing the tissue/species of interest. I'm not sure if that is really quantifiable either. ;) Good luck with your study, and by all means, review Rene J Buesa's paper and others like it. Semi-quant-IHC a tough nut to crack. Todd Sherman 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: > 9. IHC staining intensity. (louise renton) > ---------------------------------------------------------------------- > > Message: 9 > Date: Wed, 30 Nov 2005 12:13:17 +0200 > From: louise renton > Subject: [Histonet] IHC staining intensity. > To: Histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=ISO-8859-1 > > Dear all, > > I am currently in discussion with someone doing research who is using > a semi quantitative method of reporting stainig using INTENSITY of > cytoplasmic staining as a means of quantifying results. Staining for > a specific antibody is reported as ranging from zero to 3+. What I > would like to know is whether IHC intensity is influenced by the > thickness of the section. What I mean is, if staining is seen in a > whole cell, or one that has been bisected - will there not be a > variation in that one will appear darker when compared to the other? > Or is this variation minimal? Is my logic fuzzy? > > I appreciate any input on this matter > > best regards > > > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "....onwards through the fog!" > > ------------------------------ From pmarcum <@t> vet.upenn.edu Wed Nov 30 14:02:13 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Wed Nov 30 14:02:33 2005 Subject: [Histonet] Histotechs: born or made? In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328D3E2@usca0082k08.labvi sion.apogent.com> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328D3E2@usca0082k08.labvision.apogent.com> Message-ID: <6.1.1.1.2.20051130145703.01a16d30@mail.vet.upenn.edu> Love It!! He probably thought being in EM was a demotion too. Microscopy can be so misunderstood cause they can't see it. I worked with a company selling oligonucleotide sequences. We had movie company call to buy an oligo for use in a new movie as a prop. We could not make them understand it is a big word for a minuscule piece of DNA. I get scared when they say they have slide from histology of a liver or other organ and is nothing but red blood cells. Pam At 02:28 PM 11/30/2005, Morken, Tim - Labvision wrote: >One of my classmates in electron microscopy was a consultant for the Quincy >show. On one show they wanted to look for needle holes in an orange peel >using a transmission electron microscope. My friend said it couldn't be done >that way, but might be possible in a scanning EM, if the sample wasn't >altered by processing. They didn't like the SEM - not impressive enough >size-wise (it matters!). So they did it their way - putting an entire orange >in a TEM (with Sam on the EM, of course) and gave us some laughs. > > >Tim Morken > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bartlett, >Jeanine >Sent: Wednesday, November 30, 2005 10:52 AM >To: Pam Marcum; Ford Royer; Histonet >Subject: RE: [Histonet] Histotechs: born or made? > > >And wasn't it amazing how much Sam and Quincy got done in one hour!? > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam Marcum >Sent: Wednesday, November 30, 2005 1:22 PM >To: Ford Royer; 'Histonet' >Subject: RE: [Histonet] Histotechs: born or made? > >Ford, > >I don't think we are allowed to tell the new people in the field how much >fun we had in the old days. I have always loved my job however, sometimes I >have to watch my sense of humor in non-histologist/medical company as we >don't ususally see things the same way they do. > >Heck, I thought Qunicy and now CSI were sit coms at first as it was so far >from what we were doing and really funny for mistakes. All my first boss in >histology (and also a city/county coroner) wanted for several years was a >Sam like Qunicy had with all the equipment of course. He figured he could >rid of at least 5 people in the lab with one Sam. > >Pam Marcum > > > > When I was a practicing laboratory scientist (27 years ago), we would > > have some of the wildest lab parties and everyone seemed to be on the > > same page as far as having a weird sense of humor. A work day didn't > > go by without some form of laughter in our lab. Non-laboratory people > > > often asked me why this was. The only thing that I could come up with > > > is it was how we dealt with the profession that we chose. I won't go > > into details or give examples. We all know what I am talking about. > > It does take a special kind of person to this sometimes morbid (some > > would say hideous) work and I for one am glad that there are these > > types of persons to take it on. Thank you all for your dedication to > > your profession and the people that you serve - mankind. > > > > ~ Ford > > > > Ford M. Royer, MT(ASCP) > > Minneapolis, MN > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > > Morken, Tim > > - Labvision > > Sent: Wednesday, November 30, 2005 11:11 AM > > To: 'Ingles Claire'; Histonet > > Subject: [Histonet] Histotechs: born or made? > > > > The first time I walked into a histology lab it was the day after the > > 4th of July and there were 4 blackened fingers sitting on the grossing > > > bench (one guess how they got there - and it's nothing to do with > > anything Cajun!). My first thought was : 'this is going to be a > > strange job.' I've seen much stranger things since, so I think > > histotechs become strange due to exposure to unnatural sights (among > > other things!). And, of course, the pathology staff of any hospital is >infamous for their "gross" humor. > > > > > > Tim Morken > > > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > > > Claire > > Sent: Wednesday, November 30, 2005 8:55 AM > > To: Histonet > > Subject: RE: [Histonet] Re: 70% from NBF > > > > > > I have wondered the same thing many times myself. Whether it was > > naturally me or the addition of the chemicals that made me a bit > > strange. I think it may be partly both. I usually blame it on the > > chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison > > WI > > > > ________________________________ > > > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > > Llewellyn > > Sent: Tue 11/29/2005 11:15 AM > > To: Histonet > > Subject: Re: [Histonet] Re: 70% from NBF > > > > > > > > I have often wondered whether I became a histotech because I was born > > strange, or whether I became strange because of the time I spent > > training in a place like that! > > > > Bryan Llewellyn > > > > > > ----- Original Message ----- > > From: "Gayle Callis" > > To: > > Sent: Tuesday, November 29, 2005 8:23 AM > > Subject: [Histonet] Re: 70% from NBF > > > > > > > Joseph made some excellent points here > > > > > > Chloroform is an excellent clearing agent (used it back in the 60's > > > in open dip and dunk processors - O.K. so I'm old!) but no one > > > warned us about its carcinogenic nature and there were no safety > > > issue regulations then. Take his advice! > > > > > > _______________________________________________ > > 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 > > > >_______________________________________________ >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 PMonfils <@t> Lifespan.org Wed Nov 30 14:17:11 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Wed Nov 30 14:17:24 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717625@lsexch.lsmaster.lifespan.org> Yes we have come a long way since one of the pathologists on one of my first jobs referred to histotechs as "glorified salami slicers". Our supervisor however was not one to take any baloney, and promptly told the good doctor that without our salami his diagnoses would be baloney. The only line I remember from Quincy, after watching it for several years, is "Sam, get me a PAS on this - STAT!" But CSI, which I also watch frequently, really is a mix of science and science fiction. I wish we did have machines that could do everything their machines do. Some of them are really way out there. Like the episode where two people had a conversation near a pottery wheel, and the clay picked up the vibrations of their conversation, which were subsequently scanned by a laser and turned back into audible voices. :-) I even noticed a technical error related to my avocation - malacology. In one episode a dealer in illegally harvested abalone (a type of shellfish) is shot. The medical examiner says "serves him right for selling an endangered bivalve". As you may remember from your invertebrate zoology class, abalone are gastropods, not bivalves. From king.laurie <@t> marshfieldclinic.org Wed Nov 30 14:42:28 2005 From: king.laurie <@t> marshfieldclinic.org (king.laurie@marshfieldclinic.org) Date: Wed Nov 30 14:42:40 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <234001c5f5ee$942dfab0$8e0110ac@mfldclinframe.org> .....thank you for sending me off to visit my on-line dictionary, something I try to do at least once every day! Now I know what malacology is. ------Original Message------ From: "Monfils, Paul" Date: Wed Nov 30, 2005 -- 02:20:45 PM To: "'histonet@lists.utsouthwestern.edu'" Subject: RE: [Histonet] Histotechs: born or made? Yes we have come a long way since one of the pathologists on one of my first jobs referred to histotechs as "glorified salami slicers". Our supervisor however was not one to take any baloney, and promptly told the good doctor that without our salami his diagnoses would be baloney. The only line I remember from Quincy, after watching it for several years, is "Sam, get me a PAS on this - STAT!" But CSI, which I also watch frequently, really is a mix of science and science fiction. I wish we did have machines that could do everything their machines do. Some of them are really way out there. Like the episode where two people had a conversation near a pottery wheel, and the clay picked up the vibrations of their conversation, which were subsequently scanned by a laser and turned back into audible voices. :-) I even noticed a technical error related to my avocation - malacology. In one episode a dealer in illegally harvested abalone (a type of shellfish) is shot. The medical examiner says "serves him right for selling an endangered bivalve". As you may remember from your invertebrate zoology class, abalone are gastropods, not bivalves. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Wed Nov 30 14:46:45 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Wed Nov 30 14:47:17 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: I recall an episode of Quincy where Sam's diener integrity was in question. - they banished him (temporarily) to histology. I remember Quincy loudly protesting "Sam is too valuable to sit around and fill little bottles with formaldehyde all day!" I think it was the same episode where they showed someone trying to cut a block with no knife in the microtome. "Morken, Tim - Labvision" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2005 01:28 PM To: Histonet cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: RE: [Histonet] Histotechs: born or made? One of my classmates in electron microscopy was a consultant for the Quincy show. On one show they wanted to look for needle holes in an orange peel using a transmission electron microscope. My friend said it couldn't be done that way, but might be possible in a scanning EM, if the sample wasn't altered by processing. They didn't like the SEM - not impressive enough size-wise (it matters!). So they did it their way - putting an entire orange in a TEM (with Sam on the EM, of course) and gave us some laughs. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine Sent: Wednesday, November 30, 2005 10:52 AM To: Pam Marcum; Ford Royer; Histonet Subject: RE: [Histonet] Histotechs: born or made? And wasn't it amazing how much Sam and Quincy got done in one hour!? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam Marcum Sent: Wednesday, November 30, 2005 1:22 PM To: Ford Royer; 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? Ford, I don't think we are allowed to tell the new people in the field how much fun we had in the old days. I have always loved my job however, sometimes I have to watch my sense of humor in non-histologist/medical company as we don't ususally see things the same way they do. Heck, I thought Qunicy and now CSI were sit coms at first as it was so far from what we were doing and really funny for mistakes. All my first boss in histology (and also a city/county coroner) wanted for several years was a Sam like Qunicy had with all the equipment of course. He figured he could rid of at least 5 people in the lab with one Sam. Pam Marcum > When I was a practicing laboratory scientist (27 years ago), we would > have some of the wildest lab parties and everyone seemed to be on the > same page as far as having a weird sense of humor. A work day didn't > go by without some form of laughter in our lab. Non-laboratory people > often asked me why this was. The only thing that I could come up with > is it was how we dealt with the profession that we chose. I won't go > into details or give examples. We all know what I am talking about. > It does take a special kind of person to this sometimes morbid (some > would say hideous) work and I for one am glad that there are these > types of persons to take it on. Thank you all for your dedication to > your profession and the people that you serve - mankind. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minneapolis, MN > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Morken, Tim > - Labvision > Sent: Wednesday, November 30, 2005 11:11 AM > To: 'Ingles Claire'; Histonet > Subject: [Histonet] Histotechs: born or made? > > The first time I walked into a histology lab it was the day after the > 4th of July and there were 4 blackened fingers sitting on the grossing > bench (one guess how they got there - and it's nothing to do with > anything Cajun!). My first thought was : 'this is going to be a > strange job.' I've seen much stranger things since, so I think > histotechs become strange due to exposure to unnatural sights (among > other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > Claire > Sent: Wednesday, November 30, 2005 8:55 AM > To: Histonet > Subject: RE: [Histonet] Re: 70% from NBF > > > I have wondered the same thing many times myself. Whether it was > naturally me or the addition of the chemicals that made me a bit > strange. I think it may be partly both. I usually blame it on the > chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison > WI > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > Llewellyn > Sent: Tue 11/29/2005 11:15 AM > To: Histonet > Subject: Re: [Histonet] Re: 70% from NBF > > > > I have often wondered whether I became a histotech because I was born > strange, or whether I became strange because of the time I spent > training in a place like that! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, November 29, 2005 8:23 AM > Subject: [Histonet] Re: 70% from NBF > > > > Joseph made some excellent points here > > > > Chloroform is an excellent clearing agent (used it back in the 60's > > in open dip and dunk processors - O.K. so I'm old!) but no one > > warned us about its carcinogenic nature and there were no safety > > issue regulations then. Take his advice! > > > _______________________________________________ > 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 _______________________________________________ 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 funderwood <@t> mcohio.org Wed Nov 30 14:50:57 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Wed Nov 30 14:53:01 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: I'm constantly exchanging jokes with my co-worker. We are always trying to gross each other out. I'm not sure if this is normal, considering I work in a lab by myself. We must be off, Freds >>> "King, Curtis - RAS" 11/30/05 12:36PM >>> I would never make it through a day without craking up over something most people would consider IN BAD TASTE HUMOR. I Love My Job Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ford Royer Sent: Wednesday, November 30, 2005 12:28 PM To: 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? When I was a practicing laboratory scientist (27 years ago), we would have some of the wildest lab parties and everyone seemed to be on the same page as far as having a weird sense of humor. A work day didn't go by without some form of laughter in our lab. Non-laboratory people often asked me why this was. The only thing that I could come up with is it was how we dealt with the profession that we chose. I won't go into details or give examples. We all know what I am talking about. It does take a special kind of person to this sometimes morbid (some would say hideous) work and I for one am glad that there are these types of persons to take it on. Thank you all for your dedication to your profession and the people that you serve - mankind. ~ Ford Ford M. Royer, MT(ASCP) Minneapolis, MN -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Wednesday, November 30, 2005 11:11 AM To: 'Ingles Claire'; Histonet Subject: [Histonet] Histotechs: born or made? The first time I walked into a histology lab it was the day after the 4th of July and there were 4 blackened fingers sitting on the grossing bench (one guess how they got there - and it's nothing to do with anything Cajun!). My first thought was : 'this is going to be a strange job.' I've seen much stranger things since, so I think histotechs become strange due to exposure to unnatural sights (among other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Wednesday, November 30, 2005 8:55 AM To: Histonet Subject: RE: [Histonet] Re: 70% from NBF I have wondered the same thing many times myself. Whether it was naturally me or the addition of the chemicals that made me a bit strange. I think it may be partly both. I usually blame it on the chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn Sent: Tue 11/29/2005 11:15 AM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue > regulations then. Take his advice! _______________________________________________ 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 cking <@t> rallansci.com Wed Nov 30 14:54:55 2005 From: cking <@t> rallansci.com (King, Curtis - RAS) Date: Wed Nov 30 14:55:06 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <490C1EC04BA10F4891494D0ED756AC9303424257@usmi0012k03.rallansci.apogent.com> Jacki, You are correct. I was 16 years of age and in a Histology Training Program when it aired. I remember thinking is this what I am going to be doing when I am 40 (filling and labeling. WOW am I glad I stuck it out. Histology Rocks and I would not give it up even for Quincy's wage. Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie M O'Connor Sent: Wednesday, November 30, 2005 3:47 PM To: Morken, Tim - Labvision Cc: Histonet Subject: RE: [Histonet] Histotechs: born or made? I recall an episode of Quincy where Sam's diener integrity was in question. - they banished him (temporarily) to histology. I remember Quincy loudly protesting "Sam is too valuable to sit around and fill little bottles with formaldehyde all day!" I think it was the same episode where they showed someone trying to cut a block with no knife in the microtome. "Morken, Tim - Labvision" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2005 01:28 PM To: Histonet cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: RE: [Histonet] Histotechs: born or made? One of my classmates in electron microscopy was a consultant for the Quincy show. On one show they wanted to look for needle holes in an orange peel using a transmission electron microscope. My friend said it couldn't be done that way, but might be possible in a scanning EM, if the sample wasn't altered by processing. They didn't like the SEM - not impressive enough size-wise (it matters!). So they did it their way - putting an entire orange in a TEM (with Sam on the EM, of course) and gave us some laughs. