From kb2drkprk <@t> yahoo.com Thu Apr 1 00:06:24 2004 From: kb2drkprk <@t> yahoo.com (Kelly Simon) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] er/pr charging In-Reply-To: <003d01c41762$a7d36760$1d2a14ac@wchsys.org> Message-ID: <20040401060624.14769.qmail@web20909.mail.yahoo.com> According to a story by Lisa Miller in the January 2004 CAP Today, there is a new code for ER/PR and HER2/neu: "CPT 2004 separates traditional tumor morphometric analysis from semiquantitative immunohistochemistry. New code 88361 was created to report quantitative or semiquantitative immunohistochemistry for such analyses as hormone receptor and HER2/neu testing. The technical services of 88342 are incorporated into the new code." http://www.cap.org/apps/docs/cap_today/feature_stories/0104InWithNewCPT2004.html Kelly Simon, HTL (ASCP) Dynacare Laboratories Seattle, WA --- Joyce Cline wrote: > Does everyone use the CPT code 88342 for charging > the er/pr immuno's. Or is there another code that is > used for the technical component? > > ***** 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. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________ Do you Yahoo!? Yahoo! Small Business $15K Web Design Giveaway http://promotions.yahoo.com/design_giveaway/ From psanquin <@t> lugo.usc.es Thu Apr 1 01:39:46 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Lectin IHC In-Reply-To: Message-ID: <3.0.6.32.20040401093946.007c0b60@pop.lugo.usc.es> Jenny, all my encouragement after so many tests. Sure the histonetters can help you but it would be necessary to know a more detailed protocol and what organ you are studying. Don't you have problem cutting Bouin or ethanol frozen tissues? On other hand to my experience the washes buffers are not critical. Regards Pablo From bruyntjes <@t> voeding.tno.nl Thu Apr 1 02:09:35 2004 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] non-speficic binding of secundary biotinilated antibodies Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079CC8@ntexch1.voeding.tno.nl> Hi all In the past I used a rabbit anti mouse biotinilated secondary antibody with my lymphocyte-monoclonals on fresh frozen spleen and/or thymus slides of the rat. But I always had to add some ul's normal rat serum to avoid non-specific binding of this reagent to lymphocytes. Is anyone of you aware of a good secondary step in which adding of normal rat serum is not necessary anymore? Joost Bruijntjes TNO Nutrition and Food Research PO Box 360 3700 AJ Zeist The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From gudrun.lang <@t> aon.at Thu Apr 1 04:10:14 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] tissue processing Message-ID: <001a01c417d1$87b00500$eeeea8c0@SERVER> Hi We have a current problem with our routine tissue processing in the VIP. We loaded 200 capsules in the container. This time we had the half of the capsules filled with biopsy-sponges. Usually they are only about 30% of all. We do over night processing. The tissue surface was something like creamy and in the trimmed block the tissue looked wet. I was not able to get a good section (only holes in paraffin). But not all blocks were like this, most of the small biopsies worked well. My suggestion is, that the great amount of sponges is responsible for the underprocessing. I think the tissue was'nt dehydratet enough and water was taken over from one step to the next. Did anybody have the same experience? Please give me some input on this problem. thanks in advance Gudrun Lang Akh Linz, Austria From ree3 <@t> leicester.ac.uk Thu Apr 1 04:58:06 2004 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] chicken breast Message-ID: Recently we have had several mammary tumours in our flock of Leicester Blue hens.I wonder if anyone out there knows of any specific immunohistochemical markers for chicken mammary tissue/tumours?? Many thanks C. Sanders From JNocito <@t> Pathreflab.com Thu Apr 1 05:50:05 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Validation-Cyto Non-Gyn to Thin Prep In-Reply-To: <67.2570733f.2d9cc6ae@aol.com> Message-ID: Dannie, We ran side by side testing for about 5-6 specimens from each type, i.e. 5 urines, 5 breast fluids, 5 peritoneal fluids, etc. The only type we couldn't run side by side was CSF. However, our experience with CSF was that there was increased cellularity captured with the Thinprep. I guess it's up to each individual on how comfortable they feel. Hope this helps. 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 DMBCMP@aol.com Sent: Wednesday, March 31, 2004 7:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Validation-Cyto Non-Gyn to Thin Prep Hello All: The director of my lab (pathologist) and my supervisor (Cytotech) have asked me to post this question on HistoNet. Has anyone done validation for converting non-gyn cytology specimens to Thin Prep? Did you run parallel samples .....how many?,etc. Any input will be graciously welcomed. Thanks very much. Dannie Blake, HT Fresno Community Hospital Fresno, Ca. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***************************************Notice******************************************** This e-mail, including attachments, contains information that is confidential and may be legally privileged. This e-mail, including atachments, constitutes non-public information inteneded to be conveyed only to the designated recipient(s). If you are not an intended recipient,please delete this e-mail, including attachments, and notify me. The unauthorized use, dissemination, distribution or reproduction of this e-mail, including attachments, is prohibited and may be unlawful. ***************************************************************************************** From dmccaig <@t> ckha.on.ca Thu Apr 1 06:00:17 2004 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] accession numbers for multiple samples from same patient Message-ID: <3E5A3F039F0BD8118B4700C00D002024043132@CKHA9> Can I be advised how other sites label multiple specimens or where I can find the Canadian standard. Are they given different numbers ie 101-biopsy small bowel 102-biopsy cecum 103-antrum biopsy OR 101A-biopsy small bowel 101B-biopsy cecum 101C-antrum biopsy Thanks Diana McCaig, R.T. From plxja <@t> nottingham.ac.uk Thu Apr 1 06:02:44 2004 From: plxja <@t> nottingham.ac.uk (Joelle Alcock) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Microwave for antigen retrieval Message-ID: Hi In a couple of methods I still am still using the microwave for antigen retrieval. For one I microwave my paraffin sections in 1L 0.01M citrate buffer 6.4pH for 25 minutes. However I have found similar results by using the abcam enzymatic antigen retrieval kit which also takes less time in all - but care must be taken as sections are more likely to crack/disintergrate. Joelle Alcock >>> "Featherstone, Annette" 03/31/04 12:03pm >>> Are people still using the microwave for anitgen retrieval? I thought steamers and pressure cookers pretty much replaced that method. Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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 DDDeltour <@t> sig.med.navy.mil Thu Apr 1 06:10:52 2004 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] accession numbers for multiple samples from same p atient Message-ID: 101A-biopsy small bowel 101B-biopsy cecum 101C-antrum biopsy -----Original Message----- From: Diana McCaig [mailto:dmccaig@ckha.on.ca] Sent: Thursday, April 01, 2004 2:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] accession numbers for multiple samples from same patient Can I be advised how other sites label multiple specimens or where I can find the Canadian standard. Are they given different numbers ie 101-biopsy small bowel 102-biopsy cecum 103-antrum biopsy OR 101A-biopsy small bowel 101B-biopsy cecum 101C-antrum biopsy Thanks Diana McCaig, R.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. From thallada <@t> noch.org Thu Apr 1 06:25:05 2004 From: thallada <@t> noch.org (Hallada, Teri) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] er/pr charging Message-ID: I believe you must "quantitative" the results in some format. Our pathologists here don't quantitative ER/PR only Her2, so we don't use 88361 for ER/PR only Her2. Is this correct? Teri Hallada BS MT CT (ASCP) thallada@noch.org > -----Original Message----- > From: Kelly Simon [SMTP:kb2drkprk@yahoo.com] > Sent: Thursday, April 01, 2004 01:06 > To: Joyce Cline > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] er/pr charging > > According to a story by Lisa Miller in the January > 2004 CAP Today, there is a new code for ER/PR and > HER2/neu: > > "CPT 2004 separates traditional tumor morphometric > analysis from semiquantitative immunohistochemistry. > New code 88361 was created to report quantitative or > semiquantitative immunohistochemistry for such > analyses as hormone receptor and HER2/neu testing. The > technical services of 88342 are incorporated into the > new code." > > http://www.cap.org/apps/docs/cap_today/feature_stories/0104InWithNewCPT2004.html > > Kelly Simon, HTL (ASCP) > Dynacare Laboratories > Seattle, WA > > --- Joyce Cline wrote: > > Does everyone use the CPT code 88342 for charging > > the er/pr immuno's. Or is there another code that is > > used for the technical component? > > > > ***** 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. > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > __________________________________ > Do you Yahoo!? > Yahoo! Small Business $15K Web Design Giveaway > http://promotions.yahoo.com/design_giveaway/ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Statement: This message is intended only for the individual or entity to which it is addressed. It may contain privileged, confidential information which is exempt from disclosure under applicable laws. If you are not the intended recipient, please note that you are strictly prohibited from disseminating or distributing this information (other than to the intended recipient) or copying this information. If you have received this communication in error, please notify us immediately by e-mail or by telephone at (616) 842-3600. Thank you. From Jackie.O'Connor <@t> abbott.com Thu Apr 1 06:26:49 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] tissue processing Message-ID: OOooh - Flashback. I remember about 12 years ago having the exact same problem with the VIP and running over 100 biopsies with blue pads. The blue biopsy pads retain about 1 mL of solution - 2 biopsy pads per cassette - 2 mL retained solution - 100 cassettes - 200 ml of retained solution that doesn't get rinsed out well by the next solution- so Gudrun - your hypothesis is correct (per my experience). You could try a thinner biopsy pad - Surgipath's biopsy pads are about 1/2 the thickness of other vendors. At the time we just quit using biopsy pads and went back to lens paper to wrap biopsies - and the problem was gone. Also gone was a biopsy pad artifact from putting semi-fixed tissues (e.g. currettings) between blue pads. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics "Gudrun Lang" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/01/2004 04:10 AM To: "Histonetliste" cc: Subject: [Histonet] tissue processing Hi We have a current problem with our routine tissue processing in the VIP. We loaded 200 capsules in the container. This time we had the half of the capsules filled with biopsy-sponges. Usually they are only about 30% of all. We do over night processing. The tissue surface was something like creamy and in the trimmed block the tissue looked wet. I was not able to get a good section (only holes in paraffin). But not all blocks were like this, most of the small biopsies worked well. My suggestion is, that the great amount of sponges is responsible for the underprocessing. I think the tissue was'nt dehydratet enough and water was taken over from one step to the next. Did anybody have the same experience? Please give me some input on this problem. thanks in advance Gudrun Lang Akh Linz, Austria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From yichaowu <@t> hotmail.com Thu Apr 1 06:36:50 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] RE: C4d antibody on Paraffin sections Message-ID: Dear Donovan, Sorry for the late reply. Actually I learned the method from a refence,but I could not remember the exact journal and article now. The sections I used were acetone-methanol(1:1) fixed paraffin embedded sections with. I have not tried on formalin-fixed paraffin-embedded sections.But I remember that in the reference mentioned above,it reads that this method is applicable on formalin-fixed sections. The procedures are not special,just add the pretreatment with 88% formic acid.That is to say, Dewaxing; Draw a circle around the sections with PAP pen; Incubate with PBS; Incubate with 88% formic acid (diluted with ddH2O i.e. pure water) for 20 minutes in room temperature; Incubate with 10% fetal bovine serum as the blocker (or other kinds of block serum); Incubate with monoclonal mouse anti-C4d antibody (A213 from Quidel USA) at 4C overnight; Incubate with FITC-conjugated rabbit anti-mouse IgG antibody (from DAKO) for 40 mins at RT; Observe under fluorescent microscope... Besides, it is not any advertisement for those products I mentioned! :-) Just a little experience. Wish it is helpful. And thank you for your attention.You are much welcomed! Yichao WU,Ph.D Candidate Research Insititute of Nephrology 305 East Zhongshan Road Nanjing 210002,P.R.China >From: "Donovan, Mark" >To: "'yichaowu@hotmail.com'" >Subject: C4d antibody on Paraffin sections >Date: Tue, 30 Mar 2004 15:39:03 +1000 > >Dear Yichao, > >I read your response to Carmen Loiselle regarding the Quidel antibody to >C4d >with great interest. > >Could you provide me with more details of your protocol for using the >antibody on formalin fixed paraffin embedded sections. > >I have used this antibody on frozen sections but have not had any success >in >getting it to work on formalin fixed material. Your advice would be very >much appreciated. > >Regards, > >Mark Donovan >Immunohistochemistry Laboratory >Anatomical Pathology >The Alfred Hospital >Melbourne, Australia > > >THIS E-MAIL IS CONFIDENTIAL. If you have received this e-mail in error, >please notify us by return e-mail and delete the document. If you are not >the intended recipient you are hereby notified that any disclosure, >copying, >distribution or taking any action in reliance on the contents of this >information is strictly prohibited and may be unlawful. Bayside Health is >not liable for the proper and complete transmission of the information >contained in this communication or for any delay in its receipt. _________________________________________________________________ Add photos to your e-mail with MSN 8. Get 2 months FREE*. http://join.msn.com/?page=features/featuredemail From jlambrey <@t> hotmail.com Thu Apr 1 08:03:03 2004 From: jlambrey <@t> hotmail.com (Julien Lambrey de Souza) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] testing list Message-ID: This is a test. Do not consider. Thankyou. _________________________________________________________________ Help protect your entire PC with Virus Guard from [1]MSN Premium Get Two Months FREE* References 1. http://g.msn.com/8HMAENCA/2731??PS= From Rolf.Brekken <@t> UTSouthwestern.edu Thu Apr 1 08:09:04 2004 From: Rolf.Brekken <@t> UTSouthwestern.edu (Rolf Brekken) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 1 (Out of Office reply) Message-ID: I am out of the lab until Monday April 5. I'll return your message as soon as I can after then. If you need to contact someone in the lab immediately please call our lab manager, Mishel Davis, at 214 648 1462. thanks rolf >>> histonet 04/01/04 08:08 >>> 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. Mouse pancreas (Grant, Debra) 2. Re: Mouse pancreas (Jackie.O'Connor@abbott.com) 3. Loeffler's Alkaline Methylene Blue (Owen, Michael P) 4. Histonet: Loeffler's Alkaline Methylene Blue (Owen, Michael P) 5. Mouse pancreas processing schedule (Grant, Debra) 6. er/pr charging (Joyce Cline) 7. Procedure help with dermal papilla (Jenny Oblander) 8. RE: er/pr charging (Sebree Linda A.) 9. RE: Fwd: Re: [Histonet] Decalcifying mouse heads (Caroline Stott) 10. LTC4 SYNTHASE, EG2 (UniPath IHC) 11. Re: Mouse pancreas (Gayle Callis) 12. RE: Procedure help with dermal papilla (Barry R Rittman) 13. Mouse pancreas processing (Grant, Debra) 14. Lectin IHC (Amrit Samra) 15. Re: Decalcifying mouse heads (KATERINA STRATI) 16. Validation-Cyto Non-Gyn to Thin Prep (DMBCMP@aol.com) 17. ARISTAN STAINER (Ernestine Middleton) 18. Re: er/pr charging (Kelly Simon) 19. Re: Lectin IHC (Pablo S?nchez Quinteiro) 20. non-speficic binding of secundary biotinilated antibodies (Bruijntjes, J.P.) 21. tissue processing (Gudrun Lang) 22. chicken breast (Edwards, R.E.) 23. RE: Validation-Cyto Non-Gyn to Thin Prep (Joe Nocito) 24. accession numbers for multiple samples from same patient (Diana McCaig) 25. Re: Microwave for antigen retrieval (Joelle Alcock) 26. RE: accession numbers for multiple samples from same p atient (Deltour, Douglas D.(HM2)) 27. RE: er/pr charging (Hallada, Teri) 28. Re: tissue processing (Jackie.O'Connor@abbott.com) 29. RE: C4d antibody on Paraffin sections (yichao wu) 30. testing list (Julien Lambrey de Souza) ---------------------------------------------------------------------- Message: 1 Date: Wed, 31 Mar 2004 13:51:11 -0600 From: "Grant, Debra" Subject: [Histonet] Mouse pancreas To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" Hi Histonetters, I am looking for a good tissue processing schedule for mouse pancreas. I am currently using the following schedule and having trouble with sectioning the pancreas, the H&E's look cracked and parched as if my water bath is too hot. The temperature on my water bath is 42-43 degrees Celsius, I have a thermometer also that I check the temp with. I use Tissue Prep paraffin for infiltration and embedding. Fixed by the researcher in 4% PFA 24 hours rinsed in distilled water 5' x 3. 70% Alcohol- 30 min. Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour 30 min. Clearite 3- 1 hour 15 min. Clearite 3- 1 hour 15 min. Clearite 3- 1 hour 15 min. Paraffin- 1 hour Paraffin- 1 hour Paraffin- 1 hour Paraffin- 1 hour Thanks in advance! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org ------------------------------ Message: 2 Date: Wed, 31 Mar 2004 14:13:07 -0600 From: Jackie.O'Connor@abbott.com Subject: Re: [Histonet] Mouse pancreas To: "Grant, Debra" Cc: Histonet Message-ID: Content-Type: text/plain; charset="us-ascii" The processing schedule you are using would make fibroid uterus brittle. What is prosoft? Anyway - 30 minutes in two changes each of (ethanol) 70%, 95%, Absolute, Xylene, and 2 changes of paraffin at 55 minutes each should do you just fine - for just pancreas - since it is so delicate. I use the above schedule (45 minutes each station) for routine mice tissue and xenografts with beautiful results. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics "Grant, Debra" Sent by: histonet-bounces@lists.utsouthwestern.edu 03/31/2004 01:51 PM To: "Histonet" cc: Subject: [Histonet] Mouse pancreas Hi Histonetters, I am looking for a good tissue processing schedule for mouse pancreas. I am currently using the following schedule and having trouble with sectioning the pancreas, the H&E's look cracked and parched as if my water bath is too hot. The temperature on my water bath is 42-43 degrees Celsius, I have a thermometer also that I check the temp with. I use Tissue Prep paraffin for infiltration and embedding. Fixed by the researcher in 4% PFA 24 hours rinsed in distilled water 5' x 3. 70% Alcohol- 30 min. Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour 30 min. Clearite 3- 1 hour 15 min. Clearite 3- 1 hour 15 min. Clearite 3- 1 hour 15 min. Paraffin- 1 hour Paraffin- 1 hour Paraffin- 1 hour Paraffin- 1 hour Thanks in advance! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Wed, 31 Mar 2004 15:16:58 -0500 From: "Owen, Michael P" Subject: [Histonet] Loeffler's Alkaline Methylene Blue To: 'Histonet' Message-ID: Content-Type: text/plain Dear Histonet Users, I am interested in receiving all of the formulations for Loeffler's Alkaline Methylene Blue available. Its uses include visualization of Corynebacterium diphtheria and Mycobacterium leprae. I have already visited Dr. John Kiernan's Histochemistry FAQ, the FDA CFSAN BAM Online Reagents Manual, the Hardy Diagnostics Web site, and the Histonet Archives. If you know of any formulations different from the sources listed above, please send them to me at your earliest convenience. Thank you in advance for your assistance. Sincerely, The FDA Lab Rat Michael P. Owen, Regulatory Microbiologist FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov ------------------------------ Message: 4 Date: Wed, 31 Mar 2004 12:34:15 -0800 From: "Owen, Michael P" Subject: [Histonet] Histonet: Loeffler's Alkaline Methylene Blue To: 'Histonet' Message-ID: Content-Type: text/plain Dear Histonet Users, I am interested in receiving all of the formulations for Loeffler's Alkaline Methylene Blue available. Its uses include visualization of Corynebacterium diphtheria and Mycobacterium leprae. I have already visited Dr. John Kiernan's Histochemistry FAQ, the FDA CFSAN BAM Online Reagents Manual, the Hardy Diagnostics Web site, and the Histonet Archives. If you know of any formulations different from the sources listed above, please send them to me at your earliest convenience. Thank you in advance for your assistance. Sincerely, Michael P. Owen The FDA Lab Rat Michael P. Owen, Regulatory Microbiologist FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov ------------------------------ Message: 5 Date: Wed, 31 Mar 2004 14:54:55 -0600 From: "Grant, Debra" Subject: [Histonet] Mouse pancreas processing schedule To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi all, For those that wanted to know Prosoft is an alcohol substitute from Anatech and is made with: -ether and ester derivatives of propylene glycol, and propanol. Hope this helps! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org ------------------------------ Message: 6 Date: Wed, 31 Mar 2004 15:56:33 -0500 From: "Joyce Cline" Subject: [Histonet] er/pr charging To: "Histonet" Message-ID: <003d01c41762$a7d36760$1d2a14ac@wchsys.org> Content-Type: text/plain;charset="iso-8859-1" Does everyone use the CPT code 88342 for charging the er/pr immuno's. Or is there another code that is used for the technical component? ***** 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. ------------------------------ Message: 7 Date: Wed, 31 Mar 2004 15:00:46 -0600 From: Jenny Oblander Subject: [Histonet] Procedure help with dermal papilla To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi Guys, I need your help in tracing down a procedure, antibodies etc for a research project one of my investigators is doing. They would like to differentiate keratinocytes from dermal papilla fibroblasts. Does anyone have experience with TGF-beta-RII or IL-1-RI? Thanks Jenny J.Oblander, HT (A.S.C.P.) Comparative Medicine Oklahoma Medical Research Foundation MS#32 825 NE 13th St. Okc,Ok 73104 jenny-oblander@omrf.ouhsc.edu 405-271-7083 ------------------------------ Message: 8 Date: Wed, 31 Mar 2004 15:14:37 -0600 From: "Sebree Linda A." Subject: RE: [Histonet] er/pr charging To: "Joyce Cline" , "Histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" We used to use 88342 but now use 88361. 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: Joyce Cline [mailto:jcline@wchsys.org] Sent: Wednesday, March 31, 2004 2:57 PM To: Histonet Subject: [Histonet] er/pr charging Does everyone use the CPT code 88342 for charging the er/pr immuno's. Or is there another code that is used for the technical component? ***** 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Thu, 01 Apr 2004 09:52:23 +1200 From: Caroline Stott Subject: RE: Fwd: Re: [Histonet] Decalcifying mouse heads To: Histonet@lists.utsouthwestern.edu Message-ID: <5.2.1.1.0.20040401094556.020deec0@anatomy.otago.ac.nz> Content-Type: text/plain; charset="us-ascii"; format=flowed We use 10% formic acid. We use the chemical test to check if decalcification is complete. That is 5ml of ammonium hydroxide added to 5ml of the bone solution and add 5ml saturated ammonium oxalate. If the solution is milky white, there is still calcium present. So keep changing the solution every second day until it is finished. Caroline Caroline Stott Histology Service Unit Medical School University of Otago Dunedin (03) 479 7152 ------------------------------ Message: 10 Date: Wed, 31 Mar 2004 14:49:55 -0700 From: "UniPath IHC" Subject: [Histonet] LTC4 SYNTHASE, EG2 To: Message-ID: <000001c4176a$1ab3a6d0$4500a8c0@unipath02> Content-Type: text/plain; charset="us-ascii" Does anyone know where to find antibodies to LTC4 synthase or EG2? Thanks, Brianna Jackson, BS, QIHC UniPath, LLC bjackson@unipathllc.com 303-512-2220 ------------------------------ Message: 11 Date: Wed, 31 Mar 2004 15:48:28 -0700 From: Gayle Callis Subject: Re: [Histonet] Mouse pancreas To: Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040331154828.00be4610@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" >Is the Prosoft a gradient? Nothing wrong with your waterbath, it is the processing schedule. Your little mouse tissue is processed in a schedule for much bigger tissues, and is probably overexposed to dehydrants. Included is a schedule next to yours with explanation of equivalent to our schedule, cut back times in dehydrants, clearant and paraffins and if still friable, dry and cracked, cut back even more. Be sure you trim block and soak in water, you can try warm then cold, or warm then ice water if needed. What you are doing is removing too much of the bound water attached to tissue proteins along with free water in tissue spaces. >> > 70% Alcohol- 30 min. equivalant is 70% 30 min >30 min >Prosoft- 1 hour " " 80% 30 min >15 min >Prosoft- 1 hour " " 95% 15 min >15 min >Prosoft- 1 hour " " 95% 15 min >30 min >Prosoft- 1 hour " " 95% 30 min >30 min >Prosoft- 1 hour " " 100% 30 min >30 min >Prosoft- 1 hour 30 min. " " 100% 30 min >45 min or 1 hour >Clearite 3- 1 hour 15 min. >45 min or 1 hour >Clearite 3- 1 hour 15 min. >elminate >Clearite 3- 1 hour 15 min. or adjust Clearite changes to be 20, 20, and 45 min >30 min >Paraffin- 1 hour >30 min >Paraffin- 1 hour >30 min >Paraffin- 1 hour >30 min >Paraffin- 1 hour >for a total of 2 hours in paraffin at no more than 60C. >> Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 12 Date: Wed, 31 Mar 2004 16:50:28 -0600 From: "Barry R Rittman" Subject: RE: [Histonet] Procedure help with dermal papilla To: Message-ID: <566FB0B522443D43AF02D2ADBE35A6F077FD5B@UTHEVS3.mail.uthouston.edu> Content-Type: text/plain; charset="us-ascii" Jenny Why not use pan cytokeratin? Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jenny Oblander Sent: Wednesday, March 31, 2004 3:01 PM To: Histonet (E-mail) Subject: [Histonet] Procedure help with dermal papilla Hi Guys, I need your help in tracing down a procedure, antibodies etc for a research project one of my investigators is doing. They would like to differentiate keratinocytes from dermal papilla fibroblasts. Does anyone have experience with TGF-beta-RII or IL-1-RI? Thanks Jenny J.Oblander, HT (A.S.C.P.) Comparative Medicine Oklahoma Medical Research Foundation MS#32 825 NE 13th St. Okc,Ok 73104 jenny-oblander@omrf.ouhsc.edu 405-271-7083 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Wed, 31 Mar 2004 16:56:41 -0600 From: "Grant, Debra" Subject: [Histonet] Mouse pancreas processing To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" Thanks to all for the pancreas protocols, it seems I need to shorten my times in all stations of the processor. I will try this. Thanks again! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org ------------------------------ Message: 14 Date: Wed, 31 Mar 2004 15:32:20 -0800 From: "Amrit Samra" Subject: [Histonet] Lectin IHC To: Message-ID: Content-Type: text/plain; charset=US-ASCII I am doing lectin staining for frozen guinea pig tissues. I have tried different fixitives; bouin, 10% formalin, acetone, ethanol, B5 and different buffers; TBS, PBS and lectin buffer. I am still not able to get a positive staining for my known positive. The last trial I did was fixing in bouin and washing with TBS. any help would be greatly appreciated. Jenny Amrit Samra Histology Lab,the iCAPTURE Centre St.Paul's Hospital 1081 Burrard Street,Vancouver,BC Canada V6Z 1Y6 Phone: 604-682-2344 extension 62703 ------------------------------ Message: 15 Date: Wed, 31 Mar 2004 18:09:46 -0600 From: KATERINA STRATI Subject: Re: [Histonet] Decalcifying mouse heads To: Histonet@lists.utsouthwestern.edu Message-ID: <658073650c1f.650c1f658073@wiscmail.wisc.edu> Content-Type: text/plain; charset=us-ascii Thank you to everyone for useful advice Katerina Strati Lambert Lab 222 McArdle Lab 1400 University Ave. Madison, WI 53706 (608) 262-6407 kstrati@wisc.edu ----- Original Message ----- From: KATERINA STRATI Date: Tuesday, March 30, 2004 5:33 pm Subject: [Histonet] Decalcifying mouse heads > > > We are interested in decalcifying whole heads from adult mice in > order to examine histological sections throughout the oral cavity > and esophagus. Are you aware what would be appropriate for such a > large piece of tissue, or how long it would take to decalcify? In > the future we would like to have the option of performing > immunohistochemistry on our decalcified samples. Do you have any > experience with IHC on such large decalcified samples? > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 16 Date: Wed, 31 Mar 2004 20:13:18 EST From: DMBCMP@aol.com Subject: [Histonet] Validation-Cyto Non-Gyn to Thin Prep To: histonet@lists.utsouthwestern.edu Message-ID: <67.2570733f.2d9cc6ae@aol.com> Content-Type: text/plain; charset="US-ASCII" Hello All: The director of my lab (pathologist) and my supervisor (Cytotech) have asked me to post this question on HistoNet. Has anyone done validation for converting non-gyn cytology specimens to Thin Prep? Did you run parallel samples .....how many?,etc. Any input will be graciously welcomed. Thanks very much. Dannie Blake, HT Fresno Community Hospital Fresno, Ca. ------------------------------ Message: 17 Date: Wed, 31 Mar 2004 21:19:15 -0500 (EST) From: Ernestine Middleton Subject: [Histonet] ARISTAN STAINER To: "'histonet@pathology.swmed.edu'" Message-ID: <20040401021915.65884.qmail@web41607.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi; For those who are using the Aristan stainer; will you please send me you procedure for gram and wathin starry. Thank you, Ernestine Middleton, Manager, HT/HTL Montefiore Med. Ct. Bronx, NY 718-920-4157 718-547-1920 fax --------------------------------- Post your free ad now! Yahoo! Canada Personals ------------------------------ Message: 18 Date: Wed, 31 Mar 2004 22:06:24 -0800 (PST) From: Kelly Simon Subject: Re: [Histonet] er/pr charging To: Joyce Cline Cc: histonet@lists.utsouthwestern.edu Message-ID: <20040401060624.14769.qmail@web20909.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii According to a story by Lisa Miller in the January 2004 CAP Today, there is a new code for ER/PR and HER2/neu: "CPT 2004 separates traditional tumor morphometric analysis from semiquantitative immunohistochemistry. New code 88361 was created to report quantitative or semiquantitative immunohistochemistry for such analyses as hormone receptor and HER2/neu testing. The technical services of 88342 are incorporated into the new code." http://www.cap.org/apps/docs/cap_today/feature_stories/0104InWithNewCPT2004.html Kelly Simon, HTL (ASCP) Dynacare Laboratories Seattle, WA --- Joyce Cline wrote: > Does everyone use the CPT code 88342 for charging > the er/pr immuno's. Or is there another code that is > used for the technical component? > > ***** 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. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________ Do you Yahoo!? Yahoo! Small Business $15K Web Design Giveaway http://promotions.yahoo.com/design_giveaway/ ------------------------------ Message: 19 Date: Thu, 01 Apr 2004 09:39:46 +0200 From: Pablo S?nchez Quinteiro Subject: Re: [Histonet] Lectin IHC To: Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040401093946.007c0b60@pop.lugo.usc.es> Content-Type: text/plain; charset="us-ascii" Jenny, all my encouragement after so many tests. Sure the histonetters can help you but it would be necessary to know a more detailed protocol and what organ you are studying. Don't you have problem cutting Bouin or ethanol frozen tissues? On other hand to my experience the washes buffers are not critical. Regards Pablo ------------------------------ Message: 20 Date: Thu, 1 Apr 2004 10:09:35 +0200 From: "Bruijntjes, J.P." Subject: [Histonet] non-speficic binding of secundary biotinilated antibodies To: Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079CC8@ntexch1.voeding.tno.nl> Content-Type: text/plain; charset="us-ascii" Hi all In the past I used a rabbit anti mouse biotinilated secondary antibody with my lymphocyte-monoclonals on fresh frozen spleen and/or thymus slides of the rat. But I always had to add some ul's normal rat serum to avoid non-specific binding of this reagent to lymphocytes. Is anyone of you aware of a good secondary step in which adding of normal rat serum is not necessary anymore? Joost Bruijntjes TNO Nutrition and Food Research PO Box 360 3700 AJ Zeist The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html ------------------------------ Message: 21 Date: Thu, 1 Apr 2004 12:10:14 +0200 From: "Gudrun Lang" Subject: [Histonet] tissue processing To: "Histonetliste" Message-ID: <001a01c417d1$87b00500$eeeea8c0@SERVER> Content-Type: text/plain; charset="iso-8859-1" Hi We have a current problem with our routine tissue processing in the VIP. We loaded 200 capsules in the container. This time we had the half of the capsules filled with biopsy-sponges. Usually they are only about 30% of all. We do over night processing. The tissue surface was something like creamy and in the trimmed block the tissue looked wet. I was not able to get a good section (only holes in paraffin). But not all blocks were like this, most of the small biopsies worked well. My suggestion is, that the great amount of sponges is responsible for the underprocessing. I think the tissue was'nt dehydratet enough and water was taken over from one step to the next. Did anybody have the same experience? Please give me some input on this problem. thanks in advance Gudrun Lang Akh Linz, Austria ------------------------------ Message: 22 Date: Thu, 1 Apr 2004 11:58:06 +0100 From: "Edwards, R.E." Subject: [Histonet] chicken breast To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Recently we have had several mammary tumours in our flock of Leicester Blue hens.I wonder if anyone out there knows of any specific immunohistochemical markers for chicken mammary tissue/tumours?? Many thanks C. Sanders ------------------------------ Message: 23 Date: Thu, 1 Apr 2004 05:50:05 -0600 From: "Joe Nocito" Subject: RE: [Histonet] Validation-Cyto Non-Gyn to Thin Prep To: , Message-ID: Content-Type: text/plain; charset="us-ascii" Dannie, We ran side by side testing for about 5-6 specimens from each type, i.e. 5 urines, 5 breast fluids, 5 peritoneal fluids, etc. The only type we couldn't run side by side was CSF. However, our experience with CSF was that there was increased cellularity captured with the Thinprep. I guess it's up to each individual on how comfortable they feel. Hope this helps. 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 DMBCMP@aol.com Sent: Wednesday, March 31, 2004 7:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Validation-Cyto Non-Gyn to Thin Prep Hello All: The director of my lab (pathologist) and my supervisor (Cytotech) have asked me to post this question on HistoNet. Has anyone done validation for converting non-gyn cytology specimens to Thin Prep? Did you run parallel samples .....how many?,etc. Any input will be graciously welcomed. Thanks very much. Dannie Blake, HT Fresno Community Hospital Fresno, Ca. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***************************************Notice******************************************** This e-mail, including attachments, contains information that is confidential and may be legally privileged. This e-mail, including atachments, constitutes non-public information inteneded to be conveyed only to the designated recipient(s). If you are not an intended recipient,please delete this e-mail, including attachments, and notify me. The unauthorized use, dissemination, distribution or reproduction of this e-mail, including attachments, is prohibited and may be unlawful. ***************************************************************************************** ------------------------------ Message: 24 Date: Thu, 1 Apr 2004 07:00:17 -0500 From: Diana McCaig Subject: [Histonet] accession numbers for multiple samples from same patient To: histonet@pathology.swmed.edu Message-ID: <3E5A3F039F0BD8118B4700C00D002024043132@CKHA9> Content-Type: text/plain; charset="iso-8859-1" Can I be advised how other sites label multiple specimens or where I can find the Canadian standard. Are they given different numbers ie 101-biopsy small bowel 102-biopsy cecum 103-antrum biopsy OR 101A-biopsy small bowel 101B-biopsy cecum 101C-antrum biopsy Thanks Diana McCaig, R.T. ------------------------------ Message: 25 Date: Thu, 01 Apr 2004 13:02:44 +0100 From: "Joelle Alcock" Subject: Re: [Histonet] Microwave for antigen retrieval To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi In a couple of methods I still am still using the microwave for antigen retrieval. For one I microwave my paraffin sections in 1L 0.01M citrate buffer 6.4pH for 25 minutes. However I have found similar results by using the abcam enzymatic antigen retrieval kit which also takes less time in all - but care must be taken as sections are more likely to crack/disintergrate. Joelle Alcock >>> "Featherstone, Annette" 03/31/04 12:03pm >>> Are people still using the microwave for anitgen retrieval? I thought steamers and pressure cookers pretty much replaced that method. Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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 ------------------------------ Message: 26 Date: Thu, 1 Apr 2004 14:10:52 +0200 From: "Deltour, Douglas D.(HM2)" Subject: RE: [Histonet] accession numbers for multiple samples from same p atient To: 'Diana McCaig' , histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" 101A-biopsy small bowel 101B-biopsy cecum 101C-antrum biopsy -----Original Message----- From: Diana McCaig [mailto:dmccaig@ckha.on.ca] Sent: Thursday, April 01, 2004 2:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] accession numbers for multiple samples from same patient Can I be advised how other sites label multiple specimens or where I can find the Canadian standard. Are they given different numbers ie 101-biopsy small bowel 102-biopsy cecum 103-antrum biopsy OR 101A-biopsy small bowel 101B-biopsy cecum 101C-antrum biopsy Thanks Diana McCaig, R.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. ------------------------------ Message: 27 Date: Thu, 1 Apr 2004 07:25:05 -0500 From: "Hallada, Teri" Subject: RE: [Histonet] er/pr charging To: "Kelly Simon" , "Joyce Cline" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" I believe you must "quantitative" the results in some format. Our pathologists here don't quantitative ER/PR only Her2, so we don't use 88361 for ER/PR only Her2. Is this correct? Teri Hallada BS MT CT (ASCP) thallada@noch.org > -----Original Message----- > From: Kelly Simon [SMTP:kb2drkprk@yahoo.com] > Sent: Thursday, April 01, 2004 01:06 > To: Joyce Cline > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] er/pr charging > > According to a story by Lisa Miller in the January > 2004 CAP Today, there is a new code for ER/PR and > HER2/neu: > > "CPT 2004 separates traditional tumor morphometric > analysis from semiquantitative immunohistochemistry. > New code 88361 was created to report quantitative or > semiquantitative immunohistochemistry for such > analyses as hormone receptor and HER2/neu testing. The > technical services of 88342 are incorporated into the > new code." > > http://www.cap.org/apps/docs/cap_today/feature_stories/0104InWithNewCPT2004.html > > Kelly Simon, HTL (ASCP) > Dynacare Laboratories > Seattle, WA > > --- Joyce Cline wrote: > > Does everyone use the CPT code 88342 for charging > > the er/pr immuno's. Or is there another code that is > > used for the technical component? > > > > ***** 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. > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > __________________________________ > Do you Yahoo!? > Yahoo! Small Business $15K Web Design Giveaway > http://promotions.yahoo.com/design_giveaway/ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Statement: This message is intended only for the individual or entity to which it is addressed. It may contain privileged, confidential information which is exempt from disclosure under applicable laws. If you are not the intended recipient, please note that you are strictly prohibited from disseminating or distributing this information (other than to the intended recipient) or copying this information. If you have received this communication in error, please notify us immediately by e-mail or by telephone at (616) 842-3600. Thank you. ------------------------------ Message: 28 Date: Thu, 1 Apr 2004 06:26:49 -0600 From: Jackie.O'Connor@abbott.com Subject: Re: [Histonet] tissue processing To: "Gudrun Lang" Cc: Histonetliste Message-ID: Content-Type: text/plain; charset="us-ascii" OOooh - Flashback. I remember about 12 years ago having the exact same problem with the VIP and running over 100 biopsies with blue pads. The blue biopsy pads retain about 1 mL of solution - 2 biopsy pads per cassette - 2 mL retained solution - 100 cassettes - 200 ml of retained solution that doesn't get rinsed out well by the next solution- so Gudrun - your hypothesis is correct (per my experience). You could try a thinner biopsy pad - Surgipath's biopsy pads are about 1/2 the thickness of other vendors. At the time we just quit using biopsy pads and went back to lens paper to wrap biopsies - and the problem was gone. Also gone was a biopsy pad artifact from putting semi-fixed tissues (e.g. currettings) between blue pads. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics "Gudrun Lang" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/01/2004 04:10 AM To: "Histonetliste" cc: Subject: [Histonet] tissue processing Hi We have a current problem with our routine tissue processing in the VIP. We loaded 200 capsules in the container. This time we had the half of the capsules filled with biopsy-sponges. Usually they are only about 30% of all. We do over night processing. The tissue surface was something like creamy and in the trimmed block the tissue looked wet. I was not able to get a good section (only holes in paraffin). But not all blocks were like this, most of the small biopsies worked well. My suggestion is, that the great amount of sponges is responsible for the underprocessing. I think the tissue was'nt dehydratet enough and water was taken over from one step to the next. Did anybody have the same experience? Please give me some input on this problem. thanks in advance Gudrun Lang Akh Linz, Austria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 29 Date: Thu, 01 Apr 2004 20:36:50 +0800 From: "yichao wu" Subject: [Histonet] RE: C4d antibody on Paraffin sections To: M.Donovan@alfred.org.au Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Dear Donovan, Sorry for the late reply. Actually I learned the method from a refence,but I could not remember the exact journal and article now. The sections I used were acetone-methanol(1:1) fixed paraffin embedded sections with. I have not tried on formalin-fixed paraffin-embedded sections.But I remember that in the reference mentioned above,it reads that this method is applicable on formalin-fixed sections. The procedures are not special,just add the pretreatment with 88% formic acid.That is to say, Dewaxing; Draw a circle around the sections with PAP pen; Incubate with PBS; Incubate with 88% formic acid (diluted with ddH2O i.e. pure water) for 20 minutes in room temperature; Incubate with 10% fetal bovine serum as the blocker (or other kinds of block serum); Incubate with monoclonal mouse anti-C4d antibody (A213 from Quidel USA) at 4C overnight; Incubate with FITC-conjugated rabbit anti-mouse IgG antibody (from DAKO) for 40 mins at RT; Observe under fluorescent microscope... Besides, it is not any advertisement for those products I mentioned! :-) Just a little experience. Wish it is helpful. And thank you for your attention.You are much welcomed! Yichao WU,Ph.D Candidate Research Insititute of Nephrology 305 East Zhongshan Road Nanjing 210002,P.R.China >From: "Donovan, Mark" >To: "'yichaowu@hotmail.com'" >Subject: C4d antibody on Paraffin sections >Date: Tue, 30 Mar 2004 15:39:03 +1000 > >Dear Yichao, > >I read your response to Carmen Loiselle regarding the Quidel antibody to >C4d >with great interest. > >Could you provide me with more details of your protocol for using the >antibody on formalin fixed paraffin embedded sections. > >I have used this antibody on frozen sections but have not had any success >in >getting it to work on formalin fixed material. Your advice would be very >much appreciated. > >Regards, > >Mark Donovan >Immunohistochemistry Laboratory >Anatomical Pathology >The Alfred Hospital >Melbourne, Australia > > >THIS E-MAIL IS CONFIDENTIAL. If you have received this e-mail in error, >please notify us by return e-mail and delete the document. If you are not >the intended recipient you are hereby notified that any disclosure, >copying, >distribution or taking any action in reliance on the contents of this >information is strictly prohibited and may be unlawful. Bayside Health is >not liable for the proper and complete transmission of the information >contained in this communication or for any delay in its receipt. _________________________________________________________________ Add photos to your e-mail with MSN 8. Get 2 months FREE*. http://join.msn.com/?page=features/featuredemail ------------------------------ Message: 30 Date: Thu, 01 Apr 2004 14:03:03 +0000 From: "Julien Lambrey de Souza" Subject: [Histonet] testing list To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain This is a test. Do not consider. Thankyou. _________________________________________________________________ Help protect your entire PC with Virus Guard from [1]MSN Premium Get Two Months FREE* References 1. http://g.msn.com/8HMAENCA/2731??PS= ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 5, Issue 1 ************************************** From peoshel <@t> wisc.edu Thu Apr 1 08:47:09 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] chicken breast In-Reply-To: References: Message-ID: We use the ApF 1 marker from Munchausen Biotech. Phil >Recently we have had several mammary tumours in our flock >of Leicester Blue hens.I wonder if anyone out there knows >of any specific immunohistochemical markers for chicken mammary >tissue/tumours?? > >Many thanks > >C. Sanders > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From michael_lafriniere <@t> memorial.org Thu Apr 1 09:40:25 2004 From: michael_lafriniere <@t> memorial.org (LaFriniere, Mike) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] tissue processing Message-ID: We use TBS's filter perforated paper for all small biopsies -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Thursday, April 01, 2004 5:10 AM To: Histonetliste Subject: [Histonet] tissue processing Hi We have a current problem with our routine tissue processing in the VIP. We loaded 200 capsules in the container. This time we had the half of the capsules filled with biopsy-sponges. Usually they are only about 30% of all. We do over night processing. The tissue surface was something like creamy and in the trimmed block the tissue looked wet. I was not able to get a good section (only holes in paraffin). But not all blocks were like this, most of the small biopsies worked well. My suggestion is, that the great amount of sponges is responsible for the underprocessing. I think the tissue was'nt dehydratet enough and water was taken over from one step to the next. Did anybody have the same experience? Please give me some input on this problem. thanks in advance Gudrun Lang Akh Linz, Austria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joseph-galbraith <@t> uiowa.edu Thu Apr 1 09:35:20 2004 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] accession numbers for multiple samples from same patient Message-ID: <5D03ED7B9391D4119D9B0008C76B7B24030085F6@uihc-mail1.uihc.uiowa.edu> Diana: We use a single accession number with different part letters for the situation you describe. Best wishes, Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Diana McCaig Sent: Thursday, April 01, 2004 6:00 AM To: histonet@pathology.swmed.edu Subject: [Histonet] accession numbers for multiple samples from same patient Can I be advised how other sites label multiple specimens or where I can find the Canadian standard. Are they given different numbers ie 101-biopsy small bowel 102-biopsy cecum 103-antrum biopsy OR 101A-biopsy small bowel 101B-biopsy cecum 101C-antrum biopsy Thanks Diana McCaig, R.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfavara <@t> niaid.nih.gov Thu Apr 1 09:50:32 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Iba1 Message-ID: Anyone using this antibody? c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 From gcallis <@t> montana.edu Thu Apr 1 10:43:46 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Which lectin? In-Reply-To: <3.0.6.32.20040401093946.007c0b60@pop.lugo.usc.es> References: Message-ID: <3.0.6.32.20040401094346.00bc0a88@gemini.msu.montana.edu> Dear all, Please, when people discuss lectins, please specify which lectin you are attempting to stain. It may help with answers. Thanks Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From tpmorken <@t> labvision.com Thu Apr 1 10:48:20 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] chicken breast Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA217748A@usca0082k08.labvision.apogent.com> Strangley, In our lab we've only had good results with ApF 1 at certain times of the year. Tim Morken -----Original Message----- From: Philip Oshel [mailto:peoshel@wisc.edu] Sent: Thursday, April 01, 2004 6:47 AM To: Histonet@Pathology.swmed.edu Subject: Re: [Histonet] chicken breast We use the ApF 1 marker from Munchausen Biotech. Phil >Recently we have had several mammary tumours in our flock >of Leicester Blue hens.I wonder if anyone out there knows >of any specific immunohistochemical markers for chicken mammary >tissue/tumours?? > >Many thanks > >C. Sanders > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Apr 1 10:59:30 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] non-speficic binding of secundary biotinilated antibodies In-Reply-To: <3B070848E7C2204F9DEB8BCFD767728001079CC8@ntexch1.voeding.t no.nl> Message-ID: <3.0.6.32.20040401095930.00bc0a88@gemini.msu.montana.edu> We avoid rabbit antibodies, this host tends to give us far more problems with background. We always purchase our secondaries adsorbed to the host being stained. We use either goat antiMouse F(ab')2 frag of IgG adsorbed to RAT diluted in 10% goat serum OR Donkey antiMouse F(ab')2 frag of IgG, adsorbed to rat (come adsorbed to many species) from Jackson ImmunoResearch and diluted in 10% donkey serum. There is a clever way to prevent binding to B cell immunoglobulins published by e Sousa in J Exp Medicine. We have done it when staining mouse tissues, but it would also work when staining Rat (these critters are rather closely related) and you might not believe it works, but the results were very clean. I will be happy to discuss this privately. Sometimes you just have to add rat serum 1 - 5 % to the normal serum block, so it would be 10% rabbit/2.5% rat, or 10% goat/2.5% rat, etc. Serum are are heat inactivated, and normal serum blocks are applied for 30 minutes before primary. We prefer to dilute secondaries in the same normal serum block, even with rat serum if that is used. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gudrun.lang <@t> aon.at Thu Apr 1 11:00:41 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] tissue processing References: Message-ID: <001c01c4180a$ddb1c090$eeeea8c0@SERVER> Fran, We don't make them especially wet, but we store the ready capsules in formalin until the run is started. (perhaps for half an hour or less) Gudrun ----- Original Message ----- From: "Fran Lemons" To: Sent: Thursday, April 01, 2004 4:28 PM Subject: Re: [Histonet] tissue processing Did you make sure the sponges were wet when you put the tissue in them? If they aren't the reagents have a more difficult time penetrating the sponges & reaching the tissue. Were you using sponges on large pieces? Not intended for that use & detrimental to absorption. >>> "Gudrun Lang" 04/01/04 05:10AM >>> Hi We have a current problem with our routine tissue processing in the VIP. We loaded 200 capsules in the container. This time we had the half of the capsules filled with biopsy-sponges. Usually they are only about 30% of all. We do over night processing. The tissue surface was something like creamy and in the trimmed block the tissue looked wet. I was not able to get a good section (only holes in paraffin). But not all blocks were like this, most of the small biopsies worked well. My suggestion is, that the great amount of sponges is responsible for the underprocessing. I think the tissue was'nt dehydratet enough and water was taken over from one step to the next. Did anybody have the same experience? Please give me some input on this problem. thanks in advance Gudrun Lang Akh Linz, Austria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gudrun.lang <@t> aon.at Thu Apr 1 11:05:05 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] tissue processing References: <000001c417f7$950c0390$3601a8c0@brownpathology.net> Message-ID: <002701c4180b$7b192990$eeeea8c0@SERVER> Our protocol: 2 x 60 min formalin 1 x 60 min 50% alk 1 x 60 min 70% alk 1 x 60 min 80% alk 2 x 60 min 96% alk 2 x 60 min 100% alk 2 x 60 min xylolsubst. = shellsol 4 x 45 min paraplast 40 degrees in alk and xylol; 60 degrees in paraffin all with press/vacuum. And this works fine for most of our cases. Gudrun ----- Original Message ----- From: "Bonnie Whitaker" To: "'Gudrun Lang'" Sent: Thursday, April 01, 2004 4:42 PM Subject: RE: [Histonet] tissue processing What is your protocol? Are you using vacuum, pressure, mixing? It almost sounds like the sponges might not have fully exchanged solutions at each step. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Thursday, April 01, 2004 4:10 AM To: Histonetliste Subject: [Histonet] tissue processing Hi We have a current problem with our routine tissue processing in the VIP. We loaded 200 capsules in the container. This time we had the half of the capsules filled with biopsy-sponges. Usually they are only about 30% of all. We do over night processing. The tissue surface was something like creamy and in the trimmed block the tissue looked wet. I was not able to get a good section (only holes in paraffin). But not all blocks were like this, most of the small biopsies worked well. My suggestion is, that the great amount of sponges is responsible for the underprocessing. I think the tissue was'nt dehydratet enough and water was taken over from one step to the next. Did anybody have the same experience? Please give me some input on this problem. thanks in advance Gudrun Lang Akh Linz, Austria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lao_ji <@t> yahoo.com Thu Apr 1 11:09:56 2004 From: lao_ji <@t> yahoo.com (Jimmy Lao) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Automated ISH machine price? In-Reply-To: <20040401021915.65884.qmail@web41607.mail.yahoo.com> Message-ID: <20040401170956.39278.qmail@web12506.mail.yahoo.com> Hi, Our Lab is thinking to buy an automated ISH/IHC machine for slices. I found Tecan has one named genepaint for 48 slices. Anyone used this machine? How much is it? or anyone knows other made of this machine for these scale? What is the price? we don't need it for 100 or 200 slices. Thanks! Jimmy __________________________________ Do you Yahoo!? Yahoo! Small Business $15K Web Design Giveaway http://promotions.yahoo.com/design_giveaway/ From gcallis <@t> montana.edu Thu Apr 1 11:20:03 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Sponge problems in processing In-Reply-To: <001a01c417d1$87b00500$eeeea8c0@SERVER> Message-ID: <3.0.6.32.20040401102003.00bc0a88@gemini.msu.montana.edu> Gudrun, We no longer use sponges, prefer to place tissues in tissue embedding bags (Fisher) which look like tea bags or the little nylon bags (not as easy to handle, but thinner than sponges) and a tidge stiffer than tea bags. Sponges can cause artifacts in your tissues, looking like triangular holes in section. This was published by Freida Carson in Journal of Histotechnology, 1980's. Using tea bags may not be as fast during embedding, but speed there is a trade off for having to reprocess important tissue samples, ho hum tedious and time consuming while patient waits. You analysis of problem sound correct with a poor exchange of solvents through sponges. If you pack cassettes super tight in basket/cassette holder inside processor you can impede solvent flow or if sponges are crammed too tight agaist tissue then lid smashed down on tissue/sponge sandwich - this is like having too thick a tissue in a cassette. There are some clever biopsy cassettes with a folding, fine mesh inserts fitting inside a cassette. These may be worth a try to avoid sponges. I think they are available through Fisher or some other company who could provide free samples. It will be interesting to see peoples testimonials on using these. There was Histonet discussion on these sponges way back in time - it may pay to do a search for those messages in Histonet Archives. At 12:10 PM 4/1/2004 +0200, you wrote: >Hi >We have a current problem with our routine tissue processing in the VIP. We loaded 200 capsules in the container. This time we had the half of the capsules filled with biopsy-sponges. Usually they are only about 30% of all. We do over night processing. >The tissue surface was something like creamy and in the trimmed block the tissue looked wet. I was not able to get a good section (only holes in paraffin). But not all blocks were like this, most of the small biopsies worked well. >My suggestion is, that the great amount of sponges is responsible for the underprocessing. I think the tissue was'nt dehydratet enough and water was taken over from one step to the next. > >Did anybody have the same experience? Please give me some input on this problem. > >thanks in advance >Gudrun Lang >Akh Linz, Austria > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Thu Apr 1 11:23:02 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] microwave antigen retrieval done by a master immunostainer Message-ID: <3.0.6.32.20040401102302.00bc0a88@gemini.msu.montana.edu> The IHC expert/master stainer, Dr. Chris van der Loos uses a microwave for retrieval. He has a very clever device he invented for use in microwave. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From JPCOLEMA <@t> sentara.com Thu Apr 1 12:01:04 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] EGFR Message-ID: I have a clinical oncology group that is expecting a 50% positivity rate for EGFR in colon cancer patients. Is this rate of positivity familiar to anyone? Is there a clone name and procedural information that anyone can share? From la.sebree <@t> hosp.wisc.edu Thu Apr 1 12:06:39 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] EGFR Message-ID: John, There was quite a discussion of this antibody several weeks ago. You can look it up in the archives and find a wealth of information. 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: JOHN COLEMAN [mailto:JPCOLEMA@sentara.com] Sent: Thursday, April 01, 2004 12:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] EGFR I have a clinical oncology group that is expecting a 50% positivity rate for EGFR in colon cancer patients. Is this rate of positivity familiar to anyone? Is there a clone name and procedural information that anyone can share? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JCheng <@t> mrl.ubc.ca Thu Apr 1 12:06:39 2004 From: JCheng <@t> mrl.ubc.ca (Jenny Cheng) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Lectin IHC Message-ID: Hi To all, Here is more detailed descripiton of what I have done. The organism I am studying is Guinea Pig. The tissues are infiltrated with OCT and kept frozen until cut. This is the General Protocol of my lectin staining 1. Air Dry for 15 mins 2. Application of fixative (depending on the reagents used, incubation time changes) and tap water rinse 3. Block for 30mins (3X Buffer Wash) 4. Lectin Incubation for 1 hr (3X Buffer Wash) 5.Application of ABC for 30 mins(3X Buffer Wash) 6. Application of substrage for 15 mins (tap water rinse) 7. Counterstain with mayer's hemotoxylin The combinations that I have tried are ( fixitive/block/dectection method) 10% Formalin/ universal block/ avidin-biotin 10% formalin/ 0.1% BSA block/avidin-HRP or avidin-Biotin Acetone/0.1%BSA block/avidin-HRP Bouin/universal block or 0.1% BSA block/avidin-biotin I haven't been able to get a positive staining for my positive control. The lectin I been using is UEA-1 Any suggestion would be great. Jenny From zandra <@t> gwu.edu Thu Apr 1 12:15:44 2004 From: zandra <@t> gwu.edu (Alexandra de Sousa) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] bidest. water vs. sterile water Message-ID: <242c8ca2429422.2429422242c8ca@gwu.edu> Hello. I have received some freeze dried antibodies and the attached form asks that they be reconstituted with bidest. water. Is there a "real" difference between bidest. water and sterile water? Can they be treated the same for this purpose? Thanks, Alexandra From psanquin <@t> lugo.usc.es Thu Apr 1 12:48:44 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Re: Lectin IHC In-Reply-To: Message-ID: <3.0.6.32.20040401204844.007ee770@pop.lugo.usc.es> Hi Jenny, Nothing unexpected in your protocol. I usually incubate the UEA-I overnight at 4?C. It could make a difference. 15 mins of substrate (DAB?) is a long time. Staining should come off in a few minutes. I guess you use biotinylated lectin. With UEA-I I have tried both the biotinylated lectin + ABC from Sigma or the UEA-I + Peroxidase-linked antibody against UEA-I (from Dako). This works much better. You do not tell in what organ or tissue you employs UEA I. That is also relevant. For example in nervous system UEA-I only stains the olfactory bulbs. For nervous system you can see P.C. Barber, Neuroscience 30:1-9 (1989). He discusses different fixatives and protocols. Regards Pablo At 10:06 a.m. 01/04/04 -0800, you wrote: >Hi To all, >Here is more detailed descripiton of what I have done. The organism I >am studying is Guinea Pig. The tissues are infiltrated with OCT and kept >frozen until cut. >This is the General Protocol of my lectin staining >1. Air Dry for 15 mins >2. Application of fixative (depending on the reagents used, incubation >time changes) and tap water rinse >3. Block for 30mins (3X Buffer Wash) >4. Lectin Incubation for 1 hr (3X Buffer Wash) >5.Application of ABC for 30 mins(3X Buffer Wash) >6. Application of substrage for 15 mins (tap water rinse) >7. Counterstain with mayer's hemotoxylin > >The combinations that I have tried are ( fixitive/block/dectection >method) >10% Formalin/ universal block/ avidin-biotin >10% formalin/ 0.1% BSA block/avidin-HRP or avidin-Biotin >Acetone/0.1%BSA block/avidin-HRP >Bouin/universal block or 0.1% BSA block/avidin-biotin > >I haven't been able to get a positive staining for my positive control. >The lectin I been using is UEA-1 Any suggestion would be great. >Jenny > > From dmnelson <@t> iastate.edu Thu Apr 1 13:24:28 2004 From: dmnelson <@t> iastate.edu (Diane Gerjets) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] labeling multiple samples, same case Message-ID: <6.0.1.1.2.20040401131720.01b378b0@dmnelson.mail.iastate.edu> Hi Diana, 101A- cecum 101B- ileum Diane Gerjets From carl.hobbs <@t> kcl.ac.uk Thu Apr 1 13:36:35 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] re mirowave Ag retrieval Message-ID: <000b01c41820$a5312ce0$fcae8451@home> Well...I use a microwave pressure cooker! I understand that one can buy this in USA Walmarts ( be interested to know the price of it in the USA). I buy them in UK from Biogenex. Very reliable and consistent.. Started on Coplins jars and , VERY rapidly moved to rice staemers, pressure cookers...then microwave pressure cookers. I look back in no anger. From ploykasek <@t> phenopath.com Thu Apr 1 14:01:02 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] chicken breast In-Reply-To: Message-ID: I would say the "LOOF LIRPA" antibody would work nicely in this situation. Patti Loykasek PhenoPath A big fan of "C. Sanders" & his place of business "KFC"> > > Recently we have had several mammary tumours in our flock of > Leicester Blue hens.I wonder if anyone out there knows of any > specific immunohistochemical markers for chicken mammary tissue/tumours?? > Many > thanks > C. > Sanders > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gudrun.lang <@t> aon.at Thu Apr 1 14:15:59 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] tissue processing References: Message-ID: <020701c41826$262aa790$eeeea8c0@SERVER> Thank you all for your answers! It is pretty funny for me, that we have introduced the sponges in times, when others kicked them already off. But this was the first time, they made really problems. And our pathologists never complained about sponge-artefacts. Thank you again, Gudrun from Austria ----- Original Message ----- From: "S Ladd" To: "Gudrun Lang" Sent: Thursday, April 01, 2004 7:24 PM Subject: RE: [Histonet] tissue processing > Gudrun, > This very same thing happenned to us 2 weeks ago. We loaded approximately > 200 cassettes on the new VIP and about 75% of them had sponges. > Approximately 50 of the blocks were not processed at all. The 50 blocks were > all in the same area of the basket and they were all in the bottom basket > not the top! The size of the tissue did not correlate with the how well the > specimens were processed either; some big excisions processed and some > little shaves did not. We checked all of the solutions and we even checked > the formalin vials from the unprocessed cases and everything was fine. It > was a complete and total mystery. Later someone noticed that the agitation, > P/V AND temperature had been turned off on all of the stations on the > program that we had run. Having a lot of sponges probably compounded the > problem. So....try checking the P/V, agitation and temperature. What a > frustrating experience! I never thought in a million years that someone else > would have had the very same problem! > Sharron > University of South Florida > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gudrun > Lang > Sent: Thursday, April 01, 2004 5:10 AM > To: Histonetliste > Subject: [Histonet] tissue processing > > > Hi > We have a current problem with our routine tissue processing in the VIP. We > loaded 200 capsules in the container. This time we had the half of the > capsules filled with biopsy-sponges. Usually they are only about 30% of all. > We do over night processing. > The tissue surface was something like creamy and in the trimmed block the > tissue looked wet. I was not able to get a good section (only holes in > paraffin). But not all blocks were like this, most of the small biopsies > worked well. > My suggestion is, that the great amount of sponges is responsible for the > underprocessing. I think the tissue was'nt dehydratet enough and water was > taken over from one step to the next. > > Did anybody have the same experience? Please give me some input on this > problem. > > thanks in advance > Gudrun Lang > Akh Linz, Austria > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From LuckG <@t> empirehealth.org Thu Apr 1 14:17:55 2004 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] er/pr charging Message-ID: Terri, Our pathologists interpretation of ER/PR IHC is stated as either "positive or negative". This in our estimation does not meet the standard of either quantitative or semi-quantitative (which is specified in the CPT code 88361 application description. We therefore are continuing to code these as 88342's. I think that if the interpretation is stated as a raw "numerical" value (eg. 45% or 70% rather than a less empirical statement of 1+ or 3+) there is good justification to use the new 88361 code. Greg Luck Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 509.473.7077 luckg@empirehealth.org -----Original Message----- From: Hallada, Teri [mailto:thallada@noch.org] Sent: Thursday, April 01, 2004 4:25 AM To: Kelly Simon; Joyce Cline Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] er/pr charging I believe you must "quantitative" the results in some format. Our pathologists here don't quantitative ER/PR only Her2, so we don't use 88361 for ER/PR only Her2. Is this correct? Teri Hallada BS MT CT (ASCP) thallada@noch.org > -----Original Message----- > From: Kelly Simon [SMTP:kb2drkprk@yahoo.com] > Sent: Thursday, April 01, 2004 01:06 > To: Joyce Cline > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] er/pr charging > > According to a story by Lisa Miller in the January > 2004 CAP Today, there is a new code for ER/PR and > HER2/neu: > > "CPT 2004 separates traditional tumor morphometric > analysis from semiquantitative immunohistochemistry. > New code 88361 was created to report quantitative or > semiquantitative immunohistochemistry for such > analyses as hormone receptor and HER2/neu testing. The > technical services of 88342 are incorporated into the > new code." > > http://www.cap.org/apps/docs/cap_today/feature_stories/0104InWithNewCPT2004. html > > Kelly Simon, HTL (ASCP) > Dynacare Laboratories > Seattle, WA > > --- Joyce Cline wrote: > > Does everyone use the CPT code 88342 for charging > > the er/pr immuno's. Or is there another code that is > > used for the technical component? > > > > ***** 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. > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > __________________________________ > Do you Yahoo!? > Yahoo! Small Business $15K Web Design Giveaway > http://promotions.yahoo.com/design_giveaway/ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Statement: This message is intended only for the individual or entity to which it is addressed. It may contain privileged, confidential information which is exempt from disclosure under applicable laws. If you are not the intended recipient, please note that you are strictly prohibited from disseminating or distributing this information (other than to the intended recipient) or copying this information. If you have received this communication in error, please notify us immediately by e-mail or by telephone at (616) 842-3600. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Thu Apr 1 14:06:21 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Microwave Message-ID: We do various pretreatments here. The pretreatment we use most frequently is microwave pressure cook. We use a pressure cooker intended for use in the microwave. We buy the pressure cooker off the internet. I'm a big fan of steaming, too. Just fyi. Patti Loykasek PhenoPath Laboratories Seattle, WA From Luis.Chiriboga <@t> med.nyu.edu Thu Apr 1 16:18:21 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Microwave In-Reply-To: Message-ID: Would like to add........ I believe most evidence indicates that it's not the source of the heat but rather the time/temperature/buffer. pressure cookers and autoclaves work better because they can achieve temperatures above boiling as well as obtain those temps faster. Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patti Loykasek Sent: Thursday, April 01, 2004 3:06 PM To: histonet Subject: [Histonet] Microwave We do various pretreatments here. The pretreatment we use most frequently is microwave pressure cook. We use a pressure cooker intended for use in the microwave. We buy the pressure cooker off the internet. I'm a big fan of steaming, too. Just fyi. Patti Loykasek PhenoPath Laboratories Seattle, WA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stanley.Stylli <@t> mh.org.au Thu Apr 1 16:14:27 2004 From: Stanley.Stylli <@t> mh.org.au (Stylli, Stanley) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Antibody Source Message-ID: <00519F9F41D2844D9C75BEE1F6AC09AFC89ECB@rmhmail1.ssg.org.au> Dear All, Does anyone have any experience with using the following antibodies (with success) on rat tissue (especially brain) that has been formalin fixed and paraffin embedded : Caspase-3 Tenascin-R TNF-alpha Vimentin VEGF I would prefer a monoclonal but am willing to purchase a polyclonal antibody from a commercial source. Also, can you tell me if you managed to use the antibody with/without antigen retrieval. Thanks Stan From gcallis <@t> montana.edu Thu Apr 1 16:30:37 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Microwave discussion and THE book to own Message-ID: <3.0.6.32.20040401153037.00bca6f0@gemini.msu.montana.edu> Antigen unmasking, retrieval, etc, etc is discussed by the experts (two who developed methods over the years) in Antigen Retrieval Techniques, by Shi, Gu and Taylor, Eaton Publishing. This book is worth its weight in gold for educating technicians about various methods, buffers, pH, temps, how to's etc with bonus chapter for EM immuno work. Beats having to dig all this out of publications. You can also buy it from BioTechniques website, their bookstore. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From shive003 <@t> umn.edu Thu Apr 1 16:41:26 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Re: Lectin IHC Message-ID: <014e01c4183a$77b9eda0$78065486@vdl220FAC> I deleted the initial start of this discussion prematurely, but I wanted to add that back when I did lectin staining years ago, I found that not all lectins stained consistently between the various species. I reasoned that it was perhaps a variance in the glycoconjugates found on different species' cells. The lectin I wanted to work the most (UEA), I could not get to work on canine endothelium at all, for instance, though I had some success with other lectins on other cell types. Of course, I admit that maybe I had just not found the magic protocol to make the UEA work on dog tissue. However, one might consider that species variability could be the reason why one lectin works on human material but not on guinea pig or other mammals. Jan Shivers U of MN Vet Diag Lab ----- Original Message ----- From: "Pablo S?nchez Quinteiro" To: Sent: Thursday, April 01, 2004 12:48 PM Subject: [Histonet] Re: Lectin IHC Hi Jenny, Nothing unexpected in your protocol. I usually incubate the UEA-I overnight at 4?C. It could make a difference. 15 mins of substrate (DAB?) is a long time. Staining should come off in a few minutes. I guess you use biotinylated lectin. With UEA-I I have tried both the biotinylated lectin + ABC from Sigma or the UEA-I + Peroxidase-linked antibody against UEA-I (from Dako). This works much better. You do not tell in what organ or tissue you employs UEA I. That is also relevant. For example in nervous system UEA-I only stains the olfactory bulbs. For nervous system you can see P.C. Barber, Neuroscience 30:1-9 (1989). He discusses different fixatives and protocols. Regards Pablo At 10:06 a.m. 01/04/04 -0800, you wrote: >Hi To all, >Here is more detailed descripiton of what I have done. The organism I >am studying is Guinea Pig. The tissues are infiltrated with OCT and kept >frozen until cut. >This is the General Protocol of my lectin staining >1. Air Dry for 15 mins >2. Application of fixative (depending on the reagents used, incubation >time changes) and tap water rinse >3. Block for 30mins (3X Buffer Wash) >4. Lectin Incubation for 1 hr (3X Buffer Wash) >5.Application of ABC for 30 mins(3X Buffer Wash) >6. Application of substrage for 15 mins (tap water rinse) >7. Counterstain with mayer's hemotoxylin > >The combinations that I have tried are ( fixitive/block/dectection >method) >10% Formalin/ universal block/ avidin-biotin >10% formalin/ 0.1% BSA block/avidin-HRP or avidin-Biotin >Acetone/0.1%BSA block/avidin-HRP >Bouin/universal block or 0.1% BSA block/avidin-biotin > >I haven't been able to get a positive staining for my positive control. >The lectin I been using is UEA-1 Any suggestion would be great. >Jenny > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From alexander.nader <@t> wgkk.sozvers.at Fri Apr 2 00:50:21 2004 From: alexander.nader <@t> wgkk.sozvers.at (Nader, Alexander) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] tissue processing Message-ID: <4FE843CBBAC1D211A8150008C70952DAAE7D60@hk01nt05.hkh.wgkk.sozvers.at> > My suggestion is, that the great amount of sponges is responsible for > the underprocessing. I think the tissue was'nt dehydratet enough and > water was taken over from one step to the next. dear Gudrun, we also had this problems a couple of time ago, I think, you are absoletely right considering the tight packing of sponges. But there's another big problem using sponges: there can produce peculiar triangular holes, artifacts, which can be seen sometimes especially in soft tissue, f.e. breast biopsies. For that reason we changed the processing and use now perforated paper from VOGEL (Filterpapier fuer Einbettkassetten, 66x26mm) for years. If you want to know further details, please contact me (01-91021-86411). article on this topic: Arch Pathol Lab Med. 1990 Dec;114(12):1285-7. Sponge artifact in biopsy specimens. Landas SK, Bromley CM. We describe a sponge-induced artifact in histologic sections of small biopsy specimens. The artifacts are angulated, often triangular holes within the tissue. They appear to be introduced as individual sponge barbs become embedded in the perimeter of biopsy specimens during tissue processing. The artifact is generally of little importance, but in certain specimens, such as needle biopsies of the kidney or liver, it may occasionally obscure important information. Other methods, such as lens paper wrapping, may be superior in these situations. The utility of the tissue cassette sponge, in most situations, outweighs the artifact. Alexander Nader MD Institut for Pathology, Hanusch-Krankenhaus Vienna, Austria From piernoel <@t> health.nb.ca Fri Apr 2 06:28:01 2004 From: piernoel <@t> health.nb.ca (Pierre Noel) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Help with frozen section staining Message-ID: <406D5C50.65C5B84E@health.nb.ca> Hi all, We just recently notice a major problem with our frozen section staining. The cellular morphology is poor, haziness and no cellular definition. We have tried different staining techniques with different fixative but no luck. Could it be our OCT? We desperately need some input on the subject. Thank you! Pierre-Andr? Noel Histology Supervisor Bathurst Regional Hospital NB, Canada From Jamie.Dukes <@t> se.amedd.army.mil Fri Apr 2 06:52:02 2004 From: Jamie.Dukes <@t> se.amedd.army.mil (Dukes, Jamie Mr EAMC) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] need help with studying for histo exam Message-ID: <4FB3076916FCD311AF7900805FA7A678077B4A59@dasmthfdz001.amedd.army.mil> to all, Need help with studying for histo exam next July. If anyone have any pointers please feel free to point. From tflore <@t> lsuhsc.edu Fri Apr 2 06:56:59 2004 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] need help with studying for histo exam Message-ID: Jamie, where are you located? Have you purchased any of the NSH study guides? Have you gone to www.nsh.org there are a lot of study guides. Dr. Frieda Carson's Study Guide and complementing Book is also very excellent. Rosemary Velasquez & I meet once a month with approximately 10 - 12 students preparing to take the HT and others the HTL (ASCP)and the above books are our bibles. Teresa Flores LSUHSC New Orleans, LA (504)568-6042 -----Original Message----- From: Dukes, Jamie Mr EAMC [mailto:Jamie.Dukes@se.amedd.army.mil] Sent: Friday, April 02, 2004 6:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] need help with studying for histo exam to all, Need help with studying for histo exam next July. If anyone have any pointers please feel free to point. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dobbin <@t> upei.ca Fri Apr 2 06:02:23 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Help with frozen section staining In-Reply-To: <406D5C50.65C5B84E@health.nb.ca> Message-ID: <406D2C1F.19258.D3AA9@localhost> Hi Pierre-Andr? (neighbor), I don't see the OCT being the most likely culprit, unless MAYBE if you were perfusing with it. The OCT should only be surrounding your tissue sections (not entering into them). I would be more inclined to look at: a) the fixative (try brand new batch, if the fixative is faulty, your sections will not tolerate the non-isotonic and perhaps acid or basic pH of your staining solutions). And then b) the mounting medium (for clarity) and the coverslips for defects. (presumably you've ruled out the microscope as the problem?) Good luck. Greg Date sent: Fri, 02 Apr 2004 08:28:01 -0400 From: piernoel@health.nb.ca (Pierre Noel) To: histonet@lists.utsouthwestern.edu Send reply to: piernoel@health.nb.ca related fields Subject: [Histonet] Help with frozen section staining > This is a multi-part message in MIME format. > --------------5A96B0B15D35A5B00C345253 > Content-Type: text/plain; charset=iso-8859-1 > Content-Transfer-Encoding: 8bit > > Hi all, > We just recently notice a major problem with our frozen section > staining. The cellular morphology is poor, haziness and no cellular > definition. We have tried different staining techniques with different > fixative but no luck. Could it be our OCT? We desperately need some > input on the subject. Thank you! > > Pierre-Andr? Noel > Histology Supervisor > Bathurst Regional Hospital > NB, Canada > > --------------5A96B0B15D35A5B00C345253 > Content-Type: text/plain; charset="us-ascii" > MIME-Version: 1.0 > Content-Transfer-Encoding: 7bit > Content-Disposition: inline > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > --------------5A96B0B15D35A5B00C345253-- > > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Pathology Lab Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Making a living is getting; making a life is giving. From sladd <@t> hsc.usf.edu Fri Apr 2 07:11:00 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:22:45 2005 Subject: FW: [Histonet] tissue processing and sponges Message-ID: I forgot to copy this to the list yesterday. Sharron -----Original Message----- From: S Ladd [mailto:sladd@hsc.usf.edu] Sent: Thursday, April 01, 2004 12:24 PM To: Gudrun Lang Subject: RE: [Histonet] tissue processing Gudrun, This very same thing happenned to us 2 weeks ago. We loaded approximately 200 cassettes on the new VIP and about 75% of them had sponges. Approximately 50 of the blocks were not processed at all. The 50 blocks were all in the same area of the basket and they were all in the bottom basket not the top! The size of the tissue did not correlate with the how well the specimens were processed either; some big excisions processed and some little shaves did not. We checked all of the solutions and we even checked the formalin vials from the unprocessed cases and everything was fine. It was a complete and total mystery. Later someone noticed that the agitation, P/V AND temperature had been turned off on all of the stations on the program that we had run. Having a lot of sponges probably compounded the problem. So....try checking the P/V, agitation and temperature. What a frustrating experience! I never thought in a million years that someone else would have had the very same problem! Sharron University of South Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gudrun Lang Sent: Thursday, April 01, 2004 5:10 AM To: Histonetliste Subject: [Histonet] tissue processing Hi We have a current problem with our routine tissue processing in the VIP. We loaded 200 capsules in the container. This time we had the half of the capsules filled with biopsy-sponges. Usually they are only about 30% of all. We do over night processing. The tissue surface was something like creamy and in the trimmed block the tissue looked wet. I was not able to get a good section (only holes in paraffin). But not all blocks were like this, most of the small biopsies worked well. My suggestion is, that the great amount of sponges is responsible for the underprocessing. I think the tissue was'nt dehydratet enough and water was taken over from one step to the next. Did anybody have the same experience? Please give me some input on this problem. thanks in advance Gudrun Lang Akh Linz, Austria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Diane.Gladney <@t> se.amedd.army.mil Fri Apr 2 07:23:34 2004 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] need help with studying for histo exam Message-ID: <9D41AB7C56F8304F98537ABD87B249F66261B7@DASMTHGBZ001> Jamie, Are you taking the HT or the HTL exam? There are 2 web sites that you should visit for access to study material and a list of reference books: American Society for Clinical Pathology www.ascp.org National Society for Histotechnology www.nsh.org The NSH site has self examination booklets in 12 categories. Click on the Ed. Materials for this list. Also they have a list of reference books. Click on Study Aids for this list. Although I took the exam ages ago, I would recommend a thorough study of special stains, what components stain what color, what the stain demonstrates, etc. This is a tough exam and if you pass the first time, then you should be highly commended. The exam covers so much information that you need to thoroughly know all aspects of histotechnology. You are located at Eisenhower Army Medical Center aren't you? If I am correct, then you have a great bunch of pathologist. When you do your practical, have them to critique your slides. They will be of great help to you. I have had the great pleasure of working directly with most of them when they came to our facility to help us out. They are very good pathologists and are a wonderful source of information. Talk with them and see if one of them will mentor you while you study for the exam. If I can be of help, please contact me directly. Good Luck, Diane Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: Dukes, Jamie Mr EAMC [mailto:Jamie.Dukes@se.amedd.army.mil] Sent: Friday, April 02, 2004 7:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] need help with studying for histo exam to all, Need help with studying for histo exam next July. If anyone have any pointers please feel free to point. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Fri Apr 2 08:07:19 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Help with frozen section staining Message-ID: <566FB0B522443D43AF02D2ADBE35A6F077FD5C@UTHEVS3.mail.uthouston.edu> Pierre et al Hi to y'all in Canada, I have a relative in Brampton, Ontario and really enjoy my visits there. A point re OCT compound. It contains several salts including I believe some calcium. It is important therefore for some procedures such as when using certain lectins to thoroughly remove the OCT from the section to prevent interference with staining and with other procedures. This is easily taken care of with buffer/or alcohol etc. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Greg Dobbin Sent: Friday, April 02, 2004 3:02 AM To: piernoel@health.nb.ca Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Help with frozen section staining Hi Pierre-Andr? (neighbor), I don't see the OCT being the most likely culprit, unless MAYBE if you were perfusing with it. The OCT should only be surrounding your tissue sections (not entering into them). I would be more inclined to look at: a) the fixative (try brand new batch, if the fixative is faulty, your sections will not tolerate the non-isotonic and perhaps acid or basic pH of your staining solutions). And then b) the mounting medium (for clarity) and the coverslips for defects. (presumably you've ruled out the microscope as the problem?) Good luck. Greg Date sent: Fri, 02 Apr 2004 08:28:01 -0400 From: piernoel@health.nb.ca (Pierre Noel) To: histonet@lists.utsouthwestern.edu Send reply to: piernoel@health.nb.ca related fields Subject: [Histonet] Help with frozen section staining > This is a multi-part message in MIME format. > --------------5A96B0B15D35A5B00C345253 > Content-Type: text/plain; charset=iso-8859-1 > Content-Transfer-Encoding: 8bit > > Hi all, > We just recently notice a major problem with our frozen section > staining. The cellular morphology is poor, haziness and no cellular > definition. We have tried different staining techniques with different > fixative but no luck. Could it be our OCT? We desperately need some > input on the subject. Thank you! > > Pierre-Andr? Noel > Histology Supervisor > Bathurst Regional Hospital > NB, Canada > > --------------5A96B0B15D35A5B00C345253 > Content-Type: text/plain; charset="us-ascii" > MIME-Version: 1.0 > Content-Transfer-Encoding: 7bit > Content-Disposition: inline > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > --------------5A96B0B15D35A5B00C345253-- > > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Pathology Lab Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Making a living is getting; making a life is giving. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MDiCarlo <@t> KaleidaHealth.Org Fri Apr 2 09:09:42 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] bone cement Message-ID: Hello, The doctor I work for gave me a section of bone containing bone cement. Will I be able to decal, process with paraffin and cut on a sledge microtome? I don't think I could but before I speak with him I wanted to check with everyone. Thanks in advance. Peggy DiCarlo HT(ASCP) Ortho Bone Lab Buffalo General Hospital 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 joseph-galbraith <@t> uiowa.edu Fri Apr 2 10:07:24 2004 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] re mirowave Ag retrieval Message-ID: <5D03ED7B9391D4119D9B0008C76B7B2403005B1B@uihc-mail1.uihc.uiowa.edu> Carl: We get our microwave pressure cooker from NordicWare (800-328-4310 ext 629) but I have seen these in kitchen stores as well (I'll have to check Walmart). Direct from the company, they cost $30.00 US with an extra seal costing $3.00. Of course, shipping is extra. If you take care of them, the cookers last a long time (several years of once daily use) before needing to be replaced. I found a site from NordicWare also but it costs more online: http://www.nordicware.com/b2c/product_category.cfm?prod_master_cat=2&prod_cat=7 Might try Amazon.com or Everthinghome.com too. Have fun shopping. Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Carl Sent: Thursday, April 01, 2004 1:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re mirowave Ag retrieval Well...I use a microwave pressure cooker! I understand that one can buy this in USA Walmarts ( be interested to know the price of it in the USA). I buy them in UK from Biogenex. Very reliable and consistent.. Started on Coplins jars and , VERY rapidly moved to rice staemers, pressure cookers...then microwave pressure cookers. I look back in no anger. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From davenhv <@t> hotmail.com Fri Apr 2 10:31:52 2004 From: davenhv <@t> hotmail.com (Dave McClister) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] (no subject) Message-ID: Happy Friday Histonetters, I'm in search of the Alexis 688 secondary antibody. What company can I order this from? I've been searching on through catalogs and the internet most of the morning with no luck. Any help would be greatly appreciatied. Thanks Dave Medical University of South Carolina [1]davenhv@hotmail.com _________________________________________________________________ [2]Persistent heartburn? Check out Digestive Health & Wellness for information and advice. References 1. mailto:Carolinadavenhv@hotmail.com 2. http://g.msn.com/8HMBENUS/2743??PS= From davenhv <@t> hotmail.com Fri Apr 2 10:31:52 2004 From: davenhv <@t> hotmail.com (Dave McClister) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] (no subject) Message-ID: Happy Friday Histonetters, I'm in search of the Alexis 688 secondary antibody. What company can I order this from? I've been searching on through catalogs and the internet most of the morning with no luck. Any help would be greatly appreciatied. Thanks Dave Medical University of South Carolina [1]davenhv@hotmail.com _________________________________________________________________ [2]Persistent heartburn? Check out Digestive Health & Wellness for information and advice. References 1. mailto:Carolinadavenhv@hotmail.com 2. http://g.msn.com/8HMBENUS/2743??PS= From bwhitaker <@t> brownpathology.com Fri Apr 2 10:37:35 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Grossing Manual Message-ID: <000001c418d0$cdea11f0$3601a8c0@brownpathology.net> Hi All, I have never been responsible for a complete grossing manual before, but now I have been asked to prepare one. Can someone share a template of what exactly it should contain? Thanks, Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 From haldana <@t> unimoron.edu.ar Fri Apr 2 10:42:00 2004 From: haldana <@t> unimoron.edu.ar (Hernan Aldana Marcos) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] help Message-ID: <004b01c418d1$6cd757a0$a904a8c0@um.edu> I have a bad immunohistochemistry reaction. Please, Who can me explain the appearance of non specific brown DAB reaction in the tissue edges of my section? Dr. Hern?n J. Aldana Marcos Facultad de Medicina. Universidad de Mor?n Machado 914. B1708JPD. Buenos Aires. Argentina e-mail alternativo hernanjavier@yahoo.com web: http://hjaldanamarcos.bravepages.com http://histologia.bigthicketdirectory.net/main.html From dennijc <@t> vetmed.auburn.edu Fri Apr 2 10:46:02 2004 From: dennijc <@t> vetmed.auburn.edu (John C. Dennis) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Re: Lectin IHC In-Reply-To: <014e01c4183a$77b9eda0$78065486@vdl220FAC> References: <014e01c4183a$77b9eda0$78065486@vdl220FAC> Message-ID: Jan What fixative(s) did you use? In fact, my question deals with UEA in primates. The signal is strong but I was dinged for using buffered (para)formaldehyde. From what I'm gathering, choice of fixative depends on what lectin one is interested in? John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Thu, 1 Apr 2004, Jan Shivers wrote: > I deleted the initial start of this discussion prematurely, but I wanted to > add that back when I did lectin staining years ago, I found that not all > lectins stained consistently between the various species. I reasoned that > it was perhaps a variance in the glycoconjugates found on different species' > cells. The lectin I wanted to work the most (UEA), I could not get to work > on canine endothelium at all, for instance, though I had some success with > other lectins on other cell types. Of course, I admit that maybe I had just > not found the magic protocol to make the UEA work on dog tissue. However, > one might consider that species variability could be the reason why one > lectin works on human material but not on guinea pig or other mammals. > > Jan Shivers > U of MN Vet Diag Lab > > ----- Original Message ----- > From: "Pablo S?nchez Quinteiro" > To: > Sent: Thursday, April 01, 2004 12:48 PM > Subject: [Histonet] Re: Lectin IHC > > > Hi Jenny, > > Nothing unexpected in your protocol. I usually incubate the UEA-I overnight > at 4?C. It could make a difference. 15 mins of substrate (DAB?) is a long > time. Staining should come off in a few minutes. > > I guess you use biotinylated lectin. With UEA-I I have tried both the > biotinylated lectin + ABC from Sigma or the UEA-I + Peroxidase-linked > antibody against UEA-I (from Dako). This works much better. > > You do not tell in what organ or tissue you employs UEA I. That is also > relevant. For example in nervous system UEA-I only stains the olfactory > bulbs. For nervous system you can see P.C. Barber, Neuroscience 30:1-9 > (1989). He discusses different fixatives and protocols. > > Regards > > Pablo > > > At 10:06 a.m. 01/04/04 -0800, you wrote: > >Hi To all, > >Here is more detailed descripiton of what I have done. The organism I > >am studying is Guinea Pig. The tissues are infiltrated with OCT and kept > >frozen until cut. > >This is the General Protocol of my lectin staining > >1. Air Dry for 15 mins > >2. Application of fixative (depending on the reagents used, incubation > >time changes) and tap water rinse > >3. Block for 30mins (3X Buffer Wash) > >4. Lectin Incubation for 1 hr (3X Buffer Wash) > >5.Application of ABC for 30 mins(3X Buffer Wash) > >6. Application of substrage for 15 mins (tap water rinse) > >7. Counterstain with mayer's hemotoxylin > > > >The combinations that I have tried are ( fixitive/block/dectection > >method) > >10% Formalin/ universal block/ avidin-biotin > >10% formalin/ 0.1% BSA block/avidin-HRP or avidin-Biotin > >Acetone/0.1%BSA block/avidin-HRP > >Bouin/universal block or 0.1% BSA block/avidin-biotin > > > >I haven't been able to get a positive staining for my positive control. > >The lectin I been using is UEA-1 Any suggestion would be great. > >Jenny > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@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 Fri Apr 2 11:00:21 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] help Message-ID: <0E660E4E.03740857.0A1F969F@aol.com> is your tissue retrieved? does it have a high biotin level more that average ie: liver or spleen? was a rinse not well done? is this automated? could it be a machine mixing issue? new secondary antibody? higher protein content? different fixation then before? tissue necrotic on edges,dry edges? any oneor all of these could be a reason for edge effect,could you give us more info? dana From ROENN <@t> surgery.wisc.edu Fri Apr 2 11:07:39 2004 From: ROENN <@t> surgery.wisc.edu (Drew Allan Roenneburg) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] (no subject) Message-ID: Molecular Probes sells zenon products, which are alexa flouorphores. There website is www.probes.com Drew Roenneburg From ROENN <@t> surgery.wisc.edu Fri Apr 2 11:07:39 2004 From: ROENN <@t> surgery.wisc.edu (Drew Allan Roenneburg) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] (no subject) Message-ID: Molecular Probes sells zenon products, which are alexa flouorphores. There website is www.probes.com Drew Roenneburg From Barry.R.Rittman <@t> uth.tmc.edu Fri Apr 2 11:54:08 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Re: Lectin IHC Message-ID: <566FB0B522443D43AF02D2ADBE35A6F06359F0@UTHEVS3.mail.uthouston.edu> John The lectin binding may also depend on the processing as well as the fixation. We carried out lectin binding some years ago on tissue fixed in buffered formalin and processed to paraffin wax using chloroform as the intermediary agent. We could not repeat the results on tissue that was sent to us from another lab. The difference was traced to the other laboratory used xylene in their processing schedule instead of chloroform. This would suggest that some the lectins we were examining were binding to glycolipids and that some of these were removed by the xylene. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John C. Dennis Sent: Friday, April 02, 2004 10:46 AM To: Jan Shivers Cc: histonet Subject: Re: [Histonet] Re: Lectin IHC Jan What fixative(s) did you use? In fact, my question deals with UEA in primates. The signal is strong but I was dinged for using buffered (para)formaldehyde. From what I'm gathering, choice of fixative depends on what lectin one is interested in? John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Thu, 1 Apr 2004, Jan Shivers wrote: > I deleted the initial start of this discussion prematurely, but I > wanted to add that back when I did lectin staining years ago, I found > that not all lectins stained consistently between the various species. > I reasoned that it was perhaps a variance in the glycoconjugates found > on different species' cells. The lectin I wanted to work the most > (UEA), I could not get to work on canine endothelium at all, for > instance, though I had some success with other lectins on other cell > types. Of course, I admit that maybe I had just not found the magic > protocol to make the UEA work on dog tissue. However, one might > consider that species variability could be the reason why one lectin > works on human material but not on guinea pig or other mammals. > > Jan Shivers > U of MN Vet Diag Lab > > ----- Original Message ----- > From: "Pablo S?nchez Quinteiro" > To: > Sent: Thursday, April 01, 2004 12:48 PM > Subject: [Histonet] Re: Lectin IHC > > > Hi Jenny, > > Nothing unexpected in your protocol. I usually incubate the UEA-I > overnight at 4?C. It could make a difference. 15 mins of substrate > (DAB?) is a long time. Staining should come off in a few minutes. > > I guess you use biotinylated lectin. With UEA-I I have tried both the > biotinylated lectin + ABC from Sigma or the UEA-I + Peroxidase-linked > antibody against UEA-I (from Dako). This works much better. > > You do not tell in what organ or tissue you employs UEA I. That is > also relevant. For example in nervous system UEA-I only stains the > olfactory bulbs. For nervous system you can see P.C. Barber, > Neuroscience 30:1-9 (1989). He discusses different fixatives and > protocols. > > Regards > > Pablo > > > At 10:06 a.m. 01/04/04 -0800, you wrote: > >Hi To all, > >Here is more detailed descripiton of what I have done. The organism I > >am studying is Guinea Pig. The tissues are infiltrated with OCT and > >kept frozen until cut. This is the General Protocol of my lectin > >staining 1. Air Dry for 15 mins > >2. Application of fixative (depending on the reagents used, incubation > >time changes) and tap water rinse > >3. Block for 30mins (3X Buffer Wash) > >4. Lectin Incubation for 1 hr (3X Buffer Wash) > >5.Application of ABC for 30 mins(3X Buffer Wash) > >6. Application of substrage for 15 mins (tap water rinse) > >7. Counterstain with mayer's hemotoxylin > > > >The combinations that I have tried are ( fixitive/block/dectection > >method) > >10% Formalin/ universal block/ avidin-biotin > >10% formalin/ 0.1% BSA block/avidin-HRP or avidin-Biotin > >Acetone/0.1%BSA block/avidin-HRP Bouin/universal block or 0.1% BSA > >block/avidin-biotin > > > >I haven't been able to get a positive staining for my positive > >control. The lectin I been using is UEA-1 Any suggestion would be > >great. Jenny > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@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 staceylburton <@t> yahoo.com Fri Apr 2 11:41:24 2004 From: staceylburton <@t> yahoo.com (Stacey Burton) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Salary Pole Message-ID: <20040402174124.21488.qmail@web14525.mail.yahoo.com> Per request of my administrators, I am currently performing a salary pole for Anatomic Pathology Laboratory Managers and Laboratory Coordinators. I have pulled the information from salary.com for my company, however they would like to have a second source to back up the information on salary.com. Would you please email me in person or on the histonet: the Starting, the Average, and Top-out salaries of the Laboratory Managers in your areas. I do not need to know the name of your facilities however the state and city would be helpful so we recognize the difference in cost of living through out the country. Thank you, Stacey Burton, H.T. ASCP Laboratory Manager Uropath LLC San Antonio TX staceylburton@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Small Business $15K Web Design Giveaway - Enter today From gcallis <@t> montana.edu Fri Apr 2 12:15:35 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] (no subject) In-Reply-To: Message-ID: <3.0.6.32.20040402111535.00bc07e8@gemini.msu.montana.edu> Molecular Probes at www.probes.com. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jeanthebean <@t> fuse.net Fri Apr 2 05:05:43 2004 From: jeanthebean <@t> fuse.net (Jean Warren) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Histo Exam Message-ID: <007e01c418a2$717cf640$4d6ba20a@zoomtown.com> For the computer portion of the test, do not overlook the special stains for neuropathology. I just took the test in Dec. 2003 and there were 8-10 questions regarding the specialized neuro stains. For instance, the test would depict a drawing of a neuron and an arrow would point to a structure. There would be multiple choice answers to the question, " What stains this structure?" A valuable resource to me was the Frieda Carson study guide that is nothing but questions divided into different categories. I think it was 25$ from the ASCP. It includes a section with color plates. Good luck, the test is thorough, but you will feel good if you pass!! From JMacDonald <@t> mtsac.edu Fri Apr 2 15:35:36 2004 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] need help with studying for histo exam Message-ID: Jamie, Thirteen of my students have taken the exam in the last year. The feedback that I am getting is the questions are not simple recall. Many of the questions are of a troubleshooting nature. Make sure you understand the concept, not memorize the answers. The NSH self-assessment books, the ASCP study guide and Freida Carson's books are great tools to study with. If you have a copy of Sheehan's book it also provides a lot of information. There is a lot of information that you are responsible for. Don't underestimate the time required for studying. Start now. Jennifer MacDonald. -----histonet-bounces@lists.utsouthwestern.edu wrote: ----- To: histonet@lists.utsouthwestern.edu From: "Dukes, Jamie Mr EAMC" Sent by: histonet-bounces@lists.utsouthwestern.edu Date: 04/02/2004 04:52AM Subject: [Histonet] need help with studying for histo exam to all, Need help with studying for histo exam next July. If anyone have any pointers please feel free to point. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jtaylor <@t> meriter.com Fri Apr 2 15:40:19 2004 From: jtaylor <@t> meriter.com (Taylor, Jean) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] unsubscribe Message-ID: <328CBAE62F31C642B422970E879DFADC0F7C06@pcwex01.meriter.com> -----Original Message----- From: Galbraith, Joe [mailto:joseph-galbraith@uiowa.edu] Sent: Friday, April 02, 2004 10:07 To: Carl; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] re mirowave Ag retrieval Carl: We get our microwave pressure cooker from NordicWare (800-328-4310 ext 629) but I have seen these in kitchen stores as well (I'll have to check Walmart). Direct from the company, they cost $30.00 US with an extra seal costing $3.00. Of course, shipping is extra. If you take care of them, the cookers last a long time (several years of once daily use) before needing to be replaced. I found a site from NordicWare also but it costs more online: http://www.nordicware.com/b2c/product_category.cfm?prod_master_cat=2&prod_cat=7 Might try Amazon.com or Everthinghome.com too. Have fun shopping. Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Carl Sent: Thursday, April 01, 2004 1:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re mirowave Ag retrieval Well...I use a microwave pressure cooker! I understand that one can buy this in USA Walmarts ( be interested to know the price of it in the USA). I buy them in UK from Biogenex. Very reliable and consistent.. Started on Coplins jars and , VERY rapidly moved to rice staemers, pressure cookers...then microwave pressure cookers. I look back in no anger. _______________________________________________ Histonet mailing list Histonet@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 ThisisAnn <@t> aol.com Fri Apr 2 18:43:59 2004 From: ThisisAnn <@t> aol.com (ThisisAnn@aol.com) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Open Position/New Jersey Message-ID: <194.26b73051.2d9f62cf@aol.com> Labcorp, Inc., located in Raritan, NJ has an opening for a full-time HT (ASCP). The hours are Mon - Friday, 2nd shift. Responsibilities include: Some Grossing, embedding, microtomy, routine and special staining, coverslipping, etc. Anyone interested, please contact: Ann Angelo, 908-526-2400 x2912 or e-mail me at Ann_Angelo@Labcorp.com From sonyalhogg <@t> yahoo.co.nz Fri Apr 2 23:30:55 2004 From: sonyalhogg <@t> yahoo.co.nz (=?iso-8859-1?q?Sonya=20Hogg?=) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] HT Exam ? Electron Microscopy Message-ID: <20040403053055.67869.qmail@web14912.mail.yahoo.com> Hello:) I am sitting the HT exam in the next exam period and would like to know if electron microscopy is a big part of the exam. Im not too familiar with this. Thanks Sonya Hogg Find local movie times and trailers on Yahoo! Movies. http://au.movies.yahoo.com From romi <@t> stanford.edu Sat Apr 3 12:53:35 2004 From: romi <@t> stanford.edu (Romy Thomas) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Autofluorescence Message-ID: Hi Gayle, Could you please send me your reference for solving autofluorescence problems in bone & elsewhere, GFP & otherwise? Many thanks in advance Romy Thomas Post Doc Stanford University From SCheasty <@t> ahs.llumc.edu Sun Apr 4 15:02:29 2004 From: SCheasty <@t> ahs.llumc.edu (Cheasty, Sandra) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Testing Purity of Recycled Reagents Message-ID: <2E50F33F91EEDA46A77BC3B2575BB09121E057@mars.llumc.edu> How often do you test your recycled reagents, (Histo Clear and Alcohol), for purity? Every run? Once a week? Once a month? Semi annually? We have a hydrometer for testing the alcohol %, but how do you test the recycled Histo Clear? (Or xylene or xylene-substitute) Thanks, Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. From jizv66 <@t> hotmail.com Mon Apr 5 03:47:10 2004 From: jizv66 <@t> hotmail.com (Jorge Ivan Zapata Valencia) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] HELP!!! Message-ID: Hello everybody. Cordial greetings. I have been trying immunostaining for CD markers on mouse tissue samples. I have gotten some really nice slides but............sometimes the edges of the samples turn brown but the center continues completely clear. Even the negative control sometimes is completely clear and the other samples brown edges. I do not know if my problem is that I am allowing the samples to dry up, I am not completely covering samples with the antibody solutions or ............????. May one help me to find what is the problem and how to solve it?? Thank You very much. Regards, JORGE IVAN ZAPATA Universidad del Valle _________________________________________________________________ Add photos to your e-mail with MSN 8. Get 2 months FREE*. http://join.msn.com/?page=features/featuredemail From Nancy.Walker <@t> sanofi-synthelabo.com Mon Apr 5 06:18:11 2004 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] re perfusion debate Message-ID: Hello, Back to an old perfusion debate...does anyone know the relation between the pressure (mm Hg) and pump speed (ml/min) when doing a transcardiac perfusion in mouse, that is, what pump speed will create a pressure of approximately 300 mm Hg? Charles, when you say to "start the prewash at low pressure and pump up to 300 mm Hg" what do you consider to be low pressure? In the past I've done "slow" perfusions at 0.45 ml/min or "fast" perfusions at 2ml/min, I would like to calculate the corresponding pressure. Any physicists out there ...heeeeeeeeeeeeeeeeelp. have a nice day! Nancy Walker Molecular Pharmacology Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 From David.Edmondson <@t> christie-tr.nwest.nhs.uk Mon Apr 5 08:07:52 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Tissue Micro Arrays Message-ID: Hello out there, I am asked to check out Micro Arrays and wonder where the closest practitioners are. Is there anyone near Manchester UK ? We know of Vancouver and Basel but know of no brains closer to pick. Proximity has a bearing on costings, the value of a bequest being, at least in part, a limiting factor and imports would add to costs. Best wishes and thanks Dave Christie Hospital Manchester UK From dmcaloose <@t> wcs.org Mon Apr 5 09:52:47 2004 From: dmcaloose <@t> wcs.org (McAloose, Dee) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] salary survey response Message-ID: <9C26E1A5C9EE5F4B84216095CBE2E24A0778A99B@WOLF.wcs.org> Hello, I was recently interested in the type of information you are looking for and found a 2002 salary survey on that was published in the journal Laboratory Medicine and as a set of PDF files on the ASCP website. First, they answered many of the questions I had regarding salary (broken down in several ways including geographic region). Secondly, they described some issues of concern for the industry (e.g. shortage of histotechnicians/histotechnologists). Finally, it provides a published reference for salaries for both histology lab personnel and supervisors. I pulled the articles off the ASCP website (http://www.ascp.org/bor/center/center_research.asp). You can either use the link or go to the ASCP website - then CAREERS - then WAGE AND VACANCY SURVEY. You can also find the article using the following journal reference information: Ward-Cook, Kory, et.al. 2002 Wage and Vacancy Survey of Medical Laboratories Part I: Salaries Continue to Show Moderate Gains. Laboratory Medicine, Number 9 Volume 34 September 2003, pp 631-638 Ward-Cook, Kory, et.al. 2002 Wage and Vacancy Survey of Medical Laboratories Part II: Modest Easement of Staffing Shortage. Laboratory Medicine, Number 9 Volume 34 September 2003, pp 702-707 Hope this is helpful. D McAloose, VMD, Diplomate ACVP Head, Department of Pathology Wildlife Conservation Society 2300 Southern Blvd Bronx, NY 10460 phone: (718) 220-7105 fax: (718) 220-7126 email: dmcaloose@wcs.org -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Saturday, April 03, 2004 1:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 5, Issue 5 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. Salary Pole (Stacey Burton) 2. Re: (no subject) (Gayle Callis) 3. Histo Exam (Jean Warren) 4. Re: need help with studying for histo exam (Jennifer MacDonald) 5. unsubscribe (Taylor, Jean) 6. Open Position/New Jersey (ThisisAnn@aol.com) 7. HT Exam ? Electron Microscopy (Sonya Hogg) ---------------------------------------------------------------------- Message: 1 Date: Fri, 2 Apr 2004 09:41:24 -0800 (PST) From: Stacey Burton Subject: [Histonet] Salary Pole To: Histonet Message-ID: <20040402174124.21488.qmail@web14525.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Per request of my administrators, I am currently performing a salary pole for Anatomic Pathology Laboratory Managers and Laboratory Coordinators. I have pulled the information from salary.com for my company, however they would like to have a second source to back up the information on salary.com. Would you please email me in person or on the histonet: the Starting, the Average, and Top-out salaries of the Laboratory Managers in your areas. I do not need to know the name of your facilities however the state and city would be helpful so we recognize the difference in cost of living through out the country. Thank you, Stacey Burton, H.T. ASCP Laboratory Manager Uropath LLC San Antonio TX staceylburton@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Small Business $15K Web Design Giveaway - Enter today ------------------------------ Message: 2 Date: Fri, 02 Apr 2004 11:15:35 -0700 From: Gayle Callis Subject: Re: [Histonet] (no subject) To: "Dave McClister" , Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040402111535.00bc07e8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" Molecular Probes at www.probes.com. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 3 Date: Fri, 2 Apr 2004 06:05:43 -0500 From: "Jean Warren" Subject: [Histonet] Histo Exam To: Message-ID: <007e01c418a2$717cf640$4d6ba20a@zoomtown.com> Content-Type: text/plain; charset="iso-8859-1" For the computer portion of the test, do not overlook the special stains for neuropathology. I just took the test in Dec. 2003 and there were 8-10 questions regarding the specialized neuro stains. For instance, the test would depict a drawing of a neuron and an arrow would point to a structure. There would be multiple choice answers to the question, " What stains this structure?" A valuable resource to me was the Frieda Carson study guide that is nothing but questions divided into different categories. I think it was 25$ from the ASCP. It includes a section with color plates. Good luck, the test is thorough, but you will feel good if you pass!! ------------------------------ Message: 4 Date: Fri, 2 Apr 2004 13:35:36 -0800 From: Jennifer MacDonald Subject: Re: [Histonet] need help with studying for histo exam To: "Dukes, Jamie Mr EAMC" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Jamie, Thirteen of my students have taken the exam in the last year. The feedback that I am getting is the questions are not simple recall. Many of the questions are of a troubleshooting nature. Make sure you understand the concept, not memorize the answers. The NSH self-assessment books, the ASCP study guide and Freida Carson's books are great tools to study with. If you have a copy of Sheehan's book it also provides a lot of information. There is a lot of information that you are responsible for. Don't underestimate the time required for studying. Start now. Jennifer MacDonald. -----histonet-bounces@lists.utsouthwestern.edu wrote: ----- To: histonet@lists.utsouthwestern.edu From: "Dukes, Jamie Mr EAMC" Sent by: histonet-bounces@lists.utsouthwestern.edu Date: 04/02/2004 04:52AM Subject: [Histonet] need help with studying for histo exam to all, Need help with studying for histo exam next July. If anyone have any pointers please feel free to point. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Fri, 2 Apr 2004 15:40:19 -0600 From: "Taylor, Jean" Subject: [Histonet] unsubscribe To: "Galbraith, Joe" , "Carl" , Message-ID: <328CBAE62F31C642B422970E879DFADC0F7C06@pcwex01.meriter.com> Content-Type: text/plain; charset="iso-8859-1" -----Original Message----- From: Galbraith, Joe [mailto:joseph-galbraith@uiowa.edu] Sent: Friday, April 02, 2004 10:07 To: Carl; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] re mirowave Ag retrieval Carl: We get our microwave pressure cooker from NordicWare (800-328-4310 ext 629) but I have seen these in kitchen stores as well (I'll have to check Walmart). Direct from the company, they cost $30.00 US with an extra seal costing $3.00. Of course, shipping is extra. If you take care of them, the cookers last a long time (several years of once daily use) before needing to be replaced. I found a site from NordicWare also but it costs more online: http://www.nordicware.com/b2c/product_category.cfm?prod_master_cat=2&prod_cat=7 Might try Amazon.com or Everthinghome.com too. Have fun shopping. Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Carl Sent: Thursday, April 01, 2004 1:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re mirowave Ag retrieval Well...I use a microwave pressure cooker! I understand that one can buy this in USA Walmarts ( be interested to know the price of it in the USA). I buy them in UK from Biogenex. Very reliable and consistent.. Started on Coplins jars and , VERY rapidly moved to rice staemers, pressure cookers...then microwave pressure cookers. I look back in no anger. _______________________________________________ Histonet mailing list Histonet@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: 6 Date: Fri, 2 Apr 2004 19:43:59 EST From: ThisisAnn@aol.com Subject: [Histonet] Open Position/New Jersey To: histonet@lists.utsouthwestern.edu Message-ID: <194.26b73051.2d9f62cf@aol.com> Content-Type: text/plain; charset="US-ASCII" Labcorp, Inc., located in Raritan, NJ has an opening for a full-time HT (ASCP). The hours are Mon - Friday, 2nd shift. Responsibilities include: Some Grossing, embedding, microtomy, routine and special staining, coverslipping, etc. Anyone interested, please contact: Ann Angelo, 908-526-2400 x2912 or e-mail me at Ann_Angelo@Labcorp.com ------------------------------ Message: 7 Date: Sat, 3 Apr 2004 17:30:55 +1200 (NZST) From: Sonya Hogg Subject: [Histonet] HT Exam ? Electron Microscopy To: Histonet Message-ID: <20040403053055.67869.qmail@web14912.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello:) I am sitting the HT exam in the next exam period and would like to know if electron microscopy is a big part of the exam. Im not too familiar with this. Thanks Sonya Hogg Find local movie times and trailers on Yahoo! Movies. http://au.movies.yahoo.com ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 5, Issue 5 ************************************** From NSEARCY <@t> swmail.sw.org Mon Apr 5 11:56:37 2004 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Reference Work Message-ID: <04Apr5.115706cdt.119048@healthcare2.sw.org> Any reference lab perform Beta-2-Microglobulin on paraffin? Cost? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD From thoward <@t> unm.edu Mon Apr 5 12:56:45 2004 From: thoward <@t> unm.edu (Tamara Howard) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] RE: perfusion debate Message-ID: As far as the relationship of mm Hg to ml/min with a perfusion pump - aren't the innner diameter and stiffness of the tubing (and the i.d. of the cannula) going to influence the final pressure, too? I don't think adjusting final pump speed alone is going to accurately duplicate the "hanging bag" pressure. Can you check the literature and see what other groups have reported and work from there? |--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward@unm.edu |--------------------------------------------------| From cwscouten <@t> myneurolab.com Mon Apr 5 13:24:14 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] RE: perfusion debate Message-ID: It depends on where the bottleneck is. If the cardiovascular system is the main source of resistance, it will feel all 300 mm Hg. If the tubing or needle is small, it will absorb the pressure drop, and the cardiovasucalar system will get much less than the gauge pressure the perfusion bottle was pumped up to. A mouse has much more cardiovascular resistance than a rat. A pig would have much much less. The largest suitable needle and tubing (1/4" ID tubing), would insure that the animal was the main source of resistance if the flow was small (mouse or even rat), but larger animals would require larger tubing and needle. Remember, the hanging bag pressure needs to be about 13 feet above the animal to provide a pressure that will disrupt the blood brain barrier. The correct flow rate to use depends greatly on animal size and condition. The correct pressure is constant, but needs to be higher than traditional gravity flow can typically provide. So it is better to control pressure directly, rather than flow rate. For an inexpensive means of implementing this strategy, and getting no shrink perfusions, go to www.myneurolab.com, click products, click sacrifice instruments under the Histology header, click Perfusion One. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 FAX 314 522 0377 cwscouten@myneurolab.com www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tamara Howard Sent: Monday, April 05, 2004 12:57 PM To: HistoNet Subject: [Histonet] RE: perfusion debate As far as the relationship of mm Hg to ml/min with a perfusion pump - aren't the innner diameter and stiffness of the tubing (and the i.d. of the cannula) going to influence the final pressure, too? I don't think adjusting final pump speed alone is going to accurately duplicate the "hanging bag" pressure. Can you check the literature and see what other groups have reported and work from there? |--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward@unm.edu |--------------------------------------------------| _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gradice <@t> richmond.edu Mon Apr 5 13:10:39 2004 From: gradice <@t> richmond.edu (Gary Radice) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Microscopist position available, U of Richmond Message-ID: <8B456638-872C-11D8-B0A1-000393BC23F4@richmond.edu> Hello everyone, Please forward if you know someone who might be interested. Gary Radice Director of Microscopy and Imaging University of Richmond The Department of Biology at this highly selective, private, primarily undergraduate university invites applications for a Director of Microscopy and Imaging. Duties including supervising operation of a new biological imaging facility, including operation and routine maintenance of a TEM, SEM, and laser scanning confocal microscope and associated equipment. Teaching responsibilities include training and supervising faculty and student users in research and classroom activities and courses based on interests and department needs; assisting faculty and students in experimental design, specimen preparation, digital photography, and analysis of results. Qualifications: MS or PhD degree and experience with light and electron microscopy and digital imaging, strong interpersonal skills and desire and ability to work well in a team environment. This is a continuing appointment that is not eligible for tenure but may include faculty status. Applications including a letter of interest, CV, evidence of expertise in microscopy, and three letters of recommendation should be sent to Dr. Gary Radice, Department of Biology, University of Richmond, Richmond VA 23173 (gradice@richmond.edu). Review of applications will begin on April 23, 2004. The appointment is expected to begin June 1, 2004. The University of Richmond is committed to increasing the diversity of our faculty and strongly encourages applications from women and minorities. For more information on the department, resources, and teaching assignment, see (http://biology.richmond.edu/) Gary P. Radice gradice@richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond 804-289-8233 (FAX) Richmond VA 23173 http://www.richmond.edu/~gradice From neuroant <@t> hotmail.com Mon Apr 5 13:28:13 2004 From: neuroant <@t> hotmail.com (Ant Swensson) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] (no subject) Message-ID: Hi. I'm interested in taking the HT exam at the end of this year. I've been studying by myself, and working in a lab to get hands on experience. I am curious, if I need to in the future, where the schools that train for the exam are located in the USA? Thank you, Antoinette Swensson Univeristy of Washington/Harborview Medical Center Neuropathology 325 9th Ave MS 359-791 Seattle, WA 98104 (206) 731-3910 _________________________________________________________________ [1]Limited-time offer: Fast, reliable MSN 9 Dial-up Internet access FREE for 2 months! References 1. http://g.msn.com/8HMBENUS/2734??PS= From michael_lafriniere <@t> memorial.org Mon Apr 5 12:29:06 2004 From: michael_lafriniere <@t> memorial.org (LaFriniere, Mike) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Region III Meeting Message-ID: The Region III Meeting is finalizing plans for their annual meeting this year in Birmingham, Alabama. May 20th-24 The program can be found on the Alabama Society for Histotechnology (ASH) web page. The web site can be found at www.timjday.com and click on Region III, registration is welcome through the website. The hotel accommodations are at the beautiful Wynfrey attached to the Galleria Mall in Birmingham. We ask that all participants register at the Wynfrey to keep our meeting cost to a minimum! The negotiated rate for the meeting is $125.00 per room, which normally is $199.00. This is an upscale 4 star hotel! Please make your reservations at the Wynfrey as soon as possible to obtain this rate as there is less than 20 days to obtain the negotiated $125.00 per night rate! We are demonstrating an excellent program to include more than 20 vendors and 14 excellent workshops offered from speakers within our Region and expect over 100 attendees, this will hopefully help diminish our debt from last years meeting in Puerto Rico. Your attendance at the Wynfrey 1800-476-7006 is urgently requested to support Region III. Please contact the hotel for reservations and state you are with the Region III, Alabama Histology meeting! Programs have been mailed out last week to all of Region III membership as well as several State memberships! Vendors are encouraged to also use the Wynfrey to assist with our overall cost, in addition, there are a few spots left for Vendors who have not yet registered to be at the meeting and can contact me directly for additional information. Michael LaFriniere NSH Region III Director 423-495-6117 From ylee <@t> bccancer.bc.ca Mon Apr 5 18:35:44 2004 From: ylee <@t> bccancer.bc.ca (Yin Ping Lee) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Adhesive for paraffin sections Message-ID: <6BAF4D075F07D411B30900508B94CBA0C1F55F@SERVER20> Hi, Leslie, "Mount Quick Tissue Transfer Technique" is another alternative for your case. This technique allows for the transfer of tissue sections from one slide to another slide. (Therefore, the sections can be transferred to charged slides and immuno-staining can be performed as per its usual protocol.) This transfer technique is very simple to do; all you need is to purchase the "Mount Quick" medium and follow the protocol. This medium is distributed by Newcomer Supply. Tel: (608) 831-7888. Fax: (608)831-0866. Yin Ping Lee Histopathology Lab BC Cancer Agency Vancouver BC From weneng <@t> hotmail.com Mon Apr 5 18:41:06 2004 From: weneng <@t> hotmail.com (Wendy England) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Analytical Imaging Station Version 6.0 Message-ID: Hello, I was asked to search info about above systems. But I couldn't find any through my computer. Could anybody provide me the website I can look? Any help will be highly appreciated! Wendy _________________________________________________________________ MSN Toolbar provides one-click access to Hotmail from any Web page – FREE download! http://toolbar.msn.com/go/onm00200413ave/direct/01/ From michael_lafriniere <@t> memorial.org Mon Apr 5 17:26:11 2004 From: michael_lafriniere <@t> memorial.org (LaFriniere, Mike) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] RE: Region III Meeting Message-ID: The Region III Meeting is finalizing plans for their annual meeting this year in Birmingham, Alabama. May 20th-24 The program can be found on the Alabama Society for Histotechnology (ASH) web page. The web site can be found at www.timjday.com and click on Region III, registration is welcome through the website. The hotel accommodations are at the beautiful Wynfrey attached to the Galleria Mall in Birmingham. We ask that all participants register at the Wynfrey to keep our meeting cost to a minimum! The negotiated rate for the meeting is $125.00 per room, which normally is $199.00. This is an upscale 4 star hotel! Please make your reservations at the Wynfrey as soon as possible to obtain this rate as there is less than 20 days to obtain the negotiated $125.00 per night rate! We are demonstrating an excellent program to include more than 20 vendors and 14 excellent workshops offered from speakers within our Region and expect over 100 attendees, this will hopefully help diminish our debt from last years meeting in Puerto Rico. Your attendance at the Wynfrey 1800-476-7006 is urgently requested to support Region III. Please contact the hotel for reservations and state you are with the Region III, Alabama Histology meeting! Programs have been mailed out last week to all of Region III membership as well as several State memberships! Vendors are encouraged to also use the Wynfrey to assist with our overall cost, in addition, there are a few spots left for Vendors who have not yet registered to be at the meeting and can contact me directly for additional information. Michael LaFriniere NSH Region III Director 423-495-6117 From lpwenk <@t> covad.net Tue Apr 6 04:07:46 2004 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] schools References: Message-ID: <002b01c41bb6$a103dfc0$b834d445@domainnotset.invalid> The accrediting agency for lab schools is the National Accrediting Agency for Clinical Laboratory Schools (NAACLS). To find lab programs, including HT and HTL, go to http://www.naacls.org On the left side, click on "find a program" Then, enter the type of program (HT or HTL, for exam), and the state (or search for all states). (There are 27 HT and 2 HTL = 29 accredited programs in the US. Up from about 21 a couple of years ago. Still could use a lot more, in my opinion.) Peggy A. Wenk, HTL(ASCP)SLS Program Director School of Histotechnologists School of Histologic Technicians William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Ant Swensson" To: Sent: Monday, April 05, 2004 2:28 PM Subject: [Histonet] (no subject) > > Hi. I'm interested in taking the HT exam at the end of this year. > I've been studying by myself, and working in a lab to get hands on > experience. I am curious, if I need to in the future, where the > schools that train for the exam are located in the USA? > > Thank you, > > Antoinette Swensson > > Univeristy of Washington/Harborview Medical Center > Neuropathology > 325 9th Ave MS 359-791 > Seattle, WA 98104 > (206) 731-3910 > _________________________________________________________________ > > [1]Limited-time offer: Fast, reliable MSN 9 Dial-up Internet access > FREE for 2 months! > > References > > 1. http://g.msn.com/8HMBENUS/2734??PS= > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ssq5977 <@t> yahoo.com.cn Tue Apr 6 07:22:03 2004 From: ssq5977 <@t> yahoo.com.cn (=?gb2312?q?=CA=E7=C7=ED=20=C9=F2?=) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] a problem about immunostaining for immunoglobulin heavy chains Message-ID: <20040406122203.67972.qmail@web15207.mail.bjs.yahoo.com> Hello,everybody, I am intrested in immunostaining for immunoglobulin heavy chains on renal biopsy specimen ,but I have no idea about which antibodies are appropriate.Would anybody be kind enough to share me with you experience? Thank You very much. Best regards! Shuqiong Shen Jinling Hospital Nanjing 210002 P.R.China --------------------------------- Do You Yahoo!? »ÝÆÕTTÓÎÏ·¾ç£¬ÍæÓÎÏ·£¬Öд󽱣¡ From JEllin <@t> yumaregional.org Tue Apr 6 07:36:11 2004 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] a lot of stuff HELP OUT THERE!! Message-ID: Hello everyone thought that I would ask for some help here,, Currently our lab is getting a hugh over haul on everthing and I need to find out some information well were do I begin, First off our lab went live with Tamtron this past year and it has been different to say the least, but one of the ideas we are pondering is using the image module to caputre gross instead of gross dictation, only on small bx that do not require anything other than transfering it to the cassette and snapping a picture. Was wondering if anyone out there is doing this and also if they do have Tamtron how do they like the imaging module. The next item we are looking at is this processor that can process tissue in 1 hr and is any one out there doing this? This is a huge step, since we are part of the traditional way of processing over night and doing the embedding the next day. SO if anyone is attempting this please feel free to give me input. The last thing that our lab is going to under take is allow tech that are not certified and do not have 60 hrs of college to do gross,the standard is set forth in NACCLS that a person must have a minimal amount of education to do this. But our pathologist called the CAP and said that they can do transfer of specimens that can give color, size, dimension. This is were the idea of photographing specimens came amout. ANY help would greatly be appreciated. Jesus Ellin HTL Yuma Regional Medical Center From mcauliff <@t> umdnj.edu Tue Apr 6 12:09:23 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Analytical Imaging Station Version 6.0 In-Reply-To: References: Message-ID: <4072E443.1090405@umdnj.edu> Ask the manufacturer of the system. If you are looking to purchase a new image analysis system Nikon, Zeiss, Olympus etc all make such systems. In addition, there are many vendors who will build a system to meet your current and future needs. Geoff Wendy England wrote: > Hello, > I was asked to search info about above systems. But I couldn't find > any through my computer. Could anybody provide me the website I can look? > > Any help will be highly appreciated! > > Wendy > > _________________________________________________________________ > MSN Toolbar provides one-click access to Hotmail from any Web page ? > FREE download! http://toolbar.msn.com/go/onm00200413ave/direct/01/ > > > _______________________________________________ > 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 MElliott <@t> mrl.ubc.ca Tue Apr 6 09:30:40 2004 From: MElliott <@t> mrl.ubc.ca (Mark Elliott) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Human myofibroblast markers Message-ID: Was wondering what everyone is using for labelling myofibroblasts in human tissue? Preferably something that works in FFPE tissues, but frozen is OK. Thanks Mark W. Mark Elliott, PhD Research Associate, Interim Director, Histology Core Facility James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research Room 166, St. Paul's Hospital 1081 Burrard Street Vancouver, BC Canada V6Z 1Y6 From rbremer <@t> acpub.duke.edu Tue Apr 6 09:34:18 2004 From: rbremer <@t> acpub.duke.edu (Ronald E. Bremer) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] (no subject) Message-ID: <4072BFEA.3010808@acpub.duke.edu> We are intertested in purchasing a used microtome knife sharpener. I was hoping for some advice. I've noted a few American Optical models for sale for what seems a rather cheap price. I think Leica picked them up or is the same company. Are these shapreners difficult to get parts for? I'm also interested in comments on "Hacker knife sharpeners" Even new they seem rather inexpensive compared to other models. We could go new if the price was right and have GSA money if anybody could point me in the right direction. Unfortunately the right price is less than $1000, so I am not hopeful on a new model. Ron From S.Stryker <@t> lumc.nl Tue Apr 6 10:04:47 2004 From: S.Stryker <@t> lumc.nl (Stryker, S. (HLK)) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] porcine insulin Message-ID: Hello everybody, I'm hoping someone can give me any suggestions on how to detect insulin on a (paraffin embedded formalin-fixed or frozen) porcine section using the following antibodies: - primary antibody: Guinea Pig anti-insulin (Dako) - secondary antibody: Swine anti-Rabbit (Dako) According to the DAKO catalogous the anti-Rabbit should work in this combo. I have tried it over and over again, using different protocols, but somehow I just can't get the job done... Thanks in advance! Samantha s.stryker@lumc.nl Leiden University Medical Centre From siksik03 <@t> comcast.net Tue Apr 6 10:13:41 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Sponge problems in processing In-Reply-To: <3.0.6.32.20040401102003.00bc0a88@gemini.msu.montana.edu> References: <3.0.6.32.20040401102003.00bc0a88@gemini.msu.montana.edu> Message-ID: Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. From Vickroy.Jim <@t> mhsil.com Tue Apr 6 09:57:30 2004 From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Ventana Tissue Processor Message-ID: We had a processing run today that looks like the tissue was cooked especially the small GI specimens. We have a Ventana Renissance Tissue Processor, which they no long sell. Has anyone every had a problem similar to this, and is there any correlation to problems with the tissue processor? My first thoughts are that maybe the chemistry on the machine was switched, but I can't find any glaring problems. Thanks for your help James R. Vickroy BS, HT (ASCP) Technical Supervisor, Surgical Pathology 788-4046 vickroy.jim@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. From siksik03 <@t> comcast.net Tue Apr 6 10:19:33 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] microwave antigen retrieval In-Reply-To: <3.0.6.32.20040401102302.00bc0a88@gemini.msu.montana.edu> References: <3.0.6.32.20040401102302.00bc0a88@gemini.msu.montana.edu> Message-ID: Hi HistoNetters Successful antigen retrieval depends on accurate control of three factors: time, temperature and pH. Any instrument which will produce this accuracy in a reproducible way will be effective. A microwave has been shown to work well over a wide range of antigens, where other methods have required more aggressive treatment. In some cases, pressure is needed to allow for temperatures above 98?C. I can get most antigens to "work" with pH 6.0 citrate buffer at 110?C in a 3-5 minute microwave method using the Hacker/Milestone microwave. best regards, Steven Slap Microwave Consultant From Myri37 <@t> aol.com Tue Apr 6 10:18:31 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] solochrome stain Message-ID: <64E5A68B.38CAA850.0005167B@aol.com> hello i have prepared solochrome stain following this: 1 g of calcon (solochrome) 2 ml acetic acid glacial 98 ml distilled water and let it for 3 months before use i tried this stain on MMA sections of thin reconstructed and mineralized tissue, i put section of 50um in this stain for 1 hour, i didn't see anything stained on hot plate or not could any one help me ? is the stain not good ? or something else ? thank you very much for your help Myriam From Terry.Marshall <@t> rothgen.nhs.uk Tue Apr 6 10:20:47 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Sponge problems in processing Message-ID: OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From siksik03 <@t> comcast.net Tue Apr 6 10:34:30 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] re mirowave Ag retrieval In-Reply-To: <000b01c41820$a5312ce0$fcae8451@home> References: <000b01c41820$a5312ce0$fcae8451@home> Message-ID: Hi HistoNetters The Nordicware microwave pressure cooker is available at a deep discount from the Yahoo store: Nordicware Tender Cooker/Microwave Pressure Cooker 62104 shop.store.yahoo.com/dotcoms/tecoprcobyno.html However, this device does not allow for temperature control. Let the buyer beware... best regards, Steven Slap At 8:36 PM +0100 4/1/04, Carl wrote: > >Well...I use a microwave pressure cooker! I understand that one can >buy this in USA Walmarts ( be interested to know the price of it in >the USA). I buy them in UK from Biogenex. Very reliable and >consistent.. Started on Coplins jars and , VERY rapidly moved to >rice staemers, pressure cookers...then microwave pressure cookers. I >look back in no anger. From marytedo <@t> hotmail.com Tue Apr 6 11:44:57 2004 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] I need help! Message-ID: Hi Histonetters! In our lab we have received Methilyc Alcohol which we use in the tissues prossessor, but this time we don't know the Quality of it due to the quantity we have received doesn't have any label with any descripcion about it's features. I wonder if there any quality Control about Methilyc Alcohol. What I need to know if this Alcohol is 100% methilyc. I'll appreciate all you can advice me. H.T. Maria Teresa Dominguez Rio Grande, Zonal Hospital, Anatomic Pathology, Tierra del Fuego, Argentina. _________________________________________________________________ Help STOP spam with [1]the new MSN 8 and get 2 months FREE* References 1. http://g.msn.com/8HMBEN/2731??PS= From jcline <@t> wchsys.org Tue Apr 6 11:57:15 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] Ronald Bremer/knife sharpener Message-ID: <002d01c41bf8$3a14d660$1d2a14ac@wchsys.org> I have a Hacker knife sharpener for sale, it works perfectly and I even have the video that shows how to work it. I have used glass sharpeners in the past and the Hacker beats any sharpener for doing the steel knives. We are using disposables now so I have no need for the Hacker. You can email me directly if you would like. ***** 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 mckeela <@t> ucmail.uc.edu Tue Apr 6 11:56:25 2004 From: mckeela <@t> ucmail.uc.edu (Lynn McKee) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] BK and JC by IHC Message-ID: <200404061650.AWW63950@mprelay.uc.edu> Hi everyone, Is anyone doing IHC for BK and JC virus? We do INSITU but our Dr. wants to do IHC now. I found CHEMICON carries the BK but I have had no luck with JC. Thanks in advance for your help. Lynn McKee University of Cininnati From John.Garlits <@t> STJUDE.ORG Tue Apr 6 12:16:58 2004 From: John.Garlits <@t> STJUDE.ORG (Garlits, John) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] looking for a good antibody to PTH receptor Message-ID: <1E0CC447E59C974CA5C7160D2A2854EC238AD8@SJMEMXMB04.stjude.sjcrh.local> Anybody know a good one? In my search so far I seem to be finding the same antibody from Neomarkers, Upstate, and Biomeda, which is a mouse IgG clone 3D1.1. They say it works for IHC. Has anyone tried it, or a different one? I would like to find one for flow cytometry as well. John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 Phone: 901-495-3961 Fax: 901-495-2176 From John.Garlits <@t> STJUDE.ORG Tue Apr 6 12:21:42 2004 From: John.Garlits <@t> STJUDE.ORG (Garlits, John) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] baking bone slides for heat retrieval techniques? Message-ID: <1E0CC447E59C974CA5C7160D2A2854EC238AD9@SJMEMXMB04.stjude.sjcrh.local> I have tried repeatedly to use boiling or heated antigen retrieval techniques on my bone slides, but the bone always peels off. (The marrow stays.) I did hear somewhere that in this case one can heat the slides at around 55 degrees C overnight, which makes the bone adhere better to potentially survive boiling antigen retrieval. Has anyone tried this or know if this works? Thanks! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 Phone: 901-495-3961 Fax: 901-495-2176 From nmuvarak <@t> facstaff.wisc.edu Tue Apr 6 11:55:37 2004 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL E MUVARAK) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] TTC (Triphenyl Tetrazolium Chloride) Staining Message-ID: <389078384e00.384e00389078@wiscmail.wisc.edu> Hello everyone. I'm going to use TTC on perfused-ventilated mice lungs to detect any tissue damage. Has anyone used this dye before on mice, and can you share your protocol or tips on the stain? Oh, and who's a good supplier for TTC? Thanks much. Nidal E Muvarak Associate Research Specialist Vascular Tissue Biomechanics Laboratory Department of Biomedical Engineering University of Wisconsin-Madison 1550 Engineering Dr.; Rm. 2158 Madison, WI 53706-1609 Lab: (608) 265-8921; Office: (608) 265-4205; Home: (608) 256-7934; Cell: (608) 332-6068 http://vtb.bme.wisc.edu From pruegg <@t> colobio.com Tue Apr 6 13:19:05 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] I need help! In-Reply-To: Message-ID: Why don't you ask who ever sold it to you? Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mar?a Teresa Dom?nguez Sent: Tuesday, April 06, 2004 10:45 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] I need help! Hi Histonetters! In our lab we have received Methilyc Alcohol which we use in the tissues prossessor, but this time we don't know the Quality of it due to the quantity we have received doesn't have any label with any descripcion about it's features. I wonder if there any quality Control about Methilyc Alcohol. What I need to know if this Alcohol is 100% methilyc. I'll appreciate all you can advice me. H.T. Maria Teresa Dominguez Rio Grande, Zonal Hospital, Anatomic Pathology, Tierra del Fuego, Argentina. _________________________________________________________________ Help STOP spam with [1]the new MSN 8 and get 2 months FREE* References 1. http://g.msn.com/8HMBEN/2731??PS= _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Tue Apr 6 13:30:06 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] baking bone slides for heat retrieval techniques? In-Reply-To: <1E0CC447E59C974CA5C7160D2A2854EC238AD9@SJMEMXMB04.stjude.sjcrh.local> Message-ID: There have been many shared methods for adhering bone sections to slides and I have probably used them all at one time or another. In my experience it is best to avoid HIER on bone for IHC although I am sure many people do it successfully. I use enzyme digestion instead of heat retriveal for bone IHC (either Proteinase K or Pepsin in most cases). In my hands EIER is more easily controlled and less harsh on tissues than HIER. If I had to use HIER on bone I would choose a more gentle method such as steam rather than boiling. My one cent worth. Patsy Ruegg -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Garlits, John Sent: Tuesday, April 06, 2004 11:22 AM To: HistoNet Server Subject: [Histonet] baking bone slides for heat retrieval techniques? I have tried repeatedly to use boiling or heated antigen retrieval techniques on my bone slides, but the bone always peels off. (The marrow stays.) I did hear somewhere that in this case one can heat the slides at around 55 degrees C overnight, which makes the bone adhere better to potentially survive boiling antigen retrieval. Has anyone tried this or know if this works? Thanks! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 Phone: 901-495-3961 Fax: 901-495-2176 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Tue Apr 6 13:22:47 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] beta-gal on mouse tibia Message-ID: Has anyone done this on bone? Gayle was very kind to provide me with a procedure for soft tissues but I am just wondering how decalcification would effect beta-gal staining. Perhaps the samples should be stained and then processed into GMA resin without decal? Would the GMA have an adverse effect on the beta-gal whole mount staining????? Thanks for your advise. Patsy From janis-rodgers <@t> uiowa.edu Tue Apr 6 13:30:55 2004 From: janis-rodgers <@t> uiowa.edu (Rodgers, Janis) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] tinctorial staining & immuno Message-ID: <5D03ED7B9391D4119D9B0008C76B7B24062B0F8D@uihc-mail1.uihc.uiowa.edu> Hello, has anyone tried doing an immuno stain in conjunction with tinctorial stains on the same tissue slide? Thanks, Jan Rodgers From kmerriam2003 <@t> yahoo.com Tue Apr 6 13:59:11 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] porcine insulin In-Reply-To: Message-ID: <20040406185911.34902.qmail@web10003.mail.yahoo.com> I would use a biotinylated anti-guinea pig antibody. I'm not sure if the anti-rabbit antibody would bind the the GP primary antibody you are using. I'm pretty sure that Vector sells an anti-GP secondary antibody (I think they also sell a guinea pig ABC and alk phos kit, depending on the detection system you are using). Good luck, Kim Merriam' Novartis Cambridge, MA "Stryker, S. (HLK)" wrote: Hello everybody, I'm hoping someone can give me any suggestions on how to detect insulin on a (paraffin embedded formalin-fixed or frozen) porcine section using the following antibodies: - primary antibody: Guinea Pig anti-insulin (Dako) - secondary antibody: Swine anti-Rabbit (Dako) According to the DAKO catalogous the anti-Rabbit should work in this combo. I have tried it over and over again, using different protocols, but somehow I just can't get the job done... Thanks in advance! Samantha s.stryker@lumc.nl Leiden University Medical Centre _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Small Business $15K Web Design Giveaway - Enter today From amarusk1 <@t> FAIRVIEW.ORG Tue Apr 6 14:54:32 2004 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:22:45 2005 Subject: [Histonet] RUO antibodies Message-ID: Hi histonetters, For those who use RUO antibodies in a clinical setting, how do you work up these products? Is your documentation any different than when you use ASR or IVD products? Thanks for your input. Ann Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including “protected health information.” If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:ANN MARUSKA N:MARUSKA;ANN TEL;WORK:612-273-9119 ORG:;LAB EMAIL;WORK;PREF;NGW:AMARUSK1@FAIRVIEW.ORG END:VCARD From RossS <@t> BaylorHealth.edu Tue Apr 6 14:38:01 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] BK and JC by IHC Message-ID: We do BK and just started to do JC. We get JC from Oncogene(sp) 1-858-450-5558, cat# DP02 200ug. We just got it to work using Dako HighPH TR, 1:100 overnight. Then used a Ventana Detection kit to finish it off on the machine after the overnight incubation. The slides are still not perfect yet so you will definitely have to tweek this, but it gives you someplace to start. We have been doing BK for a little while. Let me know if you have any questions about that one and I'll find out. Our immuno techs here do a great job. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lynn McKee Sent: Tuesday, April 06, 2004 11:56 AM To: histonet@pathology.swmed.edu Subject: [Histonet] BK and JC by IHC Hi everyone, Is anyone doing IHC for BK and JC virus? We do INSITU but our Dr. wants to do IHC now. I found CHEMICON carries the BK but I have had no luck with JC. Thanks in advance for your help. Lynn McKee University of Cininnati _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From tpmorken <@t> labvision.com Tue Apr 6 15:24:52 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] RUO antibodies Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA21774AD@usca0082k08.labvision.apogent.com> Ann, the FDA invented the ASR designation to get around the RUO dilemmma (how to justify using a reagent marked "research use only" for diagnostics). The FDA doesn't want diagnostic labs using antibodies labeled as RUO's. The solution is to ask your vendor to supply an ASR datasheet. It will be the same antibody, just marketed differently, and will satisfy an FDA inspector. Whatevery you do, it should be worked up as an ASR - ie, you need to do your own validation studies. Tim Morken Lab Vision / Neomarkers www.labvision.com -----Original Message----- From: ANN MARUSKA [mailto:amarusk1@FAIRVIEW.ORG] Sent: Tuesday, April 06, 2004 12:55 PM To: histonet@pathology.swmed.EDU Subject: [Histonet] RUO antibodies Hi histonetters, For those who use RUO antibodies in a clinical setting, how do you work up these products? Is your documentation any different than when you use ASR or IVD products? Thanks for your input. Ann Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including "protected health information." If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> From shive003 <@t> umn.edu Tue Apr 6 15:43:21 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] porcine insulin References: <20040406185911.34902.qmail@web10003.mail.yahoo.com> Message-ID: <004b01c41c17$cca4e280$78065486@vdl220FAC> Yes, a biotinylated anti-rabbit IgG does cross-react with guinea pig antibodies. I use it all the time when I stain for Insulin, using Dako's guinea pig anti-Insulin product. Jan Shivers U of MN Vet Diag Lab ----- Original Message ----- From: "Kim Merriam" To: "Stryker, S. (HLK)" ; Sent: Tuesday, April 06, 2004 1:59 PM Subject: Re: [Histonet] porcine insulin > I would use a biotinylated anti-guinea pig antibody. I'm not sure if the anti-rabbit antibody would bind the the GP primary antibody you are using. I'm pretty sure that Vector sells an anti-GP secondary antibody (I think they also sell a guinea pig ABC and alk phos kit, depending on the detection system you are using). > > Good luck, > > Kim Merriam' > Novartis > Cambridge, MA > > "Stryker, S. (HLK)" wrote: > Hello everybody, > > I'm hoping someone can give me any suggestions on how to detect insulin on a > (paraffin embedded formalin-fixed or frozen) porcine section using the > following antibodies: > > - primary antibody: Guinea Pig anti-insulin (Dako) > - secondary antibody: Swine anti-Rabbit (Dako) > > According to the DAKO catalogous the anti-Rabbit should work in this combo. > I have tried it over and over again, using different protocols, but somehow > I just can't get the job done... > > Thanks in advance! > > Samantha > s.stryker@lumc.nl > Leiden University Medical Centre > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > --------------------------------- > Do you Yahoo!? > Yahoo! Small Business $15K Web Design Giveaway - Enter today > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From michael_lafriniere <@t> memorial.org Tue Apr 6 12:38:20 2004 From: michael_lafriniere <@t> memorial.org (LaFriniere, Mike) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Ventana processors Message-ID: Does anyone know anybody that performs PM on Ventana's processors, I am seeking private vendors. Thanks, Michael LaFriniere Pathology Manager Memorial Hospital 423-495-6117 From peptolab <@t> hamptons.com Tue Apr 6 16:43:53 2004 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Myofbroblast markers Message-ID: <003401c41c20$42578660$6e7dbd18@JEFF> Mark- I like alpha smooth muscle actin- no antigen retrieval necessary in FFPE tissues. Cross reactivity with smooth muscle should not be a problem in most systems or tissues other than gut. Some use desmin but I think actin stains more varieties/functional states of myofibroblasts than desmin. I think there is tremendous heterogeneity among myofibroblasts in different tissue- I concentrate on those in connective tissue. Additionally I believe that the ubiquitous interstitial CD34+ fibroblast-like cell found in most collagenous connective tissue and fat are myofibroblast precursers- acting as reserve cells for tissue repair and remodelling. In my surgical pathology work- some (mesenchymal) myofibroblastic tumors of skin, breast and soft tissue have maintained CD34 expression as well as actin, a neoplastic myofibroblast phenotype, though in healing wounds and other reactive lesions or even in malignant epithelial tumor stroma - indigenous fibroblasts almost always lose CD34 expression when they transform (if you believe they do) to actin+ myofibroblasts. Just some food for thought Jeff Silverman HT HTL QIHC (ASCP) Pathologists' Assistant Southside Hospital Bay Shore New York From bhewlett <@t> cogeco.ca Tue Apr 6 18:31:29 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] tinctorial staining & immuno References: <5D03ED7B9391D4119D9B0008C76B7B24062B0F8D@uihc-mail1.uihc.uiowa.edu> Message-ID: <006101c41c2f$4a3870b0$6500a8c0@mainbox> Jan, Yes! depends on the immunostain and the tinctorial stain for exact combination. What is it you are trying to demonstrate? Bryan ----- Original Message ----- From: "Rodgers, Janis" To: Sent: Tuesday, April 06, 2004 2:30 PM Subject: [Histonet] tinctorial staining & immuno Hello, has anyone tried doing an immuno stain in conjunction with tinctorial stains on the same tissue slide? Thanks, Jan Rodgers _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KAELPERS <@t> aol.com Tue Apr 6 21:20:18 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing Message-ID: <1a6.226884c3.2da4bf62@aol.com> We use these tiny screen cassettes that fit into processing cassettes, they can be purchased from Mercedes medical I think. They work really well, and another thing neat about them is they have 2 dimensions - the capsule closes 2 different directions and one side is slightly higher than the other to allow slightly bigger biopsies. lge From KAELPERS <@t> aol.com Tue Apr 6 21:27:18 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] HT Exam ? Electron Microscopy Message-ID: <1ca.1de31d13.2da4c106@aol.com> My understanding is that there is not much EM on the "HT" exam. I took mine in 2000 and I do not recall if any were on there. I would just have a basic understanding if what it is and its purpose and of course know the fixatives that one would need to use for EM that would certainly be something you would potentially run into. lge From KAELPERS <@t> aol.com Tue Apr 6 21:13:57 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Ventana Tissue Processor Message-ID: <1e4.1cf5d0c0.2da4bde5@aol.com> I have had this problem before on two different type of processors. I have researched and researched what could have happened....chemicals were placed appropriately, heat is applied only on the paraffin stations, processing times are standard...the only thing that I have came up with is that if you are processing very small delicate tissue directly after a complete change on the processor, the tissue is actually getting sucked dry by dehydration. Look at your maintenance records again and see if the processor had just been completely changed out. I just recently attended a seminar "The chemistry of processing and staining" and the lecturer referred to it as removal of the bonded water ...the bonded water is located at the cellular level (within the cell) and you do not want to remove this. If the tissue is becoming over dehydrated it is removing this bonded water as well as the free water and is causing the tissue to shrivel up and in essence probably getting cooked. I have resorted to using sponges with tiny fragile tissue and doing partial changes if I know that I will be processing delicate tissue and so far this has worked out. Another helpful hint in the event you would be cutting these tissue blocks is to soak the tissue blocks after faced in a cold glycerin solution...it will help tremendously with sectioning...although microscopically the outer edges will look "cooked". Hope this helps. lge From KAELPERS <@t> aol.com Tue Apr 6 21:39:20 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] A wonderful article on bone samples and a SAFE bone saw! Message-ID: <1d0.1ddc759c.2da4c3d8@aol.com> We are using this instrument too, has saved tremendous time and money in the lab. We love it! lge From adelaur <@t> nimr.mrc.ac.uk Wed Apr 7 05:26:31 2004 From: adelaur <@t> nimr.mrc.ac.uk (April DeLaurier) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] anti-GFP wholemounts Message-ID: Has anyone had any success with using Anti-GFP antibodies on wholemount embryonic mouse specimens stage 10.5-14.5? thanks, April -- April DeLaurier, PhD Division of Developmental Biology National Institute for Medical Research The Ridgeway, Mill Hill London NW7 1AA United Kingdom Tel: +44 208 959 3666 ext. 2095 Fax: +44 208 816 4477 E-mail: adelaur@nimr.mrc.ac.uk From StarkusL <@t> ummhc.org Wed Apr 7 06:41:22 2004 From: StarkusL <@t> ummhc.org (Starkus, Laurie) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] AFB staining on mouse tissue Message-ID: Anyone care to share their staining protocol for staining acid fast bacilli on Mouse tissue? The laboratory I am in has been using the microbiology kit which is not for paraffin embedded tissues. And they wondered why they were getting inconsistent results. I'm looking for both protocol and sources for the stain. Thanks in advance From mark.lewis <@t> thermo.com Wed Apr 7 07:26:10 2004 From: mark.lewis <@t> thermo.com (mark.lewis@thermo.com) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] cryostat decontamination ? CAP REGS Message-ID: Good morning from Pittsburgh !! Has anyone heard that the CAP regulations for cryostat decontamination are going to change ? What exactly are the current requirements from CAP for Cryostat decontamination ? Does JCAH and OSHA require something different ? Thanks for your input !! You all are always a helpful and informative group ! Have a nice day ! Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com From sladd <@t> hsc.usf.edu Wed Apr 7 07:58:24 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing In-Reply-To: Message-ID: Good point about the vacuum. Also, with or without sponges there is going to be carry over (in the tissue, in the chamber, in the hoses, clinging to the casettes etc.) I was trying to do the math... How much 95% alcohol (and ultimately how much water?) would be carried over to the xylene station, if there were 200 mL of 95% alcohol retained, each station contained 4000 mL of liquid and there were 3 100% alcohol stations in between? I got a headache trying to figure out the dilutions and I gave up. The point is, if we have an appropriate processing protocol with multiple stations of alcohols, xylenes and paraffins and we make use of vacuum and agitation and change our reagents often, then I think these fancy new processors should be able to deal with the carry over of liquid retained in the sponges. We put 200 cassettes on our processor, with 50-75% of the cassettes containing sponges and UNLESS the P/V and agitation gets mistakenly turned off, our specimens are appropriately processed. Of course, this is just my opinion and if I had the time, I would do a couple little experiments... Sharron University of South Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Tuesday, April 06, 2004 11:21 AM To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pam <@t> ategra.com Wed Apr 7 08:12:24 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Job Opportunities in Histology Latest Update 04/05/04 Message-ID: Hi Histonet, Here is the latest update on opportunities with some of my best clients throughout the US who are seeking Histology Supervisors, Histo Technologists and Histo Technicians. These are positions as direct employees of our client. There are fulltime 40 hour per week positions.. As a direct employee of one of our clients you will be provided with full benefits including Health Insurance, Vacation, Sick Pay, Relocation money and a lucrative sign-on bonus. I have part time and temporary positions as well. I have supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are some of my HOTTEST Histology Supevisory positions: 1. Colorado - Histology Lab Manager 2. Connecticut - Histology Manager 3. Connecticut - Histology Supervisor 4. Oregon - Lead Histo Tech 5. Maryland - AP Supervisor (Histology/Cytology) 6. California - Histology Technical Coordinator Here are some of my HOTTEST Histo Tech bench positions: 1. Florida - Histo Tech (full time and part time positions) 2. Florida - MOHS Tech 3. Georgia - Histo Tech 4. Pennsylvania - Histo Tech 5. Oregon - Lead Histo Tech 6. Rhode Island - MOHS Tech 7. Wisconsin - MOHS Tech 8. California - Histology Trainer 9. California - Histo Tech 10. California - Lead Histo Tech 11. Nevada - Histo Tech 12. Maryland - Histo Tech 13. Massachusetts - Histo Tech (temp) 14. New York - Histo Tech (3rd shift) If you are interested in these jobs, please CALL ME ASAP at 800 466 9919 x234. To speed things up, please also send me a copy of your resume, (if you haven't already done so). If you are interested in jobs outside the above-mentioned areas, please send me your resume as well. I have clients throughout the US. I will keep your resume confidential and will not release it to anyone without your permission (This is Ategra policy as well as my own). My services are at no charge to you. Of course, you may be happy in your present job, but it never hurts to to keep an eye open. Also, if you have friends/peers who do not have an email address, if you could pass my query & name on to them I'd be very grateful. I don't want to be a bother - I was told that you were a hands-on Histo Tech or a Lab Supervisor. If you are no longer working in a lab please send me an email and I will remove you from my list of people to contact. However, if you are interested in any of the jobs above, please call me. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Pam Barker Senior Lab Recruiter Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd. Winter Park, FL 32792 VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com To Learn More About Ategra: http://www.ategra.com -------------------------------------------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. -------------------------------------------------------------------------------------------------- From louri_c <@t> hotmail.com Wed Apr 7 08:27:44 2004 From: louri_c <@t> hotmail.com (Louri Caldwell) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Small Immunostainers Message-ID: Hi Everyone, I am looking for a used or demo small immunostainer. Any suggestions, quotes, or information would be greatly appreciated. Thank you in advance for your help. Louri _________________________________________________________________ [1]MSN Toolbar provides one-click access to Hotmail from any Web page FREE download! References 1. http://g.msn.com/8HMAENUS/2731??PS= From dmcaloose <@t> wcs.org Wed Apr 7 08:28:18 2004 From: dmcaloose <@t> wcs.org (McAloose, Dee) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] RE: Histonet Digest, Vol 5, Issue 8 Message-ID: <9C26E1A5C9EE5F4B84216095CBE2E24A059DD75F@WOLF.wcs.org> Hello, I'd like to announce an immediate opening for a full-time histotechnician in the pathology department at the Wildlife Conservation Society located at the Bronx Zoo. We provide the diagnostic pathology services to one of the largest zoological collections in the country. Applications are being accepted until May 15, 2004 or until the position is filled. Please contact Ms. Tawanda Williams (Human Resources; email: twilliams@wcs.org) if you're interested in this exciting opportunity and in becoming part of our team! Thanks for listening and we look forward to hearing from you. D McAloose, VMD, Dipl ACVP Head, Department of Pathology Wildlife Conservation Society Bronx, NY 10464 (phone) 718-220-7105 (fax) 718-220-7126 dmcaloose@wcs.org -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, April 06, 2004 1:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 5, Issue 8 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: perfusion debate (Tamara Howard) 2. RE: RE: perfusion debate (Charles Scouten) 3. Microscopist position available, U of Richmond (Gary Radice) 4. (no subject) (Ant Swensson) 5. Region III Meeting (LaFriniere, Mike) 6. Adhesive for paraffin sections (Yin Ping Lee) 7. Analytical Imaging Station Version 6.0 (Wendy England) 8. RE: Region III Meeting (LaFriniere, Mike) 9. Re: schools (Lee & Peggy Wenk) 10. a problem about immunostaining for immunoglobulin heavy chains (=?gb2312?q?