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine Sent: Wednesday, November 30, 2005 10:52 AM To: Pam Marcum; Ford Royer; Histonet Subject: RE: [Histonet] Histotechs: born or made? And wasn't it amazing how much Sam and Quincy got done in one hour!? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam Marcum Sent: Wednesday, November 30, 2005 1:22 PM To: Ford Royer; 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? Ford, I don't think we are allowed to tell the new people in the field how much fun we had in the old days. I have always loved my job however, sometimes I have to watch my sense of humor in non-histologist/medical company as we don't ususally see things the same way they do. Heck, I thought Qunicy and now CSI were sit coms at first as it was so far from what we were doing and really funny for mistakes. All my first boss in histology (and also a city/county coroner) wanted for several years was a Sam like Qunicy had with all the equipment of course. He figured he could rid of at least 5 people in the lab with one Sam. Pam Marcum > When I was a practicing laboratory scientist (27 years ago), we would > have some of the wildest lab parties and everyone seemed to be on the > same page as far as having a weird sense of humor. A work day didn't > go by without some form of laughter in our lab. Non-laboratory people > often asked me why this was. The only thing that I could come up with > is it was how we dealt with the profession that we chose. I won't go > into details or give examples. We all know what I am talking about. > It does take a special kind of person to this sometimes morbid (some > would say hideous) work and I for one am glad that there are these > types of persons to take it on. Thank you all for your dedication to > your profession and the people that you serve - mankind. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minneapolis, MN > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Morken, Tim > - Labvision > Sent: Wednesday, November 30, 2005 11:11 AM > To: 'Ingles Claire'; Histonet > Subject: [Histonet] Histotechs: born or made? > > The first time I walked into a histology lab it was the day after the > 4th of July and there were 4 blackened fingers sitting on the grossing > bench (one guess how they got there - and it's nothing to do with > anything Cajun!). My first thought was : 'this is going to be a > strange job.' I've seen much stranger things since, so I think > histotechs become strange due to exposure to unnatural sights (among > other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > Claire > Sent: Wednesday, November 30, 2005 8:55 AM > To: Histonet > Subject: RE: [Histonet] Re: 70% from NBF > > > I have wondered the same thing many times myself. Whether it was > naturally me or the addition of the chemicals that made me a bit > strange. I think it may be partly both. I usually blame it on the > chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison > WI > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > Llewellyn > Sent: Tue 11/29/2005 11:15 AM > To: Histonet > Subject: Re: [Histonet] Re: 70% from NBF > > > > I have often wondered whether I became a histotech because I was born > strange, or whether I became strange because of the time I spent > training in a place like that! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, November 29, 2005 8:23 AM > Subject: [Histonet] Re: 70% from NBF > > > > Joseph made some excellent points here > > > > Chloroform is an excellent clearing agent (used it back in the 60's > > in open dip and dunk processors - O.K. so I'm old!) but no one > > warned us about its carcinogenic nature and there were no safety > > issue regulations then. Take his advice! > > > _______________________________________________ > 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 _______________________________________________ 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 mucram11 <@t> comcast.net Wed Nov 30 14:55:59 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Wed Nov 30 14:56:35 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <113020052055.5296.438E11DF0003CEBA000014B02200734748CECE030E9D0C9A03@comcast.net> I think some of my work mates know some of your work mates!! They can be so talkitive at times when I can't see them as I am here a lone sotr of. Pam > I'm constantly exchanging jokes with my co-worker. We are always trying > to gross each other out. I'm not sure if this is normal, considering I > work in a lab by myself. > > We must be off, > Freds > > >>> "King, Curtis - RAS" 11/30/05 12:36PM >>> > I would never make it through a day without craking up over something > most > people would consider IN BAD TASTE HUMOR. I Love My Job > > Curt > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ford > Royer > Sent: Wednesday, November 30, 2005 12:28 PM > To: 'Histonet' > Subject: RE: [Histonet] Histotechs: born or made? > > > When I was a practicing laboratory scientist (27 years ago), we would > have > some of the wildest lab parties and everyone seemed to be on the same > page > as far as having a weird sense of humor. A work day didn't go by > without > some form of laughter in our lab. Non-laboratory people often asked me > why > this was. The only thing that I could come up with is it was how we > dealt > with the profession that we chose. I won't go into details or give > examples. We all know what I am talking about. It does take a special > kind > of person to this sometimes morbid (some would say hideous) work and I > for > one am glad that there are these types of persons to take it on. Thank > you > all for your dedication to your profession and the people that you > serve - > mankind. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minneapolis, MN > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, > Tim > - Labvision > Sent: Wednesday, November 30, 2005 11:11 AM > To: 'Ingles Claire'; Histonet > Subject: [Histonet] Histotechs: born or made? > > The first time I walked into a histology lab it was the day after the > 4th of > July and there were 4 blackened fingers sitting on the grossing bench > (one > guess how they got there - and it's nothing to do with anything > Cajun!). My > first thought was : 'this is going to be a strange job.' I've seen > much > stranger things since, so I think histotechs become strange due to > exposure > to unnatural sights (among other things!). And, of course, the > pathology > staff of any hospital is infamous for their "gross" humor. > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > Claire > Sent: Wednesday, November 30, 2005 8:55 AM > To: Histonet > Subject: RE: [Histonet] Re: 70% from NBF > > > I have wondered the same thing many times myself. Whether it was > naturally > me or the addition of the chemicals that made me a bit strange. I think > it > may be partly both. I usually blame it on the chemical fumes though. > :) > Claire Ingles Mohs Clinic, UW Hosp. Madison WI > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > Llewellyn > Sent: Tue 11/29/2005 11:15 AM > To: Histonet > Subject: Re: [Histonet] Re: 70% from NBF > > > > I have often wondered whether I became a histotech because I was born > strange, or whether I became strange because of the time I spent > training in > a place like that! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, November 29, 2005 8:23 AM > Subject: [Histonet] Re: 70% from NBF > > > > Joseph made some excellent points here > > > > Chloroform is an excellent clearing agent (used it back in the 60's > in > > open dip and dunk processors - O.K. so I'm old!) but no one warned us > > > about its carcinogenic nature and there were no safety issue > > regulations then. Take his advice! > > > _______________________________________________ > 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 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Nov 30 14:59:01 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 30 14:59:11 2005 Subject: [Histonet] Paraffin Block Storage In-Reply-To: <8E7AD740937B954F947F0DB4467EFEE056E967@mail.srhs-pa.org> Message-ID: <20051130205901.46950.qmail@web61215.mail.yahoo.com> We used to store our paraffin blocks in cabinets also in a room (probably 30 feets long X 6 wide X 8 high); we used to store 9 years worth of blocks and each year we disposed off the oldest to make room for the on going year. We had also extinguishers and a smoke detector. We were never cited. Did they explain you why did they cited you for? I don't see anything wrong with your description. Rene J. "Jones, Laura" wrote: Greetings to everyone in Histoland! We are hoping you all can help us with a question. We store our paraffin blocks in a small closet/room (5' x 10', we're guessing) and have had a Halon fire extinguishing system in there for years. We have apparently been cited by the PA Dept. of Health (?) for this system not being adequate. So, the question is, what type of system does everyone else have? We realize that regulations may differ from state to state, but would appreciate any input, as our safety officer seems to be puzzled as well. Thanks in advance to all the experts. Laura Jones _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From Jackie.O'Connor <@t> abbott.com Wed Nov 30 15:10:56 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Wed Nov 30 15:11:25 2005 Subject: [Histonet] Paraffin Block Storage Message-ID: Maybe it's the Halon they have a problem with - I thought those were banned because if you're in there when the halon goes off - you die. Just a little problem. Halon removes the oxygen from the air. Rene J Buesa Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2005 02:59 PM To: "Jones, Laura" , "Histonet \(E-mail\)" cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: Re: [Histonet] Paraffin Block Storage We used to store our paraffin blocks in cabinets also in a room (probably 30 feets long X 6 wide X 8 high); we used to store 9 years worth of blocks and each year we disposed off the oldest to make room for the on going year. We had also extinguishers and a smoke detector. We were never cited. Did they explain you why did they cited you for? I don't see anything wrong with your description. Rene J. "Jones, Laura" wrote: Greetings to everyone in Histoland! We are hoping you all can help us with a question. We store our paraffin blocks in a small closet/room (5' x 10', we're guessing) and have had a Halon fire extinguishing system in there for years. We have apparently been cited by the PA Dept. of Health (?) for this system not being adequate. So, the question is, what type of system does everyone else have? We realize that regulations may differ from state to state, but would appreciate any input, as our safety officer seems to be puzzled as well. Thanks in advance to all the experts. Laura Jones _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MadaryJ <@t> MedImmune.com Wed Nov 30 15:13:34 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Wed Nov 30 15:13:44 2005 Subject: [Histonet] practical joke to use Message-ID: <746FDB897740814EA52BDDCB5ED1DDBC02F6D392@medimmune4.medimmune.com> I was the recipient of a practical joke in the lab years ago. Some folks had sliced up some potatoes and placed in alcohol(like vodka, not 95), and labeled the container and it looked like the real deal. Then this one tech reached in(when we did not wear gloves), grabbed it and really studied it against the light and then popped in his mouth! He really played it up too, then I knew I was in the right job. From Charlotte.Kopczynski <@t> baycare.org Wed Nov 30 15:35:05 2005 From: Charlotte.Kopczynski <@t> baycare.org (Kopczynski, Charlotte) Date: Wed Nov 30 15:35:22 2005 Subject: [Histonet] Quincy Message-ID: <5452D66669CFC540B0452115ED26AF1F040E539D@bcexch02.bcad.baycare.org> I always thought it was funny when his room mate teased him for being sloppy.... Confidential: This electronic message and all contents contain information from BayCare Health System which may be privileged, confidential or otherwise protected from disclosure. The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify the sender and destroy the original message and all copies. From llewllew <@t> shaw.ca Wed Nov 30 15:55:50 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Wed Nov 30 15:56:13 2005 Subject: [Histonet] practical joke to use References: <746FDB897740814EA52BDDCB5ED1DDBC02F6D392@medimmune4.medimmune.com> Message-ID: <001c01c5f5f8$d44df320$690a4246@yourlk4rlmsu> Many years ago I worked in a lab where we were getting a new mortuary. The new one had been built, but the old morgue was still there with the tables etc still in place. I was the student coordinator for the med lab program as well as histo supervisor and part of the job was to choose the students and oversee the training program. Being a targeted male in a predominantly female