=CA=E7=C7=ED=20=C9=F2?=) 11. a lot of stuff HELP OUT THERE!! (Jesus Ellin) 12. Re: Analytical Imaging Station Version 6.0 (Geoff McAuliffe) 13. Human myofibroblast markers (Mark Elliott) 14. (no subject) (Ronald E. Bremer) 15. porcine insulin (Stryker, S. (HLK)) 16. Re: Sponge problems in processing (Steven E. Slap) 17. Ventana Tissue Processor (Vickroy, Jim) 18. Re: microwave antigen retrieval (Steven E. Slap) 19. solochrome stain (Myri37@aol.com) 20. RE: Sponge problems in processing (Marshall Terry Dr, Consultant Histopathologist) 21. Re: re mirowave Ag retrieval (Steven E. Slap) 22. I need help! (Mar?a Teresa Dom?nguez) 23. Ronald Bremer/knife sharpener (Joyce Cline) ---------------------------------------------------------------------- Message: 1 Date: Mon, 5 Apr 2004 11:56:45 -0600 (MDT) From: Tamara Howard Subject: [Histonet] RE: perfusion debate To: HistoNet Message-ID: Content-Type: TEXT/PLAIN; charset=US-ASCII As far as the relationship of mm Hg to ml/min with a perfusion pump - aren't the innner diameter and stiffness of the tubing (and the i.d. of the cannula) going to influence the final pressure, too? I don't think adjusting final pump speed alone is going to accurately duplicate the "hanging bag" pressure. Can you check the literature and see what other groups have reported and work from there? |--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward@unm.edu |--------------------------------------------------| ------------------------------ Message: 2 Date: Mon, 5 Apr 2004 13:24:14 -0500 From: "Charles Scouten" Subject: RE: [Histonet] RE: perfusion debate To: "Tamara Howard" , "HistoNet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" It depends on where the bottleneck is. If the cardiovascular system is the main source of resistance, it will feel all 300 mm Hg. If the tubing or needle is small, it will absorb the pressure drop, and the cardiovasucalar system will get much less than the gauge pressure the perfusion bottle was pumped up to. A mouse has much more cardiovascular resistance than a rat. A pig would have much much less. The largest suitable needle and tubing (1/4" ID tubing), would insure that the animal was the main source of resistance if the flow was small (mouse or even rat), but larger animals would require larger tubing and needle. Remember, the hanging bag pressure needs to be about 13 feet above the animal to provide a pressure that will disrupt the blood brain barrier. The correct flow rate to use depends greatly on animal size and condition. The correct pressure is constant, but needs to be higher than traditional gravity flow can typically provide. So it is better to control pressure directly, rather than flow rate. For an inexpensive means of implementing this strategy, and getting no shrink perfusions, go to www.myneurolab.com, click products, click sacrifice instruments under the Histology header, click Perfusion One. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 FAX 314 522 0377 cwscouten@myneurolab.com www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tamara Howard Sent: Monday, April 05, 2004 12:57 PM To: HistoNet Subject: [Histonet] RE: perfusion debate As far as the relationship of mm Hg to ml/min with a perfusion pump - aren't the innner diameter and stiffness of the tubing (and the i.d. of the cannula) going to influence the final pressure, too? I don't think adjusting final pump speed alone is going to accurately duplicate the "hanging bag" pressure. Can you check the literature and see what other groups have reported and work from there? |--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward@unm.edu |--------------------------------------------------| _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Mon, 5 Apr 2004 14:10:39 -0400 From: Gary Radice Subject: [Histonet] Microscopist position available, U of Richmond To: histonet@lists.utsouthwestern.edu Message-ID: <8B456638-872C-11D8-B0A1-000393BC23F4@richmond.edu> Content-Type: text/plain; charset=US-ASCII; format=flowed Hello everyone, Please forward if you know someone who might be interested. Gary Radice Director of Microscopy and Imaging University of Richmond The Department of Biology at this highly selective, private, primarily undergraduate university invites applications for a Director of Microscopy and Imaging. Duties including supervising operation of a new biological imaging facility, including operation and routine maintenance of a TEM, SEM, and laser scanning confocal microscope and associated equipment. Teaching responsibilities include training and supervising faculty and student users in research and classroom activities and courses based on interests and department needs; assisting faculty and students in experimental design, specimen preparation, digital photography, and analysis of results. Qualifications: MS or PhD degree and experience with light and electron microscopy and digital imaging, strong interpersonal skills and desire and ability to work well in a team environment. This is a continuing appointment that is not eligible for tenure but may include faculty status. Applications including a letter of interest, CV, evidence of expertise in microscopy, and three letters of recommendation should be sent to Dr. Gary Radice, Department of Biology, University of Richmond, Richmond VA 23173 (gradice@richmond.edu). Review of applications will begin on April 23, 2004. The appointment is expected to begin June 1, 2004. The University of Richmond is committed to increasing the diversity of our faculty and strongly encourages applications from women and minorities. For more information on the department, resources, and teaching assignment, see (http://biology.richmond.edu/) Gary P. Radice gradice@richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond 804-289-8233 (FAX) Richmond VA 23173 http://www.richmond.edu/~gradice ------------------------------ Message: 4 Date: Mon, 05 Apr 2004 11:28:13 -0700 From: "Ant Swensson" Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Hi. I'm interested in taking the HT exam at the end of this year. I've been studying by myself, and working in a lab to get hands on experience. I am curious, if I need to in the future, where the schools that train for the exam are located in the USA? Thank you, Antoinette Swensson Univeristy of Washington/Harborview Medical Center Neuropathology 325 9th Ave MS 359-791 Seattle, WA 98104 (206) 731-3910 _________________________________________________________________ [1]Limited-time offer: Fast, reliable MSN 9 Dial-up Internet access FREE for 2 months! References 1. http://g.msn.com/8HMBENUS/2734??PS= ------------------------------ Message: 5 Date: Mon, 5 Apr 2004 13:29:06 -0400 From: "LaFriniere, Mike" Subject: [Histonet] Region III Meeting To: Message-ID: Content-Type: text/plain; charset="us-ascii" The Region III Meeting is finalizing plans for their annual meeting this year in Birmingham, Alabama. May 20th-24 The program can be found on the Alabama Society for Histotechnology (ASH) web page. The web site can be found at www.timjday.com and click on Region III, registration is welcome through the website. The hotel accommodations are at the beautiful Wynfrey attached to the Galleria Mall in Birmingham. We ask that all participants register at the Wynfrey to keep our meeting cost to a minimum! The negotiated rate for the meeting is $125.00 per room, which normally is $199.00. This is an upscale 4 star hotel! Please make your reservations at the Wynfrey as soon as possible to obtain this rate as there is less than 20 days to obtain the negotiated $125.00 per night rate! We are demonstrating an excellent program to include more than 20 vendors and 14 excellent workshops offered from speakers within our Region and expect over 100 attendees, this will hopefully help diminish our debt from last years meeting in Puerto Rico. Your attendance at the Wynfrey 1800-476-7006 is urgently requested to support Region III. Please contact the hotel for reservations and state you are with the Region III, Alabama Histology meeting! Programs have been mailed out last week to all of Region III membership as well as several State memberships! Vendors are encouraged to also use the Wynfrey to assist with our overall cost, in addition, there are a few spots left for Vendors who have not yet registered to be at the meeting and can contact me directly for additional information. Michael LaFriniere NSH Region III Director 423-495-6117 ------------------------------ Message: 6 Date: Mon, 5 Apr 2004 16:35:44 -0700 From: Yin Ping Lee Subject: [Histonet] Adhesive for paraffin sections To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <6BAF4D075F07D411B30900508B94CBA0C1F55F@SERVER20> Content-Type: text/plain; charset="iso-8859-1" Hi, Leslie, "Mount Quick Tissue Transfer Technique" is another alternative for your case. This technique allows for the transfer of tissue sections from one slide to another slide. (Therefore, the sections can be transferred to charged slides and immuno-staining can be performed as per its usual protocol.) This transfer technique is very simple to do; all you need is to purchase the "Mount Quick" medium and follow the protocol. This medium is distributed by Newcomer Supply. Tel: (608) 831-7888. Fax: (608)831-0866. Yin Ping Lee Histopathology Lab BC Cancer Agency Vancouver BC ------------------------------ Message: 7 Date: Mon, 05 Apr 2004 16:41:06 -0700 From: "Wendy England" Subject: [Histonet] Analytical Imaging Station Version 6.0 To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed Hello, I was asked to search info about above systems. But I couldn't find any through my computer. Could anybody provide me the website I can look? Any help will be highly appreciated! Wendy _________________________________________________________________ MSN Toolbar provides one-click access to Hotmail from any Web page - FREE download! http://toolbar.msn.com/go/onm00200413ave/direct/01/ ------------------------------ Message: 8 Date: Mon, 5 Apr 2004 18:26:11 -0400 From: "LaFriniere, Mike" Subject: [Histonet] RE: Region III Meeting To: Message-ID: Content-Type: text/plain; charset="us-ascii" The Region III Meeting is finalizing plans for their annual meeting this year in Birmingham, Alabama. May 20th-24 The program can be found on the Alabama Society for Histotechnology (ASH) web page. The web site can be found at www.timjday.com and click on Region III, registration is welcome through the website. The hotel accommodations are at the beautiful Wynfrey attached to the Galleria Mall in Birmingham. We ask that all participants register at the Wynfrey to keep our meeting cost to a minimum! The negotiated rate for the meeting is $125.00 per room, which normally is $199.00. This is an upscale 4 star hotel! Please make your reservations at the Wynfrey as soon as possible to obtain this rate as there is less than 20 days to obtain the negotiated $125.00 per night rate! We are demonstrating an excellent program to include more than 20 vendors and 14 excellent workshops offered from speakers within our Region and expect over 100 attendees, this will hopefully help diminish our debt from last years meeting in Puerto Rico. Your attendance at the Wynfrey 1800-476-7006 is urgently requested to support Region III. Please contact the hotel for reservations and state you are with the Region III, Alabama Histology meeting! Programs have been mailed out last week to all of Region III membership as well as several State memberships! Vendors are encouraged to also use the Wynfrey to assist with our overall cost, in addition, there are a few spots left for Vendors who have not yet registered to be at the meeting and can contact me directly for additional information. Michael LaFriniere NSH Region III Director 423-495-6117 ------------------------------ Message: 9 Date: Tue, 6 Apr 2004 05:07:46 -0400 From: "Lee & Peggy Wenk" Subject: Re: [Histonet] schools To: "Ant Swensson" , Message-ID: <002b01c41bb6$a103dfc0$b834d445@domainnotset.invalid> Content-Type: text/plain; charset="iso-8859-1" The accrediting agency for lab schools is the National Accrediting Agency for Clinical Laboratory Schools (NAACLS). To find lab programs, including HT and HTL, go to http://www.naacls.org On the left side, click on "find a program" Then, enter the type of program (HT or HTL, for exam), and the state (or search for all states). (There are 27 HT and 2 HTL = 29 accredited programs in the US. Up from about 21 a couple of years ago. Still could use a lot more, in my opinion.) Peggy A. Wenk, HTL(ASCP)SLS Program Director School of Histotechnologists School of Histologic Technicians William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Ant Swensson" To: Sent: Monday, April 05, 2004 2:28 PM Subject: [Histonet] (no subject) > > Hi. I'm interested in taking the HT exam at the end of this year. > I've been studying by myself, and working in a lab to get hands on > experience. I am curious, if I need to in the future, where the > schools that train for the exam are located in the USA? > > Thank you, > > Antoinette Swensson > > Univeristy of Washington/Harborview Medical Center > Neuropathology > 325 9th Ave MS 359-791 > Seattle, WA 98104 > (206) 731-3910 > _________________________________________________________________ > > [1]Limited-time offer: Fast, reliable MSN 9 Dial-up Internet access > FREE for 2 months! > > References > > 1. http://g.msn.com/8HMBENUS/2734??PS= > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 10 Date: Tue, 6 Apr 2004 20:22:03 +0800 (CST) From: =?gb2312?q?=CA=E7=C7=ED=20=C9=F2?= Subject: [Histonet] a problem about immunostaining for immunoglobulin heavy chains To: histonet@lists.utsouthwestern.edu Message-ID: <20040406122203.67972.qmail@web15207.mail.bjs.yahoo.com> Content-Type: text/plain; charset=gb2312 Hello,everybody, I am intrested in immunostaining for immunoglobulin heavy chains on renal biopsy specimen ,but I have no idea about which antibodies are appropriate.Would anybody be kind enough to share me with you experience? Thank You very much. Best regards! Shuqiong Shen Jinling Hospital Nanjing 210002 P.R.China --------------------------------- Do You Yahoo!? ????TT???????????????????????? ------------------------------ Message: 11 Date: Tue, 06 Apr 2004 05:36:11 -0700 From: "Jesus Ellin" Subject: [Histonet] a lot of stuff HELP OUT THERE!! To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Hello everyone thought that I would ask for some help here,, Currently our lab is getting a hugh over haul on everthing and I need to find out some information well were do I begin, First off our lab went live with Tamtron this past year and it has been different to say the least, but one of the ideas we are pondering is using the image module to caputre gross instead of gross dictation, only on small bx that do not require anything other than transfering it to the cassette and snapping a picture. Was wondering if anyone out there is doing this and also if they do have Tamtron how do they like the imaging module. The next item we are looking at is this processor that can process tissue in 1 hr and is any one out there doing this? This is a huge step, since we are part of the traditional way of processing over night and doing the embedding the next day. SO if anyone is attempting this please feel free to give me input. The last thing that our lab is going to under take is allow tech that are not certified and do not have 60 hrs of college to do gross,the standard is set forth in NACCLS that a person must have a minimal amount of education to do this. But our pathologist called the CAP and said that they can do transfer of specimens that can give color, size, dimension. This is were the idea of photographing specimens came amout. ANY help would greatly be appreciated. Jesus Ellin HTL Yuma Regional Medical Center ------------------------------ Message: 12 Date: Tue, 06 Apr 2004 10:09:23 -0700 From: Geoff McAuliffe Subject: Re: [Histonet] Analytical Imaging Station Version 6.0 To: Wendy England Cc: histonet@pathology.swmed.edu Message-ID: <4072E443.1090405@umdnj.edu> Content-Type: text/plain; format=flowed; charset=windows-1252 Ask the manufacturer of the system. If you are looking to purchase a new image analysis system Nikon, Zeiss, Olympus etc all make such systems. In addition, there are many vendors who will build a system to meet your current and future needs. Geoff Wendy England wrote: > Hello, > I was asked to search info about above systems. But I couldn't find > any through my computer. Could anybody provide me the website I can look? > > Any help will be highly appreciated! > > Wendy > > _________________________________________________________________ > MSN Toolbar provides one-click access to Hotmail from any Web page - > FREE download! http://toolbar.msn.com/go/onm00200413ave/direct/01/ > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** ------------------------------ Message: 13 Date: Tue, 06 Apr 2004 07:30:40 -0700 From: "Mark Elliott" Subject: [Histonet] Human myofibroblast markers To: Message-ID: Content-Type: text/plain; charset=US-ASCII Was wondering what everyone is using for labelling myofibroblasts in human tissue? Preferably something that works in FFPE tissues, but frozen is OK. Thanks Mark W. Mark Elliott, PhD Research Associate, Interim Director, Histology Core Facility James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research Room 166, St. Paul's Hospital 1081 Burrard Street Vancouver, BC Canada V6Z 1Y6 ------------------------------ Message: 14 Date: Tue, 06 Apr 2004 10:34:18 -0400 From: "Ronald E. Bremer" Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Message-ID: <4072BFEA.3010808@acpub.duke.edu> Content-Type: text/plain; charset=us-ascii; format=flowed We are intertested in purchasing a used microtome knife sharpener. I was hoping for some advice. I've noted a few American Optical models for sale for what seems a rather cheap price. I think Leica picked them up or is the same company. Are these shapreners difficult to get parts for? I'm also interested in comments on "Hacker knife sharpeners" Even new they seem rather inexpensive compared to other models. We could go new if the price was right and have GSA money if anybody could point me in the right direction. Unfortunately the right price is less than $1000, so I am not hopeful on a new model. Ron ------------------------------ Message: 15 Date: Tue, 6 Apr 2004 17:04:47 +0200 From: "Stryker, S. (HLK)" Subject: [Histonet] porcine insulin To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello everybody, I'm hoping someone can give me any suggestions on how to detect insulin on a (paraffin embedded formalin-fixed or frozen) porcine section using the following antibodies: - primary antibody: Guinea Pig anti-insulin (Dako) - secondary antibody: Swine anti-Rabbit (Dako) According to the DAKO catalogous the anti-Rabbit should work in this combo. I have tried it over and over again, using different protocols, but somehow I just can't get the job done... Thanks in advance! Samantha s.stryker@lumc.nl Leiden University Medical Centre ------------------------------ Message: 16 Date: Tue, 6 Apr 2004 11:13:41 -0400 From: "Steven E. Slap" Subject: Re: [Histonet] Sponge problems in processing To: Gayle Callis , "Gudrun Lang" , Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. ------------------------------ Message: 17 Date: Tue, 6 Apr 2004 09:57:30 -0500 From: "Vickroy, Jim" Subject: [Histonet] Ventana Tissue Processor To: "Histonet \(E-mail\)" Message-ID: Content-Type: text/plain We had a processing run today that looks like the tissue was cooked especially the small GI specimens. We have a Ventana Renissance Tissue Processor, which they no long sell. Has anyone every had a problem similar to this, and is there any correlation to problems with the tissue processor? My first thoughts are that maybe the chemistry on the machine was switched, but I can't find any glaring problems. Thanks for your help James R. Vickroy BS, HT (ASCP) Technical Supervisor, Surgical Pathology 788-4046 vickroy.jim@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. ------------------------------ Message: 18 Date: Tue, 6 Apr 2004 11:19:33 -0400 From: "Steven E. Slap" Subject: Re: [Histonet] microwave antigen retrieval To: Gayle Callis , Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" ; format="flowed" Hi HistoNetters Successful antigen retrieval depends on accurate control of three factors: time, temperature and pH. Any instrument which will produce this accuracy in a reproducible way will be effective. A microwave has been shown to work well over a wide range of antigens, where other methods have required more aggressive treatment. In some cases, pressure is needed to allow for temperatures above 98?C. I can get most antigens to "work" with pH 6.0 citrate buffer at 110?C in a 3-5 minute microwave method using the Hacker/Milestone microwave. best regards, Steven Slap Microwave Consultant ------------------------------ Message: 19 Date: Tue, 06 Apr 2004 11:18:31 -0400 From: Myri37@aol.com Subject: [Histonet] solochrome stain To: histonet@pathology.swmed.edu Message-ID: <64E5A68B.38CAA850.0005167B@aol.com> Content-Type: text/plain; charset=iso-8859-1 hello i have prepared solochrome stain following this: 1 g of calcon (solochrome) 2 ml acetic acid glacial 98 ml distilled water and let it for 3 months before use i tried this stain on MMA sections of thin reconstructed and mineralized tissue, i put section of 50um in this stain for 1 hour, i didn't see anything stained on hot plate or not could any one help me ? is the stain not good ? or something else ? thank you very much for your help Myriam ------------------------------ Message: 20 Date: Tue, 6 Apr 2004 16:20:47 +0100 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Sponge problems in processing To: "Steven E. Slap" , "Gayle Callis" , "Gudrun Lang" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Tue, 6 Apr 2004 11:34:30 -0400 From: "Steven E. Slap" Subject: Re: [Histonet] re mirowave Ag retrieval To: "Carl" , Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Hi HistoNetters The Nordicware microwave pressure cooker is available at a deep discount from the Yahoo store: Nordicware Tender Cooker/Microwave Pressure Cooker 62104 shop.store.yahoo.com/dotcoms/tecoprcobyno.html However, this device does not allow for temperature control. Let the buyer beware... best regards, Steven Slap At 8:36 PM +0100 4/1/04, Carl wrote: > >Well...I use a microwave pressure cooker! I understand that one can >buy this in USA Walmarts ( be interested to know the price of it in >the USA). I buy them in UK from Biogenex. Very reliable and >consistent.. Started on Coplins jars and , VERY rapidly moved to >rice staemers, pressure cookers...then microwave pressure cookers. I >look back in no anger. ------------------------------ Message: 22 Date: Tue, 06 Apr 2004 13:44:57 -0300 From: "Mar?a Teresa Dom?nguez" Subject: [Histonet] I need help! To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Hi Histonetters! In our lab we have received Methilyc Alcohol which we use in the tissues prossessor, but this time we don't know the Quality of it due to the quantity we have received doesn't have any label with any descripcion about it's features. I wonder if there any quality Control about Methilyc Alcohol. What I need to know if this Alcohol is 100% methilyc. I'll appreciate all you can advice me. H.T. Maria Teresa Dominguez Rio Grande, Zonal Hospital, Anatomic Pathology, Tierra del Fuego, Argentina. _________________________________________________________________ Help STOP spam with [1]the new MSN 8 and get 2 months FREE* References 1. http://g.msn.com/8HMBEN/2731??PS= ------------------------------ Message: 23 Date: Tue, 6 Apr 2004 12:57:15 -0400 From: "Joyce Cline" Subject: [Histonet] Ronald Bremer/knife sharpener To: "Histonet" Message-ID: <002d01c41bf8$3a14d660$1d2a14ac@wchsys.org> Content-Type: text/plain;charset="iso-8859-1" I have a Hacker knife sharpener for sale, it works perfectly and I even have the video that shows how to work it. I have used glass sharpeners in the past and the Hacker beats any sharpener for doing the steel knives. We are using disposables now so I have no need for the Hacker. You can email me directly if you would like. ***** 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. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 5, Issue 8 ************************************** From Diane.Gladney <@t> se.amedd.army.mil Wed Apr 7 08:30:32 2004 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] cryostat decontamination ? CAP REGS Message-ID: <9D41AB7C56F8304F98537ABD87B249F66261BB@DASMTHGBZ001> Mark, The question on the CAP Inspection List is as follows: ANP.24250 Phase I Is there a documented procedure for the routine decontamination of the cryostat at defined intervals, and are decontamination records evident? We incorporated in our SOP for frozen sections/cryostat a statement at what time (intervals) the cryostate would be routinely decontaminated. We have a Log Sheet that gives the date frozen sections were cut, the date the cryostate was decontaminated, and a column for comments such as any preventative maintenance was performed (such as oiling, etc). Since we are a very small hospital, we don't get many frozen sections. We average 2 per month; therefore, we have decided that the cryostat would be cleaned and decontaminated after the completion of each frozen section case. We have rarely had more than one frozen section case in one day. The SOP gives detail instructions on how to decontaminate the cryostat. This has more than satisfied the CAP requirements. I attended a CAP Instructors Course in February and addressed this question. I explained what we do and I was given a complete " great job" from the instructor. He didn't indicate that I should be doing anything different. Thanks, Diane Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: mark.lewis@thermo.com [mailto:mark.lewis@thermo.com] Sent: Wednesday, April 07, 2004 8:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cryostat decontamination ? CAP REGS Good morning from Pittsburgh !! Has anyone heard that the CAP regulations for cryostat decontamination are going to change ? What exactly are the current requirements from CAP for Cryostat decontamination ? Does JCAH and OSHA require something different ? Thanks for your input !! You all are always a helpful and informative group ! Have a nice day ! Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Liam.Brennan <@t> bll.n-i.nhs.uk Wed Apr 7 08:32:53 2004 From: Liam.Brennan <@t> bll.n-i.nhs.uk (Brennan, Liam) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing Message-ID: We have been using these sponges for years on Sakura VIP processors with vacuum,agitation, etc without any problems of solvent carryover or artifacts in the tissue. Liam Brennan Belfast City Hospital -----Original Message----- From: S Ladd [mailto:sladd@hsc.usf.edu] Sent: 07 April 2004 13:58 To: Marshall Terry Dr, Consultant Histopathologist Cc: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Good point about the vacuum. Also, with or without sponges there is going to be carry over (in the tissue, in the chamber, in the hoses, clinging to the casettes etc.) I was trying to do the math... How much 95% alcohol (and ultimately how much water?) would be carried over to the xylene station, if there were 200 mL of 95% alcohol retained, each station contained 4000 mL of liquid and there were 3 100% alcohol stations in between? I got a headache trying to figure out the dilutions and I gave up. The point is, if we have an appropriate processing protocol with multiple stations of alcohols, xylenes and paraffins and we make use of vacuum and agitation and change our reagents often, then I think these fancy new processors should be able to deal with the carry over of liquid retained in the sponges. We put 200 cassettes on our processor, with 50-75% of the cassettes containing sponges and UNLESS the P/V and agitation gets mistakenly turned off, our specimens are appropriately processed. Of course, this is just my opinion and if I had the time, I would do a couple little experiments... Sharron University of South Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Tuesday, April 06, 2004 11:21 AM To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 Nancy.Walker <@t> sanofi-synthelabo.com Wed Apr 7 08:48:31 2004 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:46 2005 Subject: =?iso-8859-1?Q?R=E9f=2E_=3A_RE=3A_[Histonet]_RE=3A_perfusion_debate?= Message-ID: Bonjour! We tried measuring the "blood" pressure generated by our perfusion techniques but were unsuccessful. Back to calculating blood volumes, blood circulation times (using formulas from vetinarian school documents). A 35 g mouse, has a blood volume of 1.75 - 2.4 ml (blood volume= 50-80ml/kg) physiological blood flow rates are 6.3 - 8.7 ml/min and systolic blood pressure of 113 mm Hg. Conclusions: Our perfusion rate of 2 ml/min is therefore much lower than physiological flow rates. What maximum rates have been used in your experiments (without exploding the cardiovascular system)? Nancy Walker Molecular Pharmacology Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 From RossS <@t> BaylorHealth.edu Wed Apr 7 09:16:04 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing Message-ID: At my former hospital, we had a major problem with biopsies being fried once. It turned out to be a combination of things. 1. The techs had skipped a day changing that processor. 2. And most importantly a PA over the weekend had added a 3rd rack to the processor. The problem was it was a VIP 1000 and it isn't made to use 3 racks. So the top blocks were only half covered at best with solution. At the time we didn't know about the sponge carryover. What we couldn't figure out was why some of the blocks done on Friday looked just as bad as the blocks the PA had added over the weekend which were not fully covered with solution. Later when I heard about this sponge carryover it made since. There were over 120 blocks on that processor. All of them were biopsies in sponges. The solutions were already being pushed to the limit because someone had decided they could wait until Monday to change the processor. Add in the third rack and it was a recipe for disaster. Most of the blocks were severely compromised even after reprocessing. About 1/4 were unreadable. It was a mess. Sponges can be used effectively for years and years. You just have to be aware of this carryover problem and keep the processor changed properly and not overload it. The culture of the lab can't allow any skimping on changing the processor as this affects everything. I now prefer to use a much larger processor than physically necessary for biopsies so that there is a larger volume of fluid to compensate for the carryover when using sponges. Here they use the mesh cassettes for most biopsies and rarely use sponges so this is not a problem. Now mesh cassettes and air bubbles are a whole other problem that I get to deal with on occasion. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Brennan, Liam Sent: Wednesday, April 07, 2004 8:33 AM To: 'HISTONET' Subject: RE: [Histonet] Sponge problems in processing We have been using these sponges for years on Sakura VIP processors with vacuum,agitation, etc without any problems of solvent carryover or artifacts in the tissue. Liam Brennan Belfast City Hospital -----Original Message----- From: S Ladd [mailto:sladd@hsc.usf.edu] Sent: 07 April 2004 13:58 To: Marshall Terry Dr, Consultant Histopathologist Cc: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Good point about the vacuum. Also, with or without sponges there is going to be carry over (in the tissue, in the chamber, in the hoses, clinging to the casettes etc.) I was trying to do the math... How much 95% alcohol (and ultimately how much water?) would be carried over to the xylene station, if there were 200 mL of 95% alcohol retained, each station contained 4000 mL of liquid and there were 3 100% alcohol stations in between? I got a headache trying to figure out the dilutions and I gave up. The point is, if we have an appropriate processing protocol with multiple stations of alcohols, xylenes and paraffins and we make use of vacuum and agitation and change our reagents often, then I think these fancy new processors should be able to deal with the carry over of liquid retained in the sponges. We put 200 cassettes on our processor, with 50-75% of the cassettes containing sponges and UNLESS the P/V and agitation gets mistakenly turned off, our specimens are appropriately processed. Of course, this is just my opinion and if I had the time, I would do a couple little experiments... Sharron University of South Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Tuesday, April 06, 2004 11:21 AM To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , >Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding >bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh >inserts fitting inside a cassette. These may be worth a try to avoid >sponges. I think they are available through Fisher or some other >company who could provide free samples. It will be interesting to see >peoples testimonials on using these. > >There was Histonet discussion on these sponges way back in time - it >may pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From Lynne.Bell <@t> hitchcock.org Wed Apr 7 10:22:13 2004 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] cryostat decontamination ? CAP REGS Message-ID: How exactly do you decontaminate your cryostat? We currently use 100% alcohol to wipe down all the surfaces and, for obvious reasons, cannot use anything that contains water. Thanks for your help. Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From kemlo <@t> tiscali.co.uk Wed Apr 7 10:48:27 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing In-Reply-To: Message-ID: <000701c41cb7$cd37de00$ce002850@KEMLOS> I thought vacuum processing only 'sucked away' air. I mean if you reduce the air pressure above the fluids then that reduces pressure within the tissue that was preventing air escaping (as it has a positive pressure). I suppose if you reduce the Saturated Vapour Pressure of a solvent then that would cause more solvent to vapourise but wouldn't that be from the surface of the solvent rather than within the tissue? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 06 April 2004 16:21 To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 carl.hobbs <@t> kcl.ac.uk Wed Apr 7 10:54:21 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] re baking bone slides for heat retrieval techniques Message-ID: <00ac01c41cb8$97a88360$e8345c9f@Carlos> I always leave my decaled bone pwax sections overnight at 65C, and all other tissues( only one antibody doesn't like it........tinctorial methods are all fine, in my experience)with Superfrost plus slides. Double- subbed also works- still get certain amount fall-off if doing AR From laurie.colbert <@t> huntingtonhospital.com Wed Apr 7 11:01:17 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BE88@EXCHANGE1.huntingtonhospital.com> I just attended a training session for our new VIP at Sakura and was told that the function of the vacuum is in fact to only "suck away" the air bubbles that cling to the tissue. This allows the solutions to come in contact with all surfaces of the tissue. I learned something new!! Laurie Colbert Huntington Hospital Pasadena, CA -----Original Message----- From: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: Wednesday, April 07, 2004 8:48 AM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing I thought vacuum processing only 'sucked away' air. I mean if you reduce the air pressure above the fluids then that reduces pressure within the tissue that was preventing air escaping (as it has a positive pressure). I suppose if you reduce the Saturated Vapour Pressure of a solvent then that would cause more solvent to vapourise but wouldn't that be from the surface of the solvent rather than within the tissue? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 06 April 2004 16:21 To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 cwscouten <@t> myneurolab.com Wed Apr 7 11:01:39 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] cryostat decontamination ? CAP REGS Message-ID: Never decontaminate your crystat, push the button and let it decontaminate itself. Than is, if you have the right kind of cyrostat. See the link. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=475102&catdesc=Histology+Equipment&CatThreeID=732&CatOneID=4&subcatdesc=Cryostats&idsubcategory=182 If not, consider that alcohol, even 100% will not kill everything. Formalin Vapor has been mentioned on the histonet, and Perascope. We can supply Perascope. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bell, Lynne Sent: Wednesday, April 07, 2004 10:22 AM To: Gladney, Diane C Ms MACH; mark.lewis@thermo.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cryostat decontamination ? CAP REGS How exactly do you decontaminate your cryostat? We currently use 100% alcohol to wipe down all the surfaces and, for obvious reasons, cannot use anything that contains water. Thanks for your help. Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Wed Apr 7 11:10:25 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] HIK1083 Message-ID: Hi all, One of our pathologists brought me an article referencing an antibody by its clone only. He would like me to find out what antibody has a clone of: HIK1083. Familiar to anyone??? Thanks, 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 From David.Edmondson <@t> christie-tr.nwest.nhs.uk Wed Apr 7 11:18:46 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] RE: Tissue Micro Arrays Message-ID: Thanks, the site is an interesting read and provides some language to use and images to conjure with. And oportunities for training. Dave -----Original Message----- From: Thom Jensen [mailto:tissuearray@hotmail.com] Sent: 05 April 2004 16:21 To: David.Edmondson@christie-tr.nwest.nhs.uk Subject: Tissue Micro Arrays I am in the United States but I have a website to assist in instruction on creating microarrays. www.arrayworkshop.com I hope this helps. Thom Jensen >From: "Edmondson David (RBV) NHS Christie Tr" >To: "Histonet (E-mail 2)" >Subject: [Histonet] Tissue Micro Arrays >Date: Mon, 5 Apr 2004 14:07:52 +0100 > >Hello out there, > >I am asked to check out Micro Arrays and wonder where the closest >practitioners are. Is there anyone near Manchester UK ? We know of >Vancouver and Basel but know of no brains closer to pick. >Proximity has a bearing on costings, the value of a bequest being, at least >in part, a limiting factor and imports would add to costs. > >Best wishes and thanks > >Dave >Christie Hospital >Manchester UK > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ FREE pop-up blocking with the new MSN Toolbar - get it now! From David.Edmondson <@t> christie-tr.nwest.nhs.uk Wed Apr 7 11:31:48 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Myofibroblasts Message-ID: Hello I wondered that mention has been made of Calponin. We are using DAKO's cat M3556 The data sheet speaks of stromal myofibroblasts staining in some breast tumours and myoepithelial cells in breast. Best wishes David Christie Hospital Manchester UK From AliNeumann <@t> aol.com Wed Apr 7 11:11:19 2004 From: AliNeumann <@t> aol.com (AliNeumann@aol.com) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Stryker saw needed Message-ID: <679BC2BB.19E7C8C0.0AAAD598@aol.com> Hi, if anyone would like to sell a used functioning Stryker saw with blade(for autopsies including brain), please feel free to contact me. Thank you alineumann@aol.com Alice Neumann M.D. Precision Pathology PC 6429 Miller St. Unit C Arvada, CO ? 80004 303-432-7855 phone 303-525-0675 24 hour phone 303-432-7866 fax From pwg1 <@t> cdc.gov Wed Apr 7 12:30:53 2004 From: pwg1 <@t> cdc.gov (Greer, Patricia) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] HIK1083 Message-ID: By doing a google search for the antiboy clone number, I came up with this site. http://www.kanto.co.jp/hpeng/feature2.html Pat Greer Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sebree Linda A. Sent: Wednesday, April 07, 2004 12:10 PM To: Histonet (E-mail) Subject: [Histonet] HIK1083 Hi all, One of our pathologists brought me an article referencing an antibody by its clone only. He would like me to find out what antibody has a clone of: HIK1083. Familiar to anyone??? Thanks, 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Wed Apr 7 13:00:57 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] VEGF antibody Message-ID: Hi again, Wondering if anyone can recommend a good, reliable VEGF (vascular endothelial growth factor) antibody? If you've run yours on a Ventana instrument, so much the better. Thanks for the 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 From juan.gutierrez <@t> christushealth.org Wed Apr 7 13:23:07 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] HIK1083 Message-ID: Hi Linda, Idid a query and all that came up was the same clone name and a company in Tokyo that makes it. Their name is Kanto Chemicals, Tokyo, Japan. Sorry I couldn't find an address or phone #. Juan -----Original Message----- From: Sebree Linda A. [mailto:la.sebree@hosp.wisc.edu] Sent: Wed 4/7/2004 11:10 AM To: Histonet (E-mail) Cc: Subject: [Histonet] HIK1083 Hi all, One of our pathologists brought me an article referencing an antibody by its clone only. He would like me to find out what antibody has a clone of: HIK1083. Familiar to anyone??? Thanks, 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Wed Apr 7 13:33:28 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] VEGF antibody In-Reply-To: Message-ID: Linda, I recommend Santa Cruz's monoclonal VEGF, it is much better than the polyclonal. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sebree Linda A. Sent: Wednesday, April 07, 2004 12:01 PM To: Histonet (E-mail) Subject: [Histonet] VEGF antibody Hi again, Wondering if anyone can recommend a good, reliable VEGF (vascular endothelial growth factor) antibody? If you've run yours on a Ventana instrument, so much the better. Thanks for the 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DOOLEEO <@t> shands.ufl.edu Wed Apr 7 13:47:43 2004 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] CDX2 Message-ID: Dear Histonetters, Does anyone have CDX2 working on the Ventana Benchmark and would share their protocol? It says large bowel is a good control. Will small bowel work? any input would be appreciated. We have Novocastra's CDX2 (NCL-CDX2) I tried it at 1:50 for 32 min but did not get any staining but I used small bowel instead of large bowel. Elaine Dooley 352-265-0111 ext 72117 Thanks in advance From bpinkerton <@t> ccpathology.com Wed Apr 7 13:58:49 2004 From: bpinkerton <@t> ccpathology.com (Betty Pinkerton) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Lab expansion Message-ID: <028E7300440A0A45A7968235F4FD5663035B3C@exchange.ccpathology.com> Hi all Are lab is gettting ready to expand and we need some resources. Does anyone know of a lab designer? It doesn't matter where they are from. We need renovations for a larger lab. Thanks From Jim.Tunstead <@t> msnyuhealth.org Wed Apr 7 14:02:10 2004 From: Jim.Tunstead <@t> msnyuhealth.org (Tunstead, Jim) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] HIK1083 Message-ID: <1371F1DB5EE3294CA931D4321EBCB0041A9FE3@EXCEBW2K18.msnyuhealth.org> Here's the page http://www.kanto.co.jp/hpeng/feature2.html -----Original Message----- From: GUTIERREZ, JUAN [mailto:juan.gutierrez@christushealth.org] Sent: Wednesday, April 07, 2004 2:23 PM To: Sebree Linda A.; Histonet (E-mail) Subject: RE: [Histonet] HIK1083 Hi Linda, Idid a query and all that came up was the same clone name and a company in Tokyo that makes it. Their name is Kanto Chemicals, Tokyo, Japan. Sorry I couldn't find an address or phone #. Juan -----Original Message----- From: Sebree Linda A. [mailto:la.sebree@hosp.wisc.edu] Sent: Wed 4/7/2004 11:10 AM To: Histonet (E-mail) Cc: Subject: [Histonet] HIK1083 Hi all, One of our pathologists brought me an article referencing an antibody by its clone only. He would like me to find out what antibody has a clone of: HIK1083. Familiar to anyone??? Thanks, 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gudrun.lang <@t> aon.at Wed Apr 7 14:05:43 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] semithin-ultrathin Message-ID: <00b201c41cd3$53be15a0$eeeea8c0@SERVER> Hi, I've searched in the internet for a list that explains the thickness of semi-, ultra-, thin, thick sections. But I failed. My knowledge is only approximately. Perhaps someone can help me? Gudrun Lang From marytedo <@t> hotmail.com Wed Apr 7 14:09:04 2004 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Thanks for Help! Message-ID: Well, We haven't bought this Methano. I t was a "gift" from a private lab, they bought some to a new Co. and they sent to us 5 liters ina big bottle, that's why I couldn't get any information about this alcohol. Now, the problem is we don't have an Hidrometer, so we checked it with 2 ml. of Methanol in a little tube and 0,5 ml of oil, then I shaked them, and I could see little drops of oil, like an emulsion, the solution changed the colour, it turned white with drops in it. Maybe water?? The last step I did. I compair the solution with the Methanol we have kept in the lab which I know it is 98.8 % pure with 0,1% water. The drops of oil were quite bit less than in the other solution. What does it means?? I need someone who knows about chemistry soon!! Thanks again for your advices and HAPPY EASTER! for everyone. H.T. Maria Teresa Dominguez Anatomy Pathologyc, Zonal Hospital of Río Grande, Tierra del Fuego, Argentina. _________________________________________________________________ MSN 8 with [1]e-mail virus protection service: 2 months FREE* References 1. http://g.msn.com/8HMBEN/2740??PS= From tpmorken <@t> labvision.com Wed Apr 7 15:15:35 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] semithin-ultrathin Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA21774BB@usca0082k08.labvision.apogent.com> These terms are usually for electron microscopy sections or plastic light microscopy sections. In the EM lab a thick section is a one-micron plastic section for light microscopy of EM -embedded blocks. It could also be called semi-thin. Thin, or Ultra thin sections are terms used for the sections for use in the EM., ie, 400A to 800A in thickness. "Thin" can also be used for light microscopy plastic section of 1um in thickness. They are thin in relation to paraffin sections. Tim Morken Lab Vision / Neomarkers www.labvision.com -----Original Message----- From: Gudrun Lang [mailto:gudrun.lang@aon.at] Sent: Wednesday, April 07, 2004 12:06 PM To: Histonetliste Subject: [Histonet] semithin-ultrathin Hi, I've searched in the internet for a list that explains the thickness of semi-, ultra-, thin, thick sections. But I failed. My knowledge is only approximately. Perhaps someone can help me? Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eca9 <@t> georgetown.edu Wed Apr 7 15:14:45 2004 From: eca9 <@t> georgetown.edu (Eva Andersson) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] alternative to using pap pen? Message-ID: <40746135.5030209@georgetown.edu> Hi everyone, I am going to be undertaking a large immunohistochemistry staining project. To ensure that our slides do not dry out during the primary antibody incubation we have decided to use pap pen. With such a large amount of slides this may be quite a task and so I would like to know if anyone has found a good alternative to the pap pen. The slides will arrive with the samples already on them. Please let me know if you have any suggestions. Thanks From gcallis <@t> montana.edu Wed Apr 7 15:42:49 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] alternative to using pap pen? In-Reply-To: <40746135.5030209@georgetown.edu> Message-ID: <3.0.6.32.20040407144249.00bff4a0@gemini.msu.montana.edu> You never said how many slides you work with in a day? If only 30 or so, the pen is still a workable solution to your problem. We have mounted sections on hydrophobic barrier slides made for us by Erie. You can purchase ready made barrier, Plus charge slides from Erie - their selection is huge. You would have to get lab doing sectioning to buy these slides and use them to reduce work on your end. Pricey, but worth it. If you have to reduce size of area where antibody is applied for specific volume of 100 ul or less, pap pens are still worth the effort. We like ImmEdge pen from Vector, less expensive, excellent barrier and you can see it. Biogenex sells special hydrophobic barrier slides designed for their immunostainers for use on their stainers, also pricey. You may find a better deal at Erie. To keep sections from drying out, you can minimize that problem by the following. It helps to put a protein carrier i.e. very high quality bovine serum albumin, in your diluent buffer. We sometimes add 0.1% BSA, purchased from Jackson, protease and immunoglobulin free or a normal serum matching host of secondary or a nonrelated species i.e. goat, horse, swine, donkey serum (0.2% or so), etc. We add serum to all buffers and diluents. Tween 20 is also added (0.05% to aid flow of reagent over section, besides it other benefits). Good luck. At 04:14 PM 4/7/2004 -0400, you wrote: >Hi everyone, >I am going to be undertaking a large immunohistochemistry staining >project. To ensure that our slides do not dry out during the primary >antibody incubation we have decided to use pap pen. With such a large >amount of slides this may be quite a task and so I would like to know if >anyone has found a good alternative to the pap pen. The slides will >arrive with the samples already on them. >Please let me know if you have any suggestions. >Thanks > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From LBlack <@t> carilion.com Wed Apr 7 16:02:01 2004 From: LBlack <@t> carilion.com (Lisa Black) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Melanin Bleach & IHC Message-ID: Hello All, Whenever we bleach a slide prior to IHC staining, the end result is sub-optimal. Our protocol is to use 0.5% potassium permanganate and then oxalic acid for the bleach then to proceed with the automated IHC stain using Ventana's DAB I-View Kit. Most of the tissue comes off the slide and the remaining tissue has poor morphology. Does anyone have good advice for me? Thanks in advance, Lisa Black Carilion Consolidated Lab Roanoke, VA From gcallis <@t> montana.edu Wed Apr 7 16:12:06 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] semithin-ultrathin In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA21774BB@usca0082k08.labvisi on.apogent.com> Message-ID: <3.0.6.32.20040407151206.00bdfe80@gemini.msu.montana.edu> Some thoughts on section thickness: EM: Ultrathin although EM people them "thin" - 400 angstroms - 800 angstroms Semithin: 0.1 to 1 micrometer commonly referred to as the "thick section for EM" and stained with toluidine blue for light microscopy examination. Light microscopy: Thin: glycol methacrylate plastic 0.5 to 2 or 3 um Thin: paraffin 1 um to 3 um (yes, it is possible to cut 1 um paraffin sections!) Standard thickness for paraffin commonly seen in literature/routine use for light microscopy. 4 um to 6 um, some people cut at 7 to 10 um with latter often preferred for brain sections. Thick: paraffin 10 um and over up to 300 um or more. Examples: paraffin or celloidin embedded brain, brain frozen sections or vibratome sections, methylmethacrylate (plastic) embeddded bone sections cut with diamond cut off blades, ground/polished, and stained for light microscopy. It is important to think of section thickness in terms of what application is being used i.e. electron microcsopy to see ultrastructure, routine or diagnostic light microscopy, laser scanning confocal microscopy, etc. all these may limit or permit thicker section for optimal results. Consequently, a 5 um section from paraffin embedded tissue could actually be considered thin although not useful for EM. Whew, too much! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From DDittus787 <@t> aol.com Wed Apr 7 16:06:30 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] VEGF antibody Message-ID: <4B689A3D.1A1FE3E3.0A1F969F@aol.com> linda: we run zymeds on the nexes with great results. dana From DDittus787 <@t> aol.com Wed Apr 7 16:24:22 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Melanin Bleach & IHC Message-ID: <0B6B0E75.4335AFBB.0A1F969F@aol.com> lisa; instead of using the bleaching, or worrying that the dab looks like melanin, protect your tissue and the results by using alk phos fast red detection. it will stain red, is permanent and can be used on almost every antibody with the exception of ER/PR and if I recall correctly(ventana will tell me if its not) Alk Phos fast Red is a bit more sensitive for detection than dab, the pathologist will adjust i assure you and morphology is saved. dana From int09018 <@t> alphahunt.com Wed Apr 7 16:43:54 2004 From: int09018 <@t> alphahunt.com (HCS) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] looking for Fisher Microprob stain manual Message-ID: <003b01c41ce9$6c739b90$6701a8c0@hp> Dear Histonet, I am setting up my Fisher Microprob and would like to find a manual. Does anyone have a manual they would be willing to share with me? I would be happy to pay you for it too. Thanks in advance, LeRoy Brown HT(ASCP) HTL Histology Consultation Services 360-966-7300 fax 360-543-5626 www.histocs.com From bryand <@t> netbistro.com Wed Apr 7 16:46:54 2004 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Thanks for Help! References: Message-ID: <003601c41ce9$d7935050$99a5b5d0@yourlk4rlmsu41> To measure the specific gravity without a hydrometer: 1. Weigh a container that will take just over 10 mL to 4 decimal places. 2. Using a volumetric pipette, place 10 mL of the methanol into the container. 3. Weigh it again to 4 decimal places. 4. Subtract the weight of the empty container from the weight with the methanol to get the weight of 10 mL methanol. 5. Divide the weight of the methanol by 10 (move 1 decimal place to the left). 6. That is the specific gravity of the methanol. Compare it to a chart of concentrations. Bryan Llewellyn ----- Original Message ----- From: "Mar?a Teresa Dom?nguez" To: Sent: Wednesday, April 07, 2004 12:09 PM Subject: [Histonet] Thanks for Help! > > Well, > > We haven't bought this Methano. I t was a "gift" from a > private lab, they bought some to a new Co. and they sent to us 5 > liters ina big bottle, that's why I couldn't get any information about > this alcohol. > > Now, the problem is we don't have an Hidrometer, so we checked it > with 2 ml. of Methanol in a little tube and 0,5 ml of oil, then I > shaked them, and I could see little drops of oil, like an emulsion, > the solution changed the colour, it turned white with drops in it. > Maybe water?? > > The last step I did. I compair the solution with the Methanol we have > kept in the lab which I know it is 98.8 % pure with 0,1% water. The > drops of oil were quite bit less than in the other solution. What does > it means?? > > I need > someone who knows about chemistry soon!! > > Thanks again for your advices and HAPPY EASTER! for everyone. > H.T. Maria Teresa Dominguez > > Anatomy Pathologyc, > > Zonal Hospital of R?o Grande, > > Tierra del Fuego, Argentina. > _________________________________________________________________ > > MSN 8 with [1]e-mail virus protection service: 2 months FREE* > > References > > 1. http://g.msn.com/8HMBEN/2740??PS= > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From bwhitaker <@t> brownpathology.com Wed Apr 7 17:33:07 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Melanin Bleach & IHC In-Reply-To: <0B6B0E75.4335AFBB.0A1F969F@aol.com> Message-ID: <000601c41cf0$4c768120$3601a8c0@brownpathology.net> There are also other chromogens available for HRP that aren't "DAB brown". I still like AEC, but Vector has several others now. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DDittus787@aol.com Sent: Wednesday, April 07, 2004 4:24 PM To: LBlack@carilion.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Melanin Bleach & IHC lisa; instead of using the bleaching, or worrying that the dab looks like melanin, protect your tissue and the results by using alk phos fast red detection. it will stain red, is permanent and can be used on almost every antibody with the exception of ER/PR and if I recall correctly(ventana will tell me if its not) Alk Phos fast Red is a bit more sensitive for detection than dab, the pathologist will adjust i assure you and morphology is saved. dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Wed Apr 7 18:04:17 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Melanin pigment/DAB Message-ID: A red chromogen is great. I prefer the Permanent Red from Dako, as it is a cherry red & not a brick red. Better contrast from the brown melanin. It is an alk. Phos. Conjugate. Nice with an alk phos polymer detection, in my opinion. Patti Loykasek PhenoPath Laboratories Seattle, WA From katri <@t> cogeco.ca Wed Apr 7 20:41:16 2004 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Melanin Bleach & IHC References: Message-ID: <004001c41d0a$95e97500$ce989618@hala4.on.cogeco.ca> Hi Lisa, I would not recommend the melanin bleach step before immuno. I've done parallels with and without bleach and in some cases you'll lose the immuno reactivity altogether for S100 and HMB45 antibodies. Some cases did stain, but much less and it is not predictable. We use a red chromogen for the cases that are known to have endogenous pigment, therefore not confusing the positive staining with the pigment. Katri Katri Tuomala Department of Pathology St.Joseph's Health Care Hamilton, Ontario, Canada ----- Original Message ----- From: "Lisa Black" To: Sent: Wednesday, April 07, 2004 5:02 PM Subject: [Histonet] Melanin Bleach & IHC > Hello All, > Whenever we bleach a slide prior to IHC staining, the end result is > sub-optimal. Our protocol is to use 0.5% potassium permanganate and > then oxalic acid for the bleach then to proceed with the automated IHC > stain using Ventana's DAB I-View Kit. Most of the tissue comes off the > slide and the remaining tissue has poor morphology. Does anyone have > good advice for me? > Thanks in advance, > Lisa Black > Carilion Consolidated Lab > Roanoke, VA > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ESerrano <@t> imim.es Thu Apr 8 01:46:25 2004 From: ESerrano <@t> imim.es (Serrano del Pozo, Erika) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] alternative to using pap pen? Message-ID: <10708933FD0CD511BCFE000102BE5F72025FAD0C@hpserv.imim.es> Hi Eva, I think that the easier and cheaper is, after putting on the tissue the antibody to the corresponding dilution, place upon the sample a small piece of parafilm (be carefull with air bubbles). Something entertained, but is worth the trouble... Good luck, > Erika Serrano > > Centre de Regulaci? Gen?mica > Differenciation and Cancer Programme > Barcelona -SPAIN- > > > > ---------- > From: Eva Andersson > Sent: Wednesday, April 7, 2004 22:14 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] alternative to using pap pen? > > Hi everyone, > I am going to be undertaking a large immunohistochemistry staining > project. To ensure that our slides do not dry out during the primary > antibody incubation we have decided to use pap pen. With such a large > amount of slides this may be quite a task and so I would like to know if > anyone has found a good alternative to the pap pen. The slides will > arrive with the samples already on them. > Please let me know if you have any suggestions. > Thanks > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Rcartun <@t> harthosp.org Wed Apr 7 18:26:40 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] p21 Message-ID: Has anyone tried Pharmagen's mAb to p21 (clone 6B6)? If so, has it worked well on formalin-fixed, paraffin-embedded tissue? Thank you. Richard Cartun Director, Immunopathology Hartford Hospital Hartford, CT 06102 "Home of the 2004 Men's and Women's Division I College Basketball National Champions!" From S.Stryker <@t> lumc.nl Thu Apr 8 02:43:15 2004 From: S.Stryker <@t> lumc.nl (Stryker, S. (HLK)) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] summary of reactions to porcine insulin Message-ID: summary of reactions to: Hello everybody, I'm hoping someone can give me any suggestions on how to detect insulin on a (paraffin embedded formalin-fixed or frozen) porcine section using the following antibodies: - primary antibody: Guinea Pig anti-insulin (Dako) - secondary antibody: Swine anti-Rabbit (Dako) According to the DAKO catalogous the anti-Rabbit should work in this combo. I have tried it over and over again, using different protocols, but somehow I just can't get the job done... Thanks in advance! Samantha It will never work with this combination of antibodies, what you need(and what I use)is a rabbit ANTI-GUINEA PIG peroxidase conjugated, from DAKO(P0141) as your second antibody.............. Good luck Richard Edwards would use a biotinylated anti-guinea pig antibody. I'm not sure if the anti-rabbit antibody would bind the the GP primary antibody you are using. I'm pretty sure that Vector sells an anti-GP secondary antibody (I think they also sell a guinea pig ABC and alk phos kit, depending on the detection system you are using). Good luck, Kim Merriam' Novartis Cambridge, MA es, a biotinylated anti-rabbit IgG does cross-react with guinea pig antibodies. I use it all the time when I stain for Insulin, using Dako's guinea pig anti-Insulin product. Jan Shivers U of MN Vet Diag Lab Samatha, I have had great success with that antibody. I use it @1:100. I use Goat x Rabbit/Biotinylated @1:100. I run this on the Ventana but you should get similar results maually. Lin Poly-HRP goat anti-rabbit IgG would react well with guinea pig primary antibody. Kilinipath at Netherlands should be able to help. Klinipath: 31-316-250309 attention Marc or Dick. Best wishes, Jincan Samantha, I had problems years ago using an anti-rabbit secondary. I'm attaching our procedure. Robin From mikael.niku <@t> helsinki.fi Thu Apr 8 03:08:03 2004 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] alternative to using pap pen? In-Reply-To: <40746135.5030209@georgetown.edu> Message-ID: <000001c41d40$9df83b60$8c0fd680@ekk1116> Dear Eva, if you are really doing LOTS of slides, I recommend Coverplates by ThermoShandon. We moved from PAP pen to these a few years ago and really love them. The idea is that you put a sort of plastic flow-through chamber on the slide, stick it in a rack, and then you just pipette / squirt the reagents to the chamber one by one. The solutions flow through the chamber into a waste container, and the last 100 ul so stays in the chamber, so the slides cannot get dry. The constant flow appears to be very effective also, so you can significantly reduce both handling time and washing time. It's well possible to process something like 80-100 slides simultaneously. The most significant drawback is that the racks are extremely expensive at least here in Finland (a few hundred $ per a single 10-slide rack!!!!), so buying 10 or so for a single project might be a bit too much. The actual coverplates aren't too expensive (I think about $2 each). They are supposed to be disposable but were are using them until too scratched. You can by nondisposable ones but they cost a lot. On the technical side, putting very many sections on a slide might be problematic due to flow disturbances. That said, I would never go back to PAP pen! +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ + Mikael Niku + University of Helsinki, Dept. Basic Veterinary Sciences + URL: www.helsinki.fi/~mniku/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! (Gandhi) > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf > Of Eva Andersson > Sent: 7. huhtikuuta 2004 23:15 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] alternative to using pap pen? > > > Hi everyone, > I am going to be undertaking a large immunohistochemistry staining > project. To ensure that our slides do not dry out during the primary > antibody incubation we have decided to use pap pen. With such a large > amount of slides this may be quite a task and so I would like > to know if > anyone has found a good alternative to the pap pen. The slides will > arrive with the samples already on them. > Please let me know if you have any suggestions. > Thanks > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listin> fo/histonet > From AFeatherstone <@t> KaleidaHealth.Org Thu Apr 8 07:00:23 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Her 2 Staining Message-ID: What antigen retrieval methods is everyone doing for this antibody? We are currently using Citrate ph 6.0 in a steamer. Our Her 2 is not correlating with subsequent FISH. Annette Featherstone HT/MLT 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 Terry.Marshall <@t> rothgen.nhs.uk Thu Apr 8 07:24:34 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing Message-ID: When you see these "action thrillers" in airplanes, and a bullet/bomb/whatever makes a hole in the fuselage, everything rushes to the hole, not just the air. I can't envisage this process of sucking only the air or "surface air bubbles" and leaving a soggy sponge replete with its load. Under a vacuum, surely the sponge will collapse as the contained liquid runs out towards the exit. If you apply vacuum to a plastic bag containing fruit, the juice comes out, and this forms the basis of a vacuum press, one of which I possess. So there. 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 07 April 2004 16:48 To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing I thought vacuum processing only 'sucked away' air. I mean if you reduce the air pressure above the fluids then that reduces pressure within the tissue that was preventing air escaping (as it has a positive pressure). I suppose if you reduce the Saturated Vapour Pressure of a solvent then that would cause more solvent to vapourise but wouldn't that be from the surface of the solvent rather than within the tissue? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 06 April 2004 16:21 To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 LBlack <@t> carilion.com Thu Apr 8 08:17:33 2004 From: LBlack <@t> carilion.com (Lisa Black) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Re: Melanin Bleach & IHC Message-ID: Thanks to all for responding to my question. All answers were the same. Sincerely, Lisa Black Carilion Consolidated Lab Roanoke, VA From rfail <@t> toolkitmail.com Thu Apr 8 08:36:38 2004 From: rfail <@t> toolkitmail.com (rfail) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] KERMIX Message-ID: <40755566.b0.475.238573615@toolkitmail.com> I have a Pathologist requesting a broad spectrum screening cytokeratin called Kermix. Is anyone using this? Rena Fail From convmcm <@t> cc.usu.edu Thu Apr 8 09:03:18 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] AFB staining on mouse tissue In-Reply-To: Message-ID: <000001c41d72$3f0d4cd0$4a737b81@Cygnus> Your're right. if you try to use a staining procedure on tissue sections that is usually done on a smear, it won't work. Tissue staining usually requires more time, and sometimes entirely different reagents. This is what I use in my lab. Reference is AFIP, 1992 edition SOLUTIONS: Carbol Fuchsin Phenol (melted) ............... 2.5 mL 100% Ethanol .................. 5 mL Basic Fuchsin (CI 42500) ...... 0.5 g DI water ...................... 50 mL Mix in the order given. Filter before use. Methylene Blue (Stock solution) Methylene Blue (CI 52015) ..... 1.4 g DI water ...................... 100 mL Methylene Blue (working solution) Stock Methylene Blue ........... 5 mL Tap water ...................... 45 mL Acid Alcohol 70% Ethanol ................... 99 mL HCl, conc ..................... 1 mL PROCEDURE: Preheat Carbol Fuchsin in a 60 C oven then swirl well BEFORE filtering (this is my own addition to this procedure). 1. Cut sections at 4-5 um 2. Deparaffinize as usual 3. Rinse well in DI water 4. Carbol Fuchsin, 30 minutes at Room Temp 5. Rinse in running tap water to remove excess dye. 6. Dip in acid alcohol until sections turn a pale pink 7. Running tap water, 5 minutes 8. Working methylene blue, 1 or 2 dips. DO NOT OVER DO THIS STEP!!! 9. Dehydrate through graded alcohols to xylene or xyl substitute 10. Mount in the appropriate resin Hope this is helpful Connie McManus Utah Veterinary Diagnostic Laboratory Utah State University Logan, UT From ssq5977 <@t> yahoo.com.cn Thu Apr 8 09:36:23 2004 From: ssq5977 <@t> yahoo.com.cn (=?gb2312?q?=CA=E7=C7=ED=20=C9=F2?=) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] heavy - chain deposition desease Message-ID: <20040408143623.1783.qmail@web15204.mail.bjs.yahoo.com> Hello,everobody: I am very interested in the diagonosis of heavy-chain deposition disease.Firstly ,I am going to find out whether there are deposition of heavy chains along tubular and glomerular basement membranes.But I have no idea about which company's antibodyies are appropriate.Would anybody be kind enough to share me with you experieace? Thanks a lot. Beat Regards! Shuqiong Shen Research Insititute of Nephrology 305 East Zhongshan Road Nanjing 210002,P.R.China --------------------------------- Do You Yahoo!? »ÝÆÕTTÓÎÏ·¾ç£¬ÍæÓÎÏ·£¬Öд󽱣¡ From DRG <@t> Stowers-Institute.org Thu Apr 8 09:41:32 2004 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Whole mount staining wells Message-ID: Hi Histonetters, Does anyone have information on where I may purchase some multiwell plates with mesh screens for whole mount staining? Thanks in advance! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org From Jude.Carpenter <@t> vtmednet.org Thu Apr 8 09:41:23 2004 From: Jude.Carpenter <@t> vtmednet.org (Carpenter, Judith A.) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] methanol Message-ID: <1AC2A27D616B5C4D94FA103BA7F1A41D040DEB5E@FAHC12.fahc.fletcherallen.org> Does anyone have a substitute for methanol in the Wright's stain on blood smears ? Preferably non-hazardous (does not have to be hauled away for disposal). thanks Jude Jude Carpenter, BS, HTL(ASCP) Chief Technologist for Autopsy/Histology/Surgical Pathology 111 Colchester Ave. Burlington, VT 05401 jude.carpenter@vtmednet.org (802)847-5116 fax: (802)847-3509 Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. From gcallis <@t> montana.edu Thu Apr 8 10:10:25 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Melanin and red chromogens for IHC In-Reply-To: <000601c41cf0$4c768120$3601a8c0@brownpathology.net> References: <0B6B0E75.4335AFBB.0A1F969F@aol.com> Message-ID: <3.0.6.32.20040408091025.00bcf9b0@gemini.msu.montana.edu> Vector have NovaRed, HRP substrate, it is a very dark brick red and sensitive as DAB. AEC+ from DAKO or just their AEC is in ready to use form, with AEC+ being quite sensitive, although not as much so as DAB and less so than Alk Phos substrates other than NBT/BCIP (dark blue/purple). The joy of AP substrates is a pink red rather than orange red (AEC) or brick red (Vector NovaRed). You may want to try AP versus HRP for a tinctorial properties your pathologist would like best. DAKO has a new AP chromogen that competes with Vector Red, and this chromogen is permanent. Chris van der Loos has a wonderful chromogen chart in his book on Multiple Staining methods, which gives some guidelines on primary antibody concentration versus the sensitivity of chromogens. We find it extremely useful for both double staining and even single IHC staining. At 05:33 PM 4/7/2004 -0500, you wrote: >There are also other chromogens available for HRP that aren't "DAB brown". >I still like AEC, but Vector has several others now. > >Bonnie Whitaker >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >DDittus787@aol.com >Sent: Wednesday, April 07, 2004 4:24 PM >To: LBlack@carilion.com; Histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Melanin Bleach & IHC > >lisa; >instead of using the bleaching, or worrying that the dab looks like melanin, >protect your tissue and the results by using alk phos fast red detection. it >will stain red, is permanent and can be used on almost every antibody with >the exception of ER/PR and if I recall correctly(ventana will tell me if its >not) Alk Phos fast Red is a bit more sensitive for detection than dab, the >pathologist will adjust i assure you and morphology is saved. dana > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From kemlo <@t> tiscali.co.uk Thu Apr 8 10:13:44 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing In-Reply-To: Message-ID: <002701c41d7c$1ab36090$d20c2850@KEMLOS> But that's cos the air is trying to escape to equilibrate the outside reduction in air pressure. If the airplane was under water and had no air in it and the air pressure was reduced on top of the water then it would have no effect on the airplane, would it? Similarly the fruit in the bag is not under water, is it? Put the fruit under water in the bag and then reduce the pressure above the water, bet no juice comes out of the fruit except if there is trapped air and that forces some out as it exits. Bet you $10 Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 08 April 2004 13:25 To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing When you see these "action thrillers" in airplanes, and a bullet/bomb/whatever makes a hole in the fuselage, everything rushes to the hole, not just the air. I can't envisage this process of sucking only the air or "surface air bubbles" and leaving a soggy sponge replete with its load. Under a vacuum, surely the sponge will collapse as the contained liquid runs out towards the exit. If you apply vacuum to a plastic bag containing fruit, the juice comes out, and this forms the basis of a vacuum press, one of which I possess. So there. 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 07 April 2004 16:48 To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing I thought vacuum processing only 'sucked away' air. I mean if you reduce the air pressure above the fluids then that reduces pressure within the tissue that was preventing air escaping (as it has a positive pressure). I suppose if you reduce the Saturated Vapour Pressure of a solvent then that would cause more solvent to vapourise but wouldn't that be from the surface of the solvent rather than within the tissue? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 06 April 2004 16:21 To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 Terry.Marshall <@t> rothgen.nhs.uk Thu Apr 8 10:41:28 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing Message-ID: Kemlo comes back (well he would): "But that's cos the air is trying to escape to equilibrate the outside reduction in air pressure. If the airplane was under water and had no air in it and the air pressure was reduced on top of the water then it would have no effect on the airplane, would it?" Any explanations as to what this means to me please. He continues: "Similarly the fruit in the bag is not under water, is it? Put the fruit under water in the bag and then reduce the pressure above the water, bet no juice comes out of the fruit except if there is trapped air and that forces some out as it exits." No, the fruit in the bag is not under water, but soon it is under fruit juice. The first thing that happens when you apply vacuum is that the air goes, then the fruit juice exits the fruit. However, there is a difference in the two systems, in that the bag in the vacuum press collapses. The retort hopefully does not. Would this make a difference? My capacity to conceive some of these concepts is quite paltry. "Bet you $10". I don't do bets or dollars. Someone put a car sponge in a VIP and stop it at the appropriate time would you? Terry L (wishing I'd kept my mouth shut) Marshall 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 08 April 2004 16:14 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Bet you $10 Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 08 April 2004 13:25 To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing When you see these "action thrillers" in airplanes, and a bullet/bomb/whatever makes a hole in the fuselage, everything rushes to the hole, not just the air. I can't envisage this process of sucking only the air or "surface air bubbles" and leaving a soggy sponge replete with its load. Under a vacuum, surely the sponge will collapse as the contained liquid runs out towards the exit. If you apply vacuum to a plastic bag containing fruit, the juice comes out, and this forms the basis of a vacuum press, one of which I possess. So there. 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 07 April 2004 16:48 To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing I thought vacuum processing only 'sucked away' air. I mean if you reduce the air pressure above the fluids then that reduces pressure within the tissue that was preventing air escaping (as it has a positive pressure). I suppose if you reduce the Saturated Vapour Pressure of a solvent then that would cause more solvent to vapourise but wouldn't that be from the surface of the solvent rather than within the tissue? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 06 April 2004 16:21 To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 Thu Apr 8 10:54:03 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] AFB staining on mouse tissue In-Reply-To: <000001c41d72$3f0d4cd0$4a737b81@Cygnus> References: Message-ID: <3.0.6.32.20040408095403.00bcf9b0@gemini.msu.montana.edu> To avoid buying, making up and storing stock reagents, buy an AFB staining kit from Newcomer Supply, Richard Allan,Poly Scientific or another reliable source to avoid exposure to some of these chemicals. We used Kinyouns acid fast method, but others work too. Phenol must be worked with under a fume hood, and some dyes are carcinogenic. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From anicoll <@t> cellpath.co.uk Thu Apr 8 10:58:31 2004 From: anicoll <@t> cellpath.co.uk (Alastair Nicoll) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing Message-ID: <0F42D9ABA325D811BBE300902712436D0A4605@cellmail.internal.cellpath.co.uk> Further information on this item can be found here http://www.cellpath-store.co.uk/acatalog/Biopsy_Specimen_Handling.html Perth Saint -----Original Message----- From: KAELPERS@aol.com [mailto:KAELPERS@aol.com] Sent: 07 April 2004 03:20 To: Terry.Marshall@rothgen.nhs.uk; siksik03@comcast.net; gcallis@montana.edu; gudrun.lang@aon.at; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing We use these tiny screen cassettes that fit into processing cassettes, they can be purchased from Mercedes medical I think. They work really well, and another thing neat about them is they have 2 dimensions - the capsule closes 2 different directions and one side is slightly higher than the other to allow slightly bigger biopsies. lge _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo <@t> tiscali.co.uk Thu Apr 8 11:12:59 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing In-Reply-To: Message-ID: <4071A41E0000AB4E@mk-cpfrontend-3.mail.uk.tiscali.com> Explanation is easy Terry...... If you put tissue in a processor that uses vacuum embedding then the vacuum removes the air, nothing else, from the tissue. If you put a hole in an airplane that is pressurised and high up; people and things get sucked through the hole. Don't put fruit in a bag and then put it in a vacuum or it gets wet! heheheheeheh. And yes I would, wouldn't I? __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From kemlo <@t> tiscali.co.uk Thu Apr 8 11:12:59 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing In-Reply-To: Message-ID: <4071A41E0000AB4F@mk-cpfrontend-3.mail.uk.tiscali.com> Explanation is easy Terry...... If you put tissue in a processor that uses vacuum embedding then the vacuum removes the air, nothing else, from the tissue. If you put a hole in an airplane that is pressurised and high up; people and things get sucked through the hole. Don't put fruit in a bag and then put it in a vacuum or it gets wet! heheheheeheh. And yes I would, wouldn't I? __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From Terry.Marshall <@t> rothgen.nhs.uk Thu Apr 8 11:07:08 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing Message-ID: Thanks for the blatant advertising. Two points: Those capsules are so buoyant that two would serve as a life-jacket. Wow, I never realised the metal mesh inserts were so expensive, no wonder I have to argue for one each time they offer me a b..... foam pad. 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: Alastair Nicoll [mailto:anicoll@cellpath.co.uk] Sent: 08 April 2004 16:59 To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Further information on this item can be found here http://www.cellpath-store.co.uk/acatalog/Biopsy_Specimen_Handling.html Perth Saint -----Original Message----- From: KAELPERS@aol.com [mailto:KAELPERS@aol.com] Sent: 07 April 2004 03:20 To: Terry.Marshall@rothgen.nhs.uk; siksik03@comcast.net; gcallis@montana.edu; gudrun.lang@aon.at; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing We use these tiny screen cassettes that fit into processing cassettes, they can be purchased from Mercedes medical I think. They work really well, and another thing neat about them is they have 2 dimensions - the capsule closes 2 different directions and one side is slightly higher than the other to allow slightly bigger biopsies. lge _______________________________________________ Histonet mailing list Histonet@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 Terry.Marshall <@t> rothgen.nhs.uk Thu Apr 8 11:08:18 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing Message-ID: Perhaps a drawing would help:-) 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: Kemlo [mailto:kemlo@tiscali.co.uk] Sent: 08 April 2004 17:13 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Explanation is easy Terry...... If you put tissue in a processor that uses vacuum embedding then the vacuum removes the air, nothing else, from the tissue. If you put a hole in an airplane that is pressurised and high up; people and things get sucked through the hole. Don't put fruit in a bag and then put it in a vacuum or it gets wet! heheheheeheh. And yes I would, wouldn't I? __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From nfitzgi726 <@t> charter.net Thu Apr 8 11:50:09 2004 From: nfitzgi726 <@t> charter.net (nfitzgi726@charter.net) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Job opening Message-ID: <200404081650.i38Go9sY003626@mxsf18.cluster1.charter.net> Washoe Medical Center in Reno, NV ( Recently Voted one of the top 10 places to live in the United States) has a full time opening for an experienced Histotechnologist or Histotechnician. Please apply on line at WWW.Washoehealth.com or call the human resources department 775-982-4719 for more information. From histonet <@t> histonet.blogdns.com Thu Apr 8 12:09:54 2004 From: histonet <@t> histonet.blogdns.com (Judy Pariser) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] xylene substitute References: Message-ID: <003001c41d8c$503a4700$0600a8c0@IN4> Dear Eddie, I just noticed your posting dated March 15, 2004 regarding xylene substitutes. I am the Project Manager for CBG Biotech's new clearing solvent, Formula 83. Formula 83 is completely non-toxic. In clinical and laboratory tests, Formula 83 consistently outperformed xylene and other xylene substitutes. It was designed specifically for tissue processing and staining. Formula 83 dries slides faster, dissolves paraffin faster, and recycles faster than xylene. Additionally, it will not harden tissue as xylene does, and has better lipid extraction. I would be happy to share more information with you. Additionally, I can send to you a reprint of a recently published article from Health Beat, a publication of the Healthcare Division of The American Society of Safety Engineers. I can be reached at the phone number or email address below. Best regards, Judy Pariser Project Manager Email: jpariser@cbgbiotech.com Phone: 800-941-9484 Fax: 614-863-1676 Website: http://www.cbgbiotech.com ----- Original Message ----- From: "Eddie Marquez" To: Sent: Monday, March 15, 2004 4:19 PM Subject: [Histonet] xylene substitute Aloha from Hawaii, Just needed some info.-Has anybody used xylene substitute with a linear stainer; for standard H&E's? If so, how good are the results? May I also have the brand name. Thank you, Eddie from the Cancer Research _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vbaker60 <@t> yahoo.com Thu Apr 8 12:51:56 2004 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Looking for Guss Mondragon Message-ID: <20040408175156.12088.qmail@web12108.mail.yahoo.com> Hi Everyone, Guss if you're out there I need some of your sage wisdom. OR If anyone knows where Guss might be please contact me by this e-mail address. Thanks in advance. Vikki Baker Institute for Cancer Prevention Valhalla, NY 10595 __________________________________ Do you Yahoo!? Yahoo! Small Business $15K Web Design Giveaway http://promotions.yahoo.com/design_giveaway/ From laurie.colbert <@t> huntingtonhospital.com Thu Apr 8 13:06:54 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] CSH Symposium (May 13-16, 2004) - Hotel Phone Number Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BE91@EXCHANGE1.huntingtonhospital.com> The phone number that was listed for hotel reservations at the Burbank Hilton on the California Society for Histotechnology registration form is incorrect. The correct number is (800) 840-6450. We are sorry for any inconvenience this has caused. If you have any questions please contact Debbie Cobb at (818) 503-6807. From jqb7 <@t> cdc.gov Thu Apr 8 13:14:23 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] p21 Message-ID: Please share any information with the list please. Thanks! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Richard Cartun Sent: Wed 4/7/2004 7:26 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] p21 Has anyone tried Pharmagen's mAb to p21 (clone 6B6)? If so, has it worked well on formalin-fixed, paraffin-embedded tissue? Thank you. Richard Cartun Director, Immunopathology Hartford Hospital Hartford, CT 06102 "Home of the 2004 Men's and Women's Division I College Basketball National Champions!" _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Thu Apr 8 13:31:43 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:46 2005 Subject: [Histonet] Sponge problems in processing Message-ID: <494304423C63E246A5CF87A3AEEB577011B5DE@bumail01.barrynet.barry.edu> If an object is under water and the air pressure above the water is reduced, the total pressure on the object is reduced. If the object is only 17 feet down (1/2 Atm water pressure plus 1 Atm air pressure), halving the air pressure above it reduces the total pressure by 33% (1 Atm instead of 1 1/2 Atm). If the air pressure above it is reduced by 97% (to 0.03 Atm), the water boils at room temperature! If you seal the bag of fruit at normal atmospheric pressure, put the bag 17 feet underwater, and reduce the air pressure above by 50%, the fruit does not release juice because the total pressure (1 Atm) is still as high as when it was sealed. However, if you reduce the air pressure by 95%, the total pressure on the fruit will be 0.55 Atm. If you reduce the pressure to 0.55 Atm suddenly, air dissolved in the fruit juice will rupture the fruit and juice will leak out. If you reduce the pressure slowly to 0.55 Atm, the air will diffuse out without rupturing the fruit. If you slowly reduce the pressure on the fruit to 0.03 Atm, the water will diffuse out slowly, and you will have intact dehydrated fruit. If you reduce the pressure on the fruit 0.03 Atm suddenly, the rapid boiling of the juice will rupture the fruit and some juice will leak out before it, too, boils. Allen A. Smith, Ph.D. Professor of Anatomy School of Graduate Medical Sciences Barry University Miami Shores, FL (also PADI divemaster) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, April 08, 2004 11:41 AM To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Kemlo comes back (well he would): "But that's cos the air is trying to escape to equilibrate the outside reduction in air pressure. If the airplane was under water and had no air in it and the air pressure was reduced on top of the water then it would have no effect on the airplane, would it?" Any explanations as to what this means to me please. He continues: "Similarly the fruit in the bag is not under water, is it? Put the fruit under water in the bag and then reduce the pressure above the water, bet no juice comes out of the fruit except if there is trapped air and that forces some out as it exits." No, the fruit in the bag is not under water, but soon it is under fruit juice. The first thing that happens when you apply vacuum is that the air goes, then the fruit juice exits the fruit. However, there is a difference in the two systems, in that the bag in the vacuum press collapses. The retort hopefully does not. Would this make a difference? My capacity to conceive some of these concepts is quite paltry. "Bet you $10". I don't do bets or dollars. Someone put a car sponge in a VIP and stop it at the appropriate time would you? Terry L (wishing I'd kept my mouth shut) Marshall 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 08 April 2004 16:14 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Bet you $10 Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 08 April 2004 13:25 To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing When you see these "action thrillers" in airplanes, and a bullet/bomb/whatever makes a hole in the fuselage, everything rushes to the hole, not just the air. I can't envisage this process of sucking only the air or "surface air bubbles" and leaving a soggy sponge replete with its load. Under a vacuum, surely the sponge will collapse as the contained liquid runs out towards the exit. If you apply vacuum to a plastic bag containing fruit, the juice comes out, and this forms the basis of a vacuum press, one of which I possess. So there. 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 07 April 2004 16:48 To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing I thought vacuum processing only 'sucked away' air. I mean if you reduce the air pressure above the fluids then that reduces pressure within the tissue that was preventing air escaping (as it has a positive pressure). I suppose if you reduce the Saturated Vapour Pressure of a solvent then that would cause more solvent to vapourise but wouldn't that be from the surface of the solvent rather than within the tissue? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 06 April 2004 16:21 To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From RizoC <@t> chi.osu.edu Thu Apr 8 13:45:42 2004 From: RizoC <@t> chi.osu.edu (Rizo, Christian) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Thermo Shandon automatic stainer Message-ID: <3F8707A1ADC19C4FA84BC95B51CEF560094BA0EB@chi2kms03.columbuschildrens.net> Hi everybody, Can anybody share to me their experiences with a Thermo Shandon Automatic Stainer? We are trying to purchase one with a good deal. Thanks for your help. Chris Christian M. Rizo MBA, HTL (ASCP) Manager, Anatomic Pathology Childrens Hospital 700 Childrens Drive Columbus, Ohio 43205 614.722.5465 RizoC@chi.osu.edu Confidentiality Notice: This e-mail message, from Children's Hospital, Columbus, Ohio, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes pursuant to Children's Hospital's confidentiality policies. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. From Stacy_McLaughlin <@t> cooley-dickinson.org Thu Apr 8 13:52:46 2004 From: Stacy_McLaughlin <@t> cooley-dickinson.org (Stacy McLaughlin) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Sponge problems in processing Message-ID: <3D502BBF5356D31184650090275B750D0346C74D@mail.cooley-dickinson.org> Can't we all just have our fruit juice in a glass? :) -----Original Message----- From: Smith, Allen [mailto:asmith@mail.barry.edu] Sent: Thursday, April 08, 2004 1:32 PM To: Marshall Terry Dr, Consultant Histopathologist Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing If an object is under water and the air pressure above the water is reduced, the total pressure on the object is reduced. If the object is only 17 feet down (1/2 Atm water pressure plus 1 Atm air pressure), halving the air pressure above it reduces the total pressure by 33% (1 Atm instead of 1 1/2 Atm). If the air pressure above it is reduced by 97% (to 0.03 Atm), the water boils at room temperature! If you seal the bag of fruit at normal atmospheric pressure, put the bag 17 feet underwater, and reduce the air pressure above by 50%, the fruit does not release juice because the total pressure (1 Atm) is still as high as when it was sealed. However, if you reduce the air pressure by 95%, the total pressure on the fruit will be 0.55 Atm. If you reduce the pressure to 0.55 Atm suddenly, air dissolved in the fruit juice will rupture the fruit and juice will leak out. If you reduce the pressure slowly to 0.55 Atm, the air will diffuse out without rupturing the fruit. If you slowly reduce the pressure on the fruit to 0.03 Atm, the water will diffuse out slowly, and you will have intact dehydrated fruit. If you reduce the pressure on the fruit 0.03 Atm suddenly, the rapid boiling of the juice will rupture the fruit and some juice will leak out before it, too, boils. Allen A. Smith, Ph.D. Professor of Anatomy School of Graduate Medical Sciences Barry University Miami Shores, FL (also PADI divemaster) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, April 08, 2004 11:41 AM To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Kemlo comes back (well he would): "But that's cos the air is trying to escape to equilibrate the outside reduction in air pressure. If the airplane was under water and had no air in it and the air pressure was reduced on top of the water then it would have no effect on the airplane, would it?" Any explanations as to what this means to me please. He continues: "Similarly the fruit in the bag is not under water, is it? Put the fruit under water in the bag and then reduce the pressure above the water, bet no juice comes out of the fruit except if there is trapped air and that forces some out as it exits." No, the fruit in the bag is not under water, but soon it is under fruit juice. The first thing that happens when you apply vacuum is that the air goes, then the fruit juice exits the fruit. However, there is a difference in the two systems, in that the bag in the vacuum press collapses. The retort hopefully does not. Would this make a difference? My capacity to conceive some of these concepts is quite paltry. "Bet you $10". I don't do bets or dollars. Someone put a car sponge in a VIP and stop it at the appropriate time would you? Terry L (wishing I'd kept my mouth shut) Marshall 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 08 April 2004 16:14 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Bet you $10 Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 08 April 2004 13:25 To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing When you see these "action thrillers" in airplanes, and a bullet/bomb/whatever makes a hole in the fuselage, everything rushes to the hole, not just the air. I can't envisage this process of sucking only the air or "surface air bubbles" and leaving a soggy sponge replete with its load. Under a vacuum, surely the sponge will collapse as the contained liquid runs out towards the exit. If you apply vacuum to a plastic bag containing fruit, the juice comes out, and this forms the basis of a vacuum press, one of which I possess. So there. 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 07 April 2004 16:48 To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing I thought vacuum processing only 'sucked away' air. I mean if you reduce the air pressure above the fluids then that reduces pressure within the tissue that was preventing air escaping (as it has a positive pressure). I suppose if you reduce the Saturated Vapour Pressure of a solvent then that would cause more solvent to vapourise but wouldn't that be from the surface of the solvent rather than within the tissue? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 06 April 2004 16:21 To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet THIS E-MAIL MESSAGE IS INTENDED ONLY FOR THE USE OF THE INDIVIDUAL OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If the reader of this e-mail message is 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 e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. From DDittus787 <@t> aol.com Thu Apr 8 13:58:36 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Sponge problems in processing Message-ID: <2F934444.658092DE.0A1F969F@aol.com> owwwwwwwwwwwwwww my head hurts, stop using sponges pleaseeeeeeeeee. just a little humor this was getting a bit intense. Dana From Gillian.Barlow <@t> cshs.org Thu Apr 8 13:58:49 2004 From: Gillian.Barlow <@t> cshs.org (Barlow, Gillian) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Antigen retrieval for heart tissues? Message-ID: <3CFAA0108952D111A5BF00805FA6FB0F0794750E@PEDSNTAS.csmc.edu> Dear Histonetters Does anyone out there know what (if any) antigen retrieval is necessary for the following antibodies in sections of paraffin-fixed, formalin-embedded adult mouse heart: - Cx43 - alpha-actinin - beta-catenin Many thanks Gillian Gillian M. Barlow, PhD Postdoctoral Fellow Laboratory of Julie Korenberg, PhD, MD Cedars-Sinai Medical Center Davis Bldg, Lab 2007 110 George Burns Rd Los Angeles, CA 90048 Phone: (310) 423 7650 Fax: (310) 423 0302 From funderwood <@t> mcohio.org Thu Apr 8 14:47:00 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:47 2005 Subject: [BULK] - Re: [Histonet] Sponge problems in processing Message-ID: It's best to follow the lead of Elaine Benes and determine just which specimens are sponge worthy. >>> 04/08/04 02:58PM >>> owwwwwwwwwwwwwww my head hurts, stop using sponges pleaseeeeeeeeee. just a little humor this was getting a bit intense. Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From djohnson14 <@t> hotmail.com Thu Apr 8 16:18:28 2004 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Histology opening in S CA. Message-ID: Attention Histonet!, New Histology Lab in Southern Califonia has job opening for part time or fulltime histology technician. Duties include all applications for operating paraffin histology laboratory. Laboratory is processing specimens for Dermatology Practice. Please send your resume to the attention of Jane Litz, Administrator Manager, Fax: 760-724-9929. Candidate needed to fill position immediately. > Thanks >Rudy Gutierrez >Pacific Southwest Lab Equipment Inc. >Product and Sales Division >Main: 760-295-1842 >Cellular: 760-525-9071 > _________________________________________________________________ Limited-time offer: Fast, reliable MSN 9 Dial-up Internet access FREE for 2 months! http://join.msn.com/?page=dept/dialup&pgmarket=en-us&ST=1/go/onm00200361ave/direct/01/ From amosbrooks <@t> earthlink.net Thu Apr 8 17:19:03 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 In-Reply-To: <200404080720.1bbAoB5LE3NZFl50@vulture> References: <200404080720.1bbAoB5LE3NZFl50@vulture> Message-ID: <6.0.0.22.0.20040408170800.01b704e8@pop.earthlink.net> Linda, Sorry to say there's no such thing. VEGF, despite any specifications, just doesn't work properly. We've tried at least 4 different vendors with hardly any success. Either it didn't label at all, labeled in all the wrong places, or the negative control looked the same as the test tissue. Check the controls closely if you try this. It's just not a reliable antibody. Regards, Amos Brooks At 09:20 AM 4/8/2004, you wrote: >Message: 2 >Date: Wed, 7 Apr 2004 13:00:57 -0500 >From: "Sebree Linda A." >Subject: [Histonet] VEGF antibody >To: "Histonet (E-mail)" >Message-ID: > >Content-Type: text/plain; charset="iso-8859-1" > >Hi again, > >Wondering if anyone can recommend a good, reliable VEGF (vascular >endothelial growth factor) antibody? If you've run yours on a Ventana >instrument, so much the better. > >Thanks for the 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 From gudrun.lang <@t> aon.at Thu Apr 8 16:34:01 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Re: semithin-ultrathin - thank you References: Message-ID: <005f01c41db1$35901b30$eeeea8c0@SERVER> Thank you for your explanations about the thickness of sections. Gudrun Lang, Austria PS.: I read the sponge-debate with interest. ----- Original Message ----- From: "David Kelly, M.D." To: "'Gudrun Lang'" Sent: Thursday, April 08, 2004 12:04 AM Subject: RE: [Histonet] semithin-ultrathin > Tim Morken's description of these terms is accurate. If you need a > reference, the book is old and probably out of print but still an excellent > introduction to electron microscopy. Appropriately, it is entitled "An > Introduction to Diagnostic Electron Microscopy," by Bruce Mackay > (Appleton-Century-Crofts, New York, 1981). See Chapter 2 Technical > Procedures, by Paul S. Baur and Bruce Mackay, p 40 and Chapter 3 > Preparation and Interpretation of Semithin Sections, by Willard A. Burns, pp > 47-48. > > -----Original Message----- > From: Gudrun Lang [mailto:gudrun.lang@aon.at] > Sent: Wednesday, April 07, 2004 2:06 PM > To: Histonetliste > Subject: [Histonet] semithin-ultrathin > > > Hi, > I've searched in the internet for a list that explains the thickness of > semi-, ultra-, thin, thick sections. But I failed. My knowledge is only > approximately. > Perhaps someone can help me? > Gudrun Lang > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gcallis <@t> montana.edu Thu Apr 8 16:34:41 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Interesting eosinophilic artifacts Message-ID: <3.0.6.32.20040408153441.00bc20c0@gemini.msu.montana.edu> After all my years doing histo work, I have one H&E tissue section, mouse lung which shows crystalline shaped eosinophilic artifacts. I stained other lung sections from this experiment and murine reproductive tract, placenta at the same time. These have no artifacts. Some of the crystals actually look ingested by macrophages. If anyone is interested, I will be happy to drop photo privately. I have never seen artifacts like these. Baffled in Montana! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Thu Apr 8 16:48:23 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Eosinophilic artifacts posted in Histonet gallery Message-ID: <3.0.6.32.20040408154823.00ba3030@gemini.msu.montana.edu> Photo of eosinophilic crystalline structures to be posted in Histonet gallery. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From pruegg <@t> colobio.com Thu Apr 8 16:48:28 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 In-Reply-To: <6.0.0.22.0.20040408170800.01b704e8@pop.earthlink.net> Message-ID: Amos I had the same experience as you with VEGF's until I switched to monoclonal Vegf from Santa Cruz, it now appears very specific and reliable to me. I use pepsin or proteinase K digestion and stay away from HIER. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amos Brooks Sent: Thursday, April 08, 2004 4:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 Linda, Sorry to say there's no such thing. VEGF, despite any specifications, just doesn't work properly. We've tried at least 4 different vendors with hardly any success. Either it didn't label at all, labeled in all the wrong places, or the negative control looked the same as the test tissue. Check the controls closely if you try this. It's just not a reliable antibody. Regards, Amos Brooks At 09:20 AM 4/8/2004, you wrote: >Message: 2 >Date: Wed, 7 Apr 2004 13:00:57 -0500 >From: "Sebree Linda A." >Subject: [Histonet] VEGF antibody >To: "Histonet (E-mail)" >Message-ID: > >Content-Type: text/plain; charset="iso-8859-1" > >Hi again, > >Wondering if anyone can recommend a good, reliable VEGF (vascular >endothelial growth factor) antibody? If you've run yours on a Ventana >instrument, so much the better. > >Thanks for the 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Luis.Chiriboga <@t> med.nyu.edu Thu Apr 8 16:54:48 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 In-Reply-To: <6.0.0.22.0.20040408170800.01b704e8@pop.earthlink.net> Message-ID: I agree strongly with Amos and have basically gotten the same results. it's a pain in the neck (and I don't mean from looking in the scope). I have run a couple, from different manufacturers, tried all the usual tricks(automated and not) and none of them performed consistently. I figured I would just pick one and stick with and just go the brute force approach, use it until I get results that I can live with. Hope this helps, regards luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amos Brooks Sent: Thursday, April 08, 2004 6:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 Linda, Sorry to say there's no such thing. VEGF, despite any specifications, just doesn't work properly. We've tried at least 4 different vendors with hardly any success. Either it didn't label at all, labeled in all the wrong places, or the negative control looked the same as the test tissue. Check the controls closely if you try this. It's just not a reliable antibody. Regards, Amos Brooks At 09:20 AM 4/8/2004, you wrote: >Message: 2 >Date: Wed, 7 Apr 2004 13:00:57 -0500 >From: "Sebree Linda A." >Subject: [Histonet] VEGF antibody >To: "Histonet (E-mail)" >Message-ID: > >Content-Type: text/plain; charset="iso-8859-1" > >Hi again, > >Wondering if anyone can recommend a good, reliable VEGF (vascular >endothelial growth factor) antibody? If you've run yours on a Ventana >instrument, so much the better. > >Thanks for the 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Thu Apr 8 17:07:11 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 In-Reply-To: <6.0.0.22.0.20040408170800.01b704e8@pop.earthlink.net> Message-ID: Amos & all - I couldn't have said it better myself. The only difference is that we have tried >6 clones. Be very careful of the staining, read up on the expected positivity, and pick your positive controls carefully. Patti Loykasek > Linda, > Sorry to say there's no such thing. VEGF, despite any > specifications, just doesn't work properly. We've tried at least 4 > different vendors with hardly any success. Either it didn't label at all, > labeled in all the wrong places, or the negative control looked the same as > the test tissue. Check the controls closely if you try this. It's just not > a reliable antibody. > Regards, > Amos Brooks > > At 09:20 AM 4/8/2004, you wrote: >> Message: 2 >> Date: Wed, 7 Apr 2004 13:00:57 -0500 >> From: "Sebree Linda A." >> Subject: [Histonet] VEGF antibody >> To: "Histonet (E-mail)" >> Message-ID: >> >> Content-Type: text/plain; charset="iso-8859-1" >> >> Hi again, >> >> Wondering if anyone can recommend a good, reliable VEGF (vascular >> endothelial growth factor) antibody? If you've run yours on a Ventana >> instrument, so much the better. >> >> Thanks for the 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gsp26 <@t> drexel.edu Thu Apr 8 17:20:18 2004 From: gsp26 <@t> drexel.edu (gsp26@drexel.edu) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] caspase 3 and transferrin receptor for rats Message-ID: <1d855f81d8788c.1d8788c1d855f8@drexel.edu> Has anyone used caspase 3 IHC on rat paraffin embedded tissue. the same question for transferrin receptor IHC. Please advise!! From cfavara <@t> niaid.nih.gov Thu Apr 8 17:26:08 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] caspase 3 and transferrin receptor for rats Message-ID: I have done cleaved caspase 3 on FFPE mouse tissue Source: Cell Signaling #9961 c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: gsp26@drexel.edu [mailto:gsp26@drexel.edu] Sent: Thursday, April 08, 2004 4:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] caspase 3 and transferrin receptor for rats Has anyone used caspase 3 IHC on rat paraffin embedded tissue. the same question for transferrin receptor IHC. Please advise!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dixon.Leslie <@t> mayo.edu Thu Apr 8 17:34:01 2004 From: Dixon.Leslie <@t> mayo.edu (Dixon, Leslie E.) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Whipf's polychrome Message-ID: Good afternoon, I am curious if anyone has the procedure for Whipf's polychrome? Thanks in advance, Leslie From leopold <@t> mnsi.net Fri Apr 9 17:51:44 2004 From: leopold <@t> mnsi.net (Derek & Lynda Leopold) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] bcl-2 on Nexes Message-ID: <000801c41e85$3bc3fc40$3643fea9@leopold> Hi All, We're having some trouble getting our tonsil control to come up for bcl-2. After searching the archives and asking around, we are going to try citrate buffer (Declere) and a hot steamer for 1 hour. We've got the Nexes IHC. Anyone else have any tips for us? Can anyone comment on the need/non-need for DAB? Thanks Lynda Leopold From Linresearch <@t> aol.com Thu Apr 8 17:45:23 2004 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Perfusion System Message-ID: Hello, Anyone with experience using the Perfusio 1 System from MyNeuroLab, Could you give me feedback on the sytem? If someone has a better system, could you let me know? Lin From scoop <@t> mail.nih.gov Thu Apr 8 18:48:23 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] ALAS Message-ID: Hi, Does anyone know of a source for antisera to aminolevulinic acid synthase? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From scoop <@t> mail.nih.gov Thu Apr 8 18:51:06 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] lysosomal markers Message-ID: Hi, Does anyone know of an antibody/antiserum to a lysosomal marker such as Lamp 1 or Lamp 2 that cross reacts with mouse and works on formalin-fixed paraffin embedded tissue (with or without antigen retrieval)? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From katri <@t> cogeco.ca Thu Apr 8 21:23:23 2004 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] heavy - chain deposition desease References: <20040408143623.1783.qmail@web15204.mail.bjs.yahoo.com> Message-ID: <009901c41dd9$a538fa60$ce989618@hala4.on.cogeco.ca> Hi! Are you working on human or animal tissue? In human renal biopsies most commonly used technique is probably immunofluorescence on snap frozen tissue. We use FITC conjugated antibodies from DakoCytomation for heavy chains IgA, IgG and IgM in addition to C3complement, fibrinogen, kappa & lambda light chains and albumin. These can be demonstrated also on FFPE tissue, but the method is more involved and takes more time to develop to obtain reproducible results. Immunofluorescence is a long standing and a simple technique to perform... Katri Katri Tuomala Department of Pathology St.Joseph's Health Care Hamilton, Ontario, Canada ----- Original Message -----=20 From: "=CA=E7=C7=ED =C9=F2" To: Sent: Thursday, April 08, 2004 10:36 AM Subject: [Histonet] heavy - chain deposition desease > Hello,everobody: > I am very interested in the diagonosis of heavy-chain deposition disease.Firstly ,I am going to find out whether there are deposition of heavy chains along tubular and glomerular basement membranes.But I have= no idea about which company's antibodyies are appropriate.Would anybody be kind enough to share me with you experieace? > Thanks a lot. > Beat Regards! > Shuqiong Shen > Research Insititute of Nephrology > 305 East Zhongshan Road > Nanjing 210002,P.R.China > > > > > --------------------------------- > Do You Yahoo!? > =BB=DD=C6=D5TT=D3=CE=CF=B7=BE=E7=A3=AC=CD=E6=D3=CE=CF=B7=A3=AC=D6=D0=B4= =F3=BD=B1=A3=A1 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DDDeltour <@t> sig.med.navy.mil Fri Apr 9 04:52:51 2004 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? Message-ID: Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. From Barry.R.Rittman <@t> uth.tmc.edu Fri Apr 9 07:13:29 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635A00@UTHEVS3.mail.uthouston.edu> Douglas Although I do not have all the details, I do not believe that you are approaching this from the right direction. If you are only doing 800 cases a year what is occupying the rest of your time there? If you have "time on your hands" then the solution is up to you. It is not only "hands on" but also researching the literature and taking courses that will not only maintain but improve your skills. Is it possible for you to start a small project that allows you to explore new methods for demonstrating topics that you are interested in? Are you able to take courses? I was in a similar situation in England and loved it as I was able to expand my knowledge beyond the work for which I was responsible. This was not only a stimulating experience but also improved my perspective and appreciation for the routine work I had to carry out. If you have the time and facilities, I would take this as a golden opportunity to not only maintain your skills but also broaden your horizons. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Deltour, Douglas D.(HM2) Sent: Friday, April 09, 2004 4:53 AM To: Histonet (E-mail) Subject: [Histonet] Can you lose your skill? Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Apr 9 07:39:38 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? Message-ID: My advise is to look at this from a different angle. Sounds like you have a golden opportunity to expand your knowledge! First of all there might be internet courses available that will benefit you histologically. Secondly, you can review cases - slides and report, gather text books, and learn more histology/pathology than you would have an opportunity to do in a busier lab setting. I would become best friends with the pathologist and get him to work with you too! Then I'd learn transcription. The more skills the better. Next I would check out the clinical side of the lab. If possible, I'd learn phlebotomy, and how to do some of the testing. It's good to know how it all fits together. Then, I would visit all of Europe that you possibly could in your off time. That experience will provide you with memories to last your life time. As far as losing your skills, it's just like riding a bicycle. They always come back, even when they've not been used at all! Best wishes for your happiness! j:>) (who, by the way, is older than dirt and has been there/done some of that!) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Deltour, Douglas D.(HM2) Sent: Friday, April 09, 2004 5:53 AM To: Histonet (E-mail) Subject: [Histonet] Can you lose your skill? Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Jackie.O'Connor <@t> abbott.com Fri Apr 9 08:59:12 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? Message-ID: Petty Officer Deltour: I've been a histotech since 1970. My husband is a retired Navy Officer. I followed him all over the world for 28 years beginning in 1975. I've worked a gamut of places from Florida to Hawaii, never being in one place more than 3 years (he spent a lot of time in Sigonella as well - without me, however). I've worked a variety of places from an abandoned toxicology research lab in Memphis to a 600+ bed hospital in Honolulu. My skills increased with the learning experience presented at each facility. I've seen and done almost everything. It's turned me into a pretty good troubleshooter, if I do say so myself. I even took two years off to have our first two kids - but as you well know, on military pay - I needed to go back to work - I didn't lose my skills during that two year hiatus. If you look at each new job as a learning experience instead of comparing this job to the last one, you'll be a happy histotech. Try to figure out how you can improve something in this lab - and make that your legacy. Best of luck to you. Jacqueline M. O'Connor HT(ASCP) and retired Navy wife Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics "Deltour, Douglas D.(HM2)" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/09/2004 04:52 AM To: "Histonet (E-mail)" cc: Subject: [Histonet] Can you lose your skill? Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Fri Apr 9 09:03:05 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? In-Reply-To: Message-ID: <000001c41e3b$62014290$4a737b81@Cygnus> I understand your frustration in doing the routine day in and day out. When I came back into histology, the pathologist who hired me was in his -- get this -- 70's. He had NO interest in doing IHC, PCR or anything cool. He occasionally ordered a Toluidine Blue or a GMS stain and a really big day was 20 -30 H&E's. Most of the time I sat around surfing the internet, playing solitaire and doing personal stuff. I had very little to do. The pathologist finally retired at age 75 and we got our current boss, who quadrupled the number of H&E's, got me going on IHC, created a molecular lab with PCR and microarrays. This was a nightmare, but that's growth. I think you can lose skills, but it's only temporary. It's like riding a bike, you can get them back when the need arises. I gained my HT in 1973 and worked as a histotech continually until 1982. I then had a hiatus from histology when I went back to school, got my BS in biology, learned EM, microbiology, virology - cell culture and 10 years later, came back to histology. I had no problem whatsoever in cutting sections, staining (specials or H&E). I wisely did NOT let my ASCP registration lapse, even though at times it seemed like a waste of money to send in those dues each year, especially when I was in college and money was tight. Of all the diverse lab experiences I've had, I love histology the best. Believe me, if it's something you really love and enjoy (and I'm assuming you love histology or you wouldn't be so concerned about losing your skills), when the time comes to use them, you'll have them. In the mean time, keep current on technology in the field, even if you don't use it on the job. Keep going to NSH meetings, local histology meetings, etc. Read, read and read some more. You will survive. *G* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Deltour, Douglas D.(HM2) Sent: Friday, April 09, 2004 2:53 AM To: Histonet (E-mail) Subject: [Histonet] Can you lose your skill? Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BlazekL <@t> childrensdayton.org Fri Apr 9 09:38:34 2004 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? Message-ID: I agree with everyone about spending your spare time learning new and perfecting old skills and procedures. I have been a histotech since 1973 with a seven year break. When I returned to histology, within a couple of weeks it was like I had never left. I have spent the last 18 years learning every new skill I could find (including transcription). Today though, I am wondering if that was so smart as I am the only person holding down the fort. The transcriptionist is ill, one tech has the day off and who knows why the other tech didn't show up today. We have had two frozens so far with another looming and several special stains that need done. I think I need roller skates today! Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Deltour, Douglas D.(HM2)" 4/9/2004 5:52:51 AM >>> Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tracey.couse <@t> ibb.gatech.edu Fri Apr 9 11:01:09 2004 From: tracey.couse <@t> ibb.gatech.edu (Tracey Couse) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Re: caspase 3 and transferrin receptor for rats In-Reply-To: <200404090219.i392Jg3b026728@katrina.ibb.gatech.edu> Message-ID: <5.0.0.25.2.20040409105501.00a5ed40@mail.ibb.gatech.edu> I have successfully used Zymed's anti-transferrin receptor (#136800) with hier. Tracey >Message: 19 >Date: Thu, 08 Apr 2004 18:20:18 -0400 >From: gsp26@drexel.edu >Subject: [Histonet] caspase 3 and transferrin receptor for rats >To: histonet@lists.utsouthwestern.edu >Message-ID: <1d855f81d8788c.1d8788c1d855f8@drexel.edu> >Content-Type: text/plain; charset=us-ascii > > >Has anyone used caspase 3 IHC on rat paraffin embedded tissue. the same >question for transferrin receptor IHC. Please advise!! > From vbaker60 <@t> yahoo.com Fri Apr 9 10:13:09 2004 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? In-Reply-To: Message-ID: <20040409151309.99252.qmail@web12107.mail.yahoo.com> Dear Douglas, You are in the military, they send you where they need to send you, when they need to send you. The words I recollect were "I'm not to question why...". The shift in the case load number or complexity of the work they do is not always relevant, it's just procedure. Look at this as an opportunity to further yourself on YOUR OWN terms. In someways they have given you an opportunity that may never come your way again. No I haven't served in the armed forces, but I have close friends who have and sometimes also felt somewhat frustrated at the military system du jour. It has no reflection on your ability only the military needs to have a competent warm body where they need them. Do not let this deter or discourage you in any way. In the meantime, soak up all you can by reading or seeing what is available from the military in terms of courses. Italy has some fine university settings that you can also look into. The hands on experience, as far as I can tell they will never be able to automate the very basic skill each histologist holds so very valuable. Our professional integrity to do the very best we can under any circumstances. You could surprise yourself at what you can remember about a stain even if you hadn't done it for a year or for that matter 10 years when you do need to do it. It's a part of you, histology, what you decide to make of it is entirely up to you. Just like life itself. Vikki Baker Institute for Cancer Prevention Valhalla, New York __________________________________ Do you Yahoo!? Yahoo! Small Business $15K Web Design Giveaway http://promotions.yahoo.com/design_giveaway/ From mhagerty <@t> annenberg.net Fri Apr 9 10:22:04 2004 From: mhagerty <@t> annenberg.net (Hagerty, Marg) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Looking for On-the-job training Message-ID: <8E5792BDBF9CD51188CC00034795A15CB902EC@ANN6> I have a friend who really wants to become a Histotech and would like to find a hospital willing to do on-the-job training. He has appropriate education and is bright with a great work ethic. He would like to relocate to Southern California, however, is willing to go anywhere, but needs to make some money while training. If you know of any hospitals that might be hiring, I would greatly appreciate the lead. Many thanks, Marg Hagerty, HT, HTL Eisenhower Medical Center From gcallis <@t> montana.edu Fri Apr 9 10:35:15 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? No! In-Reply-To: Message-ID: <3.0.6.32.20040409093515.00bd0540@gemini.msu.montana.edu> Douglas, Listen carefully to the advice handed out by these people. Read, Read, Read, surf historelated websites, collect publications and improve your theory, methods and materials, etc and if anything, be prepared and not overwhelmed on performance of skills you are not comfortable with e.g. immunostaining or molecular biology technics. Remove the boredom and worry about losing skills - you need to be more concerned about advancing older and learning new skills which you probably will perform in less time than you think. Modern histotechnology is here. Been here, done that so many times I can't count and still do it! Even basic workshops on topics you know are fun just to see how people do things differently than you plus technical ideas/comparisons are exchanged. As for being old as dirt, yup! That's me! But still learning new and fine tuning old skills that come back to visit me after years of not doing them and ready for new challenges, the real fun in this work. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Diane.Gladney <@t> se.amedd.army.mil Fri Apr 9 10:26:34 2004 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:22:47 2005 Subject: FW: [Histonet] Can you lose your skill? Message-ID: <9D41AB7C56F8304F98537ABD87B249F66261C2@DASMTHGBZ001> I meant to send this reply to the Histonet also. Sorry for the mistake. Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: Gladney, Diane C Ms MACH Sent: Friday, April 09, 2004 8:43 AM To: 'Deltour, Douglas D.(HM2)' Subject: RE: [Histonet] Can you lose your skill? Douglas, I disagree with you. I am the wife of a retired Army NCO. We traveled all over the world and I had to take whatever lab position was available. I have worked in Blood Chemistry, and even worked in an Army Oil Analysis Lab in Panama. I couldn't get a job at the Army Hospital in Panama because of the Panama Canal Treaty...the locals had 90% hire status because of the treaty. The only reason that I got the job that I did have was because it required a security clearance and the locals could not get the clearance. My lab chemistry background made me qualified to work in a job that I had never had any kind of exposure to do. With a little training, I was highly proficient at that job. You will find that the lab science background may find you working in jobs that you never imagined. My first love has always been Histology, but over my 25 years of civil service (33 total years in lab field) I have worked several jobs in different lab environments. I never lost any of my skills. I have performed all duties in the Histology Lab. You may be a little slow at things at first if you haven't done them in a long time, but it is like riding a bicycle....you never forget. Just a little practice and you are back up to speed. Now, others may disagree with me, but each person is different. If you feel that you cannot perform certain duties just because it has been a long time, I advise you to have more confidence in yourself and your training. If you love doing Histology work, then you won't have many problems moving on to other facilities that offer more work. I know that what you are doing now is not as challenging as a large lab. Be patient; the Navy will move you somewhere else soon enough. If you are bored, ask about doing some special projects. If you can contact a pathologist at a larger facility that may be working on a special project or some research work, ask if they would send you the tissue blocks to cut and stain. I am currently working closely with a pathologist at Eisenhower Army Medical Center on a special project that he is spearheading. I work at a small Army hospital, so the project gives me new challenges. This pathologist sends me the tissue blocks which I cut and stain with the H & E stain or any other special stains that he may want performed. The only stains that I don't perform are immuno since we don't do those type of stains here (these are ship outs for us). I have worked with pathologist from the local VA Hospital on special projects, too. So, there are several ways to keep your skills up if you just look for the opportunities. Hope that this gives you more insight. I know that you must feel very frustrated. It is hard working for a pathologist just out of residency....been there, done that, and sometimes find myself doing it again! Happy Cutting, Diane Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: Deltour, Douglas D.(HM2) [mailto:DDDeltour@sig.med.navy.mil] Sent: Friday, April 09, 2004 5:53 AM To: Histonet (E-mail) Subject: [Histonet] Can you lose your skill? Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Evelyn.Flynn <@t> childrens.harvard.edu Fri Apr 9 10:14:32 2004 From: Evelyn.Flynn <@t> childrens.harvard.edu (Flynn, Evelyn) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] beta-gal on mouse tibia Message-ID: Dear Patsy, I did decalcification with EDTA (7.5%) on mouse tibias which had been previously stained with beta-gal. Some were embedded in paraffin; sections were stained with eosin. Others were embedded in JB-4. I encountered no problems. Evelyn Flynn -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Patsy Ruegg Sent: Tue 4/6/2004 2:22 PM To: Histonet@Pathology. Swmed. Edu Subject: [Histonet] beta-gal on mouse tibia Has anyone done this on bone? Gayle was very kind to provide me with a procedure for soft tissues but I am just wondering how decalcification would effect beta-gal staining. Perhaps the samples should be stained and then processed into GMA resin without decal? Would the GMA have an adverse effect on the beta-gal whole mount staining????? Thanks for your advise. Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Fri Apr 9 12:49:25 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? References: <566FB0B522443D43AF02D2ADBE35A6F0635A00@UTHEVS3.mail.uthouston.edu> Message-ID: <002301c41e5b$00068120$70494542@satx.rr.com> Doug I left Wilford Hall USAF medical center in 1986 to go to AFIP, only to return back to Wilford Hall. During the time at AFIP, I did not see a microtome for 5 years. Although I gained a wealth of training and experience in immunos, when I returned back to a hospital setting, I had to learn how to cut again. I almost ripped up my ASCP certificate. I have experienced a loss of technique myself, so I do believe you can loose your skills. Been there, done that. Good luck with fighting USN, I fought the USAF not to go to AFIP, but, we all know how that ended up. Good luck Joe Nocito BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, Texas ----- Original Message ----- From: "Barry R Rittman" To: Sent: Friday, April 09, 2004 5:13 AM Subject: RE: [Histonet] Can you lose your skill? Douglas Although I do not have all the details, I do not believe that you are approaching this from the right direction. If you are only doing 800 cases a year what is occupying the rest of your time there? If you have "time on your hands" then the solution is up to you. It is not only "hands on" but also researching the literature and taking courses that will not only maintain but improve your skills. Is it possible for you to start a small project that allows you to explore new methods for demonstrating topics that you are interested in? Are you able to take courses? I was in a similar situation in England and loved it as I was able to expand my knowledge beyond the work for which I was responsible. This was not only a stimulating experience but also improved my perspective and appreciation for the routine work I had to carry out. If you have the time and facilities, I would take this as a golden opportunity to not only maintain your skills but also broaden your horizons. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Deltour, Douglas D.(HM2) Sent: Friday, April 09, 2004 4:53 AM To: Histonet (E-mail) Subject: [Histonet] Can you lose your skill? Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@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 JWEEMS <@t> sjha.org Fri Apr 9 10:55:46 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? Message-ID: I bet its a man thing...sorry couldn't help it... Happy Spring to all! j:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Friday, April 09, 2004 1:49 PM To: Barry R Rittman; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Can you lose your skill? Doug I left Wilford Hall USAF medical center in 1986 to go to AFIP, only to return back to Wilford Hall. During the time at AFIP, I did not see a microtome for 5 years. Although I gained a wealth of training and experience in immunos, when I returned back to a hospital setting, I had to learn how to cut again. I almost ripped up my ASCP certificate. I have experienced a loss of technique myself, so I do believe you can loose your skills. Been there, done that. Good luck with fighting USN, I fought the USAF not to go to AFIP, but, we all know how that ended up. Good luck Joe Nocito BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, Texas ----- Original Message ----- From: "Barry R Rittman" To: Sent: Friday, April 09, 2004 5:13 AM Subject: RE: [Histonet] Can you lose your skill? Douglas Although I do not have all the details, I do not believe that you are approaching this from the right direction. If you are only doing 800 cases a year what is occupying the rest of your time there? If you have "time on your hands" then the solution is up to you. It is not only "hands on" but also researching the literature and taking courses that will not only maintain but improve your skills. Is it possible for you to start a small project that allows you to explore new methods for demonstrating topics that you are interested in? Are you able to take courses? I was in a similar situation in England and loved it as I was able to expand my knowledge beyond the work for which I was responsible. This was not only a stimulating experience but also improved my perspective and appreciation for the routine work I had to carry out. If you have the time and facilities, I would take this as a golden opportunity to not only maintain your skills but also broaden your horizons. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Deltour, Douglas D.(HM2) Sent: Friday, April 09, 2004 4:53 AM To: Histonet (E-mail) Subject: [Histonet] Can you lose your skill? Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From jnocito <@t> satx.rr.com Fri Apr 9 12:55:19 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] KERMIX References: <40755566.b0.475.238573615@toolkitmail.com> Message-ID: <003d01c41e5b$d326a940$70494542@satx.rr.com> where is this pathologist from? I was at the AFIP when our keratin rabbit bit the dust. I was part of the team that developed Kermix. Actually, we wanted to call it the "BOZO" antibody, but our pathologist didn't think it would be appropriate to see " The Bozo antibody was used on...." in literature, so we named it Kermix. If I remember it was a 2:1 mixture of Dako's cytokeratin cat # M701 (don't remember the clone) to Bectin-Dickinson's AE1/AE3 antibody. At the time Fluffy died, back in 1987, there wasn't many reliable pan cytokeratin antibodies available. Still would have liked it to be known as the Bozo antibody though. Joe Nocito BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, Texas ----- Original Message ----- From: "rfail" To: Sent: Thursday, April 08, 2004 6:36 AM Subject: [Histonet] KERMIX > I have a Pathologist requesting a broad spectrum screening > cytokeratin called Kermix. > Is anyone using this? > Rena Fail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Fri Apr 9 10:57:47 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? Message-ID: I left the field of histology in 1983 and then went back in 1990. I was concerned because there had been so much advancement in the field that I felt I would never catch up. I wasn't concerned over my basic technical skills. My first day I sat down at the microtome to cut some prostate blocks. They were awful! I couldn't believe how bad I was! Then I realized that the microtome, which was an old one, simply needed the block holder tightened. Voila! Beautiful sections! You don't lose the technique but you may have to work on remembering the little details. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Friday, April 09, 2004 1:49 PM To: Barry R Rittman; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Can you lose your skill? Doug I left Wilford Hall USAF medical center in 1986 to go to AFIP, only to return back to Wilford Hall. During the time at AFIP, I did not see a microtome for 5 years. Although I gained a wealth of training and experience in immunos, when I returned back to a hospital setting, I had to learn how to cut again. I almost ripped up my ASCP certificate. I have experienced a loss of technique myself, so I do believe you can loose your skills. Been there, done that. Good luck with fighting USN, I fought the USAF not to go to AFIP, but, we all know how that ended up. Good luck Joe Nocito BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, Texas ----- Original Message ----- From: "Barry R Rittman" To: Sent: Friday, April 09, 2004 5:13 AM Subject: RE: [Histonet] Can you lose your skill? Douglas Although I do not have all the details, I do not believe that you are approaching this from the right direction. If you are only doing 800 cases a year what is occupying the rest of your time there? If you have "time on your hands" then the solution is up to you. It is not only "hands on" but also researching the literature and taking courses that will not only maintain but improve your skills. Is it possible for you to start a small project that allows you to explore new methods for demonstrating topics that you are interested in? Are you able to take courses? I was in a similar situation in England and loved it as I was able to expand my knowledge beyond the work for which I was responsible. This was not only a stimulating experience but also improved my perspective and appreciation for the routine work I had to carry out. If you have the time and facilities, I would take this as a golden opportunity to not only maintain your skills but also broaden your horizons. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Deltour, Douglas D.(HM2) Sent: Friday, April 09, 2004 4:53 AM To: Histonet (E-mail) Subject: [Histonet] Can you lose your skill? Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@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 RBARNHART <@t> summithealth.org Fri Apr 9 11:06:13 2004 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? Message-ID: I agree with everyone else, learn as much as possible, it never hurts. I was hired as a phlebotomist, with no experience what so ever. I took the opportunity to learn all aspects of the lab and other areas. First I became comfortable in phlebotomy and then I learned the office, including transcription. This enabled me to be trained in histology (where I now primally reside) and I have been trained to do the grossing. I went back to school for histo tech and got my HT. I also do some of the lab billing, time cards, monthly reports, statistic, registration and I have became the computer guru. I set up and maintain the Pathology module of our computer system and I do all kinds of other fun computer based projects to occupy my time. I can do almost anything in the lab, except in the main lab running the blood test. So when pathology is slow I always have something to do, which makes some days more hectic then others (like when I am the only histo tech and have to do all the transcription myself) but it is always fun and I enjoy everything I do. I love every part of my job and it always stays interesting and I do not wake up in the morning and think "oh god I have to go to work again today". I look forward to coming to work. I have told every manager that has been here and the pathologist that if they want to teach me I will learn. Becky >>> "Linda Blazek" 04/09/04 10:38AM >>> I agree with everyone about spending your spare time learning new and perfecting old skills and procedures. I have been a histotech since 1973 with a seven year break. When I returned to histology, within a couple of weeks it was like I had never left. I have spent the last 18 years learning every new skill I could find (including transcription). Today though, I am wondering if that was so smart as I am the only person holding down the fort. The transcriptionist is ill, one tech has the day off and who knows why the other tech didn't show up today. We have had two frozens so far with another looming and several special stains that need done. I think I need roller skates today! Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Deltour, Douglas D.(HM2)" 4/9/2004 5:52:51 AM >>> Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Apr 9 11:49:06 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] eosinophilic crystals in mouse lung answer Message-ID: <3.0.6.32.20040409104906.00bc0d60@gemini.msu.montana.edu> Dear All, The question has been answered by Dr. JoAnn Schuh, and much appreciated. She wrote: Eosinophilic crystals in mouse lungs are not an artefact. The crystals form from an endogenous protein, YM-1, that is an autocrystallizing protein. These crystals in the lung, and a related crystal-forming protein (YM-2) in the gastrointestinal tract are most frequent in certain strains of mice (C57BL/6, 129, SW, etc.) and generally are associated with lung or gastrointestinal injury, respectively. FYI, I have attached a publication on the characterization of these crystals in the lungs. Best regards, JoAnn She also attached a pdf of publication and if anyone is interested via private email. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Diane.Gladney <@t> se.amedd.army.mil Fri Apr 9 11:29:11 2004 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Can you lose your skill? Message-ID: <9D41AB7C56F8304F98537ABD87B249F66261C3@DASMTHGBZ001> Douglas, I disagree with you. I am the wife of a retired Army NCO. We traveled all over the world and I had to take whatever lab position was available. I have worked in Blood Chemistry, and even worked in an Army Oil Analysis Lab in Panama. I couldn't get a job at the Army Hospital in Panama because of the Panama Canal Treaty...the locals had 90% hire status because of the treaty. The only reason that I got the job that I did have was because it required a security clearance and the locals could not get the clearance. My lab chemistry background made me qualified to work in a job that I had never had any kind of exposure to do. With a little training, I was highly proficient at that job. You will find that the lab science background may find you working in jobs that you never imagined. My first love has always been Histology, but over my 25 years of civil service (33 total years in lab field) I have worked several jobs in different lab environments. I never lost any of my skills. I have performed all duties in the Histology Lab. You may be a little slow at things at first if you haven't done them in a long time, but it is like riding a bicycle....you never forget. Just a little practice and you are back up to speed. Now, others may disagree with me, but each person is different. If you feel that you cannot perform certain duties just because it has been a long time, I advise you to have more confidence in yourself and your training. If you love doing Histology work, then you won't have many problems moving on to other facilities that offer more work. I know that what you are doing now is not as challenging as a large lab. Be patient; the Navy will move you somewhere else soon enough. If you are bored, ask about doing some special projects. If you can contact a pathologist at a larger facility that may be working on a special project or some research work, ask if they would send you the tissue blocks to cut and stain. I am currently working closely with a pathologist at Eisenhower Army Medical Center on a special project that he is spearheading. I work at a small Army hospital, so the project gives me new challenges. This pathologist sends me the tissue blocks which I cut and stain with the H & E stain or any other special stains that he may want performed. The only stains that I don't perform are immuno since we don't do those type of stains here (these are ship outs for us). I have worked with pathologist from the local VA Hospital on special projects, too. So, there are several ways to keep your skills up if you just look for the opportunities. Also, you have a great opportunity to study, study, study. I can't say this enough. I wish that I had the extra time to do this more often. Hope that this gives you more insight. I know that you must feel very frustrated. It is hard working for a pathologist just out of residency....been there, done that, and sometimes find myself doing it again! Happy Cutting, Diane Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: Friday, April 09, 2004 12:06 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Can you lose your skill? I agree with everyone else, learn as much as possible, it never hurts. I was hired as a phlebotomist, with no experience what so ever. I took the opportunity to learn all aspects of the lab and other areas. First I became comfortable in phlebotomy and then I learned the office, including transcription. This enabled me to be trained in histology (where I now primally reside) and I have been trained to do the grossing. I went back to school for histo tech and got my HT. I also do some of the lab billing, time cards, monthly reports, statistic, registration and I have became the computer guru. I set up and maintain the Pathology module of our computer system and I do all kinds of other fun computer based projects to occupy my time. I can do almost anything in the lab, except in the main lab running the blood test. So when pathology is slow I always have something to do, which makes some days more hectic then others (like when I am the only histo tech and have to do all the transcription myself) but it is always fun and I enjoy everything I do. I love every part of my job and it always stays interesting and I do not wake up in the morning and think "oh god I have to go to work again today". I look forward to coming to work. I have told every manager that has been here and the pathologist that if they want to teach me I will learn. Becky >>> "Linda Blazek" 04/09/04 10:38AM >>> I agree with everyone about spending your spare time learning new and perfecting old skills and procedures. I have been a histotech since 1973 with a seven year break. When I returned to histology, within a couple of weeks it was like I had never left. I have spent the last 18 years learning every new skill I could find (including transcription). Today though, I am wondering if that was so smart as I am the only person holding down the fort. The transcriptionist is ill, one tech has the day off and who knows why the other tech didn't show up today. We have had two frozens so far with another looming and several special stains that need done. I think I need roller skates today! Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Deltour, Douglas D.(HM2)" 4/9/2004 5:52:51 AM >>> Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@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 amos <@t> dressi.com Fri Apr 9 12:00:59 2004 From: amos <@t> dressi.com (Amy R.) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Victoria Baker Message-ID: <7B284BA8386B394DA05712C1DFD0C7AB03E1E4@01server.diversifiedrealestate.local> THANK YOU, Victoria. You made my day. Amy Do or do not. There is no try. -- Yoda, The Empire Strikes Back Message: 8 Date: Fri, 9 Apr 2004 08:13:09 -0700 (PDT) From: Victoria Baker Subject: Re: [Histonet] Can you lose your skill? To: "Deltour, Douglas D.\(HM2\)" , "Histonet \(E-mail\)" Message-ID: <20040409151309.99252.qmail@web12107.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Dear Douglas, You are in the military, they send you where they need to send you, when they need to send you. The words I recollect were "I'm not to question why...". The shift in the case load number or complexity of the work they do is not always relevant, it's just procedure. Look at this as an opportunity to further yourself on YOUR OWN terms. In someways they have given you an opportunity that may never come your way again. No I haven't served in the armed forces, but I have close friends who have and sometimes also felt somewhat frustrated at the military system du jour. It has no reflection on your ability only the military needs to have a competent warm body where they need them. Do not let this deter or discourage you in any way. In the meantime, soak up all you can by reading or seeing what is available from the military in terms of courses. Italy has some fine university settings that you can also look into. The hands on experience, as far as I can tell they will never be able to automate the very basic skill each histologist holds so very valuable. Our professional integrity to do the very best we can under any circumstances. You could surprise yourself at what you can remember about a stain even if you hadn't done it for a year or for that matter 10 years when you do need to do it. It's a part of you, histology, what you decide to make of it is entirely up to you. Just like life itself. Vikki Baker Institute for Cancer Prevention Valhalla, New York From ljb <@t> medicine.wisc.edu Fri Apr 9 13:30:19 2004 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:22:47 2005 Subject: Fwd: Re: [Histonet] Can you lose your skill? Message-ID: >>> LaCinda Burchell 04/09/04 10:17AM >>> Hi Douglas, I have changed back and forth from broad based general hospital lab experience to very specific research work over the years. I know that I have become very rusty in all sorts of areas. I've tried to combat that by keeping several excellent text books in my lab, and by keeping constant connection with resources like Histonet. A couple of years ago I had the chance to moonlight at a brand new Mohs lab. It had been three years since I'd last worked in Mohs. I was very nervous on the first day that I might have forgotten too much, or lost my skills. It turned out that I was a bit rusty, but when the first specimen was given to me I was able to switch into the right gear and do everything necessary. By the third day in the new Mohs lab I felt as though I was right back up to speed. Seven years ago I had many special stains memorized. Now I have to refer to texts in order to do most of them. I think that yes, you may forget many things in the short term, and you will feel slow and clumsy when you need to ramp up again in the future. But, if you want it bad enough you'll find your strengths again. Are the people with your new job open to/or excited about seeing any of the wonderful things you can provide for them? If you're "stuck" in a place for a while in the Navy try to be the best example of someone eager to learn anything, or figure out anything put before you. Be creative. It will be a test of patience, but initiative of that sort may lead you to the creation of new techniques and knowledge which will make you very desirable to many employers in the future. I know that the military can be very rigid when it comes to the desires of one person. As the mom of a US Sailor I wish with all my heart that this unhappy location might turn into all sorts of skills that will pad your resume should you want to shift in to the civilian world again in the future. For instance, is there a camera attached to any of the scopes? If so, be insistent that you want to become proficient with it's use. And, always listen closely to the Pathologists around you. If you ever hear them say "I wish....", see it as a chance to prove your true colors.) Try to hang in there if you must. But in the meantime, if there is ever anything that I might help you with please feel welcome to contact me. I'll do my best! Sincerely, LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 >>> "Deltour, Douglas D.(HM2)" 04/09/04 04:52AM >>> Hello everyone, I have a kind of odd question for you. I would love to have all of the experts and non-experts feedback. Let me start off by saying that I have been a Histotech since 2000 when I graduated from the school at AFIP. I was sent to a place that did 6000 cases a year. We also did special stains and autopsies. I was working there for two years when I transferred to a place that did 18,000 cases a year. Specials, autopsies, and immuno's. Now I am at a place where I do 800 cases a year. Maybe 5 special stains a year and no autopsies. I am supposed to be here for three years but I am trying to fight it. I am telling everyone that I am losing my skills being here. The people that control this place tell me otherwise. There is one pathologist here right out of residency who will not confirm that my skills will erode. He is right out of residency and would not know. Anyway do you think that my complaint is a legitimate complaint. Can you lose your skills if not used? If you have not noticed already I am in the military, Navy that is. I just need to confirm that a Histotech can lose their skill if not used. I would appreciate any feedback, advice. Thank you. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From amosbrooks <@t> earthlink.net Fri Apr 9 15:02:39 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF In-Reply-To: <200404081920.1bbLCT5eq3NZFjK0@condor> References: <200404081920.1bbLCT5eq3NZFjK0@condor> Message-ID: <6.0.0.22.0.20040409145343.01b2d9d8@pop.earthlink.net> Patsy, I'll have to check if we've tried this one when I get back Monday. If we've not tried it yet, I'll give it a whirl. I know we've tried the polyclonal before. Generally I cringe when I see a reference to Santa Cruz though. I've had a number of bum antibodies from them, VEGF polyclonal being one of them. (I'm sure they have some good ones too, at least I hope they do). When we work up a new antibody we try just about every pretreatment beginning with none at all and pick the one that works best. Thanks for the suggestion. If we haven't tried it yet, and we get it, I'll post our success or failure here. Hoping for the best, Amos At 09:20 PM 4/8/2004, you wrote: >Message: 16 >Date: Thu, 8 Apr 2004 15:48:28 -0600 >From: "Patsy Ruegg" >Subject: RE: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 >To: "Amos Brooks" , > >Message-ID: >Content-Type: text/plain; charset="US-ASCII" > >Amos I had the same experience as you with VEGF's until I switched to >monoclonal Vegf from Santa Cruz, it now appears very specific and reliable >to me. I use pepsin or proteinase K digestion and stay away from HIER. >Patsy From amosbrooks <@t> earthlink.net Fri Apr 9 15:10:15 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] BCL2 High pH retrieval In-Reply-To: <200404081920.1bbLCT5eq3NZFjK0@condor> References: <200404081920.1bbLCT5eq3NZFjK0@condor> Message-ID: <6.0.0.22.0.20040409150356.01b82c98@pop.earthlink.net> Hi, Try High pH antigen retrieval (pH 10.0). The steamer for 1/2 hour should be fine. I can't avouch for the detection kit you're stuck using with a Ventana stainer, but it should work. I would suggest a non biotin detection kit like Envision from DAKO or Mach3 from Biocare (I'm sure there are others). It seems high pH retrieval increases endogenous biotin. Amos Brooks At 09:20 PM 4/8/2004, you wrote: >Message: 22 >Date: Fri, 9 Apr 2004 18:51:44 -0400 >From: "Derek & Lynda Leopold" >Subject: [Histonet] bcl-2 on Nexes >To: >Message-ID: <000801c41e85$3bc3fc40$3643fea9@leopold> >Content-Type: text/plain; charset="iso-8859-1" > >Hi All, >We're having some trouble getting our tonsil control to come up for >bcl-2. After searching the archives and asking around, we are going to >try citrate buffer (Declere) and a hot steamer for 1 hour. We've got the >Nexes IHC. Anyone else have any tips for us? >Can anyone comment on the need/non-need for DAB? >Thanks >Lynda Leopold From pruegg <@t> colobio.com Fri Apr 9 14:38:06 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF In-Reply-To: <6.0.0.22.0.20040409145343.01b2d9d8@pop.earthlink.net> Message-ID: Amos, I totally agree with you about Santa Cruz's poly VEGF but the mono was much better. I have a customer who still wants to use the poly vegf on sheep placenta and I think it is awful but you have to give the customer what they want. I did a comparison with the poly and mono for them but they still opted for the poly because it matched their molecular studies. Oh well. To me it is all over the place but they seem to think that is expected for their samples, they think is is real and call it "an expression antibody" or something like that. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amos Brooks Sent: Friday, April 09, 2004 2:03 PM To: histonet@lists.utsouthwestern.edu Cc: pruegg@colobio.com Subject: [Histonet] VEGF Patsy, I'll have to check if we've tried this one when I get back Monday. If we've not tried it yet, I'll give it a whirl. I know we've tried the polyclonal before. Generally I cringe when I see a reference to Santa Cruz though. I've had a number of bum antibodies from them, VEGF polyclonal being one of them. (I'm sure they have some good ones too, at least I hope they do). When we work up a new antibody we try just about every pretreatment beginning with none at all and pick the one that works best. Thanks for the suggestion. If we haven't tried it yet, and we get it, I'll post our success or failure here. Hoping for the best, Amos At 09:20 PM 4/8/2004, you wrote: >Message: 16 >Date: Thu, 8 Apr 2004 15:48:28 -0600 >From: "Patsy Ruegg" >Subject: RE: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 >To: "Amos Brooks" , > >Message-ID: >Content-Type: text/plain; charset="US-ASCII" > >Amos I had the same experience as you with VEGF's until I switched to >monoclonal Vegf from Santa Cruz, it now appears very specific and reliable >to me. I use pepsin or proteinase K digestion and stay away from HIER. >Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DDittus787 <@t> aol.com Fri Apr 9 14:50:34 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] BCL2 High pH retrieval Message-ID: <6D26DFCF.293CAF0D.0A1F969F@aol.com> we have the nexes stainer , and use zymeds BCL-2 with microwave retreival 18 minutes(not nearly as much endogenous biotin or tissue loss) 32 minute incubation and i suggest a follicular lymphoma control or lymph node usually looks better than tonsil. if you need any other help don't hesitate to call me . dana 215-481-2609 From jstaruk <@t> masshistology.com Fri Apr 9 15:57:52 2004 From: jstaruk <@t> masshistology.com (Mass Histology Service) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF In-Reply-To: Message-ID: We just completed a large study on human prostate biopsies using the Santa Cruz monoclonal VEGF (C1) following pronase pre-treatment and they all came out wonderful: Very clean and the negative controls (no primary) were completely negative. Jim ____________________ James Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Friday, April 09, 2004 3:38 PM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Amos, I totally agree with you about Santa Cruz's poly VEGF but the mono was much better. I have a customer who still wants to use the poly vegf on sheep placenta and I think it is awful but you have to give the customer what they want. I did a comparison with the poly and mono for them but they still opted for the poly because it matched their molecular studies. Oh well. To me it is all over the place but they seem to think that is expected for their samples, they think is is real and call it "an expression antibody" or something like that. Patsy (snip) From pruegg <@t> colobio.com Fri Apr 9 17:17:01 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF In-Reply-To: Message-ID: Yep that was my experience, this was the most drastic difference between a monoclonal and polyclonal antibody I have ever seen. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mass Histology Service Sent: Friday, April 09, 2004 2:58 PM To: Patsy Ruegg; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF We just completed a large study on human prostate biopsies using the Santa Cruz monoclonal VEGF (C1) following pronase pre-treatment and they all came out wonderful: Very clean and the negative controls (no primary) were completely negative. Jim ____________________ James Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Friday, April 09, 2004 3:38 PM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Amos, I totally agree with you about Santa Cruz's poly VEGF but the mono was much better. I have a customer who still wants to use the poly vegf on sheep placenta and I think it is awful but you have to give the customer what they want. I did a comparison with the poly and mono for them but they still opted for the poly because it matched their molecular studies. Oh well. To me it is all over the place but they seem to think that is expected for their samples, they think is is real and call it "an expression antibody" or something like that. Patsy (snip) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Sat Apr 10 16:09:09 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF Message-ID: Jim, Did you find there was a problem with oxidation? A researcher has brought us a number of renal biopsy slides for VEGF staining, some which have been cut for a couple years and I presume stored at R. T., and I'm afraid we may have an issue with antigenicity of the epitope. What do you think? Linda Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory -----Original Message----- From: Mass Histology Service [mailto:jstaruk@masshistology.com] Sent: Fri 4/9/2004 3:57 PM To: Patsy Ruegg; Amos Brooks; histonet@lists.utsouthwestern.edu Cc: Subject: RE: [Histonet] VEGF We just completed a large study on human prostate biopsies using the Santa Cruz monoclonal VEGF (C1) following pronase pre-treatment and they all came out wonderful: Very clean and the negative controls (no primary) were completely negative. Jim ____________________ James Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Friday, April 09, 2004 3:38 PM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Amos, I totally agree with you about Santa Cruz's poly VEGF but the mono was much better. I have a customer who still wants to use the poly vegf on sheep placenta and I think it is awful but you have to give the customer what they want. I did a comparison with the poly and mono for them but they still opted for the poly because it matched their molecular studies. Oh well. To me it is all over the place but they seem to think that is expected for their samples, they think is is real and call it "an expression antibody" or something like that. Patsy (snip) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jstaruk <@t> masshistology.com Sat Apr 10 20:09:57 2004 From: jstaruk <@t> masshistology.com (Mass Histology Service) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF In-Reply-To: Message-ID: Linda, The slides were sent to us, so I have no way of knowing their age. The surgical numbers were from several years back and the glass slides were from various manufacturers. They all stained the same intensity. By the way, I don't charge for my time in answering questions from anyone who asks for help. Jim ____________________________ James E. Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sebree Linda A. Sent: Saturday, April 10, 2004 5:09 PM To: Mass Histology Service; Patsy Ruegg; Amos Brooks; histonet@lists.utsouthwestern.edu Cc: Yracheta Joseph Subject: RE: [Histonet] VEGF Jim, Did you find there was a problem with oxidation? A researcher has brought us a number of renal biopsy slides for VEGF staining, some which have been cut for a couple years and I presume stored at R. T., and I'm afraid we may have an issue with antigenicity of the epitope. What do you think? Linda Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory From amosbrooks <@t> earthlink.net Sun Apr 11 10:21:46 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF In-Reply-To: References: Message-ID: <6.0.0.22.0.20040411100816.01b28650@pop.earthlink.net> Actually no, The study we were doing was using blocks that we had cut freshly. The blocks were old (mostly) but they were generally cut and stained within a week. I have seen degradation of epitopes in old slides but not in old blocks recently cut. Truly the culprit was a very poor primary antibody. I'll certainly be looking into the monoclonal Tomorrow morning though. Hoping for the best, Amos At 04:09 PM 4/10/2004, you wrote: >Jim, > >Did you find there was a problem with oxidation? A researcher has brought >us a number of renal biopsy slides for VEGF staining, some which have been >cut for a couple years and I presume stored at R. T., and I'm afraid we >may have an issue with antigenicity of the epitope. What do you think? > >Linda Sebree >University of Wisconsin Hospital & Clinics >IHC/ISH Laboratory From miller <@t> coho.net Mon Apr 12 06:37:31 2004 From: miller <@t> coho.net (Diane G. Miller) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] About H2O2 and CD Markers References: Message-ID: <03d801c42082$8b492340$0300a8c0@desktop> Hi Jorge, Sorry for the delay in getting this to you. Just two notes for your review. Sodium Azide used in TBS buffer, plus H202 for FS only. This eliminated the air bubbles that can occur with Frozen Sections. a.. Do not use methanol on frozen sections, it destroys the antigen sites and you will get a false negative. Best Regards Diane ----- Original Message ----- From: Jorge Ivan Zapata Valencia To: histonet@lists.utsouthwestern.edu Sent: Friday, March 26, 2004 5:08 PM Subject: [Histonet] About H2O2 and CD Markers Hello everybody. I have a simple question: do someone know the effect, if any, of the different concentrations, and times, of H2O2 on the CD markers (Paraffin and/or frozen sections)? I have read that some people use 0.01%, others 0.1% and others 3% for different times, usually 10 or 30 minutes. What could happen if I use a longer time? Thank You very much. JORGE IVAN ZAPATA _________________________________________________________________ Help STOP SPAM with the new MSN 8 and get 2 months FREE* http://join.msn.com/?page=features/junkmail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nsdnz <@t> neuro.hfh.edu Mon Apr 12 08:44:17 2004 From: nsdnz <@t> neuro.hfh.edu (Danielle Zalinski) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Vibratome Considerations Message-ID: <407A9D31.70301@neuro.hfh.edu> Hello All, Our lab would like to do immunohistochem work in rat brain on vibratome sections instead of paraffin. We are able to get sections about 50-60um. Will incubation with primary antibody overnight be sufficient? Is shaking the sections an option? I was hoping some other labs that are working in vibratome sections may have some beginners advice. Thanks, Danielle Zalinski Neurosurgery Research Henry Ford Health System Detroit, MI CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. From TillRenee <@t> uams.edu Mon Apr 12 09:13:40 2004 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] whole mounts Message-ID: Hello! Does anyone have experience with whole mount slides, specifically mammary whole mounts? I have done a couple batches of rat mammary whole mounts with varying results. The protocol I follow is pretty simple. They are fixed, then defatted in acetone, rehydrated in 70% ETOH and water, stained in carmine, then dehydrated through several alcohol gradients, and cleared in xylene. Before coverslipping I also flatten the tissues by pressing them between two slides. Depending on the quality of tissue collection, most of my slides turn out well. There are usually a couple that the stain is dark and almost black instead of purple and I cannot use these. I guess my questions are, beyond just any general advice, does anyone have a different (perhaps better?) protocol and do you think the bad ones could be destained and redone? I'm of the opinion that it probably can't be done, but I keep getting asked by my boss. I'm pretty much pleased with the results myself, but if it can be done better...It seems my biggest problem is publication quality pictures to go with the data I generate off these slides. Our equipment is good, and in the microscope I can often see the structures, but some stain more darkly than others and the lighter ones just don't show up. Of course these are the ones I need good photos of. Any suggestions? Is there a better stain? Thanks, Renee' Till Arkansas Childrens Nutrition Center ------------------------------------ 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 eca9 <@t> georgetown.edu Mon Apr 12 09:31:48 2004 From: eca9 <@t> georgetown.edu (Eva Andersson) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Storage of 10M NaOH Message-ID: <407AA854.20801@georgetown.edu> Hi, Maybe not a strictly histochemistry question but maybe someone could provide me with an answer. I was just told that I shouldn't store the 10M NaOH that I use for adjusting the pH in a glass bottle. I was told that it needed to be made fresh and only kept in a plastic bottle. Why is this? How long can I keep a stock solution? If anyone has any ideas please let me know. Eva From gcallis <@t> montana.edu Mon Apr 12 09:52:15 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Vibratome sections and immunohistochemistry In-Reply-To: <407A9D31.70301@neuro.hfh.edu> Message-ID: <3.0.6.32.20040412085215.00bd9b30@gemini.msu.montana.edu> People do this all the time, and this is found in literature. A PUBMED search will help you glean a lot of knowledge, at least methods. Access this publication: Zhuo L et al, Live astrocytes visualized by green fluorescent protein in transgenic mice, Developmental Biology 187:36-42, 1997. They did 300 um vibrotome section on brain, far thicker than you want to use. Even though this is for GFP and fluorescent work, their methods and materials were excellent. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Apr 12 10:00:10 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Storage of strong bases in glass In-Reply-To: <407AA854.20801@georgetown.edu> Message-ID: <3.0.6.32.20040412090010.00bd9b30@gemini.msu.montana.edu> This was one of the first things learned in a chemistry class milleniums ago. Yes, you are correct in NOT storing 10 M sodium hydroxide in glass. This base etches the glass e.g. eats glass surface! and if you wash glass storage jar, it appears as a milky defect on glass surface that cannot be removed. Use plastic bottle or if you don't use a huge amount, a 50 ml conical tube. Set an expiration date, sometimes the solution has flocculant gunk floating on bottom, toss and make new or make up as much as you use in a year? At 10:31 AM 4/12/2004 -0400, you wrote: >Hi, >Maybe not a strictly histochemistry question but maybe someone could >provide me with an answer. >I was just told that I shouldn't store the 10M NaOH that I use for >adjusting the pH in a glass bottle. I was told that it needed to be made >fresh and only kept in a plastic bottle. Why is this? How long can I >keep a stock solution? >If anyone has any ideas please let me know. >Eva > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From peoshel <@t> wisc.edu Mon Apr 12 10:02:41 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Storage of 10M NaOH In-Reply-To: <407AA854.20801@georgetown.edu> References: <407AA854.20801@georgetown.edu> Message-ID: Eva, Glass -- silica -- is soluble in basic solutions. Not obviously, but 10M NaOH will attack the glass, and if ground-glass stoppers are used, the NaOH will "weld" the stopper to the bottle. As an interesting aside, seawater has a high enough pH, around 8, that it will dissolve low levels of silica from glass bottles, enough to interfer with studies of silica in seawater. Such as when [silica] test formation of critters like diatoms and radiolarians is the object of interest. Phil >Hi, >Maybe not a strictly histochemistry question but maybe someone could >provide me with an answer. >I was just told that I shouldn't store the 10M NaOH that I use for >adjusting the pH in a glass bottle. I was told that it needed to be >made fresh and only kept in a plastic bottle. Why is this? How long >can I keep a stock solution? >If anyone has any ideas please let me know. >Eva -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From Luis.Chiriboga <@t> med.nyu.edu Mon Apr 12 10:39:25 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF In-Reply-To: Message-ID: Have any of you tried it on mouse?????? I have stuck with the polyclonal (on mouse tissue) to avoid the species issue but it's so dirty anyway. I'm figuring I can at least try and manipulate the species issue (high dilution, absorbed secondary, blocking, biotinylating primary etc...) and get better results. Would you recommend? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Friday, April 09, 2004 6:17 PM To: Mass Histology Service; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Yep that was my experience, this was the most drastic difference between a monoclonal and polyclonal antibody I have ever seen. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mass Histology Service Sent: Friday, April 09, 2004 2:58 PM To: Patsy Ruegg; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF We just completed a large study on human prostate biopsies using the Santa Cruz monoclonal VEGF (C1) following pronase pre-treatment and they all came out wonderful: Very clean and the negative controls (no primary) were completely negative. Jim ____________________ James Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Friday, April 09, 2004 3:38 PM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Amos, I totally agree with you about Santa Cruz's poly VEGF but the mono was much better. I have a customer who still wants to use the poly vegf on sheep placenta and I think it is awful but you have to give the customer what they want. I did a comparison with the poly and mono for them but they still opted for the poly because it matched their molecular studies. Oh well. To me it is all over the place but they seem to think that is expected for their samples, they think is is real and call it "an expression antibody" or something like that. Patsy (snip) _______________________________________________ Histonet mailing list Histonet@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 Mon Apr 12 10:25:40 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] rapid or stat histo process Message-ID: <23BED360.346FA9CA.0A1F969F@aol.com> i am calling on my knowledgeable friends in histoland to tell me if you guys are charging a fee for stat or rapid processing to expedite results. thanks in advance dana From pruegg <@t> colobio.com Mon Apr 12 10:48:51 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF In-Reply-To: Message-ID: Luis, I have not tried the mono vegf on mice but I have used it successfully on rat tissue. In my hands the poly vegf is so non-specific I could not ever get it cleaned up enough to be worth anything. Patsy -----Original Message----- From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] Sent: Monday, April 12, 2004 9:39 AM To: Patsy Ruegg; Mass Histology Service; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Have any of you tried it on mouse?????? I have stuck with the polyclonal (on mouse tissue) to avoid the species issue but it's so dirty anyway. I'm figuring I can at least try and manipulate the species issue (high dilution, absorbed secondary, blocking, biotinylating primary etc...) and get better results. Would you recommend? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Friday, April 09, 2004 6:17 PM To: Mass Histology Service; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Yep that was my experience, this was the most drastic difference between a monoclonal and polyclonal antibody I have ever seen. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mass Histology Service Sent: Friday, April 09, 2004 2:58 PM To: Patsy Ruegg; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF We just completed a large study on human prostate biopsies using the Santa Cruz monoclonal VEGF (C1) following pronase pre-treatment and they all came out wonderful: Very clean and the negative controls (no primary) were completely negative. Jim ____________________ James Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Friday, April 09, 2004 3:38 PM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Amos, I totally agree with you about Santa Cruz's poly VEGF but the mono was much better. I have a customer who still wants to use the poly vegf on sheep placenta and I think it is awful but you have to give the customer what they want. I did a comparison with the poly and mono for them but they still opted for the poly because it matched their molecular studies. Oh well. To me it is all over the place but they seem to think that is expected for their samples, they think is is real and call it "an expression antibody" or something like that. Patsy (snip) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Mon Apr 12 10:46:49 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] digital microphotograph system Message-ID: Folks, I am looking to buy a digital camera to take microphotographs for my small business. I do not have a large budget for this and hope to get to most for the buck. I have several microscopes to use this camera with, most of them are Olympus models but I do have one Zeis. One of the Olympus microscopes has a video camera but it is without the power source. This scope and video camera has been in storage and I have no way of knowing how well it works without the power source. I also have a personal digital camera but it is only medium in mega pixels. Should I buy the power source for the video camera and try to get that to work? Should I buy a mount system for my personal digital camera and try that? Should I buy and completely new dig camera and microscope mounting system? Please advise. From pruegg <@t> colobio.com Mon Apr 12 11:00:08 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF In-Reply-To: Message-ID: I really doubt you would lose vegf antigenicity, usually the problem with vegf is that it is so over expressed that it stains too heavily. Perhaps old samples may look better than fresh one's in this case. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mass Histology Service Sent: Saturday, April 10, 2004 7:10 PM To: Sebree Linda A.; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Linda, The slides were sent to us, so I have no way of knowing their age. The surgical numbers were from several years back and the glass slides were from various manufacturers. They all stained the same intensity. By the way, I don't charge for my time in answering questions from anyone who asks for help. Jim ____________________________ James E. Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sebree Linda A. Sent: Saturday, April 10, 2004 5:09 PM To: Mass Histology Service; Patsy Ruegg; Amos Brooks; histonet@lists.utsouthwestern.edu Cc: Yracheta Joseph Subject: RE: [Histonet] VEGF Jim, Did you find there was a problem with oxidation? A researcher has brought us a number of renal biopsy slides for VEGF staining, some which have been cut for a couple years and I presume stored at R. T., and I'm afraid we may have an issue with antigenicity of the epitope. What do you think? Linda Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From djohnson14 <@t> hotmail.com Mon Apr 12 11:16:14 2004 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Histology OPening Message-ID: New Mohs practice in Southern Ca seeks Part time mohs tech Fax resume to Jane Lisk, practice administrator to 760-941-3986 THank you _________________________________________________________________ Persistent heartburn? Check out Digestive Health & Wellness for information and advice. http://gerd.msn.com/default.asp From cfavara <@t> niaid.nih.gov Mon Apr 12 11:17:08 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Storage of 10M NaOH Message-ID: Well I will give this a shot. I think this is strictly from a safety standpoint. We have recently undergone lab to lab evaluations by a safety officer and you would not believe what we are supposed to do. Granted we in research get away with a lot as we are not subjected to the inspections the clinical labs get. For safety purposes you should keep minimal amounts of caustic/harmful materials in plastic to avoid breakage issues, below eye level and on shelves that have a lip! If not you get a ticket!!!! Fortunately no fine yet! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Eva Andersson [mailto:eca9@georgetown.edu] Sent: Monday, April 12, 2004 8:32 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Storage of 10M NaOH Hi, Maybe not a strictly histochemistry question but maybe someone could provide me with an answer. I was just told that I shouldn't store the 10M NaOH that I use for adjusting the pH in a glass bottle. I was told that it needed to be made fresh and only kept in a plastic bottle. Why is this? How long can I keep a stock solution? If anyone has any ideas please let me know. Eva _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peoshel <@t> wisc.edu Mon Apr 12 11:35:48 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] digital microphotograph system In-Reply-To: References: Message-ID: Patsy, This question keeps popping up on the microscopy mailing list. You might try searching their archives for the full discussion: http://www.msa.microscopy.com/MicroscopyListserver/SearchMLArchive.html But briefly, what a lot of folks have done (including us) is to get a Nikon Coolpix and an eyepiece adapter. There are a couple of companies doing these, but I preferred Thales-Optem. I think MVIA is the other main company making the adapters. The Coolpix also requires a filter adapter from Nikon. For the Coolpix 5000, this is a UR-E6. Other digital cameras can work as well -- some people use an Olympus. The camera then goes in phototube or an ocular tube. The more pixels in the camera, the better. Most "personal" digital cameras don't have enough pixels. You can also buy digital camera backs to go on a C-mount (or bayonet mount) the same as film backs. These are then used as are film-camera backs **except** the metering is different. No off-the-film metering is available for the digital backs, so this may be interesting. But! Make sure your camera has an optical zoom (if you don't use a back only), and that the adapters and tubes are all correct. You may also need relay lenses. It's common to get vignetting with these setups (the optical zoom gets around this). And a stage micrometer to calibrate the true magnification. We got everything but the camera from Nation Graphics. No affiliation, but they knew what was needed to use the digital camera for photomicroscopy and had good prices. The video camera could be a good way to go, if it is a good camera, and not too old. What do you intend to use to capture the images? A computer? Video cards might be more of a problem, if so. And memory. Phil >Folks, >I am looking to buy a digital camera to take microphotographs for my small >business. I do not have a large budget for this and hope to get to most for >the buck. I have several microscopes to use this camera with, most of them >are Olympus models but I do have one Zeis. One of the Olympus microscopes >has a video camera but it is without the power source. This scope and video >camera has been in storage and I have no way of knowing how well it works >without the power source. I also have a personal digital camera but it is >only medium in mega pixels. >Should I buy the power source for the video camera and try to get that to >work? >Should I buy a mount system for my personal digital camera and try that? >Should I buy and completely new dig camera and microscope mounting system? >Please advise. -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From TJJ <@t> Stowers-Institute.org Mon Apr 12 12:26:29 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] BMP-4 antibody staining in murine tissue Message-ID: Has anybody had any success in immunostaining BMP-4 in mouse tissue? We are using EDTA decalcified, formalin fixed and paraffin embedded bone specimens, and are using DAKO's ARK kit for the mouse on mouse techniques. We have tried using trypsin digestion, epitope unmasking with citrate buffer, and no pretreatment. There are many other pretreatments to use, but before we unleash the arsenal, I thought I'd ask. Thanks for any assistance, Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From Lizbeth_Kelly <@t> hgsi.com Mon Apr 12 12:18:33 2004 From: Lizbeth_Kelly <@t> hgsi.com (Lizbeth_Kelly@hgsi.com) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] NSH Region II Symposium (June 3,4, & 5th) Message-ID: __________________ The Maryland Society of Histotechnologists (MSH) is sponsoring the Region II Symposium June 3, 4, and 1/2 day on the 5th at the Holiday Inn Select, North-Baltimore, Maryland. For information, you may contact Terri DeCarli at 410-787-4546 or Renate Jaacks at 410-879-9012. Lizbeth Kelly, HT (ASCP), QIHC Board Member, MSH From jlinda <@t> ces.clemson.edu Mon Apr 12 12:46:12 2004 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Re: digital microphotograph system Message-ID: <5.2.1.1.0.20040412133242.0288c750@mailhost.ces.clemson.edu> Patsy, Martin Microscope Co. carries a universal camera adapter that should fit all your scopes for less than $100.00. They also carry new and used microscopes and cameras. They have a website: http://www.martinmicroscope.com/ and their phone is: 864-242-3424. Ask for Bobby Martin and tell him exactly what you need and he will provide excellent advice. The really good cameras are getting less expensive each year. And a note to Philip Oshel...you are so knowledgeable on this subject...why don't you apply to do a workshop at the NSH meeting? We really need more photographic lessons. Happy hunting, Patsy! Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo/histo.htm From lesley <@t> vancouverbc.net Mon Apr 12 13:00:07 2004 From: lesley <@t> vancouverbc.net (Lesley Weston) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Storage of 10M NaOH In-Reply-To: <407AA854.20801@georgetown.edu> Message-ID: NaOH etches glass, so even dilute solutions should be stored in plastic and certainly 10M should. The reason to use fresh 10M NaOH is that it absorbs CO2 from the air, but I have kept it for some weeks without any problems. Lesley Weston. on 12/04/2004 7:31 AM, Eva Andersson at eca9@georgetown.edu wrote: > Hi, > Maybe not a strictly histochemistry question but maybe someone could > provide me with an answer. > I was just told that I shouldn't store the 10M NaOH that I use for > adjusting the pH in a glass bottle. I was told that it needed to be made > fresh and only kept in a plastic bottle. Why is this? How long can I > keep a stock solution? > If anyone has any ideas please let me know. > Eva > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Mon Apr 12 13:29:26 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] BMP-4 antibody staining in murine tissue (repost) Message-ID: Has anybody had any success in immunostaining BMP-4 in mouse tissue? We are using EDTA decalcified, formalin fixed and paraffin embedded bone specimens, and are using DAKO's ARK kit for the mouse on mouse techniques. We have tried using trypsin digestion, epitope unmasking with citrate buffer, and no pretreatment. There are many other pretreatments to use, but before we unleash the arsenal, I thought I'd ask. Thanks for any assistance, Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From CKByrne <@t> labvision.com Mon Apr 12 13:39:04 2004 From: CKByrne <@t> labvision.com (Kyle-Byrne, Carrie - Labvision) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] VEGF Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20A96E2@usca0082k08.labvision.apogent.com> Luis, I would think the quickest way to have a good idea is to check the aa alignments of human v mouse. the Santa Cruz mono antibody is raised to aa 1-140 of human VEGF. I'd suggest having a look at the mouse alignment and see what the homology is. According to Santa Cruz, it should cross react, but my own experience with Santa Cruz antibodies in general has mirrored that of many others....you just won't really know until you try it. Carrie Kyle-Byrne Lab Vision, Corp. www.labvision.com -----Original Message----- From: Patsy Ruegg [mailto:pruegg@colobio.com] Sent: Monday, April 12, 2004 8:49 AM To: Luis Chiriboga; Mass Histology Service; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Luis, I have not tried the mono vegf on mice but I have used it successfully on rat tissue. In my hands the poly vegf is so non-specific I could not ever get it cleaned up enough to be worth anything. Patsy -----Original Message----- From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] Sent: Monday, April 12, 2004 9:39 AM To: Patsy Ruegg; Mass Histology Service; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] VEGF Have any of you tried it on mouse?????? I have stuck with the polyclonal (on mouse tissue) to avoid the species issue but it's so dirty anyway. I'm figuring I can at least try and manipulate the species issue (high dilution, absorbed secondary, blocking, biotinylating primary etc...) and get better results. Would you recommend? Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MTitford <@t> aol.com Mon Apr 12 15:50:54 2004 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] HIPPA question re: research Message-ID: <0EC91BB9.64DC6DAF.00762DB1@aol.com> Has the HIPPA law effectively put an end to researchers going back through hospital records doing retrospective studies or doing research on old blocks? Mike Titford USA Pathology Mobile AL USA From amosbrooks <@t> earthlink.net Mon Apr 12 17:48:27 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Vibratome Considerations In-Reply-To: <200404121000.1bd4NO3I03NZFjw0@skylark> References: <200404121000.1bd4NO3I03NZFjw0@skylark> Message-ID: <6.0.0.22.0.20040412174128.01b3aa38@pop.earthlink.net> Danielle, The thick sections we have been staining have seemed to prefer long incubations. Overnight would probably be best. Our PGP on skin biopsies at 50 um (free floating sections) have actually had the best results when they were left over the weekend but I have a hunch that we could do better with a more concentrated antibody. The shaker table is a good idea for free floating sections (if that is your plan). If you are labeling on a mounted slide the long incubation at room temperature and the shaker would probably cause a loss of the reagent. Amos Brooks At 12:00 PM 4/12/2004, you wrote: >Message: 2 >Date: Mon, 12 Apr 2004 09:44:17 -0400 >From: Danielle Zalinski >Subject: [Histonet] Vibratome Considerations >To: HistoNet Listserve >Message-ID: <407A9D31.70301@neuro.hfh.edu> >Content-Type: text/plain > > Hello All, >Our lab would like to do immunohistochem work in rat brain on vibratome >sections instead of paraffin. We are able to get sections about >50-60um. Will incubation with primary antibody overnight be sufficient? > Is shaking the sections an option? I was hoping some other labs that >are working in vibratome sections may have some beginners advice. >Thanks, >Danielle Zalinski > >Neurosurgery Research >Henry Ford Health System >Detroit, MI From bmcmahill <@t> incytepathology.com Mon Apr 12 19:28:40 2004 From: bmcmahill <@t> incytepathology.com (Bonnie J. McMahill) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] HIPAA question re: research Message-ID: <3F7537D9A2DE6D40BA884DF1A32A08CF1F6B8A@PATH2K-SRV2.PAI.E-PATHOLOGY.COM> Mike, >From what I understand, HIPAA allows for research as long as the process goes through a IRB process that explains how PHI is to be used and protected, if the patient's consent is needed, etc. I have just gone through this process with a local university on a research project that I am going through. Most institutions are also disclosing in their patient disclaimers that patient information and/or material may be used for research purposes. I have an agreement with the institutions who are providing tissue and patient information on how I am protecting the PHI. Bonnie McMahill Histology Supervisor InCyte Pathology Spokane, WA > -----Original Message----- > From: MTitford@aol.com [SMTP:MTitford@aol.com] > Sent: Monday, April 12, 2004 1:51 PM > To: Histonet@pathology.swmed.edu > Subject: [Histonet] HIPPA question re: research > > Has the HIPPA law effectively put an end to researchers going back through hospital records doing retrospective studies or doing research on old blocks? > > Mike Titford > USA Pathology > Mobile AL USA > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emry <@t> u.washington.edu Mon Apr 12 19:44:31 2004 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] McNeal Bone Stain Message-ID: Can any one direct me to the Mc Neal...Mc Neil...Bone stain. I have seen it mentioned in distraction studies. Thank you, Trisha Seattle From michael_lafriniere <@t> memorial.org Mon Apr 12 20:46:21 2004 From: michael_lafriniere <@t> memorial.org (LaFriniere, Mike) Date: Fri Sep 16 15:22:47 2005 Subject: [Histonet] Pathology Productivity Task Force Message-ID: The NSH Productivity Task Force is now in the final stages of obtaining the results for presentation to the NSH Board of Directors from the productivity workload reporting task force. We would like to get this finalized so we can submit for approval. To complete the task force results and present hard raw averages of duties performed in an anatomic pathology department nation wide, and present final data to BOD. We would like to have 100 different laboratories participating with the raw data collection process. Should your laboratory desire to participate in the productivity collection phase, which consists of 120 questions of which will be calculated in the final national averages, please contact me via email. I will send you the documents immediately to complete and return to me by May 15, 2004. Michael LaFriniere, PA, HT(ASCP) NSH Region III Director NSH Productivity/Workload Recording Chair From lwhite <@t> lakeridgehealth.on.ca Tue Apr 13 07:10:16 2004 From: lwhite <@t> lakeridgehealth.on.ca (White, Lori) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] RE: bcl-2 on Nexes Message-ID: Hi Lynda, What manufacturer are you using for your antibody? The reason I ask is that we were using Novocastra and the antibody did not seem to like the heat on the Nexes stainer - I had to do the staining manually at room temp. I have since switched to Dako (M0887) and it's working quite nicely.... Lori -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Thursday, April 08, 2004 10:23 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 5, Issue 13 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: xylene substitute (Judy Pariser) 2. Looking for Guss Mondragon (Victoria Baker) 3. CSH Symposium (May 13-16, 2004) - Hotel Phone Number (Laurie Colbert) 4. RE: p21 (Bartlett, Jeanine) 5. RE: Sponge problems in processing (Smith, Allen) 6. Thermo Shandon automatic stainer (Rizo, Christian) 7. RE: Sponge problems in processing (Stacy McLaughlin) 8. Re: Sponge problems in processing (DDittus787@aol.com) 9. Antigen retrieval for heart tissues? (Barlow, Gillian) 10. [BULK] - Re: [Histonet] Sponge problems in processing (Fred Underwood) 11. Histology opening in S CA. (Dave Johnson) 12. Re: Histonet Digest, Vol 5, Issue 11 (Amos Brooks) 13. Re: semithin-ultrathin - thank you (Gudrun Lang) 14. Interesting eosinophilic artifacts (Gayle Callis) 15. Eosinophilic artifacts posted in Histonet gallery (Gayle Callis) 16. RE: Re: Histonet Digest, Vol 5, Issue 11 (Patsy Ruegg) 17. RE: Re: Histonet Digest, Vol 5, Issue 11 (Luis Chiriboga) 18. Re: Re: Histonet Digest, Vol 5, Issue 11 (Patti Loykasek) 19. caspase 3 and transferrin receptor for rats (gsp26@drexel.edu) 20. RE: caspase 3 and transferrin receptor for rats (Favara, Cynthia (NIH/NIAID)) 21. Whipf's polychrome (Dixon, Leslie E.) 22. bcl-2 on Nexes (Derek & Lynda Leopold) 23. Perfusion System (Linresearch@aol.com) 24. ALAS (Sharon Cooperman) 25. lysosomal markers (Sharon Cooperman) 26. Re: heavy - chain deposition desease ( Katri Tuomala) ---------------------------------------------------------------------- Message: 1 Date: Thu, 8 Apr 2004 13:09:54 -0400 From: "Judy Pariser" Subject: Re: [Histonet] xylene substitute To: "Eddie Marquez" , Message-ID: <003001c41d8c$503a4700$0600a8c0@IN4> Content-Type: text/plain; charset="iso-8859-1" Dear Eddie, I just noticed your posting dated March 15, 2004 regarding xylene substitutes. I am the Project Manager for CBG Biotech's new clearing solvent, Formula 83. Formula 83 is completely non-toxic. In clinical and laboratory tests, Formula 83 consistently outperformed xylene and other xylene substitutes. It was designed specifically for tissue processing and staining. Formula 83 dries slides faster, dissolves paraffin faster, and recycles faster than xylene. Additionally, it will not harden tissue as xylene does, and has better lipid extraction. I would be happy to share more information with you. Additionally, I can send to you a reprint of a recently published article from Health Beat, a publication of the Healthcare Division of The American Society of Safety Engineers. I can be reached at the phone number or email address below. Best regards, Judy Pariser Project Manager Email: jpariser@cbgbiotech.com Phone: 800-941-9484 Fax: 614-863-1676 Website: http://www.cbgbiotech.com ----- Original Message ----- From: "Eddie Marquez" To: Sent: Monday, March 15, 2004 4:19 PM Subject: [Histonet] xylene substitute Aloha from Hawaii, Just needed some info.-Has anybody used xylene substitute with a linear stainer; for standard H&E's? If so, how good are the results? May I also have the brand name. Thank you, Eddie from the Cancer Research _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Thu, 8 Apr 2004 10:51:56 -0700 (PDT) From: Victoria Baker Subject: [Histonet] Looking for Guss Mondragon To: HistoNet Server Message-ID: <20040408175156.12088.qmail@web12108.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi Everyone, Guss if you're out there I need some of your sage wisdom. OR If anyone knows where Guss might be please contact me by this e-mail address. Thanks in advance. Vikki Baker Institute for Cancer Prevention Valhalla, NY 10595 __________________________________ Do you Yahoo!? Yahoo! Small Business $15K Web Design Giveaway http://promotions.yahoo.com/design_giveaway/ ------------------------------ Message: 3 Date: Thu, 8 Apr 2004 11:06:54 -0700 From: "Laurie Colbert" Subject: [Histonet] CSH Symposium (May 13-16, 2004) - Hotel Phone Number To: , "Debbie Cobb (E-mail)" Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BE91@EXCHANGE1.huntingtonhospital.com> Content-Type: text/plain; charset="iso-8859-1" The phone number that was listed for hotel reservations at the Burbank Hilton on the California Society for Histotechnology registration form is incorrect. The correct number is (800) 840-6450. We are sorry for any inconvenience this has caused. If you have any questions please contact Debbie Cobb at (818) 503-6807. ------------------------------ Message: 4 Date: Thu, 8 Apr 2004 14:14:23 -0400 From: "Bartlett, Jeanine" Subject: RE: [Histonet] p21 To: "Richard Cartun" , Message-ID: Content-Type: text/plain; charset="utf-8" Please share any information with the list please. Thanks! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Richard Cartun Sent: Wed 4/7/2004 7:26 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] p21 Has anyone tried Pharmagen's mAb to p21 (clone 6B6)? If so, has it worked well on formalin-fixed, paraffin-embedded tissue? Thank you. Richard Cartun Director, Immunopathology Hartford Hospital Hartford, CT 06102 "Home of the 2004 Men's and Women's Division I College Basketball National Champions!" _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Thu, 8 Apr 2004 14:31:43 -0400 From: "Smith, Allen" Subject: RE: [Histonet] Sponge problems in processing To: "Marshall Terry Dr, Consultant Histopathologist" Cc: histonet@lists.utsouthwestern.edu Message-ID: <494304423C63E246A5CF87A3AEEB577011B5DE@bumail01.barrynet.barry.edu> Content-Type: text/plain; charset="us-ascii" If an object is under water and the air pressure above the water is reduced, the total pressure on the object is reduced. If the object is only 17 feet down (1/2 Atm water pressure plus 1 Atm air pressure), halving the air pressure above it reduces the total pressure by 33% (1 Atm instead of 1 1/2 Atm). If the air pressure above it is reduced by 97% (to 0.03 Atm), the water boils at room temperature! If you seal the bag of fruit at normal atmospheric pressure, put the bag 17 feet underwater, and reduce the air pressure above by 50%, the fruit does not release juice because the total pressure (1 Atm) is still as high as when it was sealed. However, if you reduce the air pressure by 95%, the total pressure on the fruit will be 0.55 Atm. If you reduce the pressure to 0.55 Atm suddenly, air dissolved in the fruit juice will rupture the fruit and juice will leak out. If you reduce the pressure slowly to 0.55 Atm, the air will diffuse out without rupturing the fruit. If you slowly reduce the pressure on the fruit to 0.03 Atm, the water will diffuse out slowly, and you will have intact dehydrated fruit. If you reduce the pressure on the fruit 0.03 Atm suddenly, the rapid boiling of the juice will rupture the fruit and some juice will leak out before it, too, boils. Allen A. Smith, Ph.D. Professor of Anatomy School of Graduate Medical Sciences Barry University Miami Shores, FL (also PADI divemaster) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, April 08, 2004 11:41 AM To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Kemlo comes back (well he would): "But that's cos the air is trying to escape to equilibrate the outside reduction in air pressure. If the airplane was under water and had no air in it and the air pressure was reduced on top of the water then it would have no effect on the airplane, would it?" Any explanations as to what this means to me please. He continues: "Similarly the fruit in the bag is not under water, is it? Put the fruit under water in the bag and then reduce the pressure above the water, bet no juice comes out of the fruit except if there is trapped air and that forces some out as it exits." No, the fruit in the bag is not under water, but soon it is under fruit juice. The first thing that happens when you apply vacuum is that the air goes, then the fruit juice exits the fruit. However, there is a difference in the two systems, in that the bag in the vacuum press collapses. The retort hopefully does not. Would this make a difference? My capacity to conceive some of these concepts is quite paltry. "Bet you $10". I don't do bets or dollars. Someone put a car sponge in a VIP and stop it at the appropriate time would you? Terry L (wishing I'd kept my mouth shut) Marshall 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 08 April 2004 16:14 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Bet you $10 Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 08 April 2004 13:25 To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing When you see these "action thrillers" in airplanes, and a bullet/bomb/whatever makes a hole in the fuselage, everything rushes to the hole, not just the air. I can't envisage this process of sucking only the air or "surface air bubbles" and leaving a soggy sponge replete with its load. Under a vacuum, surely the sponge will collapse as the contained liquid runs out towards the exit. If you apply vacuum to a plastic bag containing fruit, the juice comes out, and this forms the basis of a vacuum press, one of which I possess. So there. 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 07 April 2004 16:48 To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing I thought vacuum processing only 'sucked away' air. I mean if you reduce the air pressure above the fluids then that reduces pressure within the tissue that was preventing air escaping (as it has a positive pressure). I suppose if you reduce the Saturated Vapour Pressure of a solvent then that would cause more solvent to vapourise but wouldn't that be from the surface of the solvent rather than within the tissue? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 06 April 2004 16:21 To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) ------------------------------ Message: 6 Date: Thu, 8 Apr 2004 14:45:42 -0400 From: "Rizo, Christian" Subject: [Histonet] Thermo Shandon automatic stainer To: histonet@lists.utsouthwestern.edu Message-ID: <3F8707A1ADC19C4FA84BC95B51CEF560094BA0EB@chi2kms03.columbuschildrens.net> Content-Type: text/plain Hi everybody, Can anybody share to me their experiences with a Thermo Shandon Automatic Stainer? We are trying to purchase one with a good deal. Thanks for your help. Chris Christian M. Rizo MBA, HTL (ASCP) Manager, Anatomic Pathology Childrens Hospital 700 Childrens Drive Columbus, Ohio 43205 614.722.5465 RizoC@chi.osu.edu Confidentiality Notice: This e-mail message, from Children's Hospital, Columbus, Ohio, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes pursuant to Children's Hospital's confidentiality policies. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. ------------------------------ Message: 7 Date: Thu, 8 Apr 2004 14:52:46 -0400 From: Stacy McLaughlin Subject: RE: [Histonet] Sponge problems in processing To: histonet@lists.utsouthwestern.edu Message-ID: <3D502BBF5356D31184650090275B750D0346C74D@mail.cooley-dickinson.org> Content-Type: text/plain Can't we all just have our fruit juice in a glass? :) -----Original Message----- From: Smith, Allen [mailto:asmith@mail.barry.edu] Sent: Thursday, April 08, 2004 1:32 PM To: Marshall Terry Dr, Consultant Histopathologist Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing If an object is under water and the air pressure above the water is reduced, the total pressure on the object is reduced. If the object is only 17 feet down (1/2 Atm water pressure plus 1 Atm air pressure), halving the air pressure above it reduces the total pressure by 33% (1 Atm instead of 1 1/2 Atm). If the air pressure above it is reduced by 97% (to 0.03 Atm), the water boils at room temperature! If you seal the bag of fruit at normal atmospheric pressure, put the bag 17 feet underwater, and reduce the air pressure above by 50%, the fruit does not release juice because the total pressure (1 Atm) is still as high as when it was sealed. However, if you reduce the air pressure by 95%, the total pressure on the fruit will be 0.55 Atm. If you reduce the pressure to 0.55 Atm suddenly, air dissolved in the fruit juice will rupture the fruit and juice will leak out. If you reduce the pressure slowly to 0.55 Atm, the air will diffuse out without rupturing the fruit. If you slowly reduce the pressure on the fruit to 0.03 Atm, the water will diffuse out slowly, and you will have intact dehydrated fruit. If you reduce the pressure on the fruit 0.03 Atm suddenly, the rapid boiling of the juice will rupture the fruit and some juice will leak out before it, too, boils. Allen A. Smith, Ph.D. Professor of Anatomy School of Graduate Medical Sciences Barry University Miami Shores, FL (also PADI divemaster) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, April 08, 2004 11:41 AM To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Kemlo comes back (well he would): "But that's cos the air is trying to escape to equilibrate the outside reduction in air pressure. If the airplane was under water and had no air in it and the air pressure was reduced on top of the water then it would have no effect on the airplane, would it?" Any explanations as to what this means to me please. He continues: "Similarly the fruit in the bag is not under water, is it? Put the fruit under water in the bag and then reduce the pressure above the water, bet no juice comes out of the fruit except if there is trapped air and that forces some out as it exits." No, the fruit in the bag is not under water, but soon it is under fruit juice. The first thing that happens when you apply vacuum is that the air goes, then the fruit juice exits the fruit. However, there is a difference in the two systems, in that the bag in the vacuum press collapses. The retort hopefully does not. Would this make a difference? My capacity to conceive some of these concepts is quite paltry. "Bet you $10". I don't do bets or dollars. Someone put a car sponge in a VIP and stop it at the appropriate time would you? Terry L (wishing I'd kept my mouth shut) Marshall 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 08 April 2004 16:14 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing Bet you $10 Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 08 April 2004 13:25 To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing When you see these "action thrillers" in airplanes, and a bullet/bomb/whatever makes a hole in the fuselage, everything rushes to the hole, not just the air. I can't envisage this process of sucking only the air or "surface air bubbles" and leaving a soggy sponge replete with its load. Under a vacuum, surely the sponge will collapse as the contained liquid runs out towards the exit. If you apply vacuum to a plastic bag containing fruit, the juice comes out, and this forms the basis of a vacuum press, one of which I possess. So there. 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 07 April 2004 16:48 To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing I thought vacuum processing only 'sucked away' air. I mean if you reduce the air pressure above the fluids then that reduces pressure within the tissue that was preventing air escaping (as it has a positive pressure). I suppose if you reduce the Saturated Vapour Pressure of a solvent then that would cause more solvent to vapourise but wouldn't that be from the surface of the solvent rather than within the tissue? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 06 April 2004 16:21 To: Steven E. Slap; Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Sponge problems in processing OK. 200 ml. of liquid retained, with 100 blocks. However, with a vacuum processor, is that not all "sucked away"? BTW, should it sound otherwise, I hate the things (but am stuck with them). 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 06 April 2004 16:14 To: Gayle Callis; Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sponge problems in processing Hi HistoNetters I agree with Gayle, and the other posters who pointed out that sponges carry over 1ml of fluid per sponge (so, with 100 cassettes, 200 sponges, that's a lot of carryover). I have had a lot of success with the biopsy cassettes which Gayle refers to, both in microwaves and in conventional tissue processors. You can request samples from Lab Storage Systems in St. Louis by phone at (800) 345-4167 or by e-mailing Rita Lovshe at rcl@labstore.com. best regards, Steven Slap At 10:20 AM -0700 4/1/04, Gayle Callis wrote: >To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Cc: >Subject: [Histonet] Sponge problems in processing > >Gudrun, > >We no longer use sponges, prefer to place tissues in tissue embedding bags >(Fisher) which look like tea bags or the little nylon bags (not as easy to >handle, but thinner than sponges) and a tidge stiffer than tea bags. >Sponges can cause artifacts in your tissues, looking like triangular holes >in section. This was published by Freida Carson in Journal of >Histotechnology, 1980's. Using tea bags may not be as fast during >embedding, but speed there is a trade off for having to reprocess important >tissue samples, ho hum tedious and time consuming while patient waits. > >You analysis of problem sound correct with a poor exchange of solvents >through sponges. If you pack cassettes super tight in basket/cassette >holder inside processor you can impede solvent flow or if sponges are >crammed too tight agaist tissue then lid smashed down on tissue/sponge >sandwich - this is like having too thick a tissue in a cassette. > >There are some clever biopsy cassettes with a folding, fine mesh inserts >fitting inside a cassette. These may be worth a try to avoid sponges. I >think they are available through Fisher or some other company who could >provide free samples. It will be interesting to see peoples testimonials >on using these. > >There was Histonet discussion on these sponges way back in time - it may >pay to do a search for those messages in Histonet Archives. _______________________________________________ Histonet mailing list Histonet@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 The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet THIS E-MAIL MESSAGE IS INTENDED ONLY FOR THE USE OF THE INDIVIDUAL OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If the reader of this e-mail message is 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 e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. ------------------------------ Message: 8 Date: Thu, 08 Apr 2004 14:58:36 -0400 From: DDittus787@aol.com Subject: Re: [Histonet] Sponge problems in processing To: Stacy_McLaughlin@cooley-dickinson.org, histonet@lists.utsouthwestern.edu Message-ID: <2F934444.658092DE.0A1F969F@aol.com> Content-Type: text/plain; charset=iso-8859-1 owwwwwwwwwwwwwww my head hurts, stop using sponges pleaseeeeeeeeee. just a little humor this was getting a bit intense. Dana ------------------------------ Message: 9 Date: Thu, 8 Apr 2004 11:58:49 -0700 From: "Barlow, Gillian" Subject: [Histonet] Antigen retrieval for heart tissues? To: 'Histonet' Message-ID: <3CFAA0108952D111A5BF00805FA6FB0F0794750E@PEDSNTAS.csmc.edu> Content-Type: text/plain; charset="us-ascii" Dear Histonetters Does anyone out there know what (if any) antigen retrieval is necessary for the following antibodies in sections of paraffin-fixed, formalin-embedded adult mouse heart: - Cx43 - alpha-actinin - beta-catenin Many thanks Gillian Gillian M. Barlow, PhD Postdoctoral Fellow Laboratory of Julie Korenberg, PhD, MD Cedars-Sinai Medical Center Davis Bldg, Lab 2007 110 George Burns Rd Los Angeles, CA 90048 Phone: (310) 423 7650 Fax: (310) 423 0302 ------------------------------ Message: 10 Date: Thu, 08 Apr 2004 15:47:00 -0400 From: "Fred Underwood" Subject: [BULK] - Re: [Histonet] Sponge problems in processing To: Message-ID: Content-Type: text/plain; charset=US-ASCII It's best to follow the lead of Elaine Benes and determine just which specimens are sponge worthy. >>> 04/08/04 02:58PM >>> owwwwwwwwwwwwwww my head hurts, stop using sponges pleaseeeeeeeeee. just a little humor this was getting a bit intense. Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Thu, 08 Apr 2004 17:18:28 -0400 From: "Dave Johnson" Subject: [Histonet] Histology opening in S CA. To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Attention Histonet!, New Histology Lab in Southern Califonia has job opening for part time or fulltime histology technician. Duties include all applications for operating paraffin histology laboratory. Laboratory is processing specimens for Dermatology Practice. Please send your resume to the attention of Jane Litz, Administrator Manager, Fax: 760-724-9929. Candidate needed to fill position immediately. > Thanks >Rudy Gutierrez >Pacific Southwest Lab Equipment Inc. >Product and Sales Division >Main: 760-295-1842 >Cellular: 760-525-9071 > _________________________________________________________________ Limited-time offer: Fast, reliable MSN 9 Dial-up Internet access FREE for 2 months! http://join.msn.com/?page=dept/dialup&pgmarket=en-us&ST=1/go/onm00200361ave/ direct/01/ ------------------------------ Message: 12 Date: Thu, 08 Apr 2004 17:19:03 -0500 From: Amos Brooks Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 To: histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.0.20040408170800.01b704e8@pop.earthlink.net> Content-Type: text/plain; charset="us-ascii"; format=flowed Linda, Sorry to say there's no such thing. VEGF, despite any specifications, just doesn't work properly. We've tried at least 4 different vendors with hardly any success. Either it didn't label at all, labeled in all the wrong places, or the negative control looked the same as the test tissue. Check the controls closely if you try this. It's just not a reliable antibody. Regards, Amos Brooks At 09:20 AM 4/8/2004, you wrote: >Message: 2 >Date: Wed, 7 Apr 2004 13:00:57 -0500 >From: "Sebree Linda A." >Subject: [Histonet] VEGF antibody >To: "Histonet (E-mail)" >Message-ID: > >Content-Type: text/plain; charset="iso-8859-1" > >Hi again, > >Wondering if anyone can recommend a good, reliable VEGF (vascular >endothelial growth factor) antibody? If you've run yours on a Ventana >instrument, so much the better. > >Thanks for the 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 ------------------------------ Message: 13 Date: Thu, 8 Apr 2004 23:34:01 +0200 From: "Gudrun Lang" Subject: [Histonet] Re: semithin-ultrathin - thank you To: "Histonetliste" Message-ID: <005f01c41db1$35901b30$eeeea8c0@SERVER> Content-Type: text/plain; charset="iso-8859-1" Thank you for your explanations about the thickness of sections. Gudrun Lang, Austria PS.: I read the sponge-debate with interest. ----- Original Message ----- From: "David Kelly, M.D." To: "'Gudrun Lang'" Sent: Thursday, April 08, 2004 12:04 AM Subject: RE: [Histonet] semithin-ultrathin > Tim Morken's description of these terms is accurate. If you need a > reference, the book is old and probably out of print but still an excellent > introduction to electron microscopy. Appropriately, it is entitled "An > Introduction to Diagnostic Electron Microscopy," by Bruce Mackay > (Appleton-Century-Crofts, New York, 1981). See Chapter 2 Technical > Procedures, by Paul S. Baur and Bruce Mackay, p 40 and Chapter 3 > Preparation and Interpretation of Semithin Sections, by Willard A. Burns, pp > 47-48. > > -----Original Message----- > From: Gudrun Lang [mailto:gudrun.lang@aon.at] > Sent: Wednesday, April 07, 2004 2:06 PM > To: Histonetliste > Subject: [Histonet] semithin-ultrathin > > > Hi, > I've searched in the internet for a list that explains the thickness of > semi-, ultra-, thin, thick sections. But I failed. My knowledge is only > approximately. > Perhaps someone can help me? > Gudrun Lang > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 14 Date: Thu, 08 Apr 2004 15:34:41 -0600 From: Gayle Callis Subject: [Histonet] Interesting eosinophilic artifacts To: Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040408153441.00bc20c0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" After all my years doing histo work, I have one H&E tissue section, mouse lung which shows crystalline shaped eosinophilic artifacts. I stained other lung sections from this experiment and murine reproductive tract, placenta at the same time. These have no artifacts. Some of the crystals actually look ingested by macrophages. If anyone is interested, I will be happy to drop photo privately. I have never seen artifacts like these. Baffled in Montana! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 15 Date: Thu, 08 Apr 2004 15:48:23 -0600 From: Gayle Callis Subject: [Histonet] Eosinophilic artifacts posted in Histonet gallery To: Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040408154823.00ba3030@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" Photo of eosinophilic crystalline structures to be posted in Histonet gallery. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 16 Date: Thu, 8 Apr 2004 15:48:28 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 To: "Amos Brooks" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Amos I had the same experience as you with VEGF's until I switched to monoclonal Vegf from Santa Cruz, it now appears very specific and reliable to me. I use pepsin or proteinase K digestion and stay away from HIER. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amos Brooks Sent: Thursday, April 08, 2004 4:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 Linda, Sorry to say there's no such thing. VEGF, despite any specifications, just doesn't work properly. We've tried at least 4 different vendors with hardly any success. Either it didn't label at all, labeled in all the wrong places, or the negative control looked the same as the test tissue. Check the controls closely if you try this. It's just not a reliable antibody. Regards, Amos Brooks At 09:20 AM 4/8/2004, you wrote: >Message: 2 >Date: Wed, 7 Apr 2004 13:00:57 -0500 >From: "Sebree Linda A." >Subject: [Histonet] VEGF antibody >To: "Histonet (E-mail)" >Message-ID: > >Content-Type: text/plain; charset="iso-8859-1" > >Hi again, > >Wondering if anyone can recommend a good, reliable VEGF (vascular >endothelial growth factor) antibody? If you've run yours on a Ventana >instrument, so much the better. > >Thanks for the 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Thu, 08 Apr 2004 17:54:48 -0400 From: Luis Chiriboga Subject: RE: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 To: Amos Brooks , histonet@lists.utsouthwestern.edu, la.sebree@hosp.wisc.edu Message-ID: Content-Type: text/plain; charset=US-ASCII I agree strongly with Amos and have basically gotten the same results. it's a pain in the neck (and I don't mean from looking in the scope). I have run a couple, from different manufacturers, tried all the usual tricks(automated and not) and none of them performed consistently. I figured I would just pick one and stick with and just go the brute force approach, use it until I get results that I can live with. Hope this helps, regards luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amos Brooks Sent: Thursday, April 08, 2004 6:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 Linda, Sorry to say there's no such thing. VEGF, despite any specifications, just doesn't work properly. We've tried at least 4 different vendors with hardly any success. Either it didn't label at all, labeled in all the wrong places, or the negative control looked the same as the test tissue. Check the controls closely if you try this. It's just not a reliable antibody. Regards, Amos Brooks At 09:20 AM 4/8/2004, you wrote: >Message: 2 >Date: Wed, 7 Apr 2004 13:00:57 -0500 >From: "Sebree Linda A." >Subject: [Histonet] VEGF antibody >To: "Histonet (E-mail)" >Message-ID: > >Content-Type: text/plain; charset="iso-8859-1" > >Hi again, > >Wondering if anyone can recommend a good, reliable VEGF (vascular >endothelial growth factor) antibody? If you've run yours on a Ventana >instrument, so much the better. > >Thanks for the 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Thu, 08 Apr 2004 15:07:11 -0700 From: Patti Loykasek Subject: Re: [Histonet] Re: Histonet Digest, Vol 5, Issue 11 To: histonet Message-ID: Content-Type: text/plain; charset="US-ASCII" Amos & all - I couldn't have said it better myself. The only difference is that we have tried >6 clones. Be very careful of the staining, read up on the expected positivity, and pick your positive controls carefully. Patti Loykasek > Linda, > Sorry to say there's no such thing. VEGF, despite any > specifications, just doesn't work properly. We've tried at least 4 > different vendors with hardly any success. Either it didn't label at all, > labeled in all the wrong places, or the negative control looked the same as > the test tissue. Check the controls closely if you try this. It's just not > a reliable antibody. > Regards, > Amos Brooks > > At 09:20 AM 4/8/2004, you wrote: >> Message: 2 >> Date: Wed, 7 Apr 2004 13:00:57 -0500 >> From: "Sebree Linda A." >> Subject: [Histonet] VEGF antibody >> To: "Histonet (E-mail)" >> Message-ID: >> >> Content-Type: text/plain; charset="iso-8859-1" >> >> Hi again, >> >> Wondering if anyone can recommend a good, reliable VEGF (vascular >> endothelial growth factor) antibody? If you've run yours on a Ventana >> instrument, so much the better. >> >> Thanks for the 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Thu, 08 Apr 2004 18:20:18 -0400 From: gsp26@drexel.edu Subject: [Histonet] caspase 3 and transferrin receptor for rats To: histonet@lists.utsouthwestern.edu Message-ID: <1d855f81d8788c.1d8788c1d855f8@drexel.edu> Content-Type: text/plain; charset=us-ascii Has anyone used caspase 3 IHC on rat paraffin embedded tissue. the same question for transferrin receptor IHC. Please advise!! ------------------------------ Message: 20 Date: Thu, 8 Apr 2004 18:26:08 -0400 From: "Favara, Cynthia (NIH/NIAID)" Subject: RE: [Histonet] caspase 3 and transferrin receptor for rats To: "'gsp26@drexel.edu'" , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" I have done cleaved caspase 3 on FFPE mouse tissue Source: Cell Signaling #9961 c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: gsp26@drexel.edu [mailto:gsp26@drexel.edu] Sent: Thursday, April 08, 2004 4:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] caspase 3 and transferrin receptor for rats Has anyone used caspase 3 IHC on rat paraffin embedded tissue. the same question for transferrin receptor IHC. Please advise!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Thu, 8 Apr 2004 15:34:01 -0700 From: "Dixon, Leslie E." Subject: [Histonet] Whipf's polychrome To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Good afternoon, I am curious if anyone has the procedure for Whipf's polychrome? Thanks in advance, Leslie ------------------------------ Message: 22 Date: Fri, 9 Apr 2004 18:51:44 -0400 From: "Derek & Lynda Leopold" Subject: [Histonet] bcl-2 on Nexes To: Message-ID: <000801c41e85$3bc3fc40$3643fea9@leopold> Content-Type: text/plain; charset="iso-8859-1" Hi All, We're having some trouble getting our tonsil control to come up for bcl-2. After searching the archives and asking around, we are going to try citrate buffer (Declere) and a hot steamer for 1 hour. We've got the Nexes IHC. Anyone else have any tips for us? Can anyone comment on the need/non-need for DAB? Thanks Lynda Leopold ------------------------------ Message: 23 Date: Thu, 8 Apr 2004 18:45:23 EDT From: Linresearch@aol.com Subject: [Histonet] Perfusion System To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello, Anyone with experience using the Perfusio 1 System from MyNeuroLab, Could you give me feedback on the sytem? If someone has a better system, could you let me know? Lin ------------------------------ Message: 24 Date: Thu, 8 Apr 2004 19:48:23 -0400 From: Sharon Cooperman Subject: [Histonet] ALAS To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Hi, Does anyone know of a source for antisera to aminolevulinic acid synthase? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 ------------------------------ Message: 25 Date: Thu, 8 Apr 2004 19:51:06 -0400 From: Sharon Cooperman Subject: [Histonet] lysosomal markers To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Hi, Does anyone know of an antibody/antiserum to a lysosomal marker such as Lamp 1 or Lamp 2 that cross reacts with mouse and works on formalin-fixed paraffin embedded tissue (with or without antigen retrieval)? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 ------------------------------ Message: 26 Date: Thu, 8 Apr 2004 22:23:23 -0400 From: " Katri Tuomala" Subject: Re: [Histonet] heavy - chain deposition desease To: "???? ??" , Message-ID: <009901c41dd9$a538fa60$ce989618@hala4.on.cogeco.ca> Content-Type: text/plain; charset="gb2312" Hi! Are you working on human or animal tissue? In human renal biopsies most commonly used technique is probably immunofluorescence on snap frozen tissue. We use FITC conjugated antibodies from DakoCytomation for heavy chains IgA, IgG and IgM in addition to C3complement, fibrinogen, kappa & lambda light chains and albumin. These can be demonstrated also on FFPE tissue, but the method is more involved and takes more time to develop to obtain reproducible results. Immunofluorescence is a long standing and a simple technique to perform... Katri Katri Tuomala Department of Pathology St.Joseph's Health Care Hamilton, Ontario, Canada ----- Original Message ----- From: "???? ??" To: Sent: Thursday, April 08, 2004 10:36 AM Subject: [Histonet] heavy - chain deposition desease > Hello,everobody: > I am very interested in the diagonosis of heavy-chain deposition disease.Firstly ,I am going to find out whether there are deposition of heavy chains along tubular and glomerular basement membranes.But I have no idea about which company's antibodyies are appropriate.Would anybody be kind enough to share me with you experieace? > Thanks a lot. > Beat Regards! > Shuqiong Shen > Research Insititute of Nephrology > 305 East Zhongshan Road > Nanjing 210002,P.R.China > > > > > --------------------------------- > Do You Yahoo!? > ????TT???????????????????????? > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 5, Issue 13 *************************************** From psanquin <@t> lugo.usc.es Tue Apr 13 08:44:48 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Buffers for vibratome sections In-Reply-To: <014e01c4183a$77b9eda0$78065486@vdl220FAC> Message-ID: <3.0.6.32.20040413154448.007ec100@pop.lugo.usc.es> Dear listers, Before processing them I have to keep brain vibratome sections for a couple of weeks in the refrigerator. Could you suggest me a proper buffer? The buffer I use in the bath of the vibratome was Phosphate Buffer 0.1M. Thanks in advance. Pablo Sanchez From mikael.niku <@t> helsinki.fi Tue Apr 13 09:12:23 2004 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] ISH probe labelling Message-ID: <001201c42161$5795dcc0$8c0fd680@ekk1116> Dear Histonetters, I'm about to start doing some tissue in situ hybridization for a not-so-abundant mRNA target. I'm uncertain about the probe labelling method I should use. I need to use an oligonucleotide probe. I have previously used an oligo probe for a highly amplified genomic target, and cRNA probes for mRNA targets. But I don't have any experience in this particular combination. So what do you think: Will a probe with a single DIG label be sensitive enough? If not, is it feasible to order a doubly or triply DIG labeled probe, or should I try tailing instead? I will be using either the typical anti-DIG alk phos + NBT/BCIP detection system, or tyramide amplification, if necessary. With best regards, Mikael Niku +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ + Mikael Niku + University of Helsinki, Dept. Basic Veterinary Sciences + URL: www.helsinki.fi/~mniku/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! (Gandhi) From m.vankempen <@t> erasmusmc.nl Tue Apr 13 09:18:13 2004 From: m.vankempen <@t> erasmusmc.nl (m. van mkempen) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel Message-ID: <407BF6A5.9DC6D632@erasmusmc.nl> Dear all, I have a question concerning histogel!! I'm working with fetal lung and I thought histogel would be a nice way to handle then while embedding. My question is do I have to fix the histogel before processing for embedding in paraffine? How is the normal procedure for tissues's in histogel?? Thank you for all your kind answers. Regards, Marta. -- ---------------------------- *- mkempen -* MAILTO:m.vankempen@erasmusmc.nl ----------------------------------------- From sladd <@t> hsc.usf.edu Tue Apr 13 09:49:12 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel In-Reply-To: <407BF6A5.9DC6D632@erasmusmc.nl> Message-ID: Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before processing it WILL DISSOLVE! I learned this the hard way! The instructions say something vague like "process as usual". Of course, I informed Richard Allen that many of us in research do not have formalin on our tissue processors. Richard Allen said they were going to change the instructions. Once you have fixed the histogel you can process it like you would any other piece of tissue. Sharron University of South Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van mkempen Sent: Tuesday, April 13, 2004 10:18 AM To: Histonetlist Subject: [Histonet] histogel Dear all, I have a question concerning histogel!! I'm working with fetal lung and I thought histogel would be a nice way to handle then while embedding. My question is do I have to fix the histogel before processing for embedding in paraffine? How is the normal procedure for tissues's in histogel?? Thank you for all your kind answers. Regards, Marta. -- ---------------------------- *- mkempen -* MAILTO:m.vankempen@erasmusmc.nl ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Tue Apr 13 13:02:38 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Vibratome Considerations In-Reply-To: <407A9D31.70301@neuro.hfh.edu> References: <407A9D31.70301@neuro.hfh.edu> Message-ID: <407C2B3E.40602@umdnj.edu> I have done immunos on vibratome sections of rat brain. If you use a 12 well tissue culture plate (with its lid) on a shaker table overnight at room temp. you should get good results. You will need to add some Triton or Tween to the primary antibody (and probably subsequent steps) or do something to 'permeabilize' the sections (freeze-thaw or alcohol) so you get good penetration of the antibodies. If you have access to a sliding microtome with a freezing stage you could obtain sections that way. Geoff Danielle Zalinski wrote: > Hello All, >Our lab would like to do immunohistochem work in rat brain on vibratome >sections instead of paraffin. We are able to get sections about >50-60um. Will incubation with primary antibody overnight be sufficient? > Is shaking the sections an option? I was hoping some other labs that >are working in vibratome sections may have some beginners advice. >Thanks, >Danielle Zalinski > >Neurosurgery Research >Henry Ford Health System >Detroit, MI > > >CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. > >Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. >_______________________________________________ >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 ramona.tolliver <@t> yale.edu Tue Apr 13 10:14:18 2004 From: ramona.tolliver <@t> yale.edu (Ramona Tolliver) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Histology Manager Opening at Yale University Message-ID: <6.0.1.1.2.20040413111131.01e75d48@email.med.yale.edu> Good morning, Yale University has a Histology Manager (1st shift) position available from 7:00 a.m. - 3:00 p.m. If you know of anyone who may be interested in an outstanding growth opportunity in a department committed to excellence in patient care, teaching, and discovery, please forward this information on to them. Salary is commensurate with experience. Relocation assistance is available. The position is posted online at : http://websrv.its.yale.edu/hr/cgi-bin/printjob.plx?file=mp-2&job=11326 . Inquiries should be sent to ramona.tolliver@yale.edu. Thank you for your time! Ramona E. Tolliver *If you have difficulty with the link provided above, please go to www.yale.edu/jobs and search Managerial and Professional last two weeks and Pathology. The position is listed as Manager III. Ramona E. Tolliver Human Resource Manager Yale University School of Medicine Department of Pathology 310 Cedar Street New Haven, CT 06520-8023 Telephone: (203) 785-6689 Fax: (203) 785-7303 From algranth <@t> u.arizona.edu Tue Apr 13 10:28:11 2004 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel In-Reply-To: References: <407BF6A5.9DC6D632@erasmusmc.nl> Message-ID: <4.3.2.7.2.20040413082229.00cb9f10@algranth.inbox.email.arizona.edu> When I use histogel I do "process as usual" and this does not include formalin. I put the cassette containing the tissue embedded in histogel into 70% ETOH. The processor starts with 70% ETOH X 2 and the concentration of ETOH goes up from there. Haven't had any problems. Maybe I've just been lucky? Andi Grantham At 10:49 AM 4/13/2004 -0400, you wrote: >Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before >processing it WILL DISSOLVE! I learned this the hard way! The instructions >say something vague like "process as usual". Of course, I informed Richard >Allen that many of us in research do not have formalin on our tissue >processors. Richard Allen said they were going to change the instructions. >Once you have fixed the histogel you can process it like you would any other >piece of tissue. >Sharron >University of South Florida > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van >mkempen >Sent: Tuesday, April 13, 2004 10:18 AM >To: Histonetlist >Subject: [Histonet] histogel > > >Dear all, > >I have a question concerning histogel!! >I'm working with fetal lung and I thought histogel would be a nice way >to handle then while embedding. >My question is do I have to fix the histogel before processing for >embedding in paraffine? >How is the normal procedure for tissues's in histogel?? > >Thank you for all your kind answers. > >Regards, >Marta. > >-- >---------------------------- >*- mkempen -* >MAILTO:m.vankempen@erasmusmc.nl >----------------------------------------- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From cornettl <@t> hotmail.com Tue Apr 13 10:32:55 2004 From: cornettl <@t> hotmail.com (Lorraine Cornett) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Zinc formalin Message-ID: We are currentyl processing our biopsies in 10% neutral buffered formalin (short process cycle). Our pathologists miss the nuclear detail they became accustomed to when we were processing in Zinc Formalin. We seem to remember a procedure where you can "mordant" or post fix the actual slides before staining. Does anyone have this procedure that they can share with me. Thanks, Lorraine Cornett (HT ASCP) Blue Ridge Pathology Kingsport, TN 37660 423 224-5793 _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From convmcm <@t> cc.usu.edu Tue Apr 13 10:24:43 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel In-Reply-To: Message-ID: <000101c4216b$72f8c5e0$4a737b81@Cygnus> Forgive my ignorance, but what is histogel and what is it used for? thanx Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of S Ladd Sent: Tuesday, April 13, 2004 7:49 AM To: m. van mkempen Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histogel Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before processing it WILL DISSOLVE! I learned this the hard way! The instructions say something vague like "process as usual". Of course, I informed Richard Allen that many of us in research do not have formalin on our tissue processors. Richard Allen said they were going to change the instructions. Once you have fixed the histogel you can process it like you would any other piece of tissue. Sharron University of South Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van mkempen Sent: Tuesday, April 13, 2004 10:18 AM To: Histonetlist Subject: [Histonet] histogel Dear all, I have a question concerning histogel!! I'm working with fetal lung and I thought histogel would be a nice way to handle then while embedding. My question is do I have to fix the histogel before processing for embedding in paraffine? How is the normal procedure for tissues's in histogel?? Thank you for all your kind answers. Regards, Marta. -- ---------------------------- *- mkempen -* MAILTO:m.vankempen@erasmusmc.nl ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@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 sladd <@t> hsc.usf.edu Tue Apr 13 10:42:46 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel In-Reply-To: <4.3.2.7.2.20040413082229.00cb9f10@algranth.inbox.email.arizona.edu> Message-ID: Are the items you are embedding in the histogel fixed in formalin? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrea Grantham Sent: Tuesday, April 13, 2004 11:28 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histogel When I use histogel I do "process as usual" and this does not include formalin. I put the cassette containing the tissue embedded in histogel into 70% ETOH. The processor starts with 70% ETOH X 2 and the concentration of ETOH goes up from there. Haven't had any problems. Maybe I've just been lucky? Andi Grantham At 10:49 AM 4/13/2004 -0400, you wrote: >Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before >processing it WILL DISSOLVE! I learned this the hard way! The instructions >say something vague like "process as usual". Of course, I informed Richard >Allen that many of us in research do not have formalin on our tissue >processors. Richard Allen said they were going to change the instructions. >Once you have fixed the histogel you can process it like you would any other >piece of tissue. >Sharron >University of South Florida > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van >mkempen >Sent: Tuesday, April 13, 2004 10:18 AM >To: Histonetlist >Subject: [Histonet] histogel > > >Dear all, > >I have a question concerning histogel!! >I'm working with fetal lung and I thought histogel would be a nice way >to handle then while embedding. >My question is do I have to fix the histogel before processing for >embedding in paraffine? >How is the normal procedure for tissues's in histogel?? > >Thank you for all your kind answers. > >Regards, >Marta. > >-- >---------------------------- >*- mkempen -* >MAILTO:m.vankempen@erasmusmc.nl >----------------------------------------- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sladd <@t> hsc.usf.edu Tue Apr 13 10:42:45 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20A96E4@usca0082k08.labvision.apogent.com> Message-ID: Since your cells were fixed in formalin, perhaps there was enough residual formalin to sufficiently fix the histogel? You use only a few drops of histogel. My cells were not in formalin and I use about 0.5 mL of histogel. Just for fun, you could try processing a little button of histogel (without formalin fixed cells) and see if it dissolves? I use ethanol and xylene on my processor and I also start with 70% ethanol. Sharron -----Original Message----- From: Kyle-Byrne, Carrie - Labvision [mailto:CKByrne@labvision.com] Sent: Tuesday, April 13, 2004 11:23 AM To: 'S Ladd' Subject: RE: [Histonet] histogel Sharron, Interesting. I've never heard of this before. I've used Histogel for years and never had to 'fix' it in formalin. I used it to make cell pellets for use as IHC control material. I would fix the cells in formalin first then transfer to 70% EtOH and hold them until I had several to pellet. I'd decant the 70%, put on a few drops of Histogel, mix to distribute the cells, then cool the tube to harden the gel. Once solidified, I'd remove the pellet from the tube and breadloaf it like a piece of tissue, place in a cassette and onto the processor (we started in 70% EtOH) for overnight processing and embed as usual in the morning. It always worked just fine. What to you use for clearing? Carrie Kyle-Byrne Sr. Reseach Assoc., Pathology Lab Lab Vision, Corp. 47791 Westinghouse Ave. Fremont, CA 94539 ckbyrne@labvision.com www.labvision.com -----Original Message----- From: S Ladd [mailto:sladd@hsc.usf.edu] Sent: Tuesday, April 13, 2004 7:49 AM To: m. van mkempen Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histogel Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before processing it WILL DISSOLVE! I learned this the hard way! The instructions say something vague like "process as usual". Of course, I informed Richard Allen that many of us in research do not have formalin on our tissue processors. Richard Allen said they were going to change the instructions. Once you have fixed the histogel you can process it like you would any other piece of tissue. Sharron University of South Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van mkempen Sent: Tuesday, April 13, 2004 10:18 AM To: Histonetlist Subject: [Histonet] histogel Dear all, I have a question concerning histogel!! I'm working with fetal lung and I thought histogel would be a nice way to handle then while embedding. My question is do I have to fix the histogel before processing for embedding in paraffine? How is the normal procedure for tissues's in histogel?? Thank you for all your kind answers. Regards, Marta. -- ---------------------------- *- mkempen -* MAILTO:m.vankempen@erasmusmc.nl ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@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 jqb7 <@t> cdc.gov Tue Apr 13 10:43:09 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel Message-ID: We do the same and have had no problems. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Andrea Grantham Sent: Tuesday, April 13, 2004 11:28 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histogel When I use histogel I do "process as usual" and this does not include formalin. I put the cassette containing the tissue embedded in histogel into 70% ETOH. The processor starts with 70% ETOH X 2 and the concentration of ETOH goes up from there. Haven't had any problems. Maybe I've just been lucky? Andi Grantham At 10:49 AM 4/13/2004 -0400, you wrote: >Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before >processing it WILL DISSOLVE! I learned this the hard way! The >instructions say something vague like "process as usual". Of course, I >informed Richard Allen that many of us in research do not have formalin >on our tissue processors. Richard Allen said they were going to change >the instructions. Once you have fixed the histogel you can process it >like you would any other piece of tissue. Sharron >University of South Florida > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van >mkempen >Sent: Tuesday, April 13, 2004 10:18 AM >To: Histonetlist >Subject: [Histonet] histogel > > >Dear all, > >I have a question concerning histogel!! >I'm working with fetal lung and I thought histogel would be a nice way >to handle then while embedding. My question is do I have to fix the >histogel before processing for embedding in paraffine? >How is the normal procedure for tissues's in histogel?? > >Thank you for all your kind answers. > >Regards, >Marta. > >-- >---------------------------- >*- mkempen -* >MAILTO:m.vankempen@erasmusmc.nl >----------------------------------------- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Tue Apr 13 10:47:04 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel Message-ID: Yes. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of S Ladd Sent: Tuesday, April 13, 2004 11:43 AM To: Andrea Grantham Cc: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histogel Are the items you are embedding in the histogel fixed in formalin? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrea Grantham Sent: Tuesday, April 13, 2004 11:28 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histogel When I use histogel I do "process as usual" and this does not include formalin. I put the cassette containing the tissue embedded in histogel into 70% ETOH. The processor starts with 70% ETOH X 2 and the concentration of ETOH goes up from there. Haven't had any problems. Maybe I've just been lucky? Andi Grantham At 10:49 AM 4/13/2004 -0400, you wrote: >Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before >processing it WILL DISSOLVE! I learned this the hard way! The >instructions say something vague like "process as usual". Of course, I >informed Richard Allen that many of us in research do not have formalin >on our tissue processors. Richard Allen said they were going to change >the instructions. Once you have fixed the histogel you can process it >like you would any other >piece of tissue. >Sharron >University of South Florida > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van >mkempen >Sent: Tuesday, April 13, 2004 10:18 AM >To: Histonetlist >Subject: [Histonet] histogel > > >Dear all, > >I have a question concerning histogel!! >I'm working with fetal lung and I thought histogel would be a nice way >to handle then while embedding. My question is do I have to fix the >histogel before processing for embedding in paraffine? >How is the normal procedure for tissues's in histogel?? > >Thank you for all your kind answers. > >Regards, >Marta. > >-- >---------------------------- >*- mkempen -* >MAILTO:m.vankempen@erasmusmc.nl >----------------------------------------- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Tue Apr 13 11:00:05 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel In-Reply-To: Message-ID: I use histogel all the time without fixing in formalin. Like Andi I start with tissue already fixed and then placed in 70% alcohol. I embed this tissue in histogel and put it back in the 70% and then process starting with 70% alcohol, no formalin. I have not had the histogel ever dissolve. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of S Ladd Sent: Tuesday, April 13, 2004 8:49 AM To: m. van mkempen Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histogel Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before processing it WILL DISSOLVE! I learned this the hard way! The instructions say something vague like "process as usual". Of course, I informed Richard Allen that many of us in research do not have formalin on our tissue processors. Richard Allen said they were going to change the instructions. Once you have fixed the histogel you can process it like you would any other piece of tissue. Sharron University of South Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van mkempen Sent: Tuesday, April 13, 2004 10:18 AM To: Histonetlist Subject: [Histonet] histogel Dear all, I have a question concerning histogel!! I'm working with fetal lung and I thought histogel would be a nice way to handle then while embedding. My question is do I have to fix the histogel before processing for embedding in paraffine? How is the normal procedure for tissues's in histogel?? Thank you for all your kind answers. Regards, Marta. -- ---------------------------- *- mkempen -* MAILTO:m.vankempen@erasmusmc.nl ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@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 ALAbeyta <@t> salud.unm.edu Tue Apr 13 11:57:18 2004 From: ALAbeyta <@t> salud.unm.edu (Antonia Abeyta) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Rat Turbinates Message-ID: Hi all, I am planning on staining some rat turbinates with Alcian Blue, and counter-staining with Hematoxylin & Eosin. Some of the literature suggests also adding a Periodic Acid Schiff (PAS) step into the protocol. What exactly does this step change and what is the PAS step useful for? Also, if anyone has some pointers for keeping the rat turbs from floating off the slides...we are currently using charged slides but they don't seem to be working. Thanks. Antonia L. Abeyta Health Sciences Tech. III Community Environmental Health Program University of New Mexico Surge Bldg. Room 140 Albuquerque, NM 87131 (505) 272-4028 From jhabecke <@t> seattlecca.org Tue Apr 13 12:53:33 2004 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] PAS and Trichrome in mouse and canine tissue Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAA63@ldap.seattlecca.org> Hello folks, I was wondering if anyone could share procedures for PAS and Trichromes in mouse and canine tissue. Thanks, Julie Julie Randolph-Habecker, Ph.D. Pathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N, G1-300 PO Box 19023 Seattle, WA 98109-1024 Tel: (206) 288-1187 FAX: (206) 288-1345 jhabecke@fhcrc.org This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From jhabecke <@t> seattlecca.org Tue Apr 13 12:57:44 2004 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] CD4 and CD8 in FFPE mouse tissue Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAA64@ldap.seattlecca.org> Hey Folks, I know, I know, I know........Everything on the histonet archive says there are no antibodies for CD4 and CD8 in FFPE mouse tissue. BUT, I thought I would try again and see if anyone has maybe worked something our recently. If anyone has discovered antibodies that work and can share a protocol with us - Please let me know! Thanks, Julie Julie Randolph-Habecker, Ph.D. Pathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N, G1-300 PO Box 19023 Seattle, WA 98109-1024 Tel: (206) 288-1187 FAX: (206) 288-1345 jhabecke@fhcrc.org This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From watson <@t> gnf.org Tue Apr 13 12:54:03 2004 From: watson <@t> gnf.org (James Watson) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] autoflourecence in RBC Message-ID: I am experiencing autoflourecence in RBC's of brain and spinal cord. Have tried to quench this with 5%hydrogen peroxide in methanol with no success. the Hydrogen peroxide is a year old, but sigma says it has a 5 year shelf life. in the past I have limited the hydrogen peroxide to a 6 mo. shelf life. does anyone have any suggestions. James Watson HT ASCP From lu_ze <@t> sbcglobal.net Tue Apr 13 15:40:51 2004 From: lu_ze <@t> sbcglobal.net (Ze Lu) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Submit References: <200404131701.i3DH1txl223038@vmh.prodigy.net> Message-ID: <007d01c42197$9d19f980$0f1515ce@HOME2> Topic: microtome Dear histonet friends, We are seting up a histological lab. Currently we are looking for a woring microtone for paraffine block tissue sectioning. If someone can help, please contact wirh me. Thanks. Ze Lu, Dr. Optimum Therapeutics, LLC Email: optimum-t@columbus.rr.com ----- Original Message ----- From: To: Sent: Tuesday, April 13, 2004 12:01 PM Subject: Histonet Digest, Vol 5, Issue 20 > 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: histogel (S Ladd) > 2. Re: Vibratome Considerations (Geoff McAuliffe) > 3. Histology Manager Opening at Yale University (Ramona Tolliver) > 4. RE: histogel (Andrea Grantham) > 5. Zinc formalin (Lorraine Cornett) > 6. RE: histogel (Connie McManus) > 7. RE: histogel (S Ladd) > 8. RE: histogel (S Ladd) > 9. RE: histogel (Bartlett, Jeanine) > 10. RE: histogel (Bartlett, Jeanine) > 11. RE: histogel (Patsy Ruegg) > 12. Rat Turbinates (Antonia Abeyta) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Tue, 13 Apr 2004 10:49:12 -0400 > From: "S Ladd" > Subject: RE: [Histonet] histogel > To: "m. van mkempen" > Cc: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset="US-ASCII" > > Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before > processing it WILL DISSOLVE! I learned this the hard way! The instructions > say something vague like "process as usual". Of course, I informed Richard > Allen that many of us in research do not have formalin on our tissue > processors. Richard Allen said they were going to change the instructions. > Once you have fixed the histogel you can process it like you would any other > piece of tissue. > Sharron > University of South Florida > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van > mkempen > Sent: Tuesday, April 13, 2004 10:18 AM > To: Histonetlist > Subject: [Histonet] histogel > > > Dear all, > > I have a question concerning histogel!! > I'm working with fetal lung and I thought histogel would be a nice way > to handle then while embedding. > My question is do I have to fix the histogel before processing for > embedding in paraffine? > How is the normal procedure for tissues's in histogel?? > > Thank you for all your kind answers. > > Regards, > Marta. > > -- > ---------------------------- > *- mkempen -* > MAILTO:m.vankempen@erasmusmc.nl > ----------------------------------------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 2 > Date: Tue, 13 Apr 2004 11:02:38 -0700 > From: Geoff McAuliffe > Subject: Re: [Histonet] Vibratome Considerations > To: Danielle Zalinski > Cc: HistoNet Listserve > Message-ID: <407C2B3E.40602@umdnj.edu> > Content-Type: text/plain; format=flowed; charset=us-ascii > > I have done immunos on vibratome sections of rat brain. If you use a 12 > well tissue culture plate (with its lid) on a shaker table overnight at > room temp. you should get good results. You will need to add some Triton > or Tween to the primary antibody (and probably subsequent steps) or do > something to 'permeabilize' the sections (freeze-thaw or alcohol) so you > get good penetration of the antibodies. If you have access to a sliding > microtome with a freezing stage you could obtain sections that way. > > Geoff > > Danielle Zalinski wrote: > > > Hello All, > >Our lab would like to do immunohistochem work in rat brain on vibratome > >sections instead of paraffin. We are able to get sections about > >50-60um. Will incubation with primary antibody overnight be sufficient? > > Is shaking the sections an option? I was hoping some other labs that > >are working in vibratome sections may have some beginners advice. > >Thanks, > >Danielle Zalinski > > > >Neurosurgery Research > >Henry Ford Health System > >Detroit, MI > > > > > >CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. > > > >Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > -- > -- > ********************************************** > Geoff McAuliffe, Ph.D. > Neuroscience and Cell Biology > Robert Wood Johnson Medical School > 675 Hoes Lane, Piscataway, NJ 08854 > voice: (732)-235-4583; fax: -4029 > mcauliff@umdnj.edu > ********************************************** > > > > > > > ------------------------------ > > Message: 3 > Date: Tue, 13 Apr 2004 11:14:18 -0400 > From: Ramona Tolliver > Subject: [Histonet] Histology Manager Opening at Yale University > To: histonet@lists.utsouthwestern.edu > Message-ID: <6.0.1.1.2.20040413111131.01e75d48@email.med.yale.