work environment, a running joke was my interview technique for mostly female students - of the nudge, nudge, wink, wink variety. I came in one morning and the Chief Technologist attacked me verbally about leaving the old morgue in such a mess, and how he was completely disgusted. So I went to the morgue and looked. The buggers had spread ladies clothing, empty liquor bottles, pantyhose, blankets and other stuff all over it to look like an orgy. I was the butt of their jokes all day long, the sexists! Since they had draped more than three or four pairs of ladies undies around the mortuary, to this day I don't know whether to be insulted or complimented. What nobody was expecting was that right afterwards, and before it got cleaned up, an RCMP officer came into the hospital to see an autopsy on an unexpected death. He went to the old morgue instead of the new one. I understand he stood still in shock for a few minutes, then turned around and left. He was never seen again. Of course, I eventually got my revenge, but that's another story. Bryan From POWELL_SA <@t> Mercer.edu Wed Nov 30 16:01:08 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Wed Nov 30 16:01:52 2005 Subject: [Histonet] Histotechs: born or made? In-Reply-To: Message-ID: <01LW0M14PMNS8WX2CJ@Macon2.Mercer.edu> Off, Off, that is where everyone's specimen is sent, right? We used to have an "OFF" sign on our door. But that could have double meaning. I too am alone in my lab and "we" get along just fine. Shirleys -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Wednesday, November 30, 2005 3:51 PM To: froyer@bitstream.net; histonet@lists.utsouthwestern.edu; cking@rallansci.com Subject: RE: [Histonet] Histotechs: born or made? I'm constantly exchanging jokes with my co-worker. We are always trying to gross each other out. I'm not sure if this is normal, considering I work in a lab by myself. We must be off, Freds >>> "King, Curtis - RAS" 11/30/05 12:36PM >>> I would never make it through a day without craking up over something most people would consider IN BAD TASTE HUMOR. I Love My Job Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ford Royer Sent: Wednesday, November 30, 2005 12:28 PM To: 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? When I was a practicing laboratory scientist (27 years ago), we would have some of the wildest lab parties and everyone seemed to be on the same page as far as having a weird sense of humor. A work day didn't go by without some form of laughter in our lab. Non-laboratory people often asked me why this was. The only thing that I could come up with is it was how we dealt with the profession that we chose. I won't go into details or give examples. We all know what I am talking about. It does take a special kind of person to this sometimes morbid (some would say hideous) work and I for one am glad that there are these types of persons to take it on. Thank you all for your dedication to your profession and the people that you serve - mankind. ~ Ford Ford M. Royer, MT(ASCP) Minneapolis, MN -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Wednesday, November 30, 2005 11:11 AM To: 'Ingles Claire'; Histonet Subject: [Histonet] Histotechs: born or made? The first time I walked into a histology lab it was the day after the 4th of July and there were 4 blackened fingers sitting on the grossing bench (one guess how they got there - and it's nothing to do with anything Cajun!). My first thought was : 'this is going to be a strange job.' I've seen much stranger things since, so I think histotechs become strange due to exposure to unnatural sights (among other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Wednesday, November 30, 2005 8:55 AM To: Histonet Subject: RE: [Histonet] Re: 70% from NBF I have wondered the same thing many times myself. Whether it was naturally me or the addition of the chemicals that made me a bit strange. I think it may be partly both. I usually blame it on the chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn Sent: Tue 11/29/2005 11:15 AM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue > regulations then. Take his advice! _______________________________________________ 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.ingles <@t> hosp.wisc.edu Wed Nov 30 17:13:25 2005 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Wed Nov 30 17:15:22 2005 Subject: [Histonet] practical joke to use References: <746FDB897740814EA52BDDCB5ED1DDBC02F6D392@medimmune4.medimmune.com> <001c01c5f5f8$d44df320$690a4246@yourlk4rlmsu> Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE7C@uwhis-xchng2.hosp.wisc.edu> Granted I'm one of those that picks mercilessly on the guys (verbally only), but I'd love to hear the "other" story. Claire Ingles Mohs Clinic UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn Sent: Wed 11/30/2005 3:55 PM To: Histonet Subject: Re: [Histonet] practical joke to use Many years ago I worked in a lab where we were getting a new mortuary. The new one had been built, but the old morgue was still there with the tables etc still in place. I was the student coordinator for the med lab program as well as histo supervisor and part of the job was to choose the students and oversee the training program. Being a targeted male in a predominantly female work environment, a running joke was my interview technique for mostly female students - of the nudge, nudge, wink, wink variety. I came in one morning and the Chief Technologist attacked me verbally about leaving the old morgue in such a mess, and how he was completely disgusted. So I went to the morgue and looked. The buggers had spread ladies clothing, empty liquor bottles, pantyhose, blankets and other stuff all over it to look like an orgy. I was the butt of their jokes all day long, the sexists! Since they had draped more than three or four pairs of ladies undies around the mortuary, to this day I don't know whether to be insulted or complimented. What nobody was expecting was that right afterwards, and before it got cleaned up, an RCMP officer came into the hospital to see an autopsy on an unexpected death. He went to the old morgue instead of the new one. I understand he stood still in shock for a few minutes, then turned around and left. He was never seen again. Of course, I eventually got my revenge, but that's another story. Bryan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.ingles <@t> hosp.wisc.edu Wed Nov 30 17:16:11 2005 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Wed Nov 30 17:24:31 2005 Subject: [Histonet] Histotechs: born or made? References: <01LW0M14PMNS8WX2CJ@Macon2.Mercer.edu> Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE7D@uwhis-xchng2.hosp.wisc.edu> I'm so glad to hear from all of you on the subject of lab rat craziness. I know I'm not alone, and not (totally) nuts. It does kind of depress me though. I've only been a Histotech for 5 years! I guess it doesn't get any better than this! :) Claire Ingles Mohs Clinic UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Shirley Powell Sent: Wed 11/30/2005 4:01 PM To: 'Fred Underwood'; froyer@bitstream.net; histonet@lists.utsouthwestern.edu; cking@rallansci.com Subject: RE: [Histonet] Histotechs: born or made? Off, Off, that is where everyone's specimen is sent, right? We used to have an "OFF" sign on our door. But that could have double meaning. I too am alone in my lab and "we" get along just fine. Shirleys -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Wednesday, November 30, 2005 3:51 PM To: froyer@bitstream.net; histonet@lists.utsouthwestern.edu; cking@rallansci.com Subject: RE: [Histonet] Histotechs: born or made? I'm constantly exchanging jokes with my co-worker. We are always trying to gross each other out. I'm not sure if this is normal, considering I work in a lab by myself. We must be off, Freds >>> "King, Curtis - RAS" 11/30/05 12:36PM >>> I would never make it through a day without craking up over something most people would consider IN BAD TASTE HUMOR. I Love My Job Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ford Royer Sent: Wednesday, November 30, 2005 12:28 PM To: 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? When I was a practicing laboratory scientist (27 years ago), we would have some of the wildest lab parties and everyone seemed to be on the same page as far as having a weird sense of humor. A work day didn't go by without some form of laughter in our lab. Non-laboratory people often asked me why this was. The only thing that I could come up with is it was how we dealt with the profession that we chose. I won't go into details or give examples. We all know what I am talking about. It does take a special kind of person to this sometimes morbid (some would say hideous) work and I for one am glad that there are these types of persons to take it on. Thank you all for your dedication to your profession and the people that you serve - mankind. ~ Ford Ford M. Royer, MT(ASCP) Minneapolis, MN -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Wednesday, November 30, 2005 11:11 AM To: 'Ingles Claire'; Histonet Subject: [Histonet] Histotechs: born or made? The first time I walked into a histology lab it was the day after the 4th of July and there were 4 blackened fingers sitting on the grossing bench (one guess how they got there - and it's nothing to do with anything Cajun!). My first thought was : 'this is going to be a strange job.' I've seen much stranger things since, so I think histotechs become strange due to exposure to unnatural sights (among other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Wednesday, November 30, 2005 8:55 AM To: Histonet Subject: RE: [Histonet] Re: 70% from NBF I have wondered the same thing many times myself. Whether it was naturally me or the addition of the chemicals that made me a bit strange. I think it may be partly both. I usually blame it on the chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Llewellyn Sent: Tue 11/29/2005 11:15 AM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue > regulations then. Take his advice! _______________________________________________ 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 _______________________________________________ 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 azdudley <@t> hotmail.com Wed Nov 30 18:36:00 2005 From: azdudley <@t> hotmail.com (anita dudley) Date: Wed Nov 30 18:36:11 2005 Subject: [Histonet] Histology assistants In-Reply-To: <20051128130220.97265.qmail@web61224.mail.yahoo.com> Message-ID: tom do you know what the salary was for the lab assistants? thanks anita dudley, providence hosp, mobile ala >From: Rene J Buesa >To: Tom McNemar ,"'histonet@pathology.swmed.edu'" > >Subject: Re: [Histonet] Histology assistants >Date: Mon, 28 Nov 2005 05:02:20 -0800 (PST) > >Hi Tom: > The following is my experience: lab assistants are the best way of >increasing laboratory productivity and allowing the histotechs to do the >work they know and like to do. > In the last place I worked as supervisor, before retiring, a large >reference lab, the lag of uncut blocks left from days before amounted to >3-4 days work. I was able to hire 2 lab asistants and at the end of my >second month there were no uncut blocks left. > The tasks the 2 lab asistants (1 came at 5:30 AM and the other at 1:00 >PM, with an overlap of 1 hour) were the following: prepare the water baths >for the first shift of HTs, change all reagents in the autostainer and in >the tissue processors, operate the slides and cassettes numbering machines, >keep the record of the blocks assigned to each HT and file them back, take >care of drying the slides, run the H&E autostainer and the automatic >coverslipper, match paper/work with the slides, deliver the slides to the >pathologists, pull blocks for special requests and some other tasks that >would be more costly if done by the HTs (the earn more money than the lab >assistants) and that should not be done by them. Sometimes they helped in >grossing but just the tasks related to keeping records. > The lab assistants were also supposed to start getting an idea about the >histology work with the purpose to later on train in-house as a complement >to their studies, because since the beginning the idea was for them to >study as histotechs in the near future. > If you can hire lab assistants and train them well you wil see how >things will begin to run more smoothly in your lab. > Rene J. > > >Tom McNemar wrote: > I would really like some input on this..... > >How many places use assistants in Histology? > >What kind of things do they do? Gross? Enter/obtain specimens? File? > >Would really appreciate any and all thoughts. Thanks. > >Tom McNemar, HT(ASCP) >Histology Co-ordinator >Licking Memorial Hospital >Newark, Ohio 43055 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > >--------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bills <@t> icpmr.wsahs.nsw.gov.au Wed Nov 30 19:55:14 2005 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Wed Nov 30 19:55:44 2005 Subject: [Histonet] Aspergillus antibody Message-ID: <000001c5f61a$45a522c0$796a080a@wsahs.nsw.gov.au> Dear All, Is there anyone out there using an antibody