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Good morning, > > Yale University has a Histology Manager (1st shift) position available from > 7:00 a.m. - 3:00 p.m. If you know of anyone who may be interested in an > outstanding growth opportunity in a department committed to excellence in > patient care, teaching, and discovery, please forward this information on > to them. > > Salary is commensurate with experience. Relocation assistance is > available. The position is posted online at : > http://websrv.its.yale.edu/hr/cgi-bin/printjob.plx?file=mp-2&job=11326 . > > Inquiries should be sent to ramona.tolliver@yale.edu. Thank you for your time! > > > > Ramona E. Tolliver > > *If you have difficulty with the link provided above, please go to > www.yale.edu/jobs and search Managerial and Professional last two weeks and > Pathology. The position is listed as Manager III. > > > > > Ramona E. Tolliver > Human Resource Manager > Yale University School of Medicine > Department of Pathology > 310 Cedar Street > New Haven, CT 06520-8023 > Telephone: (203) 785-6689 > Fax: (203) 785-7303 > > ------------------------------ > > Message: 4 > Date: Tue, 13 Apr 2004 08:28:11 -0700 > From: Andrea Grantham > Subject: RE: [Histonet] histogel > To: histonet@lists.utsouthwestern.edu > Message-ID: > <4.3.2.7.2.20040413082229.00cb9f10@algranth.inbox.email.arizona.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > When I use histogel I do "process as usual" and this does not include > formalin. I put the cassette containing the tissue embedded in histogel > into 70% ETOH. The processor starts with 70% ETOH X 2 and the concentration > of ETOH goes up from there. Haven't had any problems. > Maybe I've just been lucky? > Andi Grantham > > > > At 10:49 AM 4/13/2004 -0400, you wrote: > >Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before > >processing it WILL DISSOLVE! I learned this the hard way! The instructions > >say something vague like "process as usual". Of course, I informed Richard > >Allen that many of us in research do not have formalin on our tissue > >processors. Richard Allen said they were going to change the instructions. > >Once you have fixed the histogel you can process it like you would any other > >piece of tissue. > >Sharron > >University of South Florida > > > >-----Original Message----- > >From: histonet-bounces@lists.utsouthwestern.edu > >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van > >mkempen > >Sent: Tuesday, April 13, 2004 10:18 AM > >To: Histonetlist > >Subject: [Histonet] histogel > > > > > >Dear all, > > > >I have a question concerning histogel!! > >I'm working with fetal lung and I thought histogel would be a nice way > >to handle then while embedding. > >My question is do I have to fix the histogel before processing for > >embedding in paraffine? > >How is the normal procedure for tissues's in histogel?? > > > >Thank you for all your kind answers. > > > >Regards, > >Marta. > > > >-- > >---------------------------- > >*- mkempen -* > >MAILTO:m.vankempen@erasmusmc.nl > >----------------------------------------- > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > > > > ------------------------------ > > Message: 5 > Date: Tue, 13 Apr 2004 11:32:55 -0400 > From: "Lorraine Cornett" > Subject: [Histonet] Zinc formalin > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; format=flowed > > We are currentyl processing our biopsies in 10% neutral buffered formalin > (short process cycle). Our pathologists miss the nuclear detail they became > accustomed to when we were processing in Zinc Formalin. We seem to remember > a procedure where you can "mordant" or post fix the actual slides before > staining. Does anyone have this procedure that they can share with me. > > Thanks, > > Lorraine Cornett (HT ASCP) > Blue Ridge Pathology > Kingsport, TN 37660 > 423 224-5793 > > _________________________________________________________________ > Is your PC infected? Get a FREE online computer virus scan from McAfee? > Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 > > > > > ------------------------------ > > Message: 6 > Date: Tue, 13 Apr 2004 09:24:43 -0600 > From: "Connie McManus" > Subject: RE: [Histonet] histogel > To: "'S Ladd'" , "'m. van mkempen'" > > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <000101c4216b$72f8c5e0$4a737b81@Cygnus> > Content-Type: text/plain; charset="us-ascii" > > Forgive my ignorance, but what is histogel and what is it used for? > thanx > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of S Ladd > Sent: Tuesday, April 13, 2004 7:49 AM > To: m. van mkempen > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] histogel > > Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before > processing it WILL DISSOLVE! I learned this the hard way! The > instructions > say something vague like "process as usual". Of course, I informed > Richard > Allen that many of us in research do not have formalin on our tissue > processors. Richard Allen said they were going to change the > instructions. > Once you have fixed the histogel you can process it like you would any > other > piece of tissue. > Sharron > University of South Florida > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van > mkempen > Sent: Tuesday, April 13, 2004 10:18 AM > To: Histonetlist > Subject: [Histonet] histogel > > > Dear all, > > I have a question concerning histogel!! > I'm working with fetal lung and I thought histogel would be a nice way > to handle then while embedding. > My question is do I have to fix the histogel before processing for > embedding in paraffine? > How is the normal procedure for tissues's in histogel?? > > Thank you for all your kind answers. > > Regards, > Marta. > > -- > ---------------------------- > *- mkempen -* > MAILTO:m.vankempen@erasmusmc.nl > ----------------------------------------- > > _______________________________________________ > Histonet mailing list > Histonet@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: 7 > Date: Tue, 13 Apr 2004 11:42:46 -0400 > From: "S Ladd" > Subject: RE: [Histonet] histogel > To: "Andrea Grantham" > Cc: Histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset="US-ASCII" > > Are the items you are embedding in the histogel fixed in formalin? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrea > Grantham > Sent: Tuesday, April 13, 2004 11:28 AM > To: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] histogel > > > When I use histogel I do "process as usual" and this does not include > formalin. I put the cassette containing the tissue embedded in histogel > into 70% ETOH. The processor starts with 70% ETOH X 2 and the concentration > of ETOH goes up from there. Haven't had any problems. > Maybe I've just been lucky? > Andi Grantham > > > > At 10:49 AM 4/13/2004 -0400, you wrote: > >Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before > >processing it WILL DISSOLVE! I learned this the hard way! The instructions > >say something vague like "process as usual". Of course, I informed Richard > >Allen that many of us in research do not have formalin on our tissue > >processors. Richard Allen said they were going to change the instructions. > >Once you have fixed the histogel you can process it like you would any > other > >piece of tissue. > >Sharron > >University of South Florida > > > >-----Original Message----- > >From: histonet-bounces@lists.utsouthwestern.edu > >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van > >mkempen > >Sent: Tuesday, April 13, 2004 10:18 AM > >To: Histonetlist > >Subject: [Histonet] histogel > > > > > >Dear all, > > > >I have a question concerning histogel!! > >I'm working with fetal lung and I thought histogel would be a nice way > >to handle then while embedding. > >My question is do I have to fix the histogel before processing for > >embedding in paraffine? > >How is the normal procedure for tissues's in histogel?? > > > >Thank you for all your kind answers. > > > >Regards, > >Marta. > > > >-- > >---------------------------- > >*- mkempen -* > >MAILTO:m.vankempen@erasmusmc.nl > >----------------------------------------- > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 8 > Date: Tue, 13 Apr 2004 11:42:45 -0400 > From: "S Ladd" > Subject: RE: [Histonet] histogel > To: "Kyle-Byrne, Carrie - Labvision" > Cc: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > Since your cells were fixed in formalin, perhaps there was enough residual > formalin to sufficiently fix the histogel? You use only a few drops of > histogel. My cells were not in formalin and I use about 0.5 mL of histogel. > Just for fun, you could try processing a little button of histogel (without > formalin fixed cells) and see if it dissolves? > I use ethanol and xylene on my processor and I also start with 70% ethanol. > Sharron > > -----Original Message----- > From: Kyle-Byrne, Carrie - Labvision [mailto:CKByrne@labvision.com] > Sent: Tuesday, April 13, 2004 11:23 AM > To: 'S Ladd' > Subject: RE: [Histonet] histogel > > > Sharron, > Interesting. I've never heard of this before. I've used Histogel for years > and never had to 'fix' it in formalin. I used it to make cell pellets for > use as IHC control material. I would fix the cells in formalin first then > transfer to 70% EtOH and hold them until I had several to pellet. I'd > decant the 70%, put on a few drops of Histogel, mix to distribute the cells, > then cool the tube to harden the gel. Once solidified, I'd remove the > pellet from the tube and breadloaf it like a piece of tissue, place in a > cassette and onto the processor (we started in 70% EtOH) for overnight > processing and embed as usual in the morning. It always worked just fine. > What to you use for clearing? > > Carrie Kyle-Byrne > Sr. Reseach Assoc., Pathology Lab > Lab Vision, Corp. > 47791 Westinghouse Ave. > Fremont, CA 94539 > ckbyrne@labvision.com > www.labvision.com > > > > -----Original Message----- > From: S Ladd [mailto:sladd@hsc.usf.edu] > Sent: Tuesday, April 13, 2004 7:49 AM > To: m. van mkempen > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] histogel > > > Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before > processing it WILL DISSOLVE! I learned this the hard way! The instructions > say something vague like "process as usual". Of course, I informed Richard > Allen that many of us in research do not have formalin on our tissue > processors. Richard Allen said they were going to change the instructions. > Once you have fixed the histogel you can process it like you would any other > piece of tissue. > Sharron > University of South Florida > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van > mkempen > Sent: Tuesday, April 13, 2004 10:18 AM > To: Histonetlist > Subject: [Histonet] histogel > > > Dear all, > > I have a question concerning histogel!! > I'm working with fetal lung and I thought histogel would be a nice way > to handle then while embedding. > My question is do I have to fix the histogel before processing for > embedding in paraffine? > How is the normal procedure for tissues's in histogel?? > > Thank you for all your kind answers. > > Regards, > Marta. > > -- > ---------------------------- > *- mkempen -* > MAILTO:m.vankempen@erasmusmc.nl > ----------------------------------------- > > _______________________________________________ > Histonet mailing list > Histonet@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: 9 > Date: Tue, 13 Apr 2004 11:43:09 -0400 > From: "Bartlett, Jeanine" > Subject: RE: [Histonet] histogel > To: "Andrea Grantham" , > > Message-ID: > > Content-Type: text/plain; charset="us-ascii" > > We do the same and have had no problems. > > Jeanine Bartlett, HT(ASCP) > Centers for Disease Control > Infectious Disease Pathology Activity > 1600 Clifton Road, MS/G-32 > Atlanta, GA 30333 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Andrea > Grantham > Sent: Tuesday, April 13, 2004 11:28 AM > To: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] histogel > > > When I use histogel I do "process as usual" and this does not include > formalin. I put the cassette containing the tissue embedded in histogel > into 70% ETOH. The processor starts with 70% ETOH X 2 and the > concentration > of ETOH goes up from there. Haven't had any problems. > Maybe I've just been lucky? > Andi Grantham > > > > At 10:49 AM 4/13/2004 -0400, you wrote: > >Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before > >processing it WILL DISSOLVE! I learned this the hard way! The > >instructions say something vague like "process as usual". Of course, I > >informed Richard Allen that many of us in research do not have formalin > > >on our tissue processors. Richard Allen said they were going to change > >the instructions. Once you have fixed the histogel you can process it > >like you would any other piece of tissue. Sharron > >University of South Florida > > > >-----Original Message----- > >From: histonet-bounces@lists.utsouthwestern.edu > >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van > >mkempen > >Sent: Tuesday, April 13, 2004 10:18 AM > >To: Histonetlist > >Subject: [Histonet] histogel > > > > > >Dear all, > > > >I have a question concerning histogel!! > >I'm working with fetal lung and I thought histogel would be a nice way > >to handle then while embedding. My question is do I have to fix the > >histogel before processing for embedding in paraffine? > >How is the normal procedure for tissues's in histogel?? > > > >Thank you for all your kind answers. > > > >Regards, > >Marta. > > > >-- > >---------------------------- > >*- mkempen -* > >MAILTO:m.vankempen@erasmusmc.nl > >----------------------------------------- > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 10 > Date: Tue, 13 Apr 2004 11:47:04 -0400 > From: "Bartlett, Jeanine" > Subject: RE: [Histonet] histogel > To: "S Ladd" , "Andrea Grantham" > > Cc: Histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset="us-ascii" > > Yes. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of S Ladd > Sent: Tuesday, April 13, 2004 11:43 AM > To: Andrea Grantham > Cc: Histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] histogel > > > Are the items you are embedding in the histogel fixed in formalin? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrea > Grantham > Sent: Tuesday, April 13, 2004 11:28 AM > To: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] histogel > > > When I use histogel I do "process as usual" and this does not include > formalin. I put the cassette containing the tissue embedded in histogel > into 70% ETOH. The processor starts with 70% ETOH X 2 and the > concentration of ETOH goes up from there. Haven't had any problems. > Maybe I've just been lucky? Andi Grantham > > > > At 10:49 AM 4/13/2004 -0400, you wrote: > >Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before > >processing it WILL DISSOLVE! I learned this the hard way! The > >instructions say something vague like "process as usual". Of course, I > >informed Richard Allen that many of us in research do not have formalin > > >on our tissue processors. Richard Allen said they were going to change > >the instructions. Once you have fixed the histogel you can process it > >like you would any > other > >piece of tissue. > >Sharron > >University of South Florida > > > >-----Original Message----- > >From: histonet-bounces@lists.utsouthwestern.edu > >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van > >mkempen > >Sent: Tuesday, April 13, 2004 10:18 AM > >To: Histonetlist > >Subject: [Histonet] histogel > > > > > >Dear all, > > > >I have a question concerning histogel!! > >I'm working with fetal lung and I thought histogel would be a nice way > >to handle then while embedding. My question is do I have to fix the > >histogel before processing for embedding in paraffine? > >How is the normal procedure for tissues's in histogel?? > > > >Thank you for all your kind answers. > > > >Regards, > >Marta. > > > >-- > >---------------------------- > >*- mkempen -* > >MAILTO:m.vankempen@erasmusmc.nl > >----------------------------------------- > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 11 > Date: Tue, 13 Apr 2004 10:00:05 -0600 > From: "Patsy Ruegg" > Subject: RE: [Histonet] histogel > To: "S Ladd" , "m. van mkempen" > > Cc: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset="us-ascii" > > I use histogel all the time without fixing in formalin. Like Andi I start > with tissue already fixed and then placed in 70% alcohol. I embed this > tissue in histogel and put it back in the 70% and then process starting with > 70% alcohol, no formalin. I have not had the histogel ever dissolve. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of S Ladd > Sent: Tuesday, April 13, 2004 8:49 AM > To: m. van mkempen > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] histogel > > > Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before > processing it WILL DISSOLVE! I learned this the hard way! The instructions > say something vague like "process as usual". Of course, I informed Richard > Allen that many of us in research do not have formalin on our tissue > processors. Richard Allen said they were going to change the instructions. > Once you have fixed the histogel you can process it like you would any other > piece of tissue. > Sharron > University of South Florida > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of m. van > mkempen > Sent: Tuesday, April 13, 2004 10:18 AM > To: Histonetlist > Subject: [Histonet] histogel > > > Dear all, > > I have a question concerning histogel!! > I'm working with fetal lung and I thought histogel would be a nice way > to handle then while embedding. > My question is do I have to fix the histogel before processing for > embedding in paraffine? > How is the normal procedure for tissues's in histogel?? > > Thank you for all your kind answers. > > Regards, > Marta. > > -- > ---------------------------- > *- mkempen -* > MAILTO:m.vankempen@erasmusmc.nl > ----------------------------------------- > > _______________________________________________ > Histonet mailing list > Histonet@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: 12 > Date: Tue, 13 Apr 2004 10:57:18 -0600 > From: "Antonia Abeyta" > Subject: [Histonet] Rat Turbinates > To: > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > Hi all, > > I am planning on staining some rat turbinates with Alcian Blue, and > counter-staining with Hematoxylin & Eosin. Some of the literature > suggests also adding a Periodic Acid Schiff (PAS) step into the > protocol. What exactly does this step change and what is the PAS step > useful for? > > Also, if anyone has some pointers for keeping the rat turbs from > floating off the slides...we are currently using charged slides but they > don't seem to be working. > > Thanks. > > Antonia L. Abeyta > Health Sciences Tech. III > Community Environmental Health Program > University of New Mexico > Surge Bldg. Room 140 > Albuquerque, NM 87131 > (505) 272-4028 > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 5, Issue 20 > *************************************** From djohnson14 <@t> hotmail.com Tue Apr 13 15:13:30 2004 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Histology Opening in Santa Monica CA Message-ID: Immediate Job opening in Santa Monica for full time histology tech. Please contact Dr Taheri at 310-922-1412 _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From cfavara <@t> niaid.nih.gov Tue Apr 13 15:32:18 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] CD4 and CD8 in FFPE mouse tissue Message-ID: Nothing here but I would also love the info! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Randolph-Habecker, Julie [mailto:jhabecke@seattlecca.org] Sent: Tuesday, April 13, 2004 11:58 AM To: Histonet (E-mail) Subject: [Histonet] CD4 and CD8 in FFPE mouse tissue Hey Folks, I know, I know, I know........Everything on the histonet archive says there are no antibodies for CD4 and CD8 in FFPE mouse tissue. BUT, I thought I would try again and see if anyone has maybe worked something our recently. If anyone has discovered antibodies that work and can share a protocol with us - Please let me know! Thanks, Julie Julie Randolph-Habecker, Ph.D. Pathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N, G1-300 PO Box 19023 Seattle, WA 98109-1024 Tel: (206) 288-1187 FAX: (206) 288-1345 jhabecke@fhcrc.org This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Tue Apr 13 15:43:33 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] MIke LaFriniere/lab survey Message-ID: <004e01c42197$fcc406a0$1d2a14ac@wchsys.org> I will participate in the survey, if you still need labs. ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From carl.hobbs <@t> kcl.ac.uk Tue Apr 13 16:02:52 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] alternative to using pap pen? Message-ID: <002501c4219a$af6b6df0$21292bd9@home> Just read the parafilm alternative.......interesting. I had a "fight" with one of the "leading lights" of immuno way back...she absolutely insisted upon the use of coverslips( parafilm equivilent?). I clearly demonstrated that without c'slips, one was able to obtain a consistent signal..ie using c'slips the signal was inconsistent. I have avoided such procedures since. Mind you, I do use c'slips for ISH with no, apparent, detrimental effect. Will check tho, very few things are sacrosanct in Histo lol. Also, whether they use the same technique now, or not, UCL/Nequas control Centre in UK never used "pap pens". As they are the reference point for quality assessment and control in UK, I wonder what their opinion is now? From cwscouten <@t> myneurolab.com Tue Apr 13 16:03:58 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Vibratome Considerations Message-ID: A primary reason to use a Vibratome(TM) is to avoid damage to cell membranes and resulting loss of cellular contents. Do not freeze and rethaw, after using a Vibratome, and you can get more sensitive staining, depending on the what sort of cellular protein you are after (in cytosol, or bound to membranes or other solids) with a Vibratome. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Geoff McAuliffe Sent: Tuesday, April 13, 2004 1:03 PM To: Danielle Zalinski Cc: HistoNet Listserve Subject: Re: [Histonet] Vibratome Considerations I have done immunos on vibratome sections of rat brain. If you use a 12 well tissue culture plate (with its lid) on a shaker table overnight at room temp. you should get good results. You will need to add some Triton or Tween to the primary antibody (and probably subsequent steps) or do something to 'permeabilize' the sections (freeze-thaw or alcohol) so you get good penetration of the antibodies. If you have access to a sliding microtome with a freezing stage you could obtain sections that way. Geoff Danielle Zalinski wrote: > Hello All, >Our lab would like to do immunohistochem work in rat brain on vibratome >sections instead of paraffin. We are able to get sections about >50-60um. Will incubation with primary antibody overnight be sufficient? > Is shaking the sections an option? I was hoping some other labs that >are working in vibratome sections may have some beginners advice. >Thanks, >Danielle Zalinski > >Neurosurgery Research >Henry Ford Health System >Detroit, MI > > >CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. > >Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. >_______________________________________________ >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 carl.hobbs <@t> kcl.ac.uk Tue Apr 13 16:20:28 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] which tissue processor...UK? Message-ID: <003a01c4219d$252ee970$21292bd9@home> Hi am negotiating for either a Shandon Citadel( basic plus fume control cage) or a Leica TP1020 with inbuilt fume control. Have got a Leica already but "fume control" is inadequate ie have to attach extract vent to an external vent, anyway. Both use charcoal filters for the xylene clearing agent. Be very grateful for any words of wisdom From cfranci <@t> rigel.com Tue Apr 13 16:55:19 2004 From: cfranci <@t> rigel.com (Christian Franci) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] PAL-E mab Message-ID: Maybe, someone out there knows where I might find a supplier of polyclonal or mAb for PAL-E an endothelial marker for "leaky" vessels. If, anyone can point me in the right direction, I'd much appreciate it. Thanks in advance, C From dcrippen <@t> buckinstitute.org Tue Apr 13 17:56:25 2004 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] anti mouse CD13 in FFPE tissue Message-ID: <4AA34A707932424EBE2D973764D226A90158EC23@inverness.buckcenter.org> Can anyone suggest a good anti-mouse CD13 (N-aminopeptidase) antibody that is known to work well in FFPE tissues??? Many thanks in advance, Danielle Crippen Morphology Core Supervisor Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen@buckinstitute.org From aostrand <@t> seattlecca.org Tue Apr 13 18:06:20 2004 From: aostrand <@t> seattlecca.org (Ostrander, Anita B) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Clean Glassware Test Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B94823B5F7@ldap.seattlecca.org> Can anyone give me the vendor name/order number of a reagent that I can buy to test for soap residue on glassware. Thank You, Anita Ostrander Pathology Manager Seattle Cancer Care Alliance Office:206-288-1347 Cell Phone: 206-226-0785 aostrand@seattlecca.org This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From brucea <@t> unimelb.edu.au Tue Apr 13 22:01:27 2004 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Histology Conference in Toronto Sept. 2004 ?? Info PLEASE Message-ID: G'Day..... I am after information RE: The Histology Conference in Toronto, Canada this September/2004. Can someone who has a weblink/info. etc. please forward on to me. Any information would be great. Thanks in advance - 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 ******************************************************************************** This electronic message and all contents contain information 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 us immediately and destroy the original message and all copies. From KAELPERS <@t> aol.com Tue Apr 13 22:21:44 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Histology Conference in Toronto Sept. 2004 ?? Info PLEASE Message-ID: <79.26e09f5c.2dae0848@aol.com> Bruce, go to the NSH web site...it has details on housing and will have the program on line April 19th. www.nsh.org click on NSH conventions. lge From balajimr <@t> drreddys.com Tue Apr 13 23:14:50 2004 From: balajimr <@t> drreddys.com (balajimr@drreddys.com) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Safranine o staining and Problem in tissues Message-ID: Can anyone give me the detailed procedure for safranine o staining for the joints (ankle. tarsal and metatarsal joints in rats). I would like to know a good decalcifying solution for the same and what is the duration of bone to be kept in the decalcifier. Secondly, of late I am facing peculiar problem now with my liver, kidney and brain sections of late. These sections are cracking after the staining (Mayer's H& E). It is something like parched earth appearance. This I am noticing when the tissue s are coming to last xylenes. I donot see the cracking before this step. This doesn't happen in all the slides. Could you please suggest some solution for this. We are using chloroform for clearing and histoplast from Shandon for infiltration. I am unable to locate where exactly is the problem. I have tried changing the makes of chloroform and xylenes used with out much help. Dr. Balaji (M. V. Sc. Path) Dept. Pre clinical safety evaluation, Discovery research, Dr. Reddys Laboratories ltd. Bollaram Road, Miyapur, Hyderabad, 500 049 Andhra Pradseh, INDIA Tel: 23045439 - Ext.432. From histol <@t> rat-hole.org Wed Apr 14 03:15:27 2004 From: histol <@t> rat-hole.org (Matt Ibbs) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Isopropanol in processing Message-ID: <5.1.0.14.2.20040414101518.009f1ec0@bigpop3.wyith.net> Dear all, I have been reading the histonet archives looking for information on processing through isopropanol instead of ethanol. I've found plenty on the subject of using isopropanol as a means of removing xylene from the schedule but hardly anything on using isopropanol as a routine dehydrating agent. The head of our department wishes to change our routine processing schedule as the price of ethanol is always rising and we have problems with taxes and new customs demands. The answer would appear to be use isopropanol routinely and we've been trying to get a good working method but find that either the small biopsies are too brittle or the larger specimens (in particular slices through material from radical prostatectomy) are insufficiently processed and fall off the slides. Presently we're thinking that isopropanol is rather more 'harsh' than ethanol resulting in the brittleness seen in small biopsies and so we have tried reducing times in alcohol in an attempt to counteract this. Of course, that results in the problem of poorly processed larger specimens. What is the answer? Should we use more short steps with small increases in concentration (50%, 70%, 80%, 90%, 100%) before going to xylene or should we use longer incubation times in fewer, stronger dilutions (70%, 90, 100%, 100%)? Any suggestions gratefully received. I look forward to hearing your opinions. From jqb7 <@t> cdc.gov Wed Apr 14 05:44:05 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Histology Conference in Toronto Sept. 2004 ?? InfoPLEASE Message-ID: If you click on "Program Preview" under NSH Convention you can already get a list of the workshops offered. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of KAELPERS@aol.com Sent: Tuesday, April 13, 2004 11:22 PM To: brucea@unimelb.edu.au; histonet@pathology.swmed.edu Subject: Re: [Histonet] Histology Conference in Toronto Sept. 2004 ?? InfoPLEASE Bruce, go to the NSH web site...it has details on housing and will have the program on line April 19th. www.nsh.org click on NSH conventions. lge _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From qiaolin_deng78 <@t> hotmail.com Wed Apr 14 07:33:42 2004 From: qiaolin_deng78 <@t> hotmail.com (dengqiaolin deng) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] (no subject) Message-ID: Dear histonetter: Can anyone share some experiences in mouse( E10-E12 ) saggital sectioning? How to embed embryos so that you can get good section? Thank you in advance! Qiaolin _________________________________________________________________ Add photos to your messages with [1]MSN 8. Get 2 months FREE*. References 1. http://g.msn.com/8HMBEN/2749??PS= From sladd <@t> hsc.usf.edu Wed Apr 14 07:34:56 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20A96E8@usca0082k08.labvision.apogent.com> Message-ID: I certainly am curious too!! This happened to me the first time I used histogel. I receive cell samples in Phosphate Buffered Saline (PBS). I believe that these particular samples were fixed in 4% paraformaldehyde. I solidify the histogel at room temperature and then after a half and hour or so, I put the samples in the fridge just to make sure. When I processed them, I put them in a 70% "holding station"... The next morning when I opened the cassettes and everything was gone...vanished...without so much as a trace. I had to tell the PI that I had lost everything!! I called Richard Allen and that is when (after at least an hour of trouble shooting on the phone) they told me it must be because I have no formalin on my processor. Ever since then, I fix the samples in formalin for at least an hour before processing. I have never had a problem since. The solution seemed simple, so I never thought more of it. Since histogel is "proprietary" I don't know exactly what is in it. However, I thought it was gelatin based. I have the AFIP "Advance Laboratory Methods in Histology and Pathology" and they have a cell block prep. protocol on page 217. They use 2% gelatin. This protocol says to "fix in formalin" for at least one hour prior to processing. After reading this, I thought this was further proof that my problem was that I hadn't fixed in formalin. I am happy to hear that many people use histogel with cells that never see formalin. Of course that leaves me wondering what went wrong that horrible day. Also, as you can imagine, after losing everything I am reluctant to eliminate the formalin step. :) For the person that was wondering what histogel was... this is from Richard Allen's web site: HistoGel? Specimen Processing Gel "No matter how small, friable, or viscous your histology or cytology specimen, HistoGel will encapsulate and retain the entire specimen during histological processing. A few drops of HistoGel will allow chemical reagents to penetrate the specimen without allowing the specimen to escape from the gel. HistoGel?s patented formulation outperforms other agarose gel mixtures or thromboplastin at a fraction of the cost. HistoGel will not retain histological stains, eliminating unwanted discoloration around specimens on slides. HistoGel is virtually unnoticeable during sectioning and will not ?pop out? of the paraffin block during cutting." Sharron -----Original Message----- From: Kyle-Byrne, Carrie - Labvision [mailto:CKByrne@labvision.com] Sent: Tuesday, April 13, 2004 4:57 PM To: 'S Ladd' Subject: RE: [Histonet] histogel There is no residual formalin left on our cells since we hold them in 70% (they go through 2 changes typically) before Histogel'ing them. I've never heard of anyone else having this problem before, so I am interested in trying to understand why you are experiencing this difficulty. What solution is your material in before putting the Histogel on it? And what temperature do you solidify the gel at before cassetting and processing? Carrie -----Original Message----- From: S Ladd [mailto:sladd@hsc.usf.edu] Sent: Tuesday, April 13, 2004 8:43 AM To: Kyle-Byrne, Carrie - Labvision Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histogel Since your cells were fixed in formalin, perhaps there was enough residual formalin to sufficiently fix the histogel? You use only a few drops of histogel. My cells were not in formalin and I use about 0.5 mL of histogel. Just for fun, you could try processing a little button of histogel (without formalin fixed cells) and see if it dissolves? I use ethanol and xylene on my processor and I also start with 70% ethanol. Sharron Histonet@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 maxim_71 <@t> mail.ru Wed Apr 14 08:58:57 2004 From: maxim_71 <@t> mail.ru (=?koi8-r?Q?=22=F0=C5=DB=CB=CF=D7=20=ED=C1=CB=D3=C9=CD=22=20?=) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet]PCR technique - Maxim Peshkov Maxim_71@mail.ru Message-ID: Dear Histonetters! Whether somebody can send me the technique of polymerase chain reaction for paraffin-embedded tissues from fixation up to final result. I don t have any literature, and similar I didn t do anything earlier. I have is only one desire, I want to learn this method and use it for routine working. Thanks You. Maxim Peshkov, Cytologist, HTL Department of biopsy and cytological researches hathological and anatomical bureau. Russia, Taganrog 347900 Maxim_71@mail.ru From sladd <@t> hsc.usf.edu Wed Apr 14 10:45:28 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] histogel In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20A96E9@usca0082k08.labvision.apogent.com> Message-ID: The PBS may have been the problem indeed. I don't need to eliminate the formalin step with what I am doing right now, however, I have told people that if they didn't want their cells to be exposed to formalin I could not use histogel. If this is not the case, there are a few people who will be very happy to hear this. Thanks for your input!! Sharron -----Original Message----- From: Kyle-Byrne, Carrie - Labvision [mailto:CKByrne@labvision.com] Sent: Wednesday, April 14, 2004 11:28 AM To: 'S Ladd' Subject: RE: [Histonet] histogel Sharron, Is it possible that there was too much carry-over of PBS? Carry-over of a bit of 70% EtOH doesn't seem to cause any problems for me, but I wonder if PBS might be a problem. hmmm....very odd indeed.... You seem to have it working for you now. Are you wanting to eliminate the fixation prior to processing step? Carrie -----Original Message----- From: S Ladd [mailto:sladd@hsc.usf.edu] Sent: Wednesday, April 14, 2004 5:35 AM To: Kyle-Byrne, Carrie - Labvision Cc: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histogel I certainly am curious too!! This happened to me the first time I used histogel. I receive cell samples in Phosphate Buffered Saline (PBS). I believe that these particular samples were fixed in 4% paraformaldehyde. I solidify the histogel at room temperature and then after a half and hour or so, I put the samples in the fridge just to make sure. When I processed them, I put them in a 70% "holding station"... The next morning when I opened the cassettes and everything was gone...vanished...without so much as a trace. I had to tell the PI that I had lost everything!! I called Richard Allen and that is when (after at least an hour of trouble shooting on the phone) they told me it must be because I have no formalin on my processor. Ever since then, I fix the samples in formalin for at least an hour before processing. I have never had a problem since. The solution seemed simple, so I never thought more of it. Since histogel is "proprietary" I don't know exactly what is in it. However, I thought it was gelatin based. I have the AFIP "Advance Laboratory Methods in Histology and Pathology" and they have a cell block prep. protocol on page 217. They use 2% gelatin. This protocol says to "fix in formalin" for at least one hour prior to processing. After reading this, I thought this was further proof that my problem was that I hadn't fixed in formalin. I am happy to hear that many people use histogel with cells that never see formalin. Of course that leaves me wondering what went wrong that horrible day. Also, as you can imagine, after losing everything I am reluctant to eliminate the formalin step. :) For the person that was wondering what histogel was... this is from Richard Allen's web site: HistoGel(tm) Specimen Processing Gel "No matter how small, friable, or viscous your histology or cytology specimen, HistoGel will encapsulate and retain the entire specimen during histological processing. A few drops of HistoGel will allow chemical reagents to penetrate the specimen without allowing the specimen to escape from the gel. HistoGel's patented formulation outperforms other agarose gel mixtures or thromboplastin at a fraction of the cost. HistoGel will not retain histological stains, eliminating unwanted discoloration around specimens on slides. HistoGel is virtually unnoticeable during sectioning and will not "pop out" of the paraffin block during cutting." Sharron -----Original Message----- From: Kyle-Byrne, Carrie - Labvision [mailto:CKByrne@labvision.com] Sent: Tuesday, April 13, 2004 4:57 PM To: 'S Ladd' Subject: RE: [Histonet] histogel There is no residual formalin left on our cells since we hold them in 70% (they go through 2 changes typically) before Histogel'ing them. I've never heard of anyone else having this problem before, so I am interested in trying to understand why you are experiencing this difficulty. What solution is your material in before putting the Histogel on it? And what temperature do you solidify the gel at before cassetting and processing? Carrie -----Original Message----- From: S Ladd [mailto:sladd@hsc.usf.edu] Sent: Tuesday, April 13, 2004 8:43 AM To: Kyle-Byrne, Carrie - Labvision Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histogel Since your cells were fixed in formalin, perhaps there was enough residual formalin to sufficiently fix the histogel? You use only a few drops of histogel. My cells were not in formalin and I use about 0.5 mL of histogel. Just for fun, you could try processing a little button of histogel (without formalin fixed cells) and see if it dissolves? I use ethanol and xylene on my processor and I also start with 70% ethanol. Sharron Histonet@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 ccrowder <@t> mail.vetmed.lsu.edu Wed Apr 14 10:58:20 2004 From: ccrowder <@t> mail.vetmed.lsu.edu (Cheryl Crowder) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Dystrophin antibody Message-ID: Hi - I have a researcher who would like to do Dystrophin IHC on formalin-fixed, paraffin embedded tissue. Does anyone know of an antibody which will work. Thank you, Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 From d.fuster <@t> ub.edu Wed Apr 14 11:17:00 2004 From: d.fuster <@t> ub.edu (Dolors Fuster) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] detach cells from a slide Message-ID: <1081959420.407d63fcc35eb@webmail1.ub.edu> Any idea to detach cells from an hematologic slide? They are dryed...stained with May Gr?nwald Giemsa...dehidrated, cleared and mounted with DPX. The idea is to use them for a DNA extraction. Thanks in advance Dolors Fuster T?cnic Especialista en Anatomia Patol?gica i Citologia Departament d'Anatomia i Embriologia Humana Facultat de Medicina (HCP) Universitat de Barcelona ICQ 14146798 ------------------------------------------------- This mail sent through IMP: http://horde.org/imp/ From Jackie.O'Connor <@t> abbott.com Wed Apr 14 11:16:44 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Decal question Message-ID: I'm working on a trial with intra-tibial injections of tumors into mice. Whole (skin and muscle trimmed) legs were fixed for about a month in 10% NBF. I used Surgipath's Decal I to decalcify (took about 6 hours - end point determined by cutting through femur with scalpel). Paraffin sections cut nicely - I used a 48C waterbath to ensure no wrinkles, picked up on positively charged slides. I let them air dry overnight, then 60C for an hour prior to H+E staining. I'm not happy. The tumors and cortical bone are lifting and falling off the slides. Any suggestions? I've been reading old posts about EDTA decalcification - since hindsight is 20/20 - would that have been a better choice?? Boo hoo. Jackie O' From jcline <@t> wchsys.org Wed Apr 14 11:26:03 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] jeff jurczak/knife sharpner Message-ID: <003801c4223d$30b90dc0$1d2a14ac@wchsys.org> I am asking $800.00, my lab is in Maryland ***** 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 juan.gutierrez <@t> christushealth.org Wed Apr 14 11:27:43 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Dystrophin antibody Message-ID: Novocastra has acouple of clones available for ffpe tissue. I have never used them before, but I have used the ones for frozens with good results. It might be worth a try. Good luck. Juan C. Gutierrez,HT(ASCP) -----Original Message----- From: Cheryl Crowder [mailto:ccrowder@mail.vetmed.lsu.edu] Sent: Wednesday, April 14, 2004 10:58 AM To: Histonet Subject: [Histonet] Dystrophin antibody Hi - I have a researcher who would like to do Dystrophin IHC on formalin-fixed, paraffin embedded tissue. Does anyone know of an antibody which will work. Thank you, Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kappeler <@t> patho.unibe.ch Wed Apr 14 11:33:52 2004 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Re: Dystrophin antibody Message-ID: <00a801c4223e$45cce870$27955c82@patho.unibe.ch> Hi Cheryl we use clone 13H6 (Novocastra, NCL-DYSA) at 1:50 after HIER in 100 mM Tris - 5% Urea, pH 9.5. Other high pH retrieval buffers may work as well. Hope this helps. Andi Kappeler Institute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "Cheryl Crowder" To: "Histonet" Sent: Wednesday, April 14, 2004 5:58 PM Subject: [Histonet] Dystrophin antibody > Hi - I have a researcher who would like to do Dystrophin IHC on > formalin-fixed, paraffin embedded tissue. Does anyone know of an antibody > which will work. Thank you, Cheryl > > > Cheryl Crowder, BA, HTL(ASCP) > Chief Technologist > Anatomic Pathology > Department of Pathobiological Sciences > School of Veterinary Medicine > Louisiana State University > Skip Bertman Drive > Baton Rouge, LA 70803 > > 225-578-9734 > FAX: 225-578-9720 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From convmcm <@t> cc.usu.edu Wed Apr 14 11:30:09 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] PAS and Trichrome in mouse and canine tissue In-Reply-To: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAA63@ldap.seattlecca.org> Message-ID: <000201c4223d$c1800ac0$4a737b81@Cygnus> Julie, I use the same PAS procedure found in the AFIP manual (the red cover) I haven't noticed any difference between it and the one found in Humason or Sheehan and Hrapchack or Ann Preece. It works great on all the animal tissues I work with. Ditto for the Masson's. Do you have a copy of these books? If not, let me know privately and I will be happy to send along my procedures. Have a nice day... Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Randolph-Habecker, Julie Sent: Tuesday, April 13, 2004 10:54 AM To: Histonet (E-mail) Subject: [Histonet] PAS and Trichrome in mouse and canine tissue Hello folks, I was wondering if anyone could share procedures for PAS and Trichromes in mouse and canine tissue. Thanks, Julie Julie Randolph-Habecker, Ph.D. Pathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N, G1-300 PO Box 19023 Seattle, WA 98109-1024 Tel: (206) 288-1187 FAX: (206) 288-1345 jhabecke@fhcrc.org This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dmikita <@t> wmcnet.org Wed Apr 14 11:43:23 2004 From: dmikita <@t> wmcnet.org (Daryl Mikita) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Microwave Staining Message-ID: Hello, Our microwave is almost dead, and we are looking at getting another microwave. I was just wondering what companies sell microwaves for staining are still out there? Also, what does everyone else use out there? Our current microwave is just a Kenmore home unit. Thanks Daryl Mikita, HT(ASCP) dmikita@wmcnet.org From David.Edmondson <@t> christie-tr.nwest.nhs.uk Wed Apr 14 11:47:46 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Microwave Staining Message-ID: Hi Daryl, Just to say that we have been using domestic microwaves for years, replacement is relatively simple and inexpensive. Matsui 650 watt and Sharp 650 watt were fine but now a Sharp 800 watt. We have always gone for the digital/numerical keypad versions, so entering the times has been staightforward. So are the lab variety any different behind the label? Dave Christie Hosp Manchester UK -----Original Message----- From: Daryl Mikita [mailto:dmikita@wmcnet.org] Sent: 14 April 2004 17:43 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave Staining Hello, Our microwave is almost dead, and we are looking at getting another microwave. I was just wondering what companies sell microwaves for staining are still out there? Also, what does everyone else use out there? Our current microwave is just a Kenmore home unit. Thanks Daryl Mikita, HT(ASCP) dmikita@wmcnet.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From d.fuster <@t> ub.edu Wed Apr 14 11:50:23 2004 From: d.fuster <@t> ub.edu (Dolors Fuster) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] detach cells from a slide....again Message-ID: <1081961423.407d6bcfa615e@webmail1.ub.edu> In my previous mail I forgot to tell you the mechanical method (scrape them off with a sharp razor blade) is made....but too slow for at least 600 slides...difficult to recover all stuff...some of recovered adheres to the eppendorf.... :o( Could the mehod not include trypsin in its procedure :o) ?? Thanks again Any idea to detach cells from an hematologic slide? They are dryed...stained with May Gr?nwald Giemsa...dehidrated, cleared and mounted with DPX. The idea is to use them for a DNA extraction. Thanks in advance _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----- Fi del missatge reenviat ----- Dolors Fuster T?cnic Especialista en Anatomia Patol?gica i Citologia Departament d'Anatomia i Embriologia Humana Facultat de Medicina (HCP) Universitat de Barcelona ICQ 14146798 ------------------------------------------------- This mail sent through IMP: http://horde.org/imp/ From convmcm <@t> cc.usu.edu Wed Apr 14 11:54:28 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Safranine o staining and Problem in tissues In-Reply-To: Message-ID: <000001c42241$2801ed10$4a737b81@Cygnus> What are you looking for in the joint staining you want? If you want something with safranine O, Movat's Pentachrome comes to mind. Not having worked with bone histology for over 20 years, I'm not the best resource for this. I do remember Movat's being a big favorite during the time I worked specifically in bone histology. It was done on decalcified bone as well a MMA embedded bones. DECALCIFIER: Decal-Stat from the Decal Corp. PO Box 236, Congers, NY 10920; Phone: 800/428-5856; fax: 845/268-383 This is a rapid decalcifier. We use it on all our tissues regardless of anmal. It works great. As for your liver, kidney and brain sections, --- I'm going out on a limb here --- but it sounds like it could be heat artifact. Are you putting your sections on a heating tray before staining them? If so, check the temperature... it may be way too hot. My water bath is set at 37 to 38 C and I keep my heating tray set to 40 C. That's pretty warm, but it works well for tissues like gut, heart, skin, ovary, uterus kidney, liver. I don't leave any tissue on this tray for more than 10 or 15 minutes and haven't had any problems like this. Generally, I don't put brain, or fetal tissues, anything frail on the tray except if they seem to need a little relaxing of the tissue. In that case, I watch the sections carefully and remove them as soon as they smooth out. For what it's worth... Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of balajimr@drreddys.com Sent: Tuesday, April 13, 2004 9:15 PM To: histonet@lists.utsouthwestern.edu Cc: histonet-bounces@lists.utsouthwestern.edu Subject: [Histonet] Safranine o staining and Problem in tissues Importance: High Can anyone give me the detailed procedure for safranine o staining for the joints (ankle. tarsal and metatarsal joints in rats). I would like to know a good decalcifying solution for the same and what is the duration of bone to be kept in the decalcifier. Secondly, of late I am facing peculiar problem now with my liver, kidney and brain sections of late. These sections are cracking after the staining (Mayer's H& E). It is something like parched earth appearance. This I am noticing when the tissue s are coming to last xylenes. I donot see the cracking before this step. This doesn't happen in all the slides. Could you please suggest some solution for this. We are using chloroform for clearing and histoplast from Shandon for infiltration. I am unable to locate where exactly is the problem. I have tried changing the makes of chloroform and xylenes used with out much help. Dr. Balaji (M. V. Sc. Path) Dept. Pre clinical safety evaluation, Discovery research, Dr. Reddys Laboratories ltd. Bollaram Road, Miyapur, Hyderabad, 500 049 Andhra Pradseh, INDIA Tel: 23045439 - Ext.432. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Wed Apr 14 12:31:47 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Decal question In-Reply-To: Message-ID: Jackie I don't think the decal method would make much difference. Lifting of cortical bone and cartilage in my experience is the norm for bone sections especially when you are trying to see bone integrated with soft tissue (?your tumor cells I assume are soft). The bone will pull away from the cells. What to do? Well it does help to carefully drain the water off the slides on edge when you pick them up and then dry them laying flat on a heat plate at say 37dc overnight, then heat less than 60c, say 55c just hot enough to melt the paraffin for several hours (?4)but with bone there is no sure fire method to keep them intact that I know of except the old coating a slide with 5% elmers glue let them airdry and use these slides to pick up the section, but then if you are doing any kind of collagen IHC the glue will interfer. This is a great method for tintorial stains on bone sections though. If you are going to do HIER you are really going to be in for lifting of the bone in my experience. I use only enzyme digestion for bone IHC because I can control it easier than HIER with less bone and cartilage lifting of the section. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie.O'Connor@abbott.com Sent: Wednesday, April 14, 2004 10:17 AM To: Histonet (E-mail); histonet-bounces@lists.utsouthwestern.edu Subject: [Histonet] Decal question I'm working on a trial with intra-tibial injections of tumors into mice. Whole (skin and muscle trimmed) legs were fixed for about a month in 10% NBF. I used Surgipath's Decal I to decalcify (took about 6 hours - end point determined by cutting through femur with scalpel). Paraffin sections cut nicely - I used a 48C waterbath to ensure no wrinkles, picked up on positively charged slides. I let them air dry overnight, then 60C for an hour prior to H+E staining. I'm not happy. The tumors and cortical bone are lifting and falling off the slides. Any suggestions? I've been reading old posts about EDTA decalcification - since hindsight is 20/20 - would that have been a better choice?? Boo hoo. Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From making <@t> nersp.nerdc.ufl.edu Wed Apr 14 12:56:19 2004 From: making <@t> nersp.nerdc.ufl.edu (Mike King) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] vibratome/frozen immuno, soap residue, autofluorescence, and bad list posting habits Message-ID: <407D7B43.9040704@nersp.nerdc.ufl.edu> re 9. RE: Vibratome Considerations (Charles Scouten) ...as for immunolabeling sensitivity differences between vibratome and frozen sections from cryoprotected tissue, is there any science to support this claim (references)? i've never seen a difference and doubt there is really any reason to expect one. especially if you need to permeabilize anyway... re Danielle Zalinski Vibratome Considerations post: ...50 micron sections of brain (vibratome or other) will label through the entire thickness depending on the fixation, permeabilization, and location of the antigen. if you're going to use the tissue for anything that demands full-thickness labeling, you should use a confocal microscope to check that the labeling is even through the entire thickness of the sections. papers in the histo literature will detail the procedures you need for your material, or post again for specifics. re 3. autoflourecence in RBC (James Watson) ...try 0.1% sodium borohydride for quenching autofluorescence; this used to be on the Molecular Probes website. please post your results if this does/doesn't work. re 13. Clean Glassware Test (Ostrander, Anita B) ...for testing for soap residue, any bartender worth his/her salt will tell you that nothing takes the head off a fine beer like a trace of soap on a poorly rinsed glass. perhaps your lab should stock a few pints of Guinness? ...and please, please, please, kind histonetters, don't include the whole digest in replies to histonet. hardly a day goes by when the digest doesn't contain at least one complete copy of all the previous day's posts because somebody didn't think before replying. we've all hit the send button too soon, but this is a real nuisance. happy histo, From gcallis <@t> montana.edu Wed Apr 14 12:58:41 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Decal question In-Reply-To: Message-ID: <3.0.6.32.20040414115841.00bbf470@gemini.msu.montana.edu> There is a better way to determine endpoint than cutting with a scalpel. You can do a weight loss, weight gain and if working with many mice, you can do one or two samples which will be relatively the same in weight, age of animal etc. and endpoint test these when doing batch decalcification. Jackie, It is not the decalcification that causes bone to fall off slides - problems with ACID decalcifiers show up as poor H&E staining. Your problem would happen with EDTA decalcified bone also. Waterbath is a bit hot, use Tissue Prep 2 to infiltrate or at least embed the bones in, harder paraffin and supports bone better. Trim block face and soak a bit on ice water (don't oversoak if bone swells out of block, not good) on top of ice block, cut with a NEW knife blade, preferred is high profile Dura Edge or EdgeRite (backwards naming??) from Richard Allan for a sturdier blade. If you want section to flatten, you can either place it on 10% alcohol cold water bath, pick up on slide and go to warm bath but don't let section float off slide, just let it stretch out in warm water. You are breaking surface tension and allowing cartilage (soft) stretch out in relation to the cortical bone (very hard). The hotter the waterbath the more likely the wrinkles will set permanently. Do not flatten with heated surface after picking up either, this can explode section. Have also used a 5% DMSO waterbath, glass insert in waterbath with 5% DMSO, let this heat up and lay sections on this. Caveat: DMSO is not to touch skin NOR be breathed, and it will cause metal parts to rust. It works, did a whole rabbit knee study with this. Some people have used Tween 20 in waterbath i.e. a detergent that allows section to flatten nicely. When you pick up on Plus charge, drain well, and then go to 37C - 40C heating platform and allow the sections to dry FLAT. We never dry at 60C, as this causes overdrying of cartilage, it curls off slide during rinses and do the latter very gently. Do not use ammonium hydroxide for bluing, use Scotts tap water or Richard Allan bluing reagent. If you still have problems you can presub clean, regular microscope slides with chrome Gelatin but use 275 bloom gelatin, the larger gelatin molecule is superior for holding boney things. Dry flat at 40C, overnight, even longer is better. We have even put a few granules of this gelatin in the waterbath as it is heating and use regular Superfrost (NOT positive charge) to pick up section. Don't overuse 275 bloom, it can cause unsightly background, or presub, then dip slide in NBF 10 dips, rinse and dry. Not knowing source of your positive charged slides, ours are made by Erie under Fisherbrand name or VWR brand name as long as they say Plus Charge. Good Luck, Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From lizchlipala <@t> premierhistology.com Wed Apr 14 12:59:59 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Decal question In-Reply-To: Message-ID: <000801c4224a$507d9bf0$74d48a80@LIZ> Jackie We cut a lot of bone sections here, we experimented quite a bit with the infiltration and paraffin embedding, and found that the harder the infiltration paraffin the better the sections would stay on the slides, no flipping of the articular cartilage. We also found that the harder paraffins made it difficult to section and obtain ribbons, so we had to make some changes to the infiltration media to find one that we were able to cut sections (get ribbons) and the sections would not flip during staining. We actually found a combination of surgipaths formula R (1/3) and richard allens paraffin 9 (2/3) worked the best with embedding in Richard allens paraffin 9. We rough trim our sections and let them sit on water and ice for about an hour or so. Then we section them and lay them flat on a hot plate, just like Patsy said, but we do not let the paraffin melt. They sit on the hot plate at about 36 degrees overnight, we do not place them in an oven to melt the paraffin, but transfer directly to staining racks and stain. We use plus slides and we rarely get any flipping. Liz Elizabeth A. Chlipala, BS, HTL(ASCP) Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie.O'Connor@abbott.com Sent: Wednesday, April 14, 2004 9:17 AM To: Histonet (E-mail); histonet-bounces@lists.utsouthwestern.edu Subject: [Histonet] Decal question I'm working on a trial with intra-tibial injections of tumors into mice. Whole (skin and muscle trimmed) legs were fixed for about a month in 10% NBF. I used Surgipath's Decal I to decalcify (took about 6 hours - end point determined by cutting through femur with scalpel). Paraffin sections cut nicely - I used a 48C waterbath to ensure no wrinkles, picked up on positively charged slides. I let them air dry overnight, then 60C for an hour prior to H+E staining. I'm not happy. The tumors and cortical bone are lifting and falling off the slides. Any suggestions? I've been reading old posts about EDTA decalcification - since hindsight is 20/20 - would that have been a better choice?? Boo hoo. Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From azdudley <@t> hotmail.com Wed Apr 14 13:41:31 2004 From: azdudley <@t> hotmail.com (anita dudley) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] silver nitrate Message-ID: does anyone know of anything that will get silver nitrate off of fingernails. thanks so much. anita dudley, providence hosp, mobile ala. _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From gcallis <@t> montana.edu Wed Apr 14 13:44:05 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Safranine o staining and Problem in tissues In-Reply-To: Message-ID: <3.0.6.32.20040414124405.00bd4628@gemini.msu.montana.edu> I sent the SafO/fast green method privately. As for your soft tissues, it sounds more like overdehydration in alcohols during processing, and possibly paraffin is too hot at infiltration, check temperature - it should be at 60C max. What is your processing schedule in terms of time per change? As for decalcification, endpoint should be determined either chemically or by some other reliable method. We have a quick, but relatively accurate method since the xray machine went to another university. It is a weight loss/weight gain method, and will work with all decalcifying agents. If you wish, I will attach to you. Routinely, we use 10 - 15% formic acid made up from 88% stock formic acid OR you can use buffered formic acid, as EDTA will extract proteoglycans which can change tinctorial quality of cartilage staining. Buffered formic acid can be made in lab using formic acid with either sodium formate or sodium citrate, these are available commercially. These recipes are found in histotechnology books under bone decalcification chapters. Buffered formic acid contains approx 4.5% formic acid with the buffer salt and is much slower to complete decalcification. Always make sure bones are fixed totally, rat joints can take as long as a week to fix in NBF, and change fixative after a couple of days to insure fixative is replenished. Clean off skin and extra muscle without damaging joints to facilitate fixation and decalcification. Another good, but fast decalcifier is 10% formic acid with 4% hydrochloric acid, and joints will take only day or so to decalcify. We prefer the slower, gentler 15% formic acid, and do endpoint testing faithfully. I suggest you also do back in Histonet archives and read discussion on decalcification as much of what is said here is already there, plus other interesting commentary. www.Histosearch.org Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From ccrowder <@t> mail.vetmed.lsu.edu Wed Apr 14 13:27:32 2004 From: ccrowder <@t> mail.vetmed.lsu.edu (Cheryl Crowder) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Dystrophin Message-ID: I want to thank all of you who replied to my query about dystrophin antibody. Not only did we get the names of vendors, but now I really look good to the researcher. You've made my day. Thank you! Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 From gcallis <@t> montana.edu Wed Apr 14 14:03:31 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] AEC+ with ARK kit Message-ID: <3.0.6.32.20040414130331.00bc5548@gemini.msu.montana.edu> Just curious, has anyone used AEC+ with the DAKO ARK kit during staining. Have a researcher who really does NOT like DAB final color, period! We are playing with Chris van der Loos's AEC recipe (quite sensitive! very similar to AEC+) and although this means tweaking antibody dilution, etc - all those good things needed for ARK'ing it seems to be working with our antibody. Hey, research is fun, and who sez we have to do it one way only! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From cwscouten <@t> myneurolab.com Wed Apr 14 14:37:46 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] vibratome/frozen immuno, soap residue, autofluorescence, and bad list posting habits Message-ID: Numerous studies have made the comparison. See a selected few below. It has been popular wisdom for 20 years that Vibratome sections, by not facturing the cell membrane, preserve the cytosol better and give more intense staining of cytosol proteins. I have personally seen differences with HRP staining, but didn't try to prove that some other variable was not responsible. I have located the studies below, and many others, that have made the comparison. I do not have access to the complete text, and do not know if all of these say the vibratome results were better. I would not expect vibratome to do better with membrane bound proteins. The science is there and done, I would compile a complete list of recent references like those below (maybe about 20-30) if somebody will volunteer to look them all up in Science Direct or the original Journals and check whether Vibratome sectioning was better, worse or the same compared to cryostat sectioning. Current concepts in neuroanatomical tracing C. K?bbert, R. Apps, I. Bechmann, J.L. Lanciego, J. Mey & S. Thanos Progress in Neurobiology, 2000, 62:4:327-351 solon@uni-muenster.de . The success of labelling can be estimated roughly, and the tissue can be sectioned with a vibratome. Cryosectioning of the tissue is not recommended, since some dye leakage occurs during the procedure. Neural tract tracing using Di-I: a review and a new method to make fast Di-I faster in human brain D. Larry Sparks, Lih-Fen Lue, Timothy A. Martin & Joseph Rogers Journal of Neuroscience Methods, 2000, 103:1:3 - 10 lsparks@mail.sunhealth.org if a fluorescent Di-I product is desired, infiltrating and embedding brain tissues in polyacrylamide prior to sectioning by vibratome eliminates the necessity of dehydrating and clearing the tissue sections prior to coverslipping ( Hayaran and Bijlani, 1992). (too thick) Although the thickness of the vibratome sections (50 ?m) was less than optimal for demonstrating individual axons, individual fiber tracts could occasionally be traced for as much as 5 mm or more, as shown here. Cryostat and other techniques were used to achieve thinner sections, but requiring freezing of the tissue, resulted in diffuse smearing of the fluorescence label Somebody let me know if this is favorable to my case. The comparison was made, but I can't get the full text to see how it came out. Triple immunofluorescence labelling of parvalbumin, calbindin-D28k and calretinin in rat and monkey brain Hartig, W. / Bruckner, G. / Brauer, K. / Seeger, G. / Bigl, V., Journal of Neuroscience Methods, Aug 1996 ...Thereafter, two brains were cut on a Vibratome (TSE, Kronberg) at 50 1lm thickness...monkey brains the tissue was cryo-protected by equili- bration...about 4 days and subsequent cryo-protection. 2.3. Tnple immu7...results were obtained with Vibratome and frozen sections and with... 53. Adenosine deaminase and histidine decarboxylase coexist in certain neurons of the rat brain Patel, B.T. / Tudball, N. / Wada, H. / Watanabe, T., Neuroscience Letters, Jan 1986 ...further 24 h at 4?C. Thick vibratome sections (50-100/~m) were obtained...required, the fixed blocks were cryo- protected with 301~:~ sucrose...and dried at 20~C for 24 h. Vibratome and cryostat sections were...immunoreactivity in rat brain vibratome sections was consistent with... Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mike King Sent: Wednesday, April 14, 2004 12:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] vibratome/frozen immuno, soap residue, autofluorescence, and bad list posting habits re 9. RE: Vibratome Considerations (Charles Scouten) ...as for immunolabeling sensitivity differences between vibratome and frozen sections from cryoprotected tissue, is there any science to support this claim (references)? i've never seen a difference and doubt there is really any reason to expect one. especially if you need to permeabilize anyway... re Danielle Zalinski Vibratome Considerations post: ...50 micron sections of brain (vibratome or other) will label through the entire thickness depending on the fixation, permeabilization, and location of the antigen. if you're going to use the tissue for anything that demands full-thickness labeling, you should use a confocal microscope to check that the labeling is even through the entire thickness of the sections. papers in the histo literature will detail the procedures you need for your material, or post again for specifics. re 3. autoflourecence in RBC (James Watson) ...try 0.1% sodium borohydride for quenching autofluorescence; this used to be on the Molecular Probes website. please post your results if this does/doesn't work. re 13. Clean Glassware Test (Ostrander, Anita B) ...for testing for soap residue, any bartender worth his/her salt will tell you that nothing takes the head off a fine beer like a trace of soap on a poorly rinsed glass. perhaps your lab should stock a few pints of Guinness? ...and please, please, please, kind histonetters, don't include the whole digest in replies to histonet. hardly a day goes by when the digest doesn't contain at least one complete copy of all the previous day's posts because somebody didn't think before replying. we've all hit the send button too soon, but this is a real nuisance. happy histo, _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Xudong_Cao <@t> brown.edu Wed Apr 14 14:39:33 2004 From: Xudong_Cao <@t> brown.edu (Xudong_Cao@brown.edu) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] PGP9.5 antibody against chicken? Message-ID: <200404141939.i3EJdXJ6002679@cepheus.services.brown.edu> Dear friends, I am looking for a PGP9.5 antibody against chicken. I checked some of the major vendor, the specificity of the PGP9.5 antibody against chicken is unknown. Anybody who has some experience would want to share some info on this? thanks. Xudong Cao, Ph.D. post-doctoral fellow Brown University Providence, RI From gcallis <@t> montana.edu Wed Apr 14 14:41:44 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] MacNeal's Tetrachrome recipe Message-ID: <3.0.6.32.20040414134144.00bc5898@gemini.msu.montana.edu> A word of advice, make up your own stain solution. We and another lab via private discussion found commercial preparation of this stain were very poor for PMMA bone section staining. MACNEALS TETRACHROME STOCK SOLUTION 0.5g methylene blue 0.8g azure a eosinate 0.1g methylene violet 250 ml methanol 250 glycerin Stir, leave at 50C for 12 hours, then 37C for 3 days. Filter, store in dark. WORKING STAINING SOLUTION 5 mls stock stain solution, 95mls distilled water. Bone is 1% formic acid etched for 30 sec to 1 minute, rinsed well with water before staining. Blot section before immersing in this stain solution. You can combine this stain with Toluidine blue method (Eurell and Sterchi) for even more brilliant results. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From algranth <@t> u.arizona.edu Wed Apr 14 14:58:17 2004 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Oil Red O questions Message-ID: <4.3.2.7.2.20040414124145.00cc0790@algranth.inbox.email.arizona.edu> Every now and then I do Oil Red O stains (sometimes 30-50 at a time) and I have a few questions that maybe the experts in histoland can help me with. First is stain residue on the slide - I make the working stain up fresh, let sit 10 minutes and filter and use right away. I use plus slides so maybe this is the problem? There is usually an even residue on the slide that I wipe away before coverslipping being careful to not wipe off the tissue. The tissue looks fine, there is staining where it should be and no residue from the stain on the tissue where it isn't supposed to be. How can this residue be reduced or eliminated or is this just the way it is? Other question is which mounting medium is the best in your opinion for Oil Red O? I use Shur/Mount and it looks great at first but I had one person who went back a couple months later to take pictures and the slides had dried out to where the fat was OK but the nuclei were not sharp. If you need to take pictures of the stain is it best to do that right away? Should I be sealing the coverslips with nail polish? Thanks. 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 cgfields <@t> lexhealth.org Wed Apr 14 15:31:51 2004 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] silver nitrate Message-ID: X-14 (Shower cleaner) will get it out of your (white) clothes and off hands. It is a strong cleaner...be careful. It smells like strong clorox. Carole Fields Lex Med Ctn W.Columbia, SC > -----Original Message----- > From: anita dudley [SMTP:azdudley@hotmail.com] > Sent: Wednesday, April 14, 2004 2:42 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] silver nitrate > > does anyone know of anything that will get silver nitrate off of > fingernails. thanks so much. anita dudley, providence hosp, mobile ala. > > _________________________________________________________________ > Is your PC infected? Get a FREE online computer virus scan from McAfee? > Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 > > > _______________________________________________ > 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 cfavara <@t> niaid.nih.gov Wed Apr 14 15:30:20 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] detach cells from a slide....again Message-ID: Proteinase K at a high conc. should do it. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Dolors Fuster [mailto:d.fuster@ub.edu] Sent: Wednesday, April 14, 2004 10:50 AM To: HISTONET Subject: [Histonet] detach cells from a slide....again In my previous mail I forgot to tell you the mechanical method (scrape them off with a sharp razor blade) is made....but too slow for at least 600 slides...difficult to recover all stuff...some of recovered adheres to the eppendorf.... :o( Could the mehod not include trypsin in its procedure :o) ?? Thanks again Any idea to detach cells from an hematologic slide? They are dryed...stained with May Gr?nwald Giemsa...dehidrated, cleared and mounted with DPX. The idea is to use them for a DNA extraction. Thanks in advance _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----- Fi del missatge reenviat ----- Dolors Fuster T?cnic Especialista en Anatomia Patol?gica i Citologia Departament d'Anatomia i Embriologia Humana Facultat de Medicina (HCP) Universitat de Barcelona ICQ 14146798 ------------------------------------------------- This mail sent through IMP: http://horde.org/imp/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sue.Kapoor <@t> uhsi.org Wed Apr 14 15:30:46 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Silver stain for Alzheimers Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F5598@khmcexch.uhsi.org> Hi, does anyone have a stain procedure they can send me for modified methenamine silver method for cerebral anyloid? THANK YOU in advance :) Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From Solverboy <@t> aol.com Wed Apr 14 15:36:01 2004 From: Solverboy <@t> aol.com (Solverboy@aol.com) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] High Quality X-ray Film Message-ID: <1061EA1C.6913B30D.0293A525@aol.com> Hello everyone, I am currently working on radioactive in-situ hybridizations that involve low abundance probes. I have been using a Mammography Film from Fuji that has a very high grain density, but the sensitivity is low. Typically i have to expose this film for 1-1.5 months to get a decent signal. Can anyone suggest an x-ray film that has a high level of contrast, dynamic range, grain density along with high sensitivity. I need to shorten my experiment times, any suggestions would be greatly appreciated. Thanks in advance, Fred Grau Dept. Neurobiology and Behavior S.U.N.Y Stony Brook From gcallis <@t> montana.edu Wed Apr 14 16:03:08 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Oil Red O questions In-Reply-To: <4.3.2.7.2.20040414124145.00cc0790@algranth.inbox.email.ari zona.edu> Message-ID: <3.0.6.32.20040414150308.00bd3d78@gemini.msu.montana.edu> Hi Andi, I have an oil red O stain that doesn't leave crud all over the slide, clean, crisp and sooooo easy to do. I am sending a copy via private email to you. You will not be unhappy with this stain, it is wonderful and far less work than you have been doing. After rinsing, just coverslip with an aqueous mounting media. It was published by Chuck Churukian, and I never looked back once I used it, NO MORE MESSY stain to deal with! Instead of nail polish that contains isopropyl alcohol, dump this out and put diluted mounting media in little fingernail polish bottle so you can use tidy little brush. This seals nicely and dries rapidly. Dilute your regular mounting media with toluene to consistency of a top coat or base coat which is thinner than regular nail polish. (Sorry, guys, a girl thing describing the different types of glamour fingernail polish stuff!) GFP was found to lose fluorescence due to nail polish that leached into aqueous media. Curiousity prompted reading of labeled ingredients on nail polish bottle. Except for the alcohol, other ingredients were not miscible with water, so I deduced the alcohol was the culprit and this was known to kill glowing GFP, probably not the eGFP of today. GFP vs nail polish is published info. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From convmcm <@t> cc.usu.edu Wed Apr 14 16:49:18 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] AEC+ with ARK kit In-Reply-To: <3.0.6.32.20040414130331.00bc5548@gemini.msu.montana.edu> Message-ID: <000001c4226a$56f39be0$4a737b81@Cygnus> I don't use the ARK kit at all. A wide variety of animal tissues pass through my lab and I have found either the DAKO LSAB2 (HRP + AEC)or their Envision polymer (HRP-AEC or DAB) works great. I've never tried the LSAB2 or Envision AP kits. I've been tempted to try them. I also like the serum-free blocker for the rabbit polyclonals I'm working with. It kills the background staining fabulously. >> Hey, research is fun, and who sez we have to do it one way only! Gayle --- HA! Isn't life great?? I think I'll go have some fun myself ... |:) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Wednesday, April 14, 2004 12:04 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] AEC+ with ARK kit Just curious, has anyone used AEC+ with the DAKO ARK kit during staining. Have a researcher who really does NOT like DAB final color, period! We are playing with Chris van der Loos's AEC recipe (quite sensitive! very similar to AEC+) and although this means tweaking antibody dilution, etc - all those good things needed for ARK'ing it seems to be working with our antibody. Gayle Callus MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Wed Apr 14 16:55:10 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] detach cells from a slide In-Reply-To: <1081959420.407d63fcc35eb@webmail1.ub.edu> Message-ID: <000501c4226b$28953a50$4a737b81@Cygnus> In cell culture, when cells need to be transferred to a new flask, a 0.25% trypsin for 15 minutes is used to detatch the cells from the flask surface. I don't know if that would work with blood cells, but you could experiment to see. If you try it, I wouldn't let the cells soak as long as 15 minutes. I'd watch the solution for cell lift off and call it good when I got most of them. just another wild and crazy notion... Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dolors Fuster Sent: Wednesday, April 14, 2004 9:17 AM To: HISTONET Subject: [Histonet] detach cells from a slide Any idea to detach cells from an hematologic slide? They are dryed...stained with May Gr?nwald Giemsa...dehidrated, cleared and mounted with DPX. The idea is to use them for a DNA extraction. Thanks in advance Dolors Fuster T?cnic Especialista en Anatomia Patol?gica i Citologia Departament d'Anatomia i Embriologia Humana Facultat de Medicina (HCP) Universitat de Barcelona ICQ 14146798 ------------------------------------------------- This mail sent through IMP: http://horde.org/imp/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KAELPERS <@t> aol.com Wed Apr 14 20:05:02 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Decal question Message-ID: <128.3f49bbb6.2daf39be@aol.com> We decal and cut femurs on all types of species. We also experience the articular cartilage wanting to flip during staining if we don't use charged slides. We are use basically the protocol mentioned previously. We us Surgipath Decal I for decalcification. Rinse thoroughly before processing. We are embedding in Paraplast embedding medium which is sturdy enough to withhold the bone specimens. Shave block until surfaced, set on ice for approximately 30-40 minutes. We also use 15 ml of Surgipaths Sta-on in our tissue baths. Section at 4 microns and place on plus slides, let air dry overnight. Rarely do we use the oven unless we need to rush a sample and then we would put it in a 60 degree oven for about 25 minutes. I don't know if this helped but it sounds like most people use similar protocols and have from time to time the same issues. lge From KAELPERS <@t> aol.com Wed Apr 14 20:24:04 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Isopropanol in processing Message-ID: <149.26adabc7.2daf3e34@aol.com> I have read articles and attended workshops in which it has been stated that isoproponal is more harsh on tissue. Has your facility considered buying a recycler to resolve the issues with purchasing of Ethanol? CBG Biotech has a unit that will recycle alcohol and xylene on the same unit. We are trialing one that recycles formalin, xylene and alcohol all in one unit. It is going to save us more than 30,000.00 a year. Propose a trial it is free. Contact eweinblatt@cbgbiotech.com for information. lge From azdudley <@t> hotmail.com Wed Apr 14 21:49:29 2004 From: azdudley <@t> hotmail.com (anita dudley) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] thanks Message-ID: thanks everyone for the info about how to remove silver nitrate. I did have gloves on, but I was making up some and I must have gotten it on the counter, I am not sure how it got on my hands. I agree tho we should always wear gloves!!!! thanks again, everyone have a nice weekend! anita dudley, providence hosp. mobile al. _________________________________________________________________ Watch LIVE baseball games on your computer with MLB.TV, included with MSN Premium! http://join.msn.com/?page=features/mlb&pgmarket=en-us/go/onm00200439ave/direct/01/ From balajimr <@t> drreddys.com Thu Apr 15 00:06:08 2004 From: balajimr <@t> drreddys.com (balajimr@drreddys.com) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] BrdU invivo dosage Message-ID: Dear histonetter: Can anyone tell me the exact dose of Bromo deoxy uridine in rats and mice for the purpose of invivo labelling for immunohistochemistry (particularly my interest is intestines and urinary bladder). We plan to carryout both intraperitonially and orally through drinking water.As for as I know the dose in mice is 100mg/kg IP at a dose volume of 20 ml/kg. Is it OK to dilute BrdU in only PBS or any other combination is required. . It will be of great help for me if you could provide me the reference literature for the same.Thanks in advance. Dr. Balaji (M. V. Sc. Path) Dept. Pre clinical safety evaluation, Discovery research, Dr. Reddys Laboratories ltd. Bollaram Road, Miyapur, Hyderabad, 500 049 Andhra Pradseh, INDIA Tel: 23045439 - Ext.432. From c.m.vanderloos <@t> amc.uva.nl Thu Apr 15 02:51:13 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] RE: PAL-E mab Message-ID: <169ae21665f7.1665f7169ae2@amc.uva.nl> Dear Christian, PAL-E antibody is available from MonoSan located in The Netherlands. Just visit www.sanbio.nl and find your local contacts to order this antibody. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Christian Franci Date Tue, 13 Apr 2004 14:55:19 -0700 To histonet@lists.utsouthwestern.edu Subject [Histonet] PAL-E mab Maybe, someone out there knows where I might find a supplier of polyclonal or mAb for PAL-E an endothelial marker for "leaky" vessels. If, anyone can point me in the right direction, I'd much appreciate it. Thanks in advance, C From histol <@t> rat-hole.org Thu Apr 15 03:14:47 2004 From: histol <@t> rat-hole.org (Matt Ibbs) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Isopropanol in processing Message-ID: <5.1.0.14.2.20040415095133.009ed340@bigpop3.wyith.net> Dear all, Thanks for the opinions. Here in Poland we have been using industrial methylated spirit (the mixture many of you describe as reagent alcohol) for years. The authorities have now decided to tax that too. That's why we need to switch to Isopropanol. Unfortunately we are not able to install a recycling machine in our facility as local regulations require that such machines are housed in bomb-proof rooms - preferably in separate buildings. In response to Gayle Callis: Thank you! I knew it could be done and I remember doing it in another department some 10 years ago. I tried to reproduce the processing protocol from memory but we have some problems. Here are two protocols we've tried so far: Protocol 1: In this first schedule I used three changes of 100% Isopropanol as experiments with only two caused the xylene to go cloudy - suggesting water remained. This problem vanished with three changes. Everything was well for two days but shortly afterwards problems emerged with small biopsies being too brittle. 1. 60% Isopropanol 1 hour 2. 80% Isopropanol 1 hour 3. 90% Isopropanol 1 hour 4. 100% Isopropanol 45 minutes 5. 100% Isopropanol 45 minutes 6. 100% Isopropanol 45 minutes 7. 2:1 Isopropanol:Xylene 30 minutes 8. 1:2 Isopropanol:Xylene 30 minutes 9. Xylene 1 hour 10. Xylene 1 hour 11. Paraffin 3 hours 12. Paraffin 3 hours Protocol 2: In order to counter the problems with small biopsies we refreshed the paraffin and removed the third change of 100% isopropanol. The time in the remaining two changes of 100% isopropanol was increased to 1 hour to allow for the reduced number of changes. Mysteriously, the clouding of xylene previously observed was no longer evident. Otherwise the schedule was unchanged. After a day or so the 80% isopropanol (now realistically nearer 70%) started to gather a cloud of slightly viscous-looking grey "mush" in the bottom of the container. Odd. This problem was fixed of course. At this point we had to process whole slices of prostate alongside our routine work - this is not uncommon so we must be able to accommodate it. The tissue was poorly processed and fell off the slides - which is annoying with extra-large blocks especially on the third or fourth attempt. ;o) Next I plan to alter xylene times as you suggested. In answer to your questions about our paraffin, it is held on the processor at around 58-60 degrees Celsius. My colleagues (a case of "we've always done it this way") insist that a small amount of bees-wax is added to soften the wax. Ideally I'd like to use four changes of paraffin but our processor cannot accommodate them. Any advice would be welcome. Now I'm going to re-search the archives as you advised. Maybe I missed something. Thanks again everyone. Matt. Matthew Ibbs Dept. of Neoplastic Pathology Karol Marcinkowski University of Medical Sciences Poznan Poland From townserl <@t> lsu.edu Thu Apr 15 07:43:54 2004 From: townserl <@t> lsu.edu (Robbie L Townsend) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] alphaMSH Message-ID: Hi histonetters. Does anyone out there know of a good alpha-MSH antibody to use on mouse brain and the appropriate dilution? I have tried one from Chemicon and one from Immunostar, but I am still getting very high background. I am using them at a very high dilution, rinse very well, and block for two hours. Any help would be greatly appreciated. Thanks Leigh Townsend Research Associate Pennington Biomedical Baton Rouge, La email- townserl@lsu.edu phone- (225)763-3001 From Ian.Bernard <@t> LACKLAND.AF.MIL Thu Apr 15 08:56:33 2004 From: Ian.Bernard <@t> LACKLAND.AF.MIL (Bernard Ian R SSgt 59 CRES/MSROP) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Isopropanol in processing Message-ID: I work in a research facility and we use Isoprapanol and have not experience any negative microscopic/tissue changes in tissue diagnosis. Hence, we continue to use Isopropanol, as it is less controlled and cheaper than ethyl alcohol. Our pathologist is fine eith it. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Matt Ibbs Sent: Thursday, April 15, 2004 3:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Isopropanol in processing Dear all, Thanks for the opinions. Here in Poland we have been using industrial methylated spirit (the mixture many of you describe as reagent alcohol) for years. The authorities have now decided to tax that too. That's why we need to switch to Isopropanol. Unfortunately we are not able to install a recycling machine in our facility as local regulations require that such machines are housed in bomb-proof rooms - preferably in separate buildings. In response to Gayle Callis: Thank you! I knew it could be done and I remember doing it in another department some 10 years ago. I tried to reproduce the processing protocol from memory but we have some problems. Here are two protocols we've tried so far: Protocol 1: In this first schedule I used three changes of 100% Isopropanol as experiments with only two caused the xylene to go cloudy - suggesting water remained. This problem vanished with three changes. Everything was well for two days but shortly afterwards problems emerged with small biopsies being too brittle. 1. 60% Isopropanol 1 hour 2. 80% Isopropanol 1 hour 3. 90% Isopropanol 1 hour 4. 100% Isopropanol 45 minutes 5. 100% Isopropanol 45 minutes 6. 100% Isopropanol 45 minutes 7. 2:1 Isopropanol:Xylene 30 minutes 8. 1:2 Isopropanol:Xylene 30 minutes 9. Xylene 1 hour 10. Xylene 1 hour 11. Paraffin 3 hours 12. Paraffin 3 hours Protocol 2: In order to counter the problems with small biopsies we refreshed the paraffin and removed the third change of 100% isopropanol. The time in the remaining two changes of 100% isopropanol was increased to 1 hour to allow for the reduced number of changes. Mysteriously, the clouding of xylene previously observed was no longer evident. Otherwise the schedule was unchanged. After a day or so the 80% isopropanol (now realistically nearer 70%) started to gather a cloud of slightly viscous-looking grey "mush" in the bottom of the container. Odd. This problem was fixed of course. At this point we had to process whole slices of prostate alongside our routine work - this is not uncommon so we must be able to accommodate it. The tissue was poorly processed and fell off the slides - which is annoying with extra-large blocks especially on the third or fourth attempt. ;o) Next I plan to alter xylene times as you suggested. In answer to your questions about our paraffin, it is held on the processor at around 58-60 degrees Celsius. My colleagues (a case of "we've always done it this way") insist that a small amount of bees-wax is added to soften the wax. Ideally I'd like to use four changes of paraffin but our processor cannot accommodate them. Any advice would be welcome. Now I'm going to re-search the archives as you advised. Maybe I missed something. Thanks again everyone. Matt. Matthew Ibbs Dept. of Neoplastic Pathology Karol Marcinkowski University of Medical Sciences Poznan Poland _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From docmichel <@t> netbulmail.com Thu Apr 15 09:04:56 2004 From: docmichel <@t> netbulmail.com (izzet oguz) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] Osteoclast demonstration Message-ID: <1975.193.255.128.130.1082037896.webmail@mail1.netbulmail.com> Hi Histonetters, Is there anybody who has been worked with an osteoclast specific antibody? We found only the histochemical TRAP method to demonstrate osteoclasts. We will be appreciated if someone offers us an antibody or a histochemical method apart from TRAP. Thank you very much... ?zzet Oguz Mersin University Medicine Faculty Histology and Embryology Department TURKEY From inka <@t> uhnres.utoronto.ca Thu Apr 15 09:29:56 2004 From: inka <@t> uhnres.utoronto.ca (Inka Tertinegg) Date: Fri Sep 16 15:22:48 2005 Subject: [Histonet] warrantee service Message-ID: <407E9C64.9AFA06B2@uhnres.utoronto.ca> Hello, Our lab is buying a new -80 freezer and I have been asked to evaluate the warrantee service of the different brands, ie how quickly and efficiently the freezer was repaired while under warrantee. We are looking at Revco, Forma and Sanyo. I was hoping that people with experience of a freezer breakdown while under warrantee could share their experience with me. Thank you -- Inka Tertinegg University of Toronto Department of Ophthalmology 399 Bathurst St., MC6-411 Toronto M5T 2S8 416 603-5800 x 2850 From inka <@t> uhnres.utoronto.ca Thu Apr 15 09:36:01 2004 From: inka <@t> uhnres.utoronto.ca (Inka Tertinegg) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] warrantee service Message-ID: <407E9DD1.CD5E8F50@uhnres.utoronto.ca> Hello, Our lab is buying a new -80 freezer and I have been asked to evaluate the warrantee service provided by the various brands, ie how quickly and efficiently the repair is made. We are looking at Revco, Forma and Sanyo. I was hoping that anyone in southern Ontario who has had one of these freezers break down on them during the warrantee period would share their experience with me. Thank you, Inka -- Inka Tertinegg University of Toronto Department of Ophthalmology 399 Bathurst St., MC6-411 Toronto M5T 2S8 416 603-5800 x 2850 From bhewlett <@t> cogeco.ca Thu Apr 15 09:34:51 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Osteoclast demonstration References: <1975.193.255.128.130.1082037896.webmail@mail1.netbulmail.com> Message-ID: <000801c422f6$d13fb910$6500a8c0@mainbox> Izzet, Zymed carry an TRAP antibody that works well on FFPE sections. The osteoclasts stain beautifully! Bryan ----- Original Message ----- From: "izzet oguz" To: Sent: Thursday, April 15, 2004 10:04 AM Subject: [Histonet] Osteoclast demonstration > Hi Histonetters, > Is there anybody who has been worked with an osteoclast specific antibody? > We found only the histochemical TRAP method to demonstrate osteoclasts. We > will be appreciated if someone offers us an antibody or a histochemical > method apart from TRAP. > Thank you very much... > ?zzet Oguz > Mersin University Medicine Faculty > Histology and Embryology Department > TURKEY > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gcallis <@t> montana.edu Thu Apr 15 09:53:41 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Oil Red O Churukian method, reposted Message-ID: <3.0.6.32.20040415085341.00bc6358@gemini.msu.montana.edu> Dear All who want this Oil Red O method, the none messy type! Enjoy Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) **************************************************************************** ***** Oil Red O/Dextrin, Churukian method Fresh tissue (not prefixed) frozen sections are immersed immediately into Neutral buffered formalin for a minimum of 10 min (can be hours/days). Rinse sections with distilled water before staining. Prefixed, with NBF should be cryoprotected in 20 - 30% sucrose, mounted on Plus Charge slides and air dried for 30 min to 1 hour, or longer to insure sections stay on slide. Do not fix frozen sections with alcohol or acetone to prevent lipid removal. Protocol: 1. Immerse dry slides directly into filtered 0.5% Oil Red O in Dextrin , stain 20 minutes 2. Rinse VERY GENTLY in running tap water 3. Counterstain with Gill II hematoxylin for 20 - 30 seconds 4. Rinse gently with water, blue in Scotts tap water type bluing solution, (NOT AMMONIA WATER), rinse gently, and coverslip with aqueous mounting media. Reagents: Dissolve 0.5 gm Oil Red O in absolute isopropyl alcohol and stir overnight. Dissolve 1 gm dextrin (bacteriological grade or TYPE III (Sigma) from corn in 100 ml distilled water Working solution is 60 mls stock Oil Red O and 40 ml 1% dextrin solution Stable for months, and reported to work on paraffin sections. Reference: Gamble and Bancroft, Theory and Practice of Histological Techniques, 5th Edition, 2001 with photos. This method also published in J of Histotechnology by Charles Churukian. Go to JOH archives for journal year/volume. From k.whalley <@t> ich.ucl.ac.uk Thu Apr 15 10:37:45 2004 From: k.whalley <@t> ich.ucl.ac.uk (Katy Whalley) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] storing sections Message-ID: <1516.194.82.240.48.1082043465.squirrel@jenner.ich.ucl.ac.uk> Hi, I apologise if this question has been asked before, but I am new to this list! I would like to know the best way to store 30 micron sections, cut on the cryostat. The sections are PFA-fixed and, after cutting, they are in PBS. I want to use them with a free floating immunofluorescence protocol, but may need to store some for later use if possible. What is the best way to store the sections before staining? Any advice would be appreciated! Katy Whalley, UCL From gcallis <@t> montana.edu Thu Apr 15 10:56:23 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Churukian Oil Red O reference In-Reply-To: References: <3.0.6.32.20040414150308.00bd3d78@gemini.msu.montana.edu> <3.0.6.32.20040414150308.00bd3d78@gemini.msu.montana.edu> Message-ID: <3.0.6.32.20040415095623.00bc6358@gemini.msu.montana.edu> C. Churukian, Lillie's Oil Red O for neutral lipids, J Histotechnology 22(4):309, 1999 For some reason, this reference was not correctly done in the Gamble Bancroft book, although the method is these. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Sue.Kapoor <@t> uhsi.org Thu Apr 15 11:19:46 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] test Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F559C@khmcexch.uhsi.org> test Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From Sue.Kapoor <@t> uhsi.org Thu Apr 15 11:53:33 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] test Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F559D@khmcexch.uhsi.org> test From mhorne <@t> upei.ca Thu Apr 15 11:49:41 2004 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] IHC on crustaceans Message-ID: <407E92F3.27028.12F932A@localhost> Hi , has anyone done C219 on FFPE sections of crustaceans? Wondering about the fixative. The literature suggests Davidson's. Anyone use anything else? Also wondering about the IHC protocol; if anyone has done C219 on 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 mcauliff <@t> umdnj.edu Thu Apr 15 15:19:00 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] storing sections In-Reply-To: <1516.194.82.240.48.1082043465.squirrel@jenner.ich.ucl.ac.uk> References: <1516.194.82.240.48.1082043465.squirrel@jenner.ich.ucl.ac.uk> Message-ID: <407EEE34.7000405@umdnj.edu> Hi Katy: I have stored 20-50 micron frozen sections of mouse or rat brain in 24/12 well tissue culture plates in PBS in the refrigerator for several weeks or longer. Immunostaining for tyrosine hydroxylase, dopamine beta-hydroxylase, GFAP and Mac-1 was not harmed. However, other antigens may not be so forgiving! The only problem I had is that mould/fungus/bacteria will eventually start to grow. I would recommend autoclaving the PBS and changing it every other week, or sooner. For longer storage you might want to try storing the sections at minus 20C in a "cryoprotectant" solution of sucrose or ethylene glycol (and glycerine?), but I have no first hand experience with that. Geoff Katy Whalley wrote: >Hi, > >I apologise if this question has been asked before, but I am new to this >list! I would like to know the best way to store 30 micron sections, cut >on the cryostat. The sections are PFA-fixed and, after cutting, they are >in PBS. I want to use them with a free floating immunofluorescence >protocol, but may need to store some for later use if possible. What is >the best way to store the sections before staining? Any advice would be >appreciated! > >Katy Whalley, UCL > >_______________________________________________ >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 Sue.Kapoor <@t> uhsi.org Thu Apr 15 12:18:26 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Alzheimer stain Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F559E@khmcexch.uhsi.org> hello histoland, Going through the archives, I found a reference to a "modified methenamine silver method for cerebral amyloid", can anyone send me a procedure for this?? THANK YOU in advance :) Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From dmcaloose <@t> wcs.org Thu Apr 15 12:43:02 2004 From: dmcaloose <@t> wcs.org (McAloose, Dee) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Job Opportunity - Wildlife Conservation Society Message-ID: <9C26E1A5C9EE5F4B84216095CBE2E24A059DD7AA@WOLF.wcs.org> Hello, I'd like to announce an immediate opening for a full-time histotechnician in the pathology department at the Wildlife Conservation Society located at the Bronx Zoo. We provide the diagnostic pathology services to one of the largest zoological collections in the country. Applications are being accepted until May 15, 2004 or until the position is filled. Please contact Ms. Tawanda Williams (Human Resources; email: twilliams@wcs.org) if you're interested in this exciting opportunity to become part of our team! Thanks for listening and we look forward to hearing from you. D McAloose, VMD, Dipl ACVP Head, Department of Pathology Wildlife Conservation Society Bronx, NY 10464 (phone) 718-220-7105 (fax) 718-220-7126 dmcaloose@wcs.org From townserl <@t> lsu.edu Thu Apr 15 13:01:53 2004 From: townserl <@t> lsu.edu (Robbie L Townsend) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Re: PFA frozen section storing Message-ID: Hi Katy We use a cryoprotectant solution in our lab. This is a long term freezing solution. The recipe: For 500ml: 150 ml Ethylene Glycol 100 ml Glycerol into 250 ml PBS (0.5M) *stir until clear Sections can be placed in this solution and stored at -16degrees. When ready to use, the sections must be rinsed with PBS very well. If you don't need to store them for very long, say 3-4 days, you may want to use a PBS with Sodium Azide solution instead of cryoprotectant.The recipe: 0.05% Sodium Azide in PBS: 0.5g Sodium Azide 1000ml PBS When ready to use, just rinse the sections about 3x10 min each in PBS. Hope this helps, Leigh Townsend Research Associate Pennington Biomedical Research Center Baton Rouge, La From ellacott <@t> ohsu.edu Thu Apr 15 13:26:33 2004 From: ellacott <@t> ohsu.edu (Kate Ellacott) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Re: alph-MSH Message-ID: Hi Leigh, The one that we tend to use is the sheep anti-alpha MSH from Chemicom, AB5087. Is this the one that you have tried? We use it at 1:6000 on free-floating perfused brain tissue. In my experience it only works in the hypothalamus (ARC) but not in the brainstem (if you know of one that works in the brainstem let me know!). From your message it sounds like you are getting a signal just with a lot of background. Have you tried increasing your blocking serum concentration? I routinely use 5% donkey serum to block for 2h at room temp and then incubate in primary antibody diluted in blocking serum overnight at 4 degrees C. What secondary are you using? You don't say if you are using HRP or fluoresence. Jackson Immunoresearch have a number of secondary antibodies that have been preabsorbed to minimize cross reactivity and background. These are good. Please feel free to e-mail me if you have any more questions as I know how frustrating this can be. Best wishes, Kate Dr. Kate Ellacott Vollum Institute Portland, OR USA Message: 1 Date: Thu, 15 Apr 2004 07:43:54 -0500 From: Robbie L Townsend Subject: [Histonet] alphaMSH To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hi histonetters. Does anyone out there know of a good alpha-MSH antibody to use on mouse brain and the appropriate dilution? I have tried one from Chemicon and one from Immunostar, but I am still getting very high background. I am using them at a very high dilution, rinse very well, and block for two hours. Any help would be greatly appreciated. Thanks Leigh Townsend Research Associate Pennington Biomedical Baton Rouge, La email- townserl@lsu.edu phone- (225)763-3001 Dr. Kate Ellacott Vollum Institute Oregon Health and Science University 3181 SW Sam Jackson Park Rd Portland OR 97239 (503) 494 4667 From carl.hobbs <@t> kcl.ac.uk Thu Apr 15 13:44:40 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] re PGP9.5 antibody against chicken Message-ID: <001201c42319$b5ed2a30$dd039a51@home> For my interest, why PGP9.5? If it's neurones and/or their processes, surely , for example the anti neurofilament, beta tubulin III and neuronal cell body( eg Hu C/D) Ab reagents are sufficient? From is135475 <@t> bcm.tmc.edu Thu Apr 15 22:41:56 2004 From: is135475 <@t> bcm.tmc.edu (Isaiah G. Schauer) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Best Marker for Proliferation? Message-ID: <026F856F-8F58-11D8-94FD-003065D60DC4@bcm.tmc.edu> Histonetters, Good day. Thanks to all of you that replied to my previous message on CD31/PECAM-1 staining. I now have another question to pose. What is the 'best' marker to use for immunohistochemical detection of cell proliferation on tissue slides from 4% paraformaldehyde-fixed, paraffin-embedded mouse xenograft tissue (human prostate epithelial cells mixed with mouse prostate stromal cells)? (Or, what is the most current, widely-accepted method) The markers I know of are Ki67, PCNA, Cdc2 and Brdu-kits (non-tritiated thymidine method, ex: Roche cat 1 299 964), although I'm open to any other experienced suggestions. Thanks for your time and advice. Sincerely, Isaiah Schauer 3rd year graduate student Lab of David R. Rowley, Ph.D. Baylor College of Medicine Department of Molecular and Cellular Biology One Baylor Plaza, room 326D Houston, TX 77030 lab: 713-798-6221 fax: 713-790-1275 3rd year graduate student Lab of David R. Rowley, Ph.D. Baylor College of Medicine Department of Molecular and Cellular Biology One Baylor Plaza, room 326D Houston, TX 77030 lab: 713-798-6221 fax: 713-790-1275 From doscwk <@t> nus.edu.sg Fri Apr 16 00:36:16 2004 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Micowave tissue processors Message-ID: Hi Histonetters, I'm confused about the different types of microwave tissue processors from Milestone and Pelco and need some advice from those of you who have these. Any pros and cons from current users would be greatly appreciated. I'm considering getting one in the near future. Our usage is mainly on processing bone specimens. Thanks in advance Julee Chan Musculoskeletal Tissue Lab National University of Singapore From soumya25 <@t> nbrc.ac.in Fri Apr 16 01:07:35 2004 From: soumya25 <@t> nbrc.ac.in (soumya) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Celloidin sections for IHC Message-ID: <002e01c42379$1d118940$3e00a8c0@nbrc.btisnet.ac.in> Hi, I'm new to this group. I need some help for doing immunostaining on celloidin embedded tissues. I recently started embedding fetal tissue in celloidin to avoid subjecting it to high temp during paraffin embedding. What I found was that the celloidin became so dark during the the DAB-Ni step that we can barely see staining in the tissue sections at all. Does anyone have any suggestions on how we can solve this problem? Any help will be greatly appreciated. Thanks, Soumya Dr. Soumya Iyengar National Brain Research Centre, NH-8, Manesar, India Dr. Soumya Iyengar Scientist National Brain Research Centre, NH-8, Manesar, Haryana - 122050, India Ph. no. 0124-2338922-8926 From alexander.nader <@t> wgkk.sozvers.at Fri Apr 16 01:24:23 2004 From: alexander.nader <@t> wgkk.sozvers.at (Nader, Alexander) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Osteoclast demonstration Message-ID: <4FE843CBBAC1D211A8150008C70952DAAE7D8D@hk01nt05.hkh.wgkk.sozvers.at> > Is there anybody who has been worked with an osteoclast > specific antibody? TRAP antibody from Zymed (Catalog-# 18-0199, clone ZY-9C5, pH 8 EDTA HIER pretreatment) demonstrates human osteoclasts nicely. Alexander Nader MD Vienna, Austria From Jackie.O'Connor <@t> abbott.com Fri Apr 16 07:13:16 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Best Marker for Proliferation? Message-ID: My vote is with Ki67. I work with human xenografts in mice, and you'll find in a lit search that Ki67 (MIB1) is the most commonly used marker for proliferation. Most manufacturers antibodies work well with HIER - tough to find one that is human specific, tho - which means you get staining in the stroma, but for the most part it's inconsequential. Jackie O'Connor "Isaiah G. Schauer" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/15/2004 10:41 PM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Best Marker for Proliferation? Histonetters, Good day. Thanks to all of you that replied to my previous message on CD31/PECAM-1 staining. I now have another question to pose. What is the 'best' marker to use for immunohistochemical detection of cell proliferation on tissue slides from 4% paraformaldehyde-fixed, paraffin-embedded mouse xenograft tissue (human prostate epithelial cells mixed with mouse prostate stromal cells)? (Or, what is the most current, widely-accepted method) The markers I know of are Ki67, PCNA, Cdc2 and Brdu-kits (non-tritiated thymidine method, ex: Roche cat 1 299 964), although I'm open to any other experienced suggestions. Thanks for your time and advice. Sincerely, Isaiah Schauer 3rd year graduate student Lab of David R. Rowley, Ph.D. Baylor College of Medicine Department of Molecular and Cellular Biology One Baylor Plaza, room 326D Houston, TX 77030 lab: 713-798-6221 fax: 713-790-1275 3rd year graduate student Lab of David R. Rowley, Ph.D. Baylor College of Medicine Department of Molecular and Cellular Biology One Baylor Plaza, room 326D Houston, TX 77030 lab: 713-798-6221 fax: 713-790-1275 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From siksik03 <@t> comcast.net Fri Apr 16 07:24:40 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Staining In-Reply-To: References: Message-ID: Hi HistoNetters Dave asks whether lab microwaves are any different behind the label. The answer is, some are, and some aren't. Many lab microwaves start life as kitchen microwaves, and have their electronics replaced. Others are built from the ground up for the lab. It's important to ask, and you get what you pay for. That being said, what all lab microwaves should provide are: ? vented cavity for safety ? accurate, reproducible, calibrated power or temperature control ? tested protocols for whatever application you need ? cycle time of 2 seconds or less or continuous power option ? agitation or stirring A NCCLS document is in the final stages which will address the use of domestic microwaves in the lab. best regards, Steven Slap Microwave Consultant At 5:47 PM +0100 4/14/04, Edmondson David (RBV) NHS Christie Tr wrote: >Hi Daryl, >Just to say that we have been using domestic microwaves for years, >replacement is relatively simple and inexpensive. Matsui 650 watt and Sharp >650 watt were fine but now a Sharp 800 watt. >We have always gone for the digital/numerical keypad versions, so entering >the times has been staightforward. >So are the lab variety any different behind the label? >Dave >Christie Hosp >Manchester UK From mprice26 <@t> juno.com Fri Apr 16 08:24:18 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Parts for Leitz 1512 Message-ID: <20040416.062425.564.44211@webmail28.nyc.untd.com> Does anyone know where I can order parts for the Leitz 1512 Microtome? I need a blade holder. Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From Jude.Carpenter <@t> vtmednet.org Fri Apr 16 08:45:29 2004 From: Jude.Carpenter <@t> vtmednet.org (Carpenter, Judith A.) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] wrist injury Message-ID: <1AC2A27D616B5C4D94FA103BA7F1A41D040DEBA9@FAHC12.fahc.fletcherallen.org> Good Morning- Does anyone have a source for a device that will open specimen bottles with minimum wrist action ? I have a Path. Asst. who is developing a wrist injury due to opening biopsy specimen containers at grossing. thanks Jude Jude Carpenter, BS, HTL(ASCP) Chief Technologist for Autopsy/Histology/Surgical Pathology 111 Colchester Ave. Burlington, VT 05401 jude.carpenter@vtmednet.org (802)847-5116 fax: (802)847-3509 Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. From psanquin <@t> lugo.usc.es Fri Apr 16 08:51:30 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Agar blocks Message-ID: <3.0.6.32.20040416155130.007d45d0@pop.lugo.usc.es> Dear histonetters, I need your help. I have several paraformaldehyde fixed brain samples (postnatal mice) to section in the vibratome. I am afraid I will not have time enough to cut them today. Could you tell me the best way to keep this agar blocks until Monday? By the way, is there any chance to undo agar-agar blocks without damaging the tissue? Thanks a lot Pablo Sanchez From JWEEMS <@t> sjha.org Fri Apr 16 08:55:24 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] wrist injury Message-ID: How about one of those under the counter openers sold in kitchen supplies. I don't know for sure, but I bet Bed, Bath, and Beyond would have them. I've seen them in catalogs. One would still have to turn the container, but maybe with alternating methods, there would be less injury. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Carpenter, Judith A. Sent: Friday, April 16, 2004 9:45 AM To: NSH Subject: [Histonet] wrist injury Good Morning- Does anyone have a source for a device that will open specimen bottles with minimum wrist action ? I have a Path. Asst. who is developing a wrist injury due to opening biopsy specimen containers at grossing. thanks Jude Jude Carpenter, BS, HTL(ASCP) Chief Technologist for Autopsy/Histology/Surgical Pathology 111 Colchester Ave. Burlington, VT 05401 jude.carpenter@vtmednet.org (802)847-5116 fax: (802)847-3509 Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From akwilliams75 <@t> hotmail.com Fri Apr 16 09:33:24 2004 From: akwilliams75 <@t> hotmail.com (kevin williams) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing and Derm Shaves Message-ID: Is anyone out there doing any processing on derm shaves with microwave processing? How is it and how are the results? Thanks. A. Kevin Williams Vermont Dermatopathology. _________________________________________________________________ [1]Get rid of annoying pop-up ads with the new MSN Toolbar FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS= From gcallis <@t> montana.edu Fri Apr 16 09:57:30 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Jar opening devices In-Reply-To: <1AC2A27D616B5C4D94FA103BA7F1A41D040DEBA9@FAHC12.fahc.fletc herallen.org> Message-ID: <3.0.6.32.20040416085730.00be1580@gemini.msu.montana.edu> Flip top or screw top? Specialty cooking stores/hardware stores have devices to open screw tops on jars used in kitchens. look like gigantic pliers. Check out Williams Sonoma, Pottery Barn, or similar store in your area. One can also wrap a rubber glove around top of screw top, to get a better grip but I don't think this reduces the pain from gripping. We have large adjustable pliers (they expand) and then old rubber glove around top, grab and torque. We do this to open EMD's xylene bottles, with lids that, I swear, are put on with superglue. Flip tops are another problem?? Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Terry.Marshall <@t> rothgen.nhs.uk Fri Apr 16 09:58:50 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing Message-ID: Sorry to but in - but on a related question - does anyone do microwave fixation in saline as described by for instance the Melbourne group, which involves bringing up to 65C in saline? Have tried it this week and the sections are even crummier than usual. When I was in Tasmania, a local private lab. did it and their sections were fine. Dr Terry L (why can't I get a good H&E) 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: kevin williams [mailto:akwilliams75@hotmail.com] Sent: 16 April 2004 15:33 To: histonet@pathology.swmed.edu Subject: [Histonet] Microwave Processing and Derm Shaves Is anyone out there doing any processing on derm shaves with microwave processing? How is it and how are the results? Thanks. A. Kevin Williams Vermont Dermatopathology. _________________________________________________________________ [1]Get rid of annoying pop-up ads with the new MSN Toolbar FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS= _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ALAbeyta <@t> salud.unm.edu Fri Apr 16 10:09:18 2004 From: ALAbeyta <@t> salud.unm.edu (Antonia Abeyta) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] cutting frozen rat turbs Message-ID: Has anyone out there cut frozen rat turbinates?? We are having trouble keeping them on the slides. Any pointers? Thanks, Antonia From histology <@t> amccares.org Fri Apr 16 10:24:58 2004 From: histology <@t> amccares.org (Histology Department) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Jar opening devices[Scanned for Viruses] Message-ID: <4F6BE54A16B5C047B2C959741629282943D099@mail.amccares.org> Jude, "OXO" the maker of the "Good Grips" line of utensils makes two styles of openers. One goes for ~$6 and the other goes for ~$12(US). They both are designed on a wedge shape that you slide over the top of the jar. Inside the wedge there are angled teeth along one edge and a metal pressure bar along the opposite side. The handle is a fairly large, oval, rubber grip. The more expensive model employs a base that can stabilize the jar on the counter. I don't use these in the grossing room, but I use them on everything else in Histology. They work really well and with little effort. At home, they are great for popping the tops off my parents home-canned pickles too. You can find them at: Bed, Bath & Beyond; Linens and Things; and most quality hardward and department stores. Good luck. Judy LaDuc, BS HTL (ASCP) Adirondack Medical Center Saranac Lake, NY From akwilliams75 <@t> hotmail.com Fri Apr 16 10:14:05 2004 From: akwilliams75 <@t> hotmail.com (kevin williams) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Linistainers and the odd aberant staining slide Message-ID: Dear All: I Have a Linistainer, I get nice staining, but evey now and then I get real bad staining on only one slide. Why? It is not the microtomy, the staining is bad all over the place, not patchy. The sections look as though they have missed the haematoxylin. Has anyone had the same problem and any intresting insights, something to placate the Pathologist. Yours faithfully, A.Kevin Williams Vermont Dermatopathology _________________________________________________________________ [1]Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. References 1. http://g.msn.com/8HMBENUS/2755??PS= From bryand <@t> netbistro.com Fri Apr 16 10:41:07 2004 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing References: Message-ID: <004101c423c9$3bd1f270$99a5b5d0@yourlk4rlmsu41> I haven't used microwave fixation, but I do remember frozen section heat fixation when I first started learning histology. Perhaps saline at 65C is a modification of this? We brought saline (or water) to the boil, took it off the heat and dropped a thin piece of fresh tissue into it for one minute. The tissue was then frozen on a clinical microtome with CO2 or a Peltier cooler and sectioned. The morphology and staining was fine, except that eosin was pinker than expected. It is important not to leave the tissue too long in the hot saline or it cooked and it was ruined. One minute was about the maximum. I've also tried fixing fresh tissue, in cassettes, in NBF at 65C in an oven for an hour. That works quite well too, at least as far as morphology and ordinary dye staining is concerned. Of course, a well sealed container is strongly advised. Bryan Llewellyn ----- Original Message ----- From: "Marshall Terry Dr,Consultant Histopathologist" To: "kevin williams" ; Sent: Friday, April 16, 2004 7:58 AM Subject: RE: [Histonet] Microwave Processing Sorry to but in - but on a related question - does anyone do microwave fixation in saline as described by for instance the Melbourne group, which involves bringing up to 65C in saline? Have tried it this week and the sections are even crummier than usual. When I was in Tasmania, a local private lab. did it and their sections were fine. Dr Terry L (why can't I get a good H&E) 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: kevin williams [mailto:akwilliams75@hotmail.com] Sent: 16 April 2004 15:33 To: histonet@pathology.swmed.edu Subject: [Histonet] Microwave Processing and Derm Shaves Is anyone out there doing any processing on derm shaves with microwave processing? How is it and how are the results? Thanks. A. Kevin Williams Vermont Dermatopathology. _________________________________________________________________ [1]Get rid of annoying pop-up ads with the new MSN Toolbar FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS= _______________________________________________ Histonet mailing list Histonet@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 juan.gutierrez <@t> christushealth.org Fri Apr 16 10:44:30 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Linistainers and the odd aberant staining slide Message-ID: Is the Linistainer the one with the bicycle chain on it? If it is, try to leave at least one space between the clips and also check to make sure the containers are pushed all the way down. Good luck, Juan -----Original Message----- From: kevin williams [mailto:akwilliams75@hotmail.com] Sent: Friday, April 16, 2004 10:14 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Linistainers and the odd aberant staining slide Dear All: I Have a Linistainer, I get nice staining, but evey now and then I get real bad staining on only one slide. Why? It is not the microtomy, the staining is bad all over the place, not patchy. The sections look as though they have missed the haematoxylin. Has anyone had the same problem and any intresting insights, something to placate the Pathologist. Yours faithfully, A.Kevin Williams Vermont Dermatopathology _________________________________________________________________ [1]Is your PC infected? Get a FREE online computer virus scan from McAfee? Security. References 1. http://g.msn.com/8HMBENUS/2755??PS= _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Fri Apr 16 10:47:52 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Not but but butt Message-ID: Aaargh. Should have been "butt in" - minus 2 Brownie points Marshall. 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: Bryan Llewellyn [mailto:bryand@netbistro.com] Sent: 16 April 2004 16:41 To: Histonet Subject: Re: [Histonet] Microwave Processing I haven't used microwave fixation, but I do remember frozen section heat fixation when I first started learning histology. Perhaps saline at 65C is a modification of this? We brought saline (or water) to the boil, took it off the heat and dropped a thin piece of fresh tissue into it for one minute. The tissue was then frozen on a clinical microtome with CO2 or a Peltier cooler and sectioned. The morphology and staining was fine, except that eosin was pinker than expected. It is important not to leave the tissue too long in the hot saline or it cooked and it was ruined. One minute was about the maximum. I've also tried fixing fresh tissue, in cassettes, in NBF at 65C in an oven for an hour. That works quite well too, at least as far as morphology and ordinary dye staining is concerned. Of course, a well sealed container is strongly advised. Bryan Llewellyn ----- Original Message ----- From: "Marshall Terry Dr,Consultant Histopathologist" To: "kevin williams" ; Sent: Friday, April 16, 2004 7:58 AM Subject: RE: [Histonet] Microwave Processing Sorry to but in - but on a related question - does anyone do microwave fixation in saline as described by for instance the Melbourne group, which involves bringing up to 65C in saline? Have tried it this week and the sections are even crummier than usual. When I was in Tasmania, a local private lab. did it and their sections were fine. Dr Terry L (why can't I get a good H&E) 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: kevin williams [mailto:akwilliams75@hotmail.com] Sent: 16 April 2004 15:33 To: histonet@pathology.swmed.edu Subject: [Histonet] Microwave Processing and Derm Shaves Is anyone out there doing any processing on derm shaves with microwave processing? How is it and how are the results? Thanks. A. Kevin Williams Vermont Dermatopathology. _________________________________________________________________ [1]Get rid of annoying pop-up ads with the new MSN Toolbar FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS= _______________________________________________ Histonet mailing list Histonet@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 kgrobert <@t> rci.rutgers.edu Fri Apr 16 11:03:11 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Re: wrist injury In-Reply-To: <004101c423c9$3bd1f270$99a5b5d0@yourlk4rlmsu41> References: <004101c423c9$3bd1f270$99a5b5d0@yourlk4rlmsu41> Message-ID: <408003BF.8060907@rci.rutgers.edu> Hmmm, how about this little gizmo from Black and Decker? The only question is, how big are the jars the tech needs to open and would they fit on here? Only one way to find out, I guess. http://www.youcansave.com/lidsoff.asp Good luck, and I hope your tech feels better! Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology and Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854 From kemlo <@t> tiscali.co.uk Fri Apr 16 11:05:44 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing In-Reply-To: Message-ID: <4071A29D0001D7EB@mk-cpfrontend-4.mail.uk.tiscali.com> Terry, you could never get a good H&E! Have you tried joining an EQA group just to see if it's you or them? Maybe you could put the sections through SurePath's stainer, they seem to have a nice haematoxylin and it uses stoichiometric staining. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From kemlo <@t> tiscali.co.uk Fri Apr 16 11:05:44 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing In-Reply-To: Message-ID: <4071A29D0001D7EC@mk-cpfrontend-4.mail.uk.tiscali.com> Terry, you could never get a good H&E! Have you tried joining an EQA group just to see if it's you or them? Maybe you could put the sections through SurePath's stainer, they seem to have a nice haematoxylin and it uses stoichiometric staining. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From mari.ann.mailhiot <@t> leica-microsystems.com Fri Apr 16 11:24:12 2004 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Parts for Leitz 1512 Message-ID: Marsha Give me a call at Leica and let me know what parts you need. There is a possiblity some parts may not be available. 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 mprice26@juno.com Sent by: To: histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] Parts for Leitz 1512 04/16/2004 08:24 AM Does anyone know where I can order parts for the Leitz 1512 Microtome? I need a blade holder. Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From Terry.Marshall <@t> rothgen.nhs.uk Fri Apr 16 11:36:10 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing Message-ID: Not true. Yours were excellent. 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: Kemlo [mailto:kemlo@tiscali.co.uk] Sent: 16 April 2004 17:06 To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Terry, you could never get a good H&E! Have you tried joining an EQA group just to see if it's you or them? Maybe you could put the sections through SurePath's stainer, they seem to have a nice haematoxylin and it uses stoichiometric staining. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From siksik03 <@t> comcast.net Fri Apr 16 11:41:47 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave tissue processors In-Reply-To: References: Message-ID: Dear HistoNetters There are considerable differences between the models of microwave tissue processors available. I would look at these features: ? How is the temperature controlled? (remote sensor vs. temperature probe?) ? How is temperature equilibrated within the solution? (magnetic stirring vs. air bubble agitation?) ? Was the unit built for the lab or is it a converted domestic microwave? ? How thorough is the User's Manual regarding procedures? ? How simple is the user's interface? How easy is it to make a serious error that would compromise the tissue? ? Can you observe the process in real time? ? Are accessories available from the manufacturer for various specific applications? Any manufacturer should be willing to provide a demonstration. Feel free to contact me directly with any specific questions. best regards, Steven Slap Microwave Consultant At 1:36 PM +0800 4/16/04, Chan Wai Kam wrote: >I'm confused about the different types of microwave tissue processors >from Milestone and Pelco and need some advice from those of you who have >these. Any pros and cons from current users would be greatly >appreciated. I'm considering getting one in the near future. Our usage >is mainly on processing bone specimens From Xudong_Cao <@t> brown.edu Fri Apr 16 11:55:03 2004 From: Xudong_Cao <@t> brown.edu (Xudong_Cao@brown.edu) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] re:PGP9.5 antibody against chicken Message-ID: <200404161655.i3GGt3B1020136@ursa.services.brown.edu> Carl, from what I figured from the literature, PGP9.5 is extensively used for study on epidermal-neuronal interations, which is what my study is essentially concentrated on. Some of the published papers suggest that PGP9.5 is more sensitive and stains all the nerve fibers (pan-neuromaker). As for neurofilament Ab, there are heavy and light chains that stain for different nerve fibers. However, I did see report using GAP-43 antibody for the same purpose as I mentioned above in epidermal-neuronal studies (not sure if it is sensitive enough to pick up the bare nerve endings in the epidermis though. As for my posting, PGP9.5 protocol works for me and it would ideal just find an antibody that works for chicken. Xudong Message: 6 Date: Thu, 15 Apr 2004 19:44:40 +0100 From: "Carl" Subject: [Histonet] re PGP9.5 antibody against chicken To: Message-ID: <001201c42319$b5ed2a30$dd039a51@home> Content-Type: text/plain; charset="iso-8859-1" For my interest, why PGP9.5? If it's neurones and/or their processes, surely , for example the anti neurofilament, beta tubulin III and neuronal cell body( eg Hu C/D) Ab reagents are sufficient? From jcline <@t> wchsys.org Fri Apr 16 11:55:20 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Leica 1512 Message-ID: <008401c423d3$9e697700$1d2a14ac@wchsys.org> Hello Marsha, I have a regular steel knife holder and a diposable holder. I have a couple of 1512's that I don't use. Contact me at jcline@wchsys.org. ***** 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 jcline <@t> wchsys.org Fri Apr 16 12:00:00 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] derm shaves Message-ID: <009101c423d4$41839d80$1d2a14ac@wchsys.org> Hello A. Kevin Williams, I have processed skin shaves in the past, but one was a possible melanoma and our paths decided not to repeat this again. (we had one ruined because not enough additional time was added to the program to cover a larger skin shaving) ***** 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 LBlack <@t> carilion.com Fri Apr 16 12:21:39 2004 From: LBlack <@t> carilion.com (Lisa Black) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Jar opening devices[Scanned for Viruses] Message-ID: Black & Decker has an electronic jar opener. It costs $50. I found them at Wal-Mart, Sears, and Target. From gcallis <@t> montana.edu Fri Apr 16 13:14:40 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] cutting frozen rat turbs,, a reply In-Reply-To: Message-ID: <3.0.6.32.20040416121440.00be1580@gemini.msu.montana.edu> I am presuming you mean UNDECALCIFIED rat turbinates? We do mouse undecalcified turbinates, but need to sue the tape transfer system, Cryojane from Instrumedics OR you crush delicate turbinate bone, do not get a flat section that adheres to slide surface hence falling off. It requires a d profile (some like c profile) tungsten carbide knife in extremely sharp condition in order to cut decently through bone. Without these tools, we get terrible sections. At 09:09 AM 4/16/2004 -0600, you wrote: >Has anyone out there cut frozen rat turbinates?? We are having trouble >keeping them on the slides. Any pointers? > >Thanks, >Antonia > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Fri Apr 16 13:16:25 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Jar opening devices[Scanned for Viruses] In-Reply-To: <4F6BE54A16B5C047B2C959741629282943D099@mail.amccares.org> Message-ID: <3.0.6.32.20040416121625.00be1580@gemini.msu.montana.edu> Lovely thing about these Good Grips tools, they are designed with ergonomics in mind, for arthritic hands. I have seen them at Target also. Gayle Callis From mrsgbd2001 <@t> yahoo.com Fri Apr 16 13:17:07 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] cutting frozen rat turbs In-Reply-To: Message-ID: <20040416181707.65811.qmail@web13305.mail.yahoo.com> I have been cutting frozen rat turbinates for a couple weeks. I have not actually stained them yet, but plan to next week. I was told by my PI to pick up the sections on charged slides and then allow them to dry overnight at room temp. Then the next morning they are put in -80 freezer until ready to stain. She has had luck with this method, but I have not actually tried to stain them, as I already said. Anyway, maybe it the way you cryopreserved your tissue. I let you know how my stains go. Gareth Davis Research Assistant Tennessee State University Antonia Abeyta wrote: Has anyone out there cut frozen rat turbinates?? We are having trouble keeping them on the slides. Any pointers? Thanks, Antonia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Tax Center - File online by April 15th From portera203 <@t> yahoo.com Fri Apr 16 13:24:40 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Parts for Reichert-Jung 2030 Microtome Message-ID: <20040416182440.22507.qmail@web40912.mail.yahoo.com> Our lab is looking for a used/refurbished low profile disposable blade holder for a Reichert Jung 2030 Rotary Microtome. If any one out there knows where we could obtain one your assistance would be greatly appreciated. I have already checked with Marianne at Leica, she informed me that they no longer make or carry this part. We actually only need the top half that is tan with a pressure plate on the front with two screws. If you need a photo to help you out i have one and could send it to your direct address to help in identification. Thanks in advance to all for any help. Amy S.Porter, HT(ASCP) Michigan State University Department of Physiology Division of Human Pathology College of Human Medicine portera203@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Tax Center - File online by April 15th From yangpw <@t> umich.edu Fri Apr 16 13:53:18 2004 From: yangpw <@t> umich.edu (yangpw@umich.edu) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] mouse vs. rat in situ; overfixed vs. poorfixed Message-ID: <1082141598.40802b9e7a10f@mail.umich.edu> Dear All, Recently I have tried radioactive in situ hybridization on mouse brain tissue with mouse-cfos riboprobe. I found that with same in situ protocol, the signal of mouse is lighter than the signal of rat-cfos on rat tissue (to reach the similar intensity, I need to lay down the film on mouse tissue longer time as twice much as on rat tissue.) I am not sure if it is normal for mouse tissue get lighter signal or the rat in situ protocol just is not optimal for mouse. Any suggestion? If I need to modify my rat protocol for mouse, what aspect should I change? The second question is kind of interesting. Still about in situ, I tried two in situ on mouse brain tissue (the tissues are two copies from same animals, and I use same in situ protocol). The staining solutions are all the same except the 4% PFA I used to fix the tissue. In the first staining I used a three-month old PFA, and in the second staining I made fresh PFA. The signal of second staining is still kind of light but much more intense than the signal from first staining. My conclusion is in the first staining the tissue was poor fixed because stale PFA was degraded. So to improve the staining, I will try fresh PFA with longer fixing time. But one of my collegue has a exactly contrary opinion. He thinks in the first staining, the tissue was over fixed becuase the old PFA became more concentrated. So he suggest using fresh PFA and shorter fixing time. By the way, the PFA was stored in a caped container in fridge. Well, same fact but totally different viewpoints. Any comment? THanks. Pengwei Yang Biopsychology Program University of Michigan From carl.hobbs <@t> kcl.ac.uk Fri Apr 16 14:05:34 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] re:PGP9.5 antibody against chicken Message-ID: <000e01c423e5$cbbb4ff0$c4039a51@home> Thanks,Xudong. Interesting point re sensitivity. I will check whether my PGP9.5 Ab reagent works on chick next week. Yes re ati NFs, your right for that caveat. However, personally I check a few other Abs , to ensure that I am picking up everything eg partic anti beta tubulin III. The GAP 43 Ab reagents I have work beautifully on chick. All require Ag heat retrieval, in my hands Thanks again Carl From pruegg <@t> colobio.com Fri Apr 16 14:10:56 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] QIHC exam Message-ID: Folks, I am in the process of updating my presentation on "Taking the QIHC Exam" and there are a couple of new projects for 2004 that I need to demonstrat by microphotographs and do not have. If you would be willing to share your digital images of tumors stained for CK7 and CK20 and anything including muscle stained for MSA I would really appreciate it. I will of course credit any use of others microphotographs. Thank you all for considering this request. Patsy From cfavara <@t> niaid.nih.gov Fri Apr 16 14:16:16 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] mouse vs. rat in situ; overfixed vs. poorfixed Message-ID: I had one researcher that did a lot of in-situ with chromagens. We fix in NBF and the conclusion was the longer the fixation time the better the signal. Oh yes this was in mice. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: yangpw@umich.edu [mailto:yangpw@umich.edu] Sent: Friday, April 16, 2004 12:53 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] mouse vs. rat in situ; overfixed vs. poorfixed Dear All, Recently I have tried radioactive in situ hybridization on mouse brain tissue with mouse-cfos riboprobe. I found that with same in situ protocol, the signal of mouse is lighter than the signal of rat-cfos on rat tissue (to reach the similar intensity, I need to lay down the film on mouse tissue longer time as twice much as on rat tissue.) I am not sure if it is normal for mouse tissue get lighter signal or the rat in situ protocol just is not optimal for mouse. Any suggestion? If I need to modify my rat protocol for mouse, what aspect should I change? The second question is kind of interesting. Still about in situ, I tried two in situ on mouse brain tissue (the tissues are two copies from same animals, and I use same in situ protocol). The staining solutions are all the same except the 4% PFA I used to fix the tissue. In the first staining I used a three-month old PFA, and in the second staining I made fresh PFA. The signal of second staining is still kind of light but much more intense than the signal from first staining. My conclusion is in the first staining the tissue was poor fixed because stale PFA was degraded. So to improve the staining, I will try fresh PFA with longer fixing time. But one of my collegue has a exactly contrary opinion. He thinks in the first staining, the tissue was over fixed becuase the old PFA became more concentrated. So he suggest using fresh PFA and shorter fixing time. By the way, the PFA was stored in a caped container in fridge. Well, same fact but totally different viewpoints. Any comment? THanks. Pengwei Yang Biopsychology Program University of Michigan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Fri Apr 16 14:31:20 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Micowave tissue processors Message-ID: It is commonly believed that the value of a microwave, and its biggest drawback, is the heat. Heat makes every reaction run faster. Microwaves heat. Microwaves facilitate reactions. Qed. The drawback part is that the heat is unevenly distributed, changes rapidly, and too much heat destroys the tissue. The Pelco makes other claims, and offers research support. They use a water pad to absorb the microwaves, and circulate it to keep the inside and the tissue cool. A microwave without heat! What could be the point? Dr. Day has divided tissue samples, run immunostains part the traditional way, part a much shorter protocol using the "cold" microwave, and part the short protocol, but without the microwave. Reaction in the microwave progressed beyond even the traditional controls, excellent staining was seen, with the "cold" microwave, but not the sample treated identically except no microwaving. One idea would be that kicking the molecule once, in a way that would build up heat, but heat is not allowed to build up, is something like a micro stirrer. Anyway, solid evidence shows it works. Other advantages are you never have overheated tissue, and can leave the microwave on coninuously while the tissue is in without fear of overheating. Microwave-assisted Immunostaining: A New Approach Yields Fast and Consistent Results Teresa Elena Mu?oz, Richard T. Giberson*, Richard Demaree, Jonathan R. Day Department of Biological Sciences, California State University, Chico, Chico CA 95929-0515, USA In Press or published in Neuroscience Methods, 2004. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=472001&catdesc=Histology+Equipment&CatThreeID=714&CatOneID=4&subcatdesc=Microwave+Reaction+Facilitation&idsubcategory=204 Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Chan Wai Kam Sent: Friday, April 16, 2004 12:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Micowave tissue processors Hi Histonetters, I'm confused about the different types of microwave tissue processors from Milestone and Pelco and need some advice from those of you who have these. Any pros and cons from current users would be greatly appreciated. I'm considering getting one in the near future. Our usage is mainly on processing bone specimens. Thanks in advance Julee Chan Musculoskeletal Tissue Lab National University of Singapore _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Fri Apr 16 14:32:06 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] re Best Marker for Proliferation? Message-ID: <001d01c423e9$80f72850$c4039a51@home> IMHO.....agreed Ki67( tho be careful of the Ab reagent you use - I was informed of this by Jorge Luogo and he referred me to Catoretti's Ab site for info). I would steer clear of PCNA. BrdU incorp. , if you can , is invaluable. Anti phosph H3( cells in mitosis) also invaluable . From carl.hobbs <@t> kcl.ac.uk Fri Apr 16 14:33:01 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Fw: re:PGP9.5 antibody against chicken Message-ID: <002301c423e9$a19316a0$c4039a51@home> Resending in case of HTML setting defaulting. Apologies ----- Original Message ----- From: Carl To: histonet@lists.utsouthwestern.edu Sent: Friday, April 16, 2004 8:05 PM Subject: re:PGP9.5 antibody against chicken Thanks,Xudong. Interesting point re sensitivity. I will check whether my PGP9.5 Ab reagent works on chick next week. Yes re ati NFs, your right for that caveat. However, personally I check a few other Abs , to ensure that I am picking up everything eg partic anti beta tubulin III. The GAP 43 Ab reagents I have work beautifully on chick. All require Ag heat retrieval, in my hands Thanks again Carl From pruegg <@t> colobio.com Fri Apr 16 14:24:26 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] QIHC exam In-Reply-To: Message-ID: Gary just corrected my wordage (is wordage a word?), I guess what I really need are photomicrographs, not microphotographs. I beg your pardon. Patsy Microphotographs are small pictures of something macroscopic; photomicrographs are large pictures of something microscopic. Gary Gill -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Friday, April 16, 2004 1:11 PM To: Histonet@Pathology. Swmed. Edu Cc: Ihcrg@Yahoogroups. Com Subject: [Histonet] QIHC exam Folks, I am in the process of updating my presentation on "Taking the QIHC Exam" and there are a couple of new projects for 2004 that I need to demonstrat by microphotographs and do not have. If you would be willing to share your digital images of tumors stained for CK7 and CK20 and anything including muscle stained for MSA I would really appreciate it. I will of course credit any use of others microphotographs. Thank you all for considering this request. Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Fri Apr 16 15:39:31 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing In-Reply-To: <004101c423c9$3bd1f270$99a5b5d0@yourlk4rlmsu41> Message-ID: <000001c423f2$ec219c60$4a737b81@Cygnus> Has anyone read the latest issue of the NSH's In Action??? Front page story deals with this issue of microwaves domestic vs lab. The NCCLS has just come out with some guidelines dealing with this. I suggest a little visit to http://www.nccls.org and check out these guidelines. for what it's worth.... Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 "Where am I going and why am I in the handbasket???" -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: Friday, April 16, 2004 8:41 AM To: Histonet Subject: Re: [Histonet] Microwave Processing I haven't used microwave fixation, but I do remember frozen section heat fixation when I first started learning histology. Perhaps saline at 65C is a modification of this? We brought saline (or water) to the boil, took it off the heat and dropped a thin piece of fresh tissue into it for one minute. The tissue was then frozen on a clinical microtome with CO2 or a Peltier cooler and sectioned. The morphology and staining was fine, except that eosin was pinker than expected. It is important not to leave the tissue too long in the hot saline or it cooked and it was ruined. One minute was about the maximum. I've also tried fixing fresh tissue, in cassettes, in NBF at 65C in an oven for an hour. That works quite well too, at least as far as morphology and ordinary dye staining is concerned. Of course, a well sealed container is strongly advised. Bryan Llewellyn ----- Original Message ----- From: "Marshall Terry Dr,Consultant Histopathologist" To: "kevin williams" ; Sent: Friday, April 16, 2004 7:58 AM Subject: RE: [Histonet] Microwave Processing Sorry to but in - but on a related question - does anyone do microwave fixation in saline as described by for instance the Melbourne group, which involves bringing up to 65C in saline? Have tried it this week and the sections are even crummier than usual. When I was in Tasmania, a local private lab. did it and their sections were fine. Dr Terry L (why can't I get a good H&E) 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: kevin williams [mailto:akwilliams75@hotmail.com] Sent: 16 April 2004 15:33 To: histonet@pathology.swmed.edu Subject: [Histonet] Microwave Processing and Derm Shaves Is anyone out there doing any processing on derm shaves with microwave processing? How is it and how are the results? Thanks. A. Kevin Williams Vermont Dermatopathology. _________________________________________________________________ [1]Get rid of annoying pop-up ads with the new MSN Toolbar FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS= _______________________________________________ Histonet mailing list Histonet@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 cbryan <@t> seattlecca.org Fri Apr 16 16:40:04 2004 From: cbryan <@t> seattlecca.org (Bryan, Clara) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] EBBER Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948782AC2@ldap.seattlecca.org> The company that we ordered the EBER probe no longer exist. Does anyone have a source for this probe? I will like prompt response and some information as to were to ordered from. Thank you. Clara Bryan Pathology Department Seattle Cancer Care Alliance 206-288-7083 206-288-1355 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From sebres <@t> comcast.net Fri Apr 16 17:08:11 2004 From: sebres <@t> comcast.net (sebres) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] mouse vs. rat in situ; overfixed vs. poorfixed References: <1082141598.40802b9e7a10f@mail.umich.edu> Message-ID: <000401c423ff$4fcc2cb0$0300a8c0@RESLAPTOP> 1) A number of factors could cause the actual abundance of this mRNA in your tissues to be lower in the mouse, under your conditions, even if the probe works comparably well across species (and typically the sequence is pretty close between rat & mouse), e.g. age, stress, experience. 2) Fixation should improve the anatomical resolution of your signal, but I wouldn't think less fixation would reduce the signal. Some protocols call for eliminating fixation altogether. But high concentration of fixative will definitely interfere with the penetration of probe into the tissue, especially for ribo's. So I suspect that your colleague is on target. Of course the proof will be empirical comparison! Are you using PFA/PBS? Good luck! (BTW, I'm an alum of the UM biopsych program [back in the dark ages when we used to be called psychobio]--please convey my regards!) Susan Bachus, Psychology Dept., George Mason University ----- Original Message ----- From: To: Sent: Friday, April 16, 2004 2:53 PM Subject: [Histonet] mouse vs. rat in situ; overfixed vs. poorfixed > Dear All, > > Recently I have tried radioactive in situ hybridization on mouse brain tissue > with mouse-cfos riboprobe. I found that with same in situ protocol, the signal > of mouse is lighter than the signal of rat-cfos on rat tissue (to reach the > similar intensity, I need to lay down the film on mouse tissue longer time as > twice much as on rat tissue.) I am not sure if it is normal for mouse tissue > get lighter signal or the rat in situ protocol just is not optimal for mouse. > Any suggestion? If I need to modify my rat protocol for mouse, what aspect > should I change? > > The second question is kind of interesting. Still about in situ, I tried two in > situ on mouse brain tissue (the tissues are two copies from same animals, and I > use same in situ protocol). The staining solutions are all the same except the > 4% PFA I used to fix the tissue. In the first staining I used a three-month old > PFA, and in the second staining I made fresh PFA. The signal of second staining > is still kind of light but much more intense than the signal from first > staining. My conclusion is in the first staining the tissue was poor fixed > because stale PFA was degraded. So to improve the staining, I will try fresh > PFA with longer fixing time. But one of my collegue has a exactly contrary > opinion. He thinks in the first staining, the tissue was over fixed becuase the > old PFA became more concentrated. So he suggest using fresh PFA and shorter > fixing time. By the way, the PFA was stored in a caped container in fridge. > Well, same fact but totally different viewpoints. Any comment? > > THanks. > > Pengwei Yang > > Biopsychology Program > University of Michigan > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jincan <@t> itsa.ucsf.edu Sat Apr 17 00:04:43 2004 From: jincan <@t> itsa.ucsf.edu (jincan) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] mouse vs. rat in situ; overfixed vs. poorfixed Message-ID: <5.1.1.6.0.20040416220307.00b31280@itsa.ucsf.edu> Dear Dr. Yang, I am not familiar with differences of ISH on mouse and rat tissues. However, on old vs fresh paraformaldehyde, the old one has more power of cross link, ie more likely to cause over-fixation. Because, upon storage, PFD condenses to poly PFD, a chemical reaction occur easily at neutral pH at 4oC. Best regards, James Guo At 02:53 PM 4/16/2004 -0400, you wrote: >Dear All, > >Recently I have tried radioactive in situ hybridization on mouse brain tissue >with mouse-cfos riboprobe. I found that with same in situ protocol, the >signal >of mouse is lighter than the signal of rat-cfos on rat tissue (to reach the >similar intensity, I need to lay down the film on mouse tissue longer time as >twice much as on rat tissue.) I am not sure if it is normal for mouse tissue >get lighter signal or the rat in situ protocol just is not optimal for mouse. >Any suggestion? If I need to modify my rat protocol for mouse, what aspect >should I change? > >The second question is kind of interesting. Still about in situ, I tried >two in >situ on mouse brain tissue (the tissues are two copies from same animals, >and I >use same in situ protocol). The staining solutions are all the same except the >4% PFA I used to fix the tissue. In the first staining I used a >three-month old >PFA, and in the second staining I made fresh PFA. The signal of second >staining >is still kind of light but much more intense than the signal from first >staining. My conclusion is in the first staining the tissue was poor fixed >because stale PFA was degraded. So to improve the staining, I will try fresh >PFA with longer fixing time. But one of my collegue has a exactly contrary >opinion. He thinks in the first staining, the tissue was over fixed >becuase the >old PFA became more concentrated. So he suggest using fresh PFA and shorter >fixing time. By the way, the PFA was stored in a caped container in fridge. >Well, same fact but totally different viewpoints. Any comment? > >THanks. > >Pengwei Yang > >Biopsychology Program >University of Michigan > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Edmondson <@t> christie-tr.nwest.nhs.uk Sat Apr 17 06:36:17 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] EBBER Message-ID: Hi Clara, We have been using a DAKO EBER probe, though it's a PNA probe. Appears to be fine but we have never compared. Dave Christie Hosp Manchester UK -----Original Message----- From: Bryan, Clara [mailto:cbryan@seattlecca.org] Sent: 16 April 2004 22:40 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] EBBER The company that we ordered the EBER probe no longer exist. Does anyone have a source for this probe? I will like prompt response and some information as to were to ordered from. Thank you. Clara Bryan Pathology Department Seattle Cancer Care Alliance 206-288-7083 206-288-1355 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Edmondson <@t> christie-tr.nwest.nhs.uk Sat Apr 17 06:42:49 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Linistainers and the odd aberant staining slide Message-ID: Kevin, When our clips detatch, partially, then sometimes they cause chaos on the machine with slides everywhere and sometimes they just come to the end looking horrid. Those mounting will pop them in acid alcohol before setting them back in the chain at Haematoxylin stage. If the clip looks past its best it is ditched. Bother the cost. Dave Christie Manchester UK -----Original Message----- From: kevin williams [mailto:akwilliams75@hotmail.com] Sent: 16 April 2004 16:14 To: histonet@pathology.swmed.edu Subject: [Histonet] Linistainers and the odd aberant staining slide Dear All: I Have a Linistainer, I get nice staining, but evey now and then I get real bad staining on only one slide. Why? It is not the microtomy, the staining is bad all over the place, not patchy. The sections look as though they have missed the haematoxylin. Has anyone had the same problem and any intresting insights, something to placate the Pathologist. Yours faithfully, A.Kevin Williams Vermont Dermatopathology _________________________________________________________________ [1]Is your PC infected? Get a FREE online computer virus scan from McAfee? Security. References 1. http://g.msn.com/8HMBENUS/2755??PS= _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From k.whalley <@t> ich.ucl.ac.uk Sat Apr 17 07:34:56 2004 From: k.whalley <@t> ich.ucl.ac.uk (Katy Whalley) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] tissue slicer Message-ID: <4823.194.82.240.48.1082205296.squirrel@jenner.ich.ucl.ac.uk> Hi, We are looking for a device which can be used to cut tissue quickly into slices of an even thickness. We're not sure yet exactly how thick these will be but something in the range 30-300 microns is likely. My supervisor has in mind something in which several blades are attached to a holder that keeps them the correct distance apart, so that all the slices are cut at once. Has anyone ever used/ seen this kind of thing, or anything else which would do the job? thanks, Katy, UCL From dlcowie <@t> prodigy.net Sat Apr 17 09:11:53 2004 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] pretreat for her2neu Message-ID: <20040417141153.17114.qmail@web80509.mail.yahoo.com> hello histonetters, I'm hoping someone can help me with this. We are using the ventana benchmark for all our immunos. The benchmark calls for using the cell conditioning 2 as a pretreatment for her2neu. It is very expensive (about 950.00 for 1 litre) It only has a shelf life of 2 months. Our problem is that it is expiring before we can use hardly any of it. We are looking at trying to use cell marques Declere as the de paraffinization and pretreat step off line and then continuing with the protocol on the benchmark from that point on. The Cell condition 2 is a citrate buffer, as is the Declere. My question is, has anyone out there who is using benchmark had any experience of this or of using another alternative? Any help would be appreciated. Dawn Cowie HT (ASCP) Pensacola Path Florida From mrsgbd2001 <@t> yahoo.com Sat Apr 17 11:22:42 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] cutting frozen rat turbs,, a reply In-Reply-To: <3.0.6.32.20040416121440.00be1580@gemini.msu.montana.edu> Message-ID: <20040417162242.77791.qmail@web13306.mail.yahoo.com> Hi Gayle, We decalcified our rat skulls in Regular Cal Immuno for two weeks and then cryopreserved in 10%, 20% and 30% washes of sucrose. Then we washed in OCT:H2O for 45 minutes, while oscillating at 100rpm. Then froze the tissue (on dry ice) and "embedded" in OCT. My cuts have come out beautiful, I even impressed myself. I figured I would get terrible sections through the turbinates, but I don't. Gareth Davis Research Assistant Tennessee State University Gayle Callis wrote: I am presuming you mean UNDECALCIFIED rat turbinates? We do mouse undecalcified turbinates, but need to sue the tape transfer system, Cryojane from Instrumedics OR you crush delicate turbinate bone, do not get a flat section that adheres to slide surface hence falling off. It requires a d profile (some like c profile) tungsten carbide knife in extremely sharp condition in order to cut decently through bone. Without these tools, we get terrible sections. At 09:09 AM 4/16/2004 -0600, you wrote: >Has anyone out there cut frozen rat turbinates?? We are having trouble >keeping them on the slides. Any pointers? > >Thanks, >Antonia > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Tax Center - File online by April 15th From Zorbutt <@t> aol.com Sat Apr 17 12:44:26 2004 From: Zorbutt <@t> aol.com (Zorbutt@aol.com) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] p16 Message-ID: <142.27290a7e.2db2c6fa@aol.com> Hi Has anyone used the p16 INK4a research kit by dako, or does anyone know of a p16 antibody that is available (not sold as a research kit). From KHays <@t> mbhs.org Sat Apr 17 14:59:25 2004 From: KHays <@t> mbhs.org (KHays@mbhs.org) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] need information on leica CM 3050s Message-ID: Kathy Tedford-Hays HT (ASCP) Technical Specialist, Histology Dept (601)-968-3070 ext 7398 Baptist Medical Center 1225 North State Street Jackson, MS 39202 i am looking for positive and negative feedback from users that have the leica CM 3050s cryostat. we are looking into doing a demo in the next week or so and want your expert advice on this machine please. thank you for all of your replies concerning this matter. From epitek <@t> yahoo.com Sat Apr 17 15:19:17 2004 From: epitek <@t> yahoo.com (Elaine Pitek) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] unsubscribe In-Reply-To: Message-ID: <20040417201917.7833.qmail@web41601.mail.yahoo.com> Nick Kirk wrote:Try http://members.pgonline.com/~bryand/StainsFile/ or http://www.nottingham.ac.uk/pathology/default.html or http://medlib.med.utah.edu/WebPath/HISTHTML/MANUALS/MANUALS.html Nick Kirk Head BMS Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sonya Hogg Sent: 22 January 2004 06:40 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Website help I am looking for websites relating to histology special staining techniques and am having difficulty does anyone have any good links?? Thanks http://personals.yahoo.com.au - Yahoo! Personals New people, new possibilities. FREE for a limited time. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Elaine Pitek APMG/Ameripath 408-399-5050 x 123 --------------------------------- Do you Yahoo!? Yahoo! Photos: High-quality 4x6 digital prints for 25¢ From siksik03 <@t> comcast.net Sat Apr 17 16:54:19 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing In-Reply-To: References: Message-ID: Hi Terry & HistoNetters I have done lots of successful large organ stabilization in saline at 65?C for 20-35 minutes (depending on the size of the organ. However, it is important to emphasize that the purpose of this step usually is to firm up the tissue so that it can be subsequently cut up into thinner pieces for processing, which must include a fixation step, either in the microwave or on a conventional processor, in formalin or some non-formalin fixative. The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- a procedure I have not tried myself. best regards, Steven Slap Microwave Consultant At 3:58 PM +0100 4/16/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Sorry to but in - but on a related question - >does anyone do microwave fixation in saline as >described by for instance the Melbourne group, >which involves bringing up to 65C in saline? >Have tried it this week and the sections are even crummier than usual. >When I was in Tasmania, a local private lab. did >it and their sections were fine. From siksik03 <@t> comcast.net Sat Apr 17 16:46:24 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing and Derm Shaves In-Reply-To: References: Message-ID: Hi HistoNetters Derm shaves are excellent candidates for microwave processing. In fact, several of the earliest adopters of the method over ten years ago were major dermpath labs. The reputable manufactureres should be able to provide good references. Most lab microwaves can process shaves as though they were conventional biopsies. Skin punches process well, too, but require more time. best regards, Steven Slap Microwave Consultant At 2:33 PM +0000 4/16/04, kevin williams wrote: > > Is anyone out there doing any processing on derm shaves with microwave > processing? How is it and how are the results? > Thanks. > > A. Kevin Williams > Vermont Dermatopathology. From JColCLEFA <@t> aol.com Sun Apr 18 15:26:55 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 28 Message-ID: Biocare offers the p16 antibody in a cocktail with another prostate appropriate antibody --K903(34bE12) or P504s for doublestaining of prostates. I think they might sell it solo too. From Robert.Fauck <@t> ccdhb.org.nz Sun Apr 18 17:19:38 2004 From: Robert.Fauck <@t> ccdhb.org.nz (Robert Fauck) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] MLH1 and MSH2 anti bodies Message-ID: <9952FA35C61B4F4F90DC0B2D58FE68A960E5B6@WN0NTEML01.hiq.net.nz> Dear Histonetters Could somebody give me some advice for a good protocol for the above antibodies? We are trialing at the moment them from BD and with not optimum results. We use pH 6 citric buffer. Thanking you in advance for your help. Robert Fauck Wellington Hospital New Zealand C&C DHB Secure Mail Server. ******************************************************************************** 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. (AC_S001) No Viruses were detected in this message. ******************************************************************************** From carl.hobbs <@t> kcl.ac.uk Mon Apr 19 02:37:31 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] re PGP 9. Message-ID: <004501c425e1$2c4d9dd0$e8345c9f@Carlos> Xudong, I use the Ab reagent from Ultraclone. This rabbit polyclonal works very well on pwax sections of human, mouse and chick, in my hands From kappeler <@t> patho.unibe.ch Mon Apr 19 03:17:57 2004 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Re: Dystrophin antibody References: Message-ID: <00f801c425e6$d2e18f80$27955c82@patho.unibe.ch> Hi Steven Urea is a denaturing reagent which found its way into IHC-labs probably from molecular biology. There it was used in DNA sequencing gels, before the arrival of the automated capillyry sequencing machines used today. Its 'task' was to help keep denatured DNA in the single strand conformation. I've seen the recipe for this 'buffer' (it's not really a buffer, because at pH 9.5 the buffering capacity of tris is close to zero) published a couple of years ago (would have to unearth the reference ...) - we tried it, and it worked quite well, at least for some antibodies. What it actually DOES on our tissue sections is beyond my knowledge, however with many of all these retrieval recipes we do probably not exactly know, what all the ingredients in buffers, etc. do. As with other high pH retrieval solutions, urea-containing buffer also reveals endogenous biotin, so you have to do an avidin-biotin block at least for such citical tissue samples as kidney, liver, etc. or use a non-biotin dependent visualization system. Hope this helps. Andi ----- Original Message ----- From: To: "Andi Kappeler" Sent: Wednesday, April 14, 2004 9:04 PM Subject: Re: [Histonet] Re: Dystrophin antibody > Andi, > > If I may ask, what is Urea used for? > > > > > "Andi Kappeler" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 04/14/2004 11:33 AM > > > To: "Histonet" > cc: > Subject: [Histonet] Re: Dystrophin antibody > > > Hi Cheryl > we use clone 13H6 (Novocastra, NCL-DYSA) at 1:50 after HIER in 100 mM Tris > - > 5% Urea, pH 9.5. Other high pH retrieval buffers may work as well. Hope > this > helps. > > Andi Kappeler > Institute of Pathology, University of Bern, Switzerland > > ----- Original Message ----- > From: "Cheryl Crowder" > To: "Histonet" > Sent: Wednesday, April 14, 2004 5:58 PM > Subject: [Histonet] Dystrophin antibody > > > > Hi - I have a researcher who would like to do Dystrophin IHC on > > formalin-fixed, paraffin embedded tissue. Does anyone know of an > antibody > > which will work. Thank you, Cheryl > > > > > > Cheryl Crowder, BA, HTL(ASCP) > > Chief Technologist > > Anatomic Pathology > > Department of Pathobiological Sciences > > School of Veterinary Medicine > > Louisiana State University > > Skip Bertman Drive > > Baton Rouge, LA 70803 > > > > 225-578-9734 > > FAX: 225-578-9720 > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@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 abright <@t> brightinstruments.com Mon Apr 19 06:04:00 2004 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] tissue slicer Message-ID: Dear Katy, Our Cryostat Model OTF5000 will section at the range of thicknesses you specify, but only one section after another, I have not seen a device that can section the whole specimen at the desired thickness in one cut, also I cannot see how it would be possible to section by this method accurately. Please let me have your contact details to enable me to explain how I can assist you. 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: Katy Whalley [mailto:k.whalley@ich.ucl.ac.uk] Sent: 17 April 2004 13:35 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] tissue slicer Hi, We are looking for a device which can be used to cut tissue quickly into slices of an even thickness. We're not sure yet exactly how thick these will be but something in the range 30-300 microns is likely. My supervisor has in mind something in which several blades are attached to a holder that keeps them the correct distance apart, so that all the slices are cut at once. Has anyone ever used/ seen this kind of thing, or anything else which would do the job? thanks, Katy, UCL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AFeatherstone <@t> KaleidaHealth.Org Mon Apr 19 05:47:32 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Tdt IHC Message-ID: Can anyone give me a quick protocol for their Tdt stain, Dilution, pretreat, vendor? Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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 rfail <@t> toolkitmail.com Mon Apr 19 06:14:58 2004 From: rfail <@t> toolkitmail.com (rfail) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Tdt IHC Message-ID: <4083b4b2.142.6f9e.1764108600@toolkitmail.com> Annette, DAKO,1:40, high pH HIER Rena Fail HT(ASCP)QIHC Can anyone give me a quick protocol for their Tdt stain, > Dilution, pretreat, vendor? > Annette Featherstone HT/MLT > Kaleida Health > Buffalo General Hospital > 100 High St > Buffalo NY 14203 > > > > > 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 kgrobert <@t> rci.rutgers.edu Mon Apr 19 08:57:19 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] tissue slicer In-Reply-To: <4823.194.82.240.48.1082205296.squirrel@jenner.ich.ucl.ac.uk> References: <4823.194.82.240.48.1082205296.squirrel@jenner.ich.ucl.ac.uk> Message-ID: <4083DABF.8040403@rci.rutgers.edu> Hmmm, sounds like a really tiny egg slicer. :o) The only thing that I could think of-and it doesn't match your required thickness-was this: http://www.emsdiasum.com/microscopy/products/preparation/slice.aspx#69010 This will help you make accurate freehand sections of 1mm thick, but again you can't make all the slices at once. Good luck- Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Rutgers University Katy Whalley wrote: >Hi, > >We are looking for a device which can be used to cut tissue quickly into >slices of an even thickness. We're not sure yet exactly how thick these >will be but something in the range 30-300 microns is likely. My supervisor >has in mind something in which several blades are attached to a holder >that keeps them the correct distance apart, so that all the slices are cut >at once. Has anyone ever used/ seen this kind of thing, or anything else >which would do the job? > >thanks, >Katy, UCL > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From DonnaWillis <@t> texashealth.org Mon Apr 19 09:48:27 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing Message-ID: <5C6CBCCEB04B894C8BD15B312487F7B201A33071@ftwex01.txhealth.org> Terry, Saline will not fix your tissue. It will only stabilize it. If you do not post fix in formalin after stabilizing in saline then your tissue will be fixed in the alcohol when processed. Saline works great to firm up the tissue to assist in dissection. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr, Consultant Histopathologist Sent: Friday, April 16, 2004 9:59 AM To: kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Sorry to but in - but on a related question - does anyone do microwave fixation in saline as described by for instance the Melbourne group, which involves bringing up to 65C in saline? Have tried it this week and the sections are even crummier than usual. When I was in Tasmania, a local private lab. did it and their sections were fine. Dr Terry L (why can't I get a good H&E) 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: kevin williams [mailto:akwilliams75@hotmail.com] Sent: 16 April 2004 15:33 To: histonet@pathology.swmed.edu Subject: [Histonet] Microwave Processing and Derm Shaves Is anyone out there doing any processing on derm shaves with microwave processing? How is it and how are the results? Thanks. A. Kevin Williams Vermont Dermatopathology. _________________________________________________________________ [1]Get rid of annoying pop-up ads with the new MSN Toolbar FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS= _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From Terry.Marshall <@t> rothgen.nhs.uk Mon Apr 19 09:47:09 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Microwave Processing Message-ID: Thanks Steven. It was the Leong method to which I referred. I am totally befuddled over fixation in the microwave. Surely in the Leong method (for blocks), the saline is irrelevent other than as a carrier and buffer medium, and it is heat fixation? Firming up breasts in the microwave seems fun - but the breasts in Yorkshire are bigger than the microwaves:-) 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 17 April 2004 22:54 To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hi Terry & HistoNetters I have done lots of successful large organ stabilization in saline at 65?C for 20-35 minutes (depending on the size of the organ. However, it is important to emphasize that the purpose of this step usually is to firm up the tissue so that it can be subsequently cut up into thinner pieces for processing, which must include a fixation step, either in the microwave or on a conventional processor, in formalin or some non-formalin fixative. The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- a procedure I have not tried myself. best regards, Steven Slap Microwave Consultant At 3:58 PM +0100 4/16/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Sorry to but in - but on a related question - >does anyone do microwave fixation in saline as >described by for instance the Melbourne group, >which involves bringing up to 65C in saline? >Have tried it this week and the sections are even crummier than usual. >When I was in Tasmania, a local private lab. did >it and their sections were fine. From mcauliff <@t> umdnj.edu Mon Apr 19 13:08:26 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] tissue slicer In-Reply-To: <4823.194.82.240.48.1082205296.squirrel@jenner.ich.ucl.ac.uk> References: <4823.194.82.240.48.1082205296.squirrel@jenner.ich.ucl.ac.uk> Message-ID: <4084159A.1000208@umdnj.edu> Hi Katy: You could buy a Vibratome, a device with a vibrating blade that will cut fixed or unfixed tissue at a thickness you select. I think there are several models and vendors. Or, you could make an inexpensive device for little more than pocket change. Buy some high-quality double-edge razor blades and some material to use for spacing the blades. For 1 mm or more use square aluminum rod, for 0.5 mm or less use "shim stock". A well-stocked hardware store or maching shop will have these items. Use "super-glue" to glue up a "blade-spacer-blade-spacer-blade ..." tool with as many blades as your project demands. One 'application' of the tool to the sample will give you uniform and reproducable slices. Be sure to cut off or mask the edge of the blade not in use so you won't cut yourself. Geoff Katy Whalley wrote: >Hi, > >We are looking for a device which can be used to cut tissue quickly into >slices of an even thickness. We're not sure yet exactly how thick these >will be but something in the range 30-300 microns is likely. My supervisor >has in mind something in which several blades are attached to a holder >that keeps them the correct distance apart, so that all the slices are cut >at once. Has anyone ever used/ seen this kind of thing, or anything else >which would do the job? > >thanks, >Katy, UCL > >_______________________________________________ >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 Terry.Marshall <@t> rothgen.nhs.uk Mon Apr 19 10:18:33 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] tissue slicer Message-ID: Always wondered what a vibratome was. I've got a wifatome. 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: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: 19 April 2004 19:08 To: k.whalley@ich.ucl.ac.uk Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] tissue slicer Hi Katy: You could buy a Vibratome, a device with a vibrating blade that will cut fixed or unfixed tissue at a thickness you select. I think there are several models and vendors. Or, you could make an inexpensive device for little more than pocket change. Buy some high-quality double-edge razor blades and some material to use for spacing the blades. For 1 mm or more use square aluminum rod, for 0.5 mm or less use "shim stock". A well-stocked hardware store or maching shop will have these items. Use "super-glue" to glue up a "blade-spacer-blade-spacer-blade ..." tool with as many blades as your project demands. One 'application' of the tool to the sample will give you uniform and reproducable slices. Be sure to cut off or mask the edge of the blade not in use so you won't cut yourself. Geoff Katy Whalley wrote: >Hi, > >We are looking for a device which can be used to cut tissue quickly into >slices of an even thickness. We're not sure yet exactly how thick these >will be but something in the range 30-300 microns is likely. My supervisor >has in mind something in which several blades are attached to a holder >that keeps them the correct distance apart, so that all the slices are cut >at once. Has anyone ever used/ seen this kind of thing, or anything else >which would do the job? > >thanks, >Katy, UCL > >_______________________________________________ >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 gcallis <@t> montana.edu Mon Apr 19 10:20:50 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] cutting frozen rat turbs,, a reply In-Reply-To: Message-ID: <3.0.6.32.20040419092050.00bdcd30@gemini.msu.montana.edu> We cut decalcified turbinates without using Cryojane. Turbinates are decalcified with (from 1.25% up to 14% tetrasodium EDTA in Dulbeccos PBS containing 20% sucrose, pH is adjusted to 7.6 You could use formic acid IF you rinse turbinates well and cryoprotect after acid solution. EDTA decalcification is done at 4C, formic acid is done at RT. It takes a LONG time, and we do a weight loss,weight gain endpoint check with mouse and hamster heads taking as long as 2 weeks, sometimes more. Patience! If you decalcify with formic acid and after the water rinse, you may need to cryoprotect maybe a bit longer than overnight with 20 - 30% sucrose in PBS at 4C to insure the cryoprotectant is infiltrated well into bone, etc. I think you could even add OCT to this cryoprotectant mixture without problems. I helps to vacuum samples to pull out air bubbles in rat nose before going to 4C. Turbinates are snap frozen embedded in OCT by floating a petri dish on a layer of liquid nitrogen, and using a Peel away mold, the one that is 22 X 22, nose facing down to accomodate longer specimen. Cryosectioning is done with a high profile AccuEdge blade - very sharp and very sturdy. Sectioning is carried out at colder temps, -26C with sections picked up on Plus charge, air dried at RT before IHC. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jqb7 <@t> cdc.gov Mon Apr 19 10:27:32 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] tissue slicer Message-ID: That is so bad! :) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Monday, April 19, 2004 11:19 AM To: Geoff McAuliffe; k.whalley@ich.ucl.ac.uk Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] tissue slicer Always wondered what a vibratome was. I've got a wifatome. 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: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: 19 April 2004 19:08 To: k.whalley@ich.ucl.ac.uk Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] tissue slicer Hi Katy: You could buy a Vibratome, a device with a vibrating blade that will cut fixed or unfixed tissue at a thickness you select. I think there are several models and vendors. Or, you could make an inexpensive device for little more than pocket change. Buy some high-quality double-edge razor blades and some material to use for spacing the blades. For 1 mm or more use square aluminum rod, for 0.5 mm or less use "shim stock". A well-stocked hardware store or maching shop will have these items. Use "super-glue" to glue up a "blade-spacer-blade-spacer-blade ..." tool with as many blades as your project demands. One 'application' of the tool to the sample will give you uniform and reproducable slices. Be sure to cut off or mask the edge of the blade not in use so you won't cut yourself. Geoff Katy Whalley wrote: >Hi, > >We are looking for a device which can be used to cut tissue quickly >into slices of an even thickness. We're not sure yet exactly how thick >these will be but something in the range 30-300 microns is likely. My >supervisor has in mind something in which several blades are attached >to a holder that keeps them the correct distance apart, so that all the >slices are cut at once. Has anyone ever used/ seen this kind of thing, >or anything else which would do the job? > >thanks, >Katy, UCL > >_______________________________________________ >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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Mon Apr 19 10:55:08 2004 From: jkiernan <@t> uwo.ca (J. A. Kiernan) Date: Fri Sep 16 15:22:49 2005 Subject: Urea. (Re: [Histonet] Re: Dystrophin antibody) References: <00f801c425e6$d2e18f80$27955c82@patho.unibe.ch> Message-ID: <4083F65C.5904FB0D@uwo.ca> Urea was among the first substances to be used for what we now call antigen retrieval, by Hausen & Dreyer (1982). They put hydrated paraffin sections into 3M aqueous urea (that's 18%w/v) for 5 minutes prior to immunofluorescent staining. Their tissues had been prepared by freeze-substitution into ethanol, and urea treatment would not reactivate antigens after aldehyde fixation. Evidently the hydrogen-bonding actions of urea could not break the covalent bonds formed by formaldehyde or glutaraldehyde, which is the presumed mode of action of the hot water used in modern antigen retrieval procedures. Urea has been combined with hot water to enhance real antigen retrieval in formaldehyde-fixed tissue. Shi et al (1993) microwaved slides in 5% aqueous urea and got better results than with lead thiocyanate, an earlier retrieval reagent. (Hausen & Dreyer also commented that longer times than 5 min or higher concentrations of urea caused detachment of sections from slides.) Refs: Hausen P., Dreyer, C. 1982. Urea reactivates antigens in paraffin sections. Stain Technol. 57: 321-324. Shi, S.R. et 4 al. 1993. Antigen retrieval using citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections. J. Histochem. Cytochem. 41: 1599-1604. -- ------------------------------- 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 _______________________________ Andi Kappeler wrote: > > Hi Steven > > Urea is a denaturing reagent which found its way into IHC-labs probably from > molecular biology. There it was used in DNA sequencing gels, before the > arrival of the automated capillyry sequencing machines used today. Its > 'task' was to help keep denatured DNA in the single strand conformation. > I've seen the recipe for this 'buffer' (it's not really a buffer, because at > pH 9.5 the buffering capacity of tris is close to zero) published a couple > of years ago (would have to unearth the reference ...) - we tried it, and it > worked quite well, at least for some antibodies. What it actually DOES on > our tissue sections is beyond my knowledge, however with many of all these > retrieval recipes we do probably not exactly know, what all the ingredients > in buffers, etc. do. As with other high pH retrieval solutions, > urea-containing buffer also reveals endogenous biotin, so you have to do an > avidin-biotin block at least for such citical tissue samples as kidney, > liver, etc. or use a non-biotin dependent visualization system. Hope this > helps. > Andi > > ----- Original Message ----- > From: > To: "Andi Kappeler" > Sent: Wednesday, April 14, 2004 9:04 PM > Subject: Re: [Histonet] Re: Dystrophin antibody > > > Andi, > > > > If I may ask, what is Urea used for? > > > > > > > > > > "Andi Kappeler" > > Sent by: histonet-bounces@lists.utsouthwestern.edu > > 04/14/2004 11:33 AM > > > > > > To: "Histonet" > > cc: > > Subject: [Histonet] Re: Dystrophin antibody > > > > > > Hi Cheryl > > we use clone 13H6 (Novocastra, NCL-DYSA) at 1:50 after HIER in 100 mM Tris > > - > > 5% Urea, pH 9.5. Other high pH retrieval buffers may work as well. Hope > > this > > helps. > > > > Andi Kappeler > > Institute of Pathology, University of Bern, Switzerland > > > > ----- Original Message ----- > > From: "Cheryl Crowder" > > To: "Histonet" > > Sent: Wednesday, April 14, 2004 5:58 PM > > Subject: [Histonet] Dystrophin antibody > > > > > > > Hi - I have a researcher who would like to do Dystrophin IHC on > > > formalin-fixed, paraffin embedded tissue. Does anyone know of an > > antibody > > > which will work. Thank you, Cheryl > > > > > > > > > Cheryl Crowder, BA, HTL(ASCP) > > > Chief Technologist > > > Anatomic Pathology > > > Department of Pathobiological Sciences > > > School of Veterinary Medicine > > > Louisiana State University > > > Skip Bertman Drive > > > Baton Rouge, LA 70803 > > > > > > 225-578-9734 > > > FAX: 225-578-9720 > > > > > > > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@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 dennijc <@t> vetmed.auburn.edu Mon Apr 19 11:26:49 2004 From: dennijc <@t> vetmed.auburn.edu (John C. Dennis) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] BrdU invivo dosage In-Reply-To: References: Message-ID: Dear Dr Balaji I use 50mg/kg body weight. I have a reference, not at hand, but will round it up if you will remind me. John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Thu, 15 Apr 2004 balajimr@drreddys.com wrote: > Dear histonetter: > > Can anyone tell me the exact dose of Bromo deoxy uridine in > rats and mice for the purpose of invivo labelling for immunohistochemistry > (particularly my interest is intestines and urinary bladder). We plan to > carryout both intraperitonially and orally through drinking water.As for > as I know the dose in mice is 100mg/kg IP at a dose volume of 20 ml/kg. Is > it OK to dilute BrdU in only PBS or any other combination is required. . > It will be of great help for me if you could provide me the reference > literature for the same.Thanks in advance. > > > Dr. Balaji (M. V. Sc. Path) > Dept. Pre clinical safety evaluation, > Discovery research, > Dr. Reddys Laboratories ltd. Bollaram Road, > Miyapur, Hyderabad, 500 049 > Andhra Pradseh, INDIA > > Tel: 23045439 - Ext.432. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gudrun.lang <@t> aon.at Mon Apr 19 11:33:35 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] IF on frozen sections Message-ID: <003301c4262c$0fe2b9f0$eeeea8c0@SERVER> A question about immunfluorescence on frozen skin sections: We mount the skin with buffer on the sample-table to freeze it. After sectioning we let the cuts airdry and do the IF (Immunglobulins). Is there any reason, why we cannot use OCT for mounting? I think sectioning would be easier. Perhaps someone can help me? Lang Gudrun From spoulos <@t> saa.ars.usda.gov Mon Apr 19 11:53:12 2004 From: spoulos <@t> saa.ars.usda.gov (Sylvia Poulos) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] BrdU invivo dosage Message-ID: I used a dose of 30 mg/kg in newborn and weaning rats. I did an IP injection 1 hour before sacrificing the pups and got decent labelling though you may want to consider allowing more time for incorporation in older animals. I did dilute in PBS (just make sure yours does not contain sodium azide as a preservative. If you need a reference it is: Poulos SP, Sisk M, Hausman DB, Azain MJ, Hausman GJ. Pre- and postnatal dietary conjugated linoleic acid alters adipose development, body weight gain and body composition in Sprague-Dawley rats. J Nutr. 2001 Oct;131(10):2722-31. Good luck! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) From james.zimmerman <@t> pharma.novartis.com Mon Apr 19 11:33:37 2004 From: james.zimmerman <@t> pharma.novartis.com (james.zimmerman@pharma.novartis.com) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Grimelius Stain Message-ID: Hi, Does anyone have a tried and true method for the Grimelius Stain ? Thanks, JPZ From TMcNemar <@t> lmhealth.org Mon Apr 19 12:22:43 2004 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Specimen exception list.... Message-ID: <90092A4ED388D7119575006008F7112049CB8A@NT_EXCHANGE> I have a question regarding which specimens are sent to Pathology and which are not. Our "Exception List" is essentially the same as the "Gross Only" list and is approved by the Medical Staff. Our policy does include the statement..."this list may be extended to include other specimens as requested by the physician." When a speciem is not sent, a notation is made on the surgery record that a specimen was "not sent per physician". So that is my question. Do you require your physicians to adhere strictly to the list and send all other specimens to Pathology? Do you allow them any discretion in what they send or don't send? Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 From RITA.ANGEL <@t> UC.EDU Mon Apr 19 12:24:11 2004 From: RITA.ANGEL <@t> UC.EDU (Rita Angel) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] fixation for frozen tissue Message-ID: <5.1.0.14.2.20040419131349.00b2ac70@ucmail3.uc.edu> Hi all, I have several questions about fixing frozen tissue. In the past, we've always frozen tissue fresh from the animal into frozen embedding media and then immersing into liquid nitrogen. I have an investigator that brought frozen tissue that he first fixed in formalin, then froze. I'm unable to get sections because the tissue is soft. It seems there is still moisture in the tissue, so I melted the block & blotted off the tissue, then re-froze. I'm still not able to get sections although it doesn't seem as mushy now. It still seems like the tissue is wet.Does anyone have any suggestions? He was also asking about a protocol for sucrose, and I was wondering if anyone could get one to me? Also why do people use this procedure, and when do you need to use it? Thanks for all your help, Rita Angel, HT From cornettl <@t> hotmail.com Mon Apr 19 12:48:49 2004 From: cornettl <@t> hotmail.com (Lorraine Cornett) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Biopsy processing schedule Message-ID: Could someone please share with me your processing protocol for your biopsy specimens. We have been using Zinc Formalin on our processors, we were noticing a lot of "crunchy" colon polyps, etc. We tried going back to 10% NBF, but the Dr.'s prefer the nuclear detail that Zinc Formalin provides, especially when the 10% NBF did not make a noticeable difference in the "crunchiness". Does anyone have an idea to use 10% NBF and Zinc Formalin on the processor, knowing that somehow the cassettes will need to be rinsed before the Zinc Formalin? All ideas and thoughts are appreciated. Thanks, Lorraine Cornett, HT (ASCP) Blue Ridge Pathology Kingsport,TN 37660 423 224-6767 fax 423 224-5349 _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From TMcNemar <@t> lmhealth.org Mon Apr 19 12:37:12 2004 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] Specimens returned to patients.... Message-ID: <90092A4ED388D7119575006008F7112049CB8B@NT_EXCHANGE> Another question.... What specimens do you return to patients? We used to give them anything they asked for with their physician's consent. A few yeas ago, we stopped give them anything that had to go into formalin. About the only thing we give back to them now are gallstones and a question has been raised about that. I would appreciate your input. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 From peoshel <@t> wisc.edu Mon Apr 19 12:09:49 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] tissue slicer In-Reply-To: References: Message-ID: Bad vibes, man. But, Geoff, your device would work, but as you note, not for less than 1000 micron slices, definitely not 30 - 300 micron, and there'd be more tissue damage from compression. A vibratome is pretty much the only choice, there is no "all slices at once" instrument that I've seen. Phil >Always wondered what a vibratome was. >I've got a wifatome. > >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: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] >Sent: 19 April 2004 19:08 >To: k.whalley@ich.ucl.ac.uk >Cc: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] tissue slicer > > >Hi Katy: > > You could buy a Vibratome, a device with a vibrating blade that will >cut fixed or unfixed tissue at a thickness you select. I think there are >several models and vendors. > Or, you could make an inexpensive device for little more than pocket >change. Buy some high-quality double-edge razor blades and some material >to use for spacing the blades. For 1 mm or more use square aluminum rod, >for 0.5 mm or less use "shim stock". A well-stocked hardware store or >maching shop will have these items. Use "super-glue" to glue up a >"blade-spacer-blade-spacer-blade ..." tool with as many blades as your >project demands. One 'application' of the tool to the sample will give >you uniform and reproducable slices. Be sure to cut off or mask the >edge of the blade not in use so you won't cut yourself. > >Geoff > >Katy Whalley wrote: > >>Hi, >> >>We are looking for a device which can be used to cut tissue quickly into >>slices of an even thickness. We're not sure yet exactly how thick these >>will be but something in the range 30-300 microns is likely. My supervisor >>has in mind something in which several blades are attached to a holder >>that keeps them the correct distance apart, so that all the slices are cut >>at once. Has anyone ever used/ seen this kind of thing, or anything else >>which would do the job? >> >>thanks, >>Katy, UCL -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From funderwood <@t> mcohio.org Mon Apr 19 12:55:23 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:49 2005 Subject: [Histonet] fixation for frozen tissue Message-ID: Hi Rita. I find that washing the tissue in tap water after taking it out of the formalin makes frozen sectioning easier. Fred >>> Rita Angel 04/19/04 01:24PM >>> Hi all, I have several questions about fixing frozen tissue. In the past, we've always frozen tissue fresh from the animal into frozen embedding media and then immersing into liquid nitrogen. I have an investigator that brought frozen tissue that he first fixed in formalin, then froze. I'm unable to get sections because the tissue is soft. It seems there is still moisture in the tissue, so I melted the block & blotted off the tissue, then re-froze. I'm still not able to get sections although it doesn't seem as mushy now. It still seems like the tissue is wet.Does anyone have any suggestions? He was also asking about a protocol for sucrose, and I was wondering if anyone could get one to me? Also why do people use this procedure, and when do you need to use it? Thanks for all your help, Rita Angel, HT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GauchV <@t> mail.amc.edu Mon Apr 19 12:53:57 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Specimens returned to patients.... Message-ID: Tom, We are in the middle of changing our policy on foreign body releases as well. We used to release all foreign bodies as long as the patient signed for them, gave proof of ID, etc. ..then we also stopped releasing anything that had been in formalin. Now our Risk Management Department is trying to have us release nothing with the exception of law enforcement agencies and court subpoenas. We are working on the policy as we speak...it looks as if our exempt tissue list will come into play as well . If I can be of any help to you, please feel free to contact me at this e-mail address.. Vicki Gauch AMCH Albany, NY >>> Tom McNemar 4/19/2004 1:37:12 PM >>> Another question.... What specimens do you return to patients? We used to give them anything they asked for with their physician's consent. A few yeas ago, we stopped give them anything that had to go into formalin. About the only thing we give back to them now are gallstones and a question has been raised about that. I would appreciate your input. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Mon Apr 19 12:58:32 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Biopsy processing schedule Message-ID: Have one of the stations on the processor filled with tap water to rinse the tissue prior to going into the zinc formalin. Change this station daily. I have done this in the past and it worked well. Fred >>> "Lorraine Cornett" 04/19/04 01:48PM >>> Could someone please share with me your processing protocol for your biopsy specimens. We have been using Zinc Formalin on our processors, we were noticing a lot of "crunchy" colon polyps, etc. We tried going back to 10% NBF, but the Dr.'s prefer the nuclear detail that Zinc Formalin provides, especially when the 10% NBF did not make a noticeable difference in the "crunchiness". Does anyone have an idea to use 10% NBF and Zinc Formalin on the processor, knowing that somehow the cassettes will need to be rinsed before the Zinc Formalin? All ideas and thoughts are appreciated. Thanks, Lorraine Cornett, HT (ASCP) Blue Ridge Pathology Kingsport,TN 37660 423 224-6767 fax 423 224-5349 _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee? Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Apr 19 13:27:32 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Mounting skin with OCT In-Reply-To: <003301c4262c$0fe2b9f0$eeeea8c0@SERVER> Message-ID: <3.0.6.32.20040419122732.00bbcad0@gemini.msu.montana.edu> Gudrun, Yes, you are correct. You can embed also skin in OCT, snap freeze then mount block on block with OCT for sectioning. We do all samples for cryosectioning this way. OCT surrounds the sample for support during sectioning and sectioning is easy, The section passes under an antiroll device nicely. Frozen OCT also is easily captured by brush bristles for brush technic, our favorite way. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From akwilliams75 <@t> hotmail.com Mon Apr 19 13:57:08 2004 From: akwilliams75 <@t> hotmail.com (kevin williams) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] MSDS and a lack of information Message-ID: Dear All: When requesting MSDS sheets I am getting less info than I need and I find my self undecided on the numbers to put in the Flamability, Reactivity, Health and Special Notice sections on my diamonds. Is anone aware of a good athorative web site on the subject? Thanks, A. Kevin Williams _________________________________________________________________ [1]Lose those love handles! MSN Fitness shows you two moves to slim your waist. References 1. http://g.msn.com/8HMAENUS/2743??PS= From juan.gutierrez <@t> christushealth.org Mon Apr 19 13:55:29 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Specimens returned to patients.... Message-ID: Texas law only allows teeth and in-vitro cultures that have not been exposed to pathogens to be releaseddirectly to patients. Anything else has to be released to an approved and/or licensed professional. I think Texas follows OSHA guidelines for thi, so you might want to check with them. When a patient asks for something back I just tell them that OSHA wont allow it and they go away scared. :) Juan -----Original Message----- From: Tom McNemar [mailto:TMcNemar@lmhealth.org] Sent: Monday, April 19, 2004 12:37 PM To: 'Histonet (histonet@pathology.swmed.edu)' Subject: [Histonet] Specimens returned to patients.... Another question.... What specimens do you return to patients? We used to give them anything they asked for with their physician's consent. A few yeas ago, we stopped give them anything that had to go into formalin. About the only thing we give back to them now are gallstones and a question has been raised about that. I would appreciate your input. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Mon Apr 19 17:16:49 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] vibratome or frozen sections for immuno In-Reply-To: References: Message-ID: <40844FD1.8040606@umdnj.edu> Re: the Vibratome v. frozen sections for immunostaining discussion. I looked up the papers Charles mentions below. The first two deal with tract tracing and thus do not pertain to the original question about penetration of antibodies/immunostaining reagents into 50 micron sections. The second two papers use both Vibratome and frozen sections. Hartig et al., J. Neurosci Meth. 67:89-95, 1996 finds no difference between their results with the two types of sections. Patel et al., Neurosci. Lett. 63:185-190, 1986 uses both types of sections, but their Vibratome sections are 50-100 microns while the cryostat sections are 2-6 microns. This does not seem like a valid comparison to me. However, they did not mention any difference in results between one type of section and the other. I think confusions arises in the intended purpose of the sections. Certainly for immuno some sort of permeabilization is usually used to insure that the very large antibody molecules penetrate into the cell cytoplasm. This is usually freeze-thaw (usually after appropriate cryoprotection), Triton, Tween, buffered ethanol treatment or some combination of these. However, for neuronal tract tracing large molecules need not penetrate the cell membrane, the HRP (or whatever tracer) is already in the cytoplasm and only DAB and peroxide, relatively small molecules, need to penetrate the cell. At the same time, one does not want the tracer to leak out of the cytoplasm. Perhaps Vibratome sections are more appropriate for that task but I suspect other variables would need to be considered as well Geoff Charles Scouten wrote: > Numerous studies have made the comparison. See a selected few below. > >It has been popular wisdom for 20 years that Vibratome sections, by not facturing the cell membrane, preserve the cytosol better and give more intense staining of cytosol proteins. I have personally seen differences with HRP staining, but didn't try to prove that some other variable was not responsible. > >I have located the studies below, and many others, that have made the comparison. I do not have access to the complete text, and do not know if all of these say the vibratome results were better. I would not expect vibratome to do better with membrane bound proteins. > >The science is there and done, I would compile a complete list of recent references like those below (maybe about 20-30) if somebody will volunteer to look them all up in Science Direct or the original Journals and check whether Vibratome sectioning was better, worse or the same compared to cryostat sectioning. > > >Current concepts in neuroanatomical tracing >C. K?bbert, R. Apps, I. Bechmann, J.L. Lanciego, J. Mey & S. Thanos Progress in Neurobiology, 2000, 62:4:327-351 >solon@uni-muenster.de >. The success of labelling can be estimated roughly, and the tissue can be sectioned with a vibratome. Cryosectioning of the tissue is not recommended, since some dye leakage occurs during the procedure. > > >Neural tract tracing using Di-I: a review and a new method to make fast Di-I faster in human brain >D. Larry Sparks, Lih-Fen Lue, Timothy A. Martin & Joseph Rogers Journal of Neuroscience Methods, 2000, 103:1:3 - 10 >lsparks@mail.sunhealth.org >if a fluorescent Di-I product is desired, infiltrating and embedding brain tissues in polyacrylamide prior to sectioning by vibratome eliminates the necessity of dehydrating and clearing the tissue sections prior to coverslipping ( Hayaran and Bijlani, 1992). >(too thick) >Although the thickness of the vibratome sections (50 ?m) was less than optimal for demonstrating individual axons, individual fiber tracts could occasionally be traced for as much as 5 mm or more, as shown here. Cryostat and other techniques were used to achieve thinner sections, but requiring freezing of the tissue, resulted in diffuse smearing of the fluorescence label > >Somebody let me know if this is favorable to my case. The comparison was made, but I can't get the full text to see how it came out. > > > Triple immunofluorescence labelling of parvalbumin, calbindin-D28k and calretinin in rat and monkey brain >Hartig, W. / Bruckner, G. / Brauer, K. / Seeger, G. / Bigl, V., Journal of Neuroscience Methods, Aug 1996 >...Thereafter, two brains were cut on a Vibratome (TSE, Kronberg) at 50 1lm thickness...monkey brains the tissue was cryo-protected by equili- bration...about 4 days and subsequent cryo-protection. 2.3. Tnple immu7...results were obtained with Vibratome and frozen sections and with... > > > 53. Adenosine deaminase and histidine decarboxylase coexist in certain neurons of the rat brain >Patel, B.T. / Tudball, N. / Wada, H. / Watanabe, T., Neuroscience Letters, Jan 1986 >...further 24 h at 4?C. Thick vibratome sections (50-100/~m) were obtained...required, the fixed blocks were cryo- protected with 301~:~ sucrose...and dried at 20~C for 24 h. Vibratome and cryostat sections were...immunoreactivity in rat brain vibratome sections was consistent with... > >Cordially, >Charles W. Scouten, Ph.D. >myNeuroLab.com >5918 Evergreen Blvd. >St. Louis, MO 63134 >Ph: 314 522 0300 >FAX 314 522 0377 >cwscouten@myneurolab.com >http://www.myneurolab.com > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mike King >Sent: Wednesday, April 14, 2004 12:56 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] vibratome/frozen immuno, soap residue, autofluorescence, and bad list posting habits > >re 9. RE: Vibratome Considerations (Charles Scouten) ...as for immunolabeling sensitivity differences between vibratome and frozen sections from cryoprotected tissue, is there any science to support this claim (references)? i've never seen a difference and doubt there is really any reason to expect one. especially if you need to permeabilize anyway... > >re Danielle Zalinski Vibratome Considerations post: >...50 micron sections of brain (vibratome or other) will label through the entire thickness depending on the fixation, permeabilization, and location of the antigen. if you're going to use the tissue for anything that demands full-thickness labeling, you should use a confocal microscope to check that the labeling is even through the entire thickness of the sections. papers in the histo literature will detail the procedures you need for your material, or post again for specifics. > > > > < other stuff deleted in the interest of brevity. -- -- ********************************************** 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 AInman <@t> stmarygj.com Mon Apr 19 14:41:31 2004 From: AInman <@t> stmarygj.com (Anna Inman) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] In-Situ Hybridization HPV Typing Message-ID: <013351202ADA9E42B7B48602D5DC5A8718FE98@smg-exchange.exchange.co.sclhs.net> Can anyone give me some information on: In-Situ Hybridization - HPV typing. We want to compare the Ventana method with the Digene method for HPV testing *high risk & low risk/costs/differences in methodology/any info you are willing to share would be most helpful! Thank You in advance. Anna Inman SMH Pathology ainman@smg.co.sclhs.net From carl.hobbs <@t> kcl.ac.uk Mon Apr 19 15:02:40 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] re Terry's fixation problem Message-ID: <000f01c42649$452f1c80$58a79951@home> A vibratome can be demonstrated by the Anne Summer's rep......perhaps the wifatome has one ? ;-) Re the size of Yorkshire chick breasts..surely a pathological fixation with hypertrophied anterio- thoracic protuberances, Terry? It would be interesting to try an avian cross-reactive antidystrophin Ab to see if there is abnormal patterning? From Jackie.O'Connor <@t> abbott.com Mon Apr 19 15:01:05 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] MSDS and a lack of information Message-ID: This link is pretty good. http://www.ilpi.com/msds/index.html#Manufacturers Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics 847.938.4919 "kevin williams" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/19/2004 01:57 PM To: histonet@pathology.swmed.edu cc: Subject: [Histonet] MSDS and a lack of information Dear All: When requesting MSDS sheets I am getting less info than I need and I find my self undecided on the numbers to put in the Flamability, Reactivity, Health and Special Notice sections on my diamonds. Is anone aware of a good athorative web site on the subject? Thanks, A. Kevin Williams _________________________________________________________________ [1]Lose those love handles! MSN Fitness shows you two moves to slim your waist. References 1. http://g.msn.com/8HMAENUS/2743??PS= _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Mon Apr 19 15:06:04 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] re Urea disclosure of endog biotin Message-ID: <001301c42649$be99b8f0$58a79951@home> Mmmmm...I have to "disclose" that citric acid M/Wing at pH6 does lead to endog biotin disclosure yet, when I do same with TRIS adjusted to pH10, I lose practically all endog biotin. (chick tissue being a perfect example) From rjr6 <@t> psu.edu Mon Apr 19 15:07:06 2004 From: rjr6 <@t> psu.edu (Roberta Horner) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] MSDS and a lack of information In-Reply-To: Message-ID: <007901c42649$e3d4aad0$8861ba92@padlspsu.psu.edu> I like the site from the University of Vermont http://hazard.com/msds/ when I need a MSDS form. Roberta Horner HT/HTL Animal Diagnostic Lab Penn State University -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kevin williams Sent: Monday, April 19, 2004 2:57 PM To: histonet@pathology.swmed.edu Subject: [Histonet] MSDS and a lack of information Dear All: When requesting MSDS sheets I am getting less info than I need and I find my self undecided on the numbers to put in the Flamability, Reactivity, Health and Special Notice sections on my diamonds. Is anone aware of a good athorative web site on the subject? Thanks, A. Kevin Williams _________________________________________________________________ [1]Lose those love handles! MSN Fitness shows you two moves to slim your waist. References 1. http://g.msn.com/8HMAENUS/2743??PS= _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SCheasty <@t> ahs.llumc.edu Mon Apr 19 15:16:04 2004 From: SCheasty <@t> ahs.llumc.edu (Cheasty, Sandra) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Sheets of Slide Labels that work in a LaserJet Printer Message-ID: <2E50F33F91EEDA46A77BC3B2575BB09121E08C@mars.llumc.edu> Hello all, Does anyone have a source of blank slide labels that come on individual sheets that fit in a LaserJet printer? Thanks, Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. From carl.hobbs <@t> kcl.ac.uk Mon Apr 19 15:25:57 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] re lead isothiocyanate Message-ID: <001e01c4264c$8637f280$58a79951@home> Blew the cobwebs there, John K. I remember reeling with horror at the thought of using it ( I did....once). Was I justified in my fears? It didn't do anything for my MIB1s/p53s , I have to disclose.Neither did Urea........ NB Must admit, Urea never did anything for me with other Ags( except strip my sections off). Also,how come so many of us have best results with different high temp AR solutions, yet with same( it seems) Ab reagents? From garygill <@t> dcla.com Mon Apr 19 15:34:43 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Sheets of Slide Labels that work in a LaserJet Pri nter Message-ID: http://labelarts.com/ -----Original Message----- From: Cheasty, Sandra [mailto:SCheasty@ahs.llumc.edu] Sent: Monday, April 19, 2004 3:16 PM To: HistoNet (E-mail) Subject: [Histonet] Sheets of Slide Labels that work in a LaserJet Printer Hello all, Does anyone have a source of blank slide labels that come on individual sheets that fit in a LaserJet printer? Thanks, Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Mon Apr 19 15:34:05 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Sheets of Slide Labels that work in a LaserJet Printer Message-ID: VWR - They're called "PolyPaper Laser Labels, Catalog # 51280-116. They're actually a Nalge Nunc product (6314-0010) These are 7/8 x 7/8" labels that fit up to 5 lines of text nicely. Jackie O' "Cheasty, Sandra" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/19/2004 03:16 PM To: "HistoNet \(E-mail\)" cc: Subject: [Histonet] Sheets of Slide Labels that work in a LaserJet Printer Hello all, Does anyone have a source of blank slide labels that come on individual sheets that fit in a LaserJet printer? Thanks, Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcheng <@t> mrl.ubc.ca Mon Apr 19 15:39:20 2004 From: jcheng <@t> mrl.ubc.ca (Jenny Cheng) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] CD34 and CD59 Message-ID: Hi to all, Does anyone know which manufacture supply CD34 and CD59 antibody that is known to react with guinea pig tissues?? Thank you From emry <@t> u.washington.edu Mon Apr 19 15:44:17 2004 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] brdu protocols Message-ID: Hi, Can anyone out there direct me to a brdu protocol? Do all of them take 12 hours? Thanks, Trisha Seattle From cfranci <@t> rigel.com Mon Apr 19 16:39:40 2004 From: cfranci <@t> rigel.com (Christian Franci) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Peeling Message-ID: We've been routinely sending out blocks for sectioning (we're a bit short handed at the moment) with out any problems until recently. The slides I got back looked fine. After HIER, though, the tissues on them just kept peeling off as I was staining them.... So, I had them sent out again and the sections put on super charged slides. Well, while the tissues didn't peel off as much, I still lost a fair amount... Is there any way to prevent this? Any way to reduce peeling after HIER? Or, am I *@#!! out of luck? My samples represent too much work to watch them being lost. Thanks in advance! Cheers C From Xudong_Cao <@t> brown.edu Mon Apr 19 17:09:50 2004 From: Xudong_Cao <@t> brown.edu (Xudong_Cao@brown.edu) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] RE: PGP9.5 Message-ID: <200404192209.i3JM9oB1029759@ursa.services.brown.edu> Carl, thank you very much for the info. do you have a protocol that you can share with me? many thanks. Xudong From RFail <@t> Charleston.net Mon Apr 19 16:50:51 2004 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] IF on frozen sections In-Reply-To: <003301c4262c$0fe2b9f0$eeeea8c0@SERVER> Message-ID: <001701c42658$64834ac0$de11a6a5@rena> We routinely embed skins in OCT and snap freeze in Liquid nitrogen cooled isopentane Rena Fail -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Monday, April 19, 2004 12:34 PM To: Histonetliste Subject: [Histonet] IF on frozen sections A question about immunfluorescence on frozen skin sections: We mount the skin with buffer on the sample-table to freeze it. After sectioning we let the cuts airdry and do the IF (Immunglobulins). Is there any reason, why we cannot use OCT for mounting? I think sectioning would be easier. Perhaps someone can help me? Lang Gudrun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From katri <@t> cogeco.ca Mon Apr 19 18:50:59 2004 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Peeling References: Message-ID: <007701c42669$2fd92cc0$ce989618@hala4.on.cogeco.ca> Hi Christian, Find out what they have in their waterbath, when floating out your sections. Ask them to use just distilled water with no additives (gelatin ect.). We used to have tap water with sprinkled gelatin in it and I kept losing my sections, particularly after HIER. Since I switched to plain distilled water I hardly ever lose any sections as long as you drain the water well from underneath the section... Good luck, I know how frustrating it is. Katri Katri Tuomala Department of Pathology St.Joseph's Health Care Hamilton, Ontario, Canada ----- Original Message ----- From: "Christian Franci" To: Sent: Monday, April 19, 2004 5:39 PM Subject: [Histonet] Peeling > We've been routinely sending out blocks for sectioning (we're a bit short > handed at the moment) with out any problems until recently. > The slides I got back looked fine. After HIER, though, the tissues on them > just kept peeling off as I was staining them.... > So, I had them sent out again and the sections put on super charged slides. > Well, while the tissues didn't peel off as much, I still lost a fair > amount... > Is there any way to prevent this? > Any way to reduce peeling after HIER? > Or, am I *@#!! out of luck? > My samples represent too much work to watch them being lost. > Thanks in advance! > Cheers > C > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akwilliams75 <@t> hotmail.com Mon Apr 19 19:22:18 2004 From: akwilliams75 <@t> hotmail.com (kevin williams) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] MSDS and a lack of information (Responses) Message-ID: Dear All Here are the responses. >From: Jackie.O'Connor@abbott.com > >This link is pretty good. > >http://www.ilpi.com/msds/index.html#Manufacturers I like the site from the University of Vermont [1]http://hazard.com/msds/ when I need a MSDS form. Roberta Horner HT/HTL Animal Diagnostic Lab Penn State University _________________________________________________________________ [2]Watch LIVE baseball games on your computer with MLB.TV, included with MSN Premium! References 1. http://64.4.14.250/cgi-bin/linkrd?_lang=EN&lah=f197fd2b80655eda904460125ac0dcc4&lat=1082419661&hm___action=http%3a%2f%2fhazard%2ecom%2fmsds%2f 2. http://g.msn.com/8HMAENUS/2740??PS= From tomers <@t> shaw.ca Mon Apr 19 20:01:14 2004 From: tomers <@t> shaw.ca (Tom Wells) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Training programs in other countries (long) Message-ID: <006601c42672$faee6070$6701a8c0@laptop> I am seeking information from those Histotechnologists that were trained in countries other than Canada or the US. I want to know how Histotechnology is taught in those countries. In Canada, for example, Histotechnology is part of the general technology training that all laboratory technology students receive. In the US, Histotechnology is a separate subject (I believe) and does not include Hematology, Microbiology, Clinical Chemistry, etc. The training is separate from that of the rest of medical technology. I would welcome more information from you American trained techs as well. How is Histotechnology training handled in other countries and what are the reasons for the particular setup? History, Politics or Logic ;-) The reason that I am seeking this information is that there is a feeling in some camps that Histotechnology is somehow "less" of a subject than the other tradional Medical Laboratory subjects and as such should be relegated to a separate program much like laboratory aids are. This opinion is generally expressed by people who have no current experience in Anatomical Pathology and were largely trained before the advent of Immunohistochemistry and FISH, etc. I need as much ammunition as possible to battle these people. Any comments would be appreciated. Even those of you that may agree with the separation of Histotechnology philosophy. Thanks for you help in this. Tom Wells Histotechnology Instructor British Columbia Institute of Technology Burnaby, BC Canada From rowani.mohdrawi <@t> student.adelaide.edu.au Mon Apr 19 20:25:30 2004 From: rowani.mohdrawi <@t> student.adelaide.edu.au (Rowani) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Blade Mouse Brain AND Dissection References: Message-ID: <00ec01c42676$5eb197f0$87997f81@LaptopRowani> I like to cut small specific areas of mouse brain (fixed or fresh) for various purposes including ultrastructure studies. Could anyone suggest the best blade that will result in minimum tissue distortion. Rowani From JColCLEFA <@t> aol.com Mon Apr 19 20:53:30 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 29 Message-ID: <1d4.1f67b5e8.2db5dc9a@aol.com> Our protocol for TDT is Citrate buffer pH 6.0 @60c for 25 minutes in a water bath, H2O2 block, caseinate block, cell marque polyclonal tdt concentrate at a 1:50 dilution, 30 minutes RT primary antibody, and we use a combined control of tonsil, thymus and lymphoblastic lymphoma.(the tonsil is included to make sure we aren't getting false positives and should NOT stain. ) We use goat biotinylated link, (streptavidin biotin secondary reagent kit,) hrp label, DAB chromogen. No protein digestion. In working up the antibody , we found staining of negative cases was our biggest problem , and was caused by routine heat induced epitope retrieval methods. (25 min in citrate 6.0 @95C and EDTA pH 8.0 15- 25 min both caused negative cases and normal tonsil to stain. ) Questions? just ask J. Coleman QIHC From JColCLEFA <@t> aol.com Mon Apr 19 21:00:51 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 29 Message-ID: <60.3db7a9c7.2db5de53@aol.com> I routinely use OCT for frozen skins (also Heart bxs and kidney bxs) for IF staining and it's been working fine for me for years. We use FITC conjugated antibodies from DAKO and get great results with minimum background. From ekaplan <@t> squ.edu.om Mon Apr 19 23:27:39 2004 From: ekaplan <@t> squ.edu.om (Evelyn Kaplan) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Grimelius Stain In-Reply-To: Message-ID: Hi James, The Grimelius (1968) method in Bancroft and Gamble, 2002, page 367 is one I have used successfully. Regards, Evelyn Kaplan, Dept of Pathology, College of Medicine and Health Sciences, Sultan Qaboos University, Oman -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of james.zimmerman@pharma.novartis.com Sent: Monday, April 19, 2004 8:34 PM To: Histonet@Pathology.swmed.edu Subject: [Histonet] Grimelius Stain Hi, Does anyone have a tried and true method for the Grimelius Stain ? Thanks, JPZ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nick.kirk3 <@t> btopenworld.com Tue Apr 20 00:23:30 2004 From: nick.kirk3 <@t> btopenworld.com (=?iso-8859-1?q?NICK=20KIRK?=) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Training programs in other countries (long) In-Reply-To: <006601c42672$faee6070$6701a8c0@laptop> Message-ID: <20040420052330.63577.qmail@web86311.mail.ukl.yahoo.com> Tom In the UK we initially take a multi-disciplinary approach, with a BSc (Hons) in Biomedical Science.This is then followed by a period of in house training where the individual completes a training portfolio in their own particular discipline. Once complete and when their training officer is happy that they have reached the required standard, they are externally assessed and if they meet the requirements set by the Health Professions Council they are admitted onto the Biomedical Scientists register and are then allowed to practice as a Biomedical Scientist in their particular discipline. Many then go onto do a Masters degree and in some cases MPhils and PhDs. The professional body in the UK, the Institute of Biomedical Science (IBMS), has also developed a set of professional examinations called Higher Specialist Certificates which are also discipline specific. If you need more information on any of these areas, may I suggest you look at the IBMS website at www.ibms.org as their is much on it to explain the process further. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England Tom Wells wrote: I am seeking information from those Histotechnologists that were trained in countries other than Canada or the US. I want to know how Histotechnology is taught in those countries. In Canada, for example, Histotechnology is part of the general technology training that all laboratory technology students receive. In the US, Histotechnology is a separate subject (I believe) and does not include Hematology, Microbiology, Clinical Chemistry, etc. The training is separate from that of the rest of medical technology. I would welcome more information from you American trained techs as well. How is Histotechnology training handled in other countries and what are the reasons for the particular setup? History, Politics or Logic ;-) The reason that I am seeking this information is that there is a feeling in some camps that Histotechnology is somehow "less" of a subject than the other tradional Medical Laboratory subjects and as such should be relegated to a separate program much like laboratory aids are. This opinion is generally expressed by people who have no current experience in Anatomical Pathology and were largely trained before the advent of Immunohistochemistry and FISH, etc. I need as much ammunition as possible to battle these people. Any comments would be appreciated. Even those of you that may agree with the separation of Histotechnology philosophy. Thanks for you help in this. Tom Wells Histotechnology Instructor British Columbia Institute of Technology Burnaby, BC Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From marktarango <@t> earthlink.net Tue Apr 20 01:28:18 2004 From: marktarango <@t> earthlink.net (Mark Adam Tarango) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] IF on frozen sections In-Reply-To: <200404191000.1bfC894383NZFji0@eagle> Message-ID: <000401c426a0$abb0cc90$2702a8c0@markie> >>Date: Mon, 19 Apr 2004 18:33:35 +0200 >>From: "Gudrun Lang" >>Subject: [Histonet] IF on frozen sections >>To: "Histonetliste" >>Message-ID: <003301c4262c$0fe2b9f0$eeeea8c0@SERVER> >>Content-Type: text/plain; charset="iso-8859-1" >>A question about immunfluorescence on frozen skin sections: >>We mount the skin with buffer on the sample-table to freeze it. After >>sectioning we let the cuts airdry and do the IF (Immunglobulins). >>Is there any reason, why we cannot use OCT for mounting? I think >>sectioning would be easier. >>Perhaps someone can help me? >>Lang Gudrun I use OCT for cutting skin for IF staining. My stains come out just fine.... Mark Tarango From ESerrano <@t> imim.es Tue Apr 20 02:02:40 2004 From: ESerrano <@t> imim.es (Serrano del Pozo, Erika) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] fixation for frozen tissue Message-ID: <10708933FD0CD511BCFE000102BE5F72025FAD10@HPSERV> Hi Rita, I only use fixation before freeze a tissue when I can?t handle it when it?s fresh (diaphragm), and I perform a sucrose treatment after the fixation. Here is what I do: fixation: paraformaldehid 4% at 4?C (minimum time o/n) fast washed in PBS 0,1M criproteccion (I): sucrose 15% (75g sucrose - 1ml NaN3 10% - to 500ml PBS) o/n at 4?C; shaking crioproteccion (II): sucrose 30% (150g sucrose - 2ml NaN3 10% - to 1000ml PBS) shaking at 4?C; several changes (minimum 3) until the samples are at heart deposited of the container (I don?t know why, but this not always happens; don?t worry, it also works) fast washed in PBS 0,1M frezee in OCT with liquid nitrogen Wishing you a "sweet" procedure. Erika Serrano Centre de Regulaci? Gen?mica Differenciation and Cancer Programme Barcelona -SPAIN- > ---------- > From: Rita Angel > Sent: Monday, April 19, 2004 19:24 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] fixation for frozen tissue > > Hi all, > > I have several questions about fixing frozen tissue. In the past, we've > always frozen tissue fresh from the animal into frozen embedding media and > > then immersing into liquid nitrogen. > > I have an investigator that brought frozen tissue that he first fixed in > formalin, then froze. I'm unable to get sections because the tissue is > soft. It seems there is still moisture in the tissue, so I melted the > block > & blotted off the tissue, then re-froze. I'm still not able to get > sections > although it doesn't seem as mushy now. It still seems like the tissue is > wet.Does anyone have any suggestions? > > He was also asking about a protocol for sucrose, and I was wondering if > anyone could get one to me? Also why do people use this procedure, and > when > do you need to use it? > > Thanks for all your help, > > Rita Angel, HT > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From carl.hobbs <@t> kcl.ac.uk Tue Apr 20 02:17:08 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] re PGP9.5 Message-ID: <001101c426a7$7dcc4730$e8345c9f@Carlos> Sent you the protocol.....realise you have the method working well, anyway. From histoembriyo <@t> netbulmail.com Tue Apr 20 03:07:35 2004 From: histoembriyo <@t> netbulmail.com (izzet oguz) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] osteoclast demonstration Message-ID: <3711.193.255.128.130.1082448455.webmail@mail1.netbulmail.com> Hi Histonetters, There was a problem about my subscription and with an another mail address I would like to repeat my question. Is there anybody worked with osteoclast antibody? We found Trap assay but it does not work with formalin fixed tissues so anybody who suggests an antibody which detects osteoclast in formalin fixed rat bone (not human as Zymed antibody) will be appreciated.. Thanks in advance.. ?zzet O?uz Mersin University Medicine Faculty Histology and Embryology Department TURKEY izzetoguz@yahoo.com From k.whalley <@t> ich.ucl.ac.uk Tue Apr 20 03:29:17 2004 From: k.whalley <@t> ich.ucl.ac.uk (Katy Whalley) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] tissue slicer In-Reply-To: References: Message-ID: <2068.194.82.240.48.1082449757.squirrel@jenner.ich.ucl.ac.uk> Hi, Thanks to everyone for your advice and suggestions. It seems I might have to re-think the thickness of the slices required - in retrospect, I was probably aiming too low anyway. Basically, the reason we want to cut them quickly is in order to culture the slices later, so I'm not sure a vibratome or cryostat would be appropriate as I think the tissue would have to be embedded/ frozen first. Overall the 'egg-slicer' or matrix type of device seem like they may be the best option, but I may try to make my own, as described by Geoff, to cut costs a bit. Katy Bad vibes, man. > > But, Geoff, your device would work, but as you note, not for less > than 1000 micron slices, definitely not 30 - 300 micron, and there'd > be more tissue damage from compression. A vibratome is pretty much > the only choice, there is no "all slices at once" instrument that > I've seen. > > Phil > >>Always wondered what a vibratome was. >>I've got a wifatome. >> >>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: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] >>Sent: 19 April 2004 19:08 >>To: k.whalley@ich.ucl.ac.uk >>Cc: histonet@lists.utsouthwestern.edu >>Subject: Re: [Histonet] tissue slicer >> >> >>Hi Katy: >> >> You could buy a Vibratome, a device with a vibrating blade that will >>cut fixed or unfixed tissue at a thickness you select. I think there are >>several models and vendors. >> Or, you could make an inexpensive device for little more than pocket >>change. Buy some high-quality double-edge razor blades and some material >>to use for spacing the blades. For 1 mm or more use square aluminum rod, >>for 0.5 mm or less use "shim stock". A well-stocked hardware store or >>maching shop will have these items. Use "super-glue" to glue up a >>"blade-spacer-blade-spacer-blade ..." tool with as many blades as your >>project demands. One 'application' of the tool to the sample will give >>you uniform and reproducable slices. Be sure to cut off or mask the >>edge of the blade not in use so you won't cut yourself. >> >>Geoff >> >>Katy Whalley wrote: >> >>>Hi, >>> >>>We are looking for a device which can be used to cut tissue quickly into >>>slices of an even thickness. We're not sure yet exactly how thick these >>>will be but something in the range 30-300 microns is likely. My >>> supervisor >>>has in mind something in which several blades are attached to a holder >>>that keeps them the correct distance apart, so that all the slices are >>> cut >>>at once. Has anyone ever used/ seen this kind of thing, or anything else >>>which would do the job? >>> >>>thanks, >>>Katy, UCL > > -- > Philip Oshel > Supervisor, BBPIC microscopy facility > Department of Animal Sciences > University of Wisconsin > 1675 Observatory Drive > Madison, WI 53706 - 1284 > voice: (608) 263-4162 > fax: (608) 262-5157 (dept. fax) > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From kappeler <@t> patho.unibe.ch Tue Apr 20 03:28:51 2004 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:22:50 2005 Subject: Urea. (Re: [Histonet] Re: Dystrophin antibody) References: <00f801c425e6$d2e18f80$27955c82@patho.unibe.ch> <4083F65C.5904FB0D@uwo.ca> Message-ID: <006301c426b1$83049900$27955c82@patho.unibe.ch> Thanks John for giving us this insight into the history of urea. I wasn't aware of Hausen & Dreyer - and could not tell off hands where I originally had the urea recipe from, but it must have been Shi et al.. Wish I had my references a little bit better organized... Andi Kappeler ----- Original Message ----- From: "J. A. Kiernan" To: "Andi Kappeler" ; Sent: Monday, April 19, 2004 5:55 PM Subject: Urea. (Re: [Histonet] Re: Dystrophin antibody) > Urea was among the first substances to be used for > what we now call antigen retrieval, by Hausen & Dreyer > (1982). They put hydrated paraffin sections into > 3M aqueous urea (that's 18%w/v) for 5 minutes prior > to immunofluorescent staining. Their tissues had been > prepared by freeze-substitution into ethanol, and > urea treatment would not reactivate antigens after > aldehyde fixation. Evidently the hydrogen-bonding > actions of urea could not break the covalent bonds > formed by formaldehyde or glutaraldehyde, which is > the presumed mode of action of the hot water used > in modern antigen retrieval procedures. > > Urea has been combined with hot water to enhance real > antigen retrieval in formaldehyde-fixed tissue. > Shi et al (1993) microwaved slides in 5% aqueous > urea and got better results than with lead thiocyanate, > an earlier retrieval reagent. > > (Hausen & Dreyer also commented that longer times than > 5 min or higher concentrations of urea caused detachment > of sections from slides.) > > Refs: > Hausen P., Dreyer, C. 1982. Urea reactivates antigens > in paraffin sections. Stain Technol. 57: 321-324. > Shi, S.R. et 4 al. 1993. Antigen retrieval using citrate > buffer or urea solution for immunohistochemical demonstration > of androgen receptor in formalin-fixed paraffin sections. > J. Histochem. Cytochem. 41: 1599-1604. > -- > ------------------------------- > 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 > _______________________________ > Andi Kappeler wrote: > > > > Hi Steven > > > > Urea is a denaturing reagent which found its way into IHC-labs probably from > > molecular biology. There it was used in DNA sequencing gels, before the > > arrival of the automated capillyry sequencing machines used today. Its > > 'task' was to help keep denatured DNA in the single strand conformation. > > I've seen the recipe for this 'buffer' (it's not really a buffer, because at > > pH 9.5 the buffering capacity of tris is close to zero) published a couple > > of years ago (would have to unearth the reference ...) - we tried it, and it > > worked quite well, at least for some antibodies. What it actually DOES on > > our tissue sections is beyond my knowledge, however with many of all these > > retrieval recipes we do probably not exactly know, what all the ingredients > > in buffers, etc. do. As with other high pH retrieval solutions, > > urea-containing buffer also reveals endogenous biotin, so you have to do an > > avidin-biotin block at least for such citical tissue samples as kidney, > > liver, etc. or use a non-biotin dependent visualization system. Hope this > > helps. > > Andi > > > > ----- Original Message ----- > > From: > > To: "Andi Kappeler" > > Sent: Wednesday, April 14, 2004 9:04 PM > > Subject: Re: [Histonet] Re: Dystrophin antibody > > > > > Andi, > > > > > > If I may ask, what is Urea used for? > > > > > > > > > > > > > > > "Andi Kappeler" > > > Sent by: histonet-bounces@lists.utsouthwestern.edu > > > 04/14/2004 11:33 AM > > > > > > > > > To: "Histonet" > > > cc: > > > Subject: [Histonet] Re: Dystrophin antibody > > > > > > > > > Hi Cheryl > > > we use clone 13H6 (Novocastra, NCL-DYSA) at 1:50 after HIER in 100 mM Tris > > > - > > > 5% Urea, pH 9.5. Other high pH retrieval buffers may work as well. Hope > > > this > > > helps. > > > > > > Andi Kappeler > > > Institute of Pathology, University of Bern, Switzerland > > > > > > ----- Original Message ----- > > > From: "Cheryl Crowder" > > > To: "Histonet" > > > Sent: Wednesday, April 14, 2004 5:58 PM > > > Subject: [Histonet] Dystrophin antibody > > > > > > > > > > Hi - I have a researcher who would like to do Dystrophin IHC on > > > > formalin-fixed, paraffin embedded tissue. Does anyone know of an > > > antibody > > > > which will work. Thank you, Cheryl > > > > > > > > > > > > Cheryl Crowder, BA, HTL(ASCP) > > > > Chief Technologist > > > > Anatomic Pathology > > > > Department of Pathobiological Sciences > > > > School of Veterinary Medicine > > > > Louisiana State University > > > > Skip Bertman Drive > > > > Baton Rouge, LA 70803 > > > > > > > > 225-578-9734 > > > > FAX: 225-578-9720 > > > > > > > > > > > > > > > > _______________________________________________ > > > > Histonet mailing list > > > > Histonet@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 doscwk <@t> nus.edu.sg Tue Apr 20 03:57:32 2004 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Microwave tissue processors Message-ID: Hi Histonetters, Thanks to all who responded to my query. Julee Chan Musculoskeletal Tissue Lab National University of Singapore From Luis.Chiriboga <@t> med.nyu.edu Mon Apr 19 15:32:01 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] NYSHS 2004 Meeting Message-ID: The 2004 New York State Histotechnology Society annual meeting will be held at the Holiday Inn in beautiful Saratoga Springs, New York from Friday April 30th through Saturday May 1st. The meeting is open to all. Onsite registration is available. An e-mail mini program is available upon request by replying to this message. For a program and registration packet, please contact Judy LaDuc at 518-897-2247 or jaladuc@capital.net From c.m.vanderloos <@t> amc.uva.nl Tue Apr 20 06:39:34 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] RE: Urea disclosure of endog biotin Message-ID: <2a52ef2a6498.2a64982a52ef@amc.uva.nl> Hi Carl, In my hands on human kidney FFPE sections it's just the other way around: after citrate 6.0 retrieval there is hardly any endog. biotin (like with untreated sections) but after EDTA 9.0 there is lots of endog. biotin around again! Who ever said that IHC is a simple and straightforward technique??? Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Carl Date Mon, 19 Apr 2004 21:06:04 +0100 To histonet@lists.utsouthwestern.edu Subject [Histonet] re Urea disclosure of endog biotin Mmmmm...I have to "disclose" that citric acid M/Wing at pH6 does lead to endog biotin disclosure yet, when I do same with TRIS adjusted to pH10, I lose practically all endog biotin. (chick tissue being a perfect example) From rschoon <@t> email.unc.edu Tue Apr 20 06:44:28 2004 From: rschoon <@t> email.unc.edu (Robert Schoonhoven) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] brdu protocols References: Message-ID: <40850D1C.9030107@email.unc.edu> My procedure takes about 3.5 hours and has been in several publications. I'd be happy to send you a pdf file of the procedure if you like. Robert Schoonhoven UNC-CH Dir. IHC & IA Core Center for Environmental Health & Susceptibility P. Emry wrote: >Hi, > >Can anyone out there direct me to a brdu protocol? Do all of them take 12 >hours? > >Thanks, > >Trisha >Seattle > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From soumya25 <@t> nbrc.ac.in Tue Apr 20 06:54:22 2004 From: soumya25 <@t> nbrc.ac.in (soumya) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Celloidin and IHC Message-ID: <003401c426ce$38af2f60$3e00a8c0@nbrc.btisnet.ac.in> Thanks for all the information. Soumya From Tbarnhart <@t> primecare.org Tue Apr 20 06:52:19 2004 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Hairy cell marker Message-ID: <1779904B5E82D511914C00D0B793339205BFD801@exchangent> What antibody are people using to mark Hairy cell lymphomas? We would like to get rid of our TRAP stain and replace it with a immunohistochemical marker. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND tbarnhart@primecare.org Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. From japoteete <@t> saintfrancis.com Tue Apr 20 07:28:50 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Training programs in other countries (long) Message-ID: For some of us who trained just a few years after Columbus discovered America, Histology was part of our training in our School of Medical Technology, and the graduates who passed the registry bear the title of MT(ASCP). Unfortunately for those going into the program now, Histology is just a one day observation, if at all. It's a shame, because Histology needs all the exposure we can get. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Tom Wells [SMTP:tomers@shaw.ca] > Sent: Monday, April 19, 2004 8:01 PM > To: histonet > Subject: [Histonet] Training programs in other countries (long) > > I am seeking information from those Histotechnologists that were trained > in countries other than Canada or the US. I want to know how > Histotechnology is taught in those countries. > > In Canada, for example, Histotechnology is part of the general technology > training that all laboratory technology students receive. > > In the US, Histotechnology is a separate subject (I believe) and does not > include Hematology, Microbiology, Clinical Chemistry, etc. The training is > separate from that of the rest of medical technology. I would welcome more > information from you American trained techs as well. > > How is Histotechnology training handled in other countries and what are > the reasons for the particular setup? History, Politics or Logic ;-) > > The reason that I am seeking this information is that there is a feeling > in some camps that Histotechnology is somehow "less" of a subject than the > other tradional Medical Laboratory subjects and as such should be > relegated to a separate program much like laboratory aids are. This > opinion is generally expressed by people who have no current experience in > Anatomical Pathology and were largely trained before the advent of > Immunohistochemistry and FISH, etc. I need as much ammunition as possible > to battle these people. Any comments would be appreciated. Even those of > you that may agree with the separation of Histotechnology philosophy. > > Thanks for you help in this. > Tom Wells > Histotechnology Instructor > British Columbia Institute of Technology > Burnaby, BC > Canada > _______________________________________________ > 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 japoteete <@t> saintfrancis.com Tue Apr 20 07:33:46 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Hairy cell marker Message-ID: We use DAKO's clone DBA.44, order #0880 with great results. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Barnhart, Tammy [SMTP:Tbarnhart@primecare.org] > Sent: Tuesday, April 20, 2004 6:52 AM > To: Histonet (E-mail) > Subject: [Histonet] Hairy cell marker > > What antibody are people using to mark Hairy cell lymphomas? We would > like > to get rid of our TRAP stain and replace it with a immunohistochemical > marker. > > Tammy Barnhart, BS, HTL(ASCP) > Anatomic Pathology Supervisor > St. Alexius Medical Center > Bismarck, ND > tbarnhart@primecare.org > > > > Confidentiality Notice:This e-mail message is for sole use of intended > recipient(s) and may contain confidential and privileged information. Any > unauthorized review, use, disclosure, distribution, or copying is > prohibited. If you are not the intended recipient, please contact the > sender > by replying to this e-mail and destroy/delete all copies of this e-mail > message. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From juan.gutierrez <@t> christushealth.org Tue Apr 20 07:55:17 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] IF on frozen sections Message-ID: I've always used OCT on my skins with no adverse effect. It's so much easier. Happy cutting, Juan -----Original Message----- From: Gudrun Lang [mailto:gudrun.lang@aon.at] Sent: Monday, April 19, 2004 11:34 AM To: Histonetliste Subject: [Histonet] IF on frozen sections A question about immunfluorescence on frozen skin sections: We mount the skin with buffer on the sample-table to freeze it. After sectioning we let the cuts airdry and do the IF (Immunglobulins). Is there any reason, why we cannot use OCT for mounting? I think sectioning would be easier. Perhaps someone can help me? Lang Gudrun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sue.Kapoor <@t> uhsi.org Tue Apr 20 08:18:05 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Reichert-Jung 2030 Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F55A6@khmcexch.uhsi.org> Can anyone in histoland help me? I have a Reichert-Jung 2030 Biocut microtome that suddenly stoped advancing, I think something inside slipped, does anyone know how to open this microtome? Thank you in advance! Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From making <@t> nersp.nerdc.ufl.edu Tue Apr 20 08:38:08 2004 From: making <@t> nersp.nerdc.ufl.edu (Mike King) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] tissue slicer Message-ID: <408527C0.6030109@nersp.nerdc.ufl.edu> re: Katy Whalley wrote: ...We are looking for a device which can be used to cut tissue quickly into slices of an even thickness. We're not sure yet exactly how thick these will be but something in the range 30-300 microns is likely. Katy, see http://www.sd-instruments.com/MX-TS.HTM for a slicer that is advertised as being able to cut sections as thin as 200 um from fresh (live) tissue chunks. it isn't cheap, but it might do what you need. From DKUMISKI <@t> mail.mcg.edu Tue Apr 20 08:40:37 2004 From: DKUMISKI <@t> mail.mcg.edu (Donna Kumiski) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Mesentery tissue Message-ID: I am looking for a protocol to stain and mount mesentery. In a past discussion I asked about mounting the tissue but I need more details about the staining. I would like to do an H&E on the tissue then mount it on slides. These slides will be used in a Histology course for first year medical students Donna ---------------------------------------------- Donna Kumiski HT (ASCP) Medical College of Georgia Dept. of Cellular Biology and Anatomy CB 1202 Augusta, Georgia 30912-2000 (706) 721-6278 dkumiski@mail.mcg.edu ----------------------------------------------- From DELONG_CYNTHIA_A <@t> LILLY.COM Tue Apr 20 08:41:20 2004 From: DELONG_CYNTHIA_A <@t> LILLY.COM (Cynthia A Delong) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] PGP that cross reacts in mouse Message-ID: Hello all, Does anyone know of an antibody for PGP that cross reacts in mouse. I have someone that wants this and the only PGP that I can find is only specific in human. Thank you all for your time Cindy Cynthia A DeLong LRL - Corporate Center DC0533 - 48B/3042 355 E Merrill St Indianapolis, IN 46225 phone: 317-276-7635 fax: 317-277-6146 From mcauliff <@t> umdnj.edu Tue Apr 20 12:04:54 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] tissue slicer In-Reply-To: <2068.194.82.240.48.1082449757.squirrel@jenner.ich.ucl.ac.uk> References: <2068.194.82.240.48.1082449757.squirrel@jenner.ich.ucl.ac.uk> Message-ID: <40855836.1010604@umdnj.edu> Almost forgot ........... There is a device called a "Stadie-Riggs tissue slicer" that will slice fresh tissue fairly thin, a few hundred microns. Thomas Scientific sells them, I don't know if anyone else does. A few hundred dollars in US currency, I think. Geoff Katy Whalley wrote: >Hi, > >Thanks to everyone for your advice and suggestions. It seems I might have >to re-think the thickness of the slices required - in retrospect, I was >probably aiming too low anyway. Basically, the reason we want to cut them >quickly is in order to culture the slices later, so I'm not sure a >vibratome or cryostat would be appropriate as I think the tissue would >have to be embedded/ frozen first. Overall the 'egg-slicer' or matrix type >of device seem like they may be the best option, but I may try to make my >own, as described by Geoff, to cut costs a bit. > >Katy > > Bad vibes, man. > > >>But, Geoff, your device would work, but as you note, not for less >>than 1000 micron slices, definitely not 30 - 300 micron, and there'd >>be more tissue damage from compression. A vibratome is pretty much >>the only choice, there is no "all slices at once" instrument that >>I've seen. >> >>Phil >> >> >> >>>Always wondered what a vibratome was. >>>I've got a wifatome. >>> >>>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: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] >>>Sent: 19 April 2004 19:08 >>>To: k.whalley@ich.ucl.ac.uk >>>Cc: histonet@lists.utsouthwestern.edu >>>Subject: Re: [Histonet] tissue slicer >>> >>> >>>Hi Katy: >>> >>> You could buy a Vibratome, a device with a vibrating blade that will >>>cut fixed or unfixed tissue at a thickness you select. I think there are >>>several models and vendors. >>> Or, you could make an inexpensive device for little more than pocket >>>change. Buy some high-quality double-edge razor blades and some material >>>to use for spacing the blades. For 1 mm or more use square aluminum rod, >>>for 0.5 mm or less use "shim stock". A well-stocked hardware store or >>>maching shop will have these items. Use "super-glue" to glue up a >>>"blade-spacer-blade-spacer-blade ..." tool with as many blades as your >>>project demands. One 'application' of the tool to the sample will give >>>you uniform and reproducable slices. Be sure to cut off or mask the >>>edge of the blade not in use so you won't cut yourself. >>> >>>Geoff >>> >>>Katy Whalley wrote: >>> >>> >>> >>>>Hi, >>>> >>>>We are looking for a device which can be used to cut tissue quickly into >>>>slices of an even thickness. We're not sure yet exactly how thick these >>>>will be but something in the range 30-300 microns is likely. My >>>>supervisor >>>>has in mind something in which several blades are attached to a holder >>>>that keeps them the correct distance apart, so that all the slices are >>>>cut >>>>at once. Has anyone ever used/ seen this kind of thing, or anything else >>>>which would do the job? >>>> >>>>thanks, >>>>Katy, UCL >>>> >>>> >>-- >>Philip Oshel >>Supervisor, BBPIC microscopy facility >>Department of Animal Sciences >>University of Wisconsin >>1675 Observatory Drive >>Madison, WI 53706 - 1284 >>voice: (608) 263-4162 >>fax: (608) 262-5157 (dept. 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 > > > -- -- ********************************************** 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 siksik03 <@t> comcast.net Tue Apr 20 09:17:53 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Grimelius Stain In-Reply-To: References: Message-ID: HistoNetters If you have a microwave with temperature control, there is a Grimelius method on pp. 198-200 of the newest edition of Kok & Boon (Microwaves for the Art of Microscopy) which should do the trick. I believe it can also be found in previous editions. best regards, Steven Slap Microwave Consultant At 12:33 PM -0400 4/19/04, james.zimmerman@pharma.novartis.com wrote: >Hi, > > > >Does anyone have a tried and true method for the Grimelius Stain ? > > > >Thanks, > >JPZ From gudrun.lang <@t> aon.at Tue Apr 20 09:21:02 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] IF on frozen sections References: Message-ID: <008e01c426e2$b5b92290$eeeea8c0@SERVER> Thank you all for the answers! This list is really, really helpfull. Gudrun ----- Original Message ----- From: "GUTIERREZ, JUAN" To: "Gudrun Lang" ; "Histonetliste" Sent: Tuesday, April 20, 2004 2:55 PM Subject: RE: [Histonet] IF on frozen sections I've always used OCT on my skins with no adverse effect. It's so much easier. Happy cutting, Juan -----Original Message----- From: Gudrun Lang [mailto:gudrun.lang@aon.at] Sent: Monday, April 19, 2004 11:34 AM To: Histonetliste Subject: [Histonet] IF on frozen sections A question about immunfluorescence on frozen skin sections: We mount the skin with buffer on the sample-table to freeze it. After sectioning we let the cuts airdry and do the IF (Immunglobulins). Is there any reason, why we cannot use OCT for mounting? I think sectioning would be easier. Perhaps someone can help me? Lang Gudrun _______________________________________________ Histonet mailing list Histonet@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 siksik03 <@t> comcast.net Tue Apr 20 09:33:30 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] tissue slicer In-Reply-To: References: Message-ID: Hi HistoNetters Another option would be the McIlwain tissue chopper (yes, I'm showing my age), still available at http://www.campden-inst.com/chopper.html. It is limited in tissue thicknesses to thicker slices than a Vibratome?, but is still much better than my lovely Lisatome can do. Steven At 12:09 PM -0500 4/19/04, Philip Oshel wrote: >Bad vibes, man. > >But, Geoff, your device would work, but as you >note, not for less than 1000 micron slices, >definitely not 30 - 300 micron, and there'd be >more tissue damage from compression. A vibratome >is pretty much the only choice, there is no "all >slices at once" instrument that I've seen. > >Phil From siksik03 <@t> comcast.net Tue