to aspergillus species? If so how do you do it (dilutions, manually or by machine)? Do any antibody work better than others? Is it available for FFPE specimens. Any info would be greatly appreciated. Thanks Bill Sinai Chief Hospital Scientist Tissue Pathology ICPMR Westmead Hospital Westmead NSW 2145 Phone 02 9845 7774 Fax 02 9687 2330 __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From abilger <@t> ptd.net Wed Nov 30 20:21:04 2005 From: abilger <@t> ptd.net (Andrea C. Bilger) Date: Wed Nov 30 20:22:30 2005 Subject: [Histonet] cassette labeler Message-ID: <044c01c5f61d$e1de64a0$0300a8c0@andrea> Have any of you had experience with the Thermo Electron Carousel Microwriter cassette labeler? Our lab has had one for about a month now and the PA's are having issues with the cassettes. They seem to close with some difficulty. We had a couple that didn't get closed completely and tissue floated out. Have any of you had problems and how did you resolve them? Thanks for your help. Andrea From Eric <@t> ategra.com Wed Nov 30 18:40:56 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Thu Dec 1 07:56:57 2005 Subject: [Histonet] Immediate openings for HistoTechs Bench, Supervisors, and Managers............. Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. This is a follow up e-mail with the newest jobs listed first(1-5) (I have a few travel/temp HistoTech positions as well - see below). Most of my Histo Techs positions are clinical, although I do have a few Research Histo Tech positions as well. Here are some of my Hottest jobs: 1. Virginia Opening for a Histotech(bench) No weekends, No Call, Top dollar 2. Florida(Greater Florida area) Openings for Bench Histotech No weekends, No call 3. Ohio Opening for Histotechs(bench, NightShift and Dayshift) No Weekends, No Call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 4. Ohio Opening for CytoTech No Weekends, No Call, Top Dollar 5. Tennessee Opening for a Histotech Great Location, No Call, No Weekends, Top Dollar 6.Iowa Openings for a HistoTech(Bench) No weekends, No call, and Top Dollar. 7. Ohio(Dayton Area) HistoTech Manager No Weekends, No call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 8. Colorado (Greater Denver area) Opening for Several Bench Positions(Night Shift) No weekends, No call, Top Dollar 9. Georgia Openings for Histotech(Temp to Perm) No Weekends, No Call, Top Dollar, Excellent Benefits if you are interested, please call me today at 1-800-466-9919 ext 223 10. Pennsylvania(Philadelphia area) Openings for HistoTech(Bench) No Weekends, No Call, Top Dollar 11.Rhode Island Openings for Histotech(Bench) No weekends, No call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 12. California( Southern) HistoTech Supervisor(2nd shift) Openings for Histotechs(Bench,2nd shift) Great Location, No weekends, No call, Top Dollar 13. California(Southern) Openings for HistoTechs(Bench) Great Location, No weekends, No call, Top Dollar 14. Pennsylvania(Multiple jobs in greater Pittsburgh area) HistoTech opening(Bench) if you are interested, please call me today at 1-800-466-9919 ext 223 15. Illinois(Multiple jobs in Greater Illinois area) Opening for Histotech(Bench) 16. Michigan(Multiple jobs in Greater Michigan area) Openings for Histotechs(Bench) 17. Washington State(Eastern) Opening for Lead HistoTech(Bench) No weekends, No call if you are interested, please call me today at 1-800-466-9919 ext 223 18. Oklahoma Opening for Histotech(Bench) No weekends, No call. 19. Massachusetts (Several Opportunities in the Boston Area) Part time Histo Tech(Permanent) No weekends, No call 20. New Mexico openings for Supervisor and Bench Techs No Weekends, No Call, Excellent Benefits.. 21. Nebraska Openings for Histo Techs(Bench) No Weekends, No Call, Top Dollar 22. Virginia(Western Virginia) Opening for Bench Histotech No weekends, No call if you are interested, please call me today at 1-800-466-9919 ext 223 23. Ohio( Southern) Opening for Bench HistoTech No weekends, No call 24. Missouri(Greater Missouri area) Opening for Bench Histotech No weekends, No call 25. Wisconsin(Eastern area) Opening for a Bench Histotech No weekends, No call 26. Florida(Greater Florida area) Openings for Bench Histotech No weekends, No call The clients are currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 --------------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu ---------------------------------------------------------------- ---------------------------------------------------------------- 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 brucea <@t> unimelb.edu.au Tue Nov 29 19:15:35 2005 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Tue Dec 6 13:49:27 2005 Subject: [Histonet] NBF fixed tissue and subsequent first step for dehydration. In-Reply-To: <20051128015304.17579.qmail@web30608.mail.mud.yahoo.com> References: <20051128015304.17579.qmail@web30608.mail.mud.yahoo.com> Message-ID: Lorraine, In our Lab. we fix in 70% ETOH without any detrimental effects to morphology & then start processing in this also.....am happy to help in any way. Cheers, Bruce in OZ -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA. AUSTRALIA 3010 Nobody Can Make You Feel Inferior Without YOUR Permission - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING I am happy and content because I think I am. Q: What's a specimen? A: An Italian astronaut. ******************************************************************************** This email and any attachments may contain personal information or information that is otherwise privileged, confidential or otherwise protected from disclosure/subject of copyright. Any use, disclosure or copying of any part of it is prohibited. The University does not warrant that this email or any attachments are free from viruses or defects. Please check any attachments for viruses and defects before opening them. If this email is received in error please delete it and notify us by return email. The University does not necessarily share or endorse the views expressed in this email. From kj49505 <@t> yahoo.com Wed Nov 30 19:16:50 2005 From: kj49505 <@t> yahoo.com (karen johnson) Date: Tue Dec 6 13:49:30 2005 Subject: [Histonet] Mart 1 Message-ID: <20051201011650.86833.qmail@web52305.mail.yahoo.com> I work in a small dermatology lab and want to start to do MART-1 on frozen sections especially for lentigo maligna. I need a procedure that is easy, and pretty quick. Any help would be appreciated. THANKS __________________________________ Start your day with Yahoo! - Make it your home page! http://www.yahoo.com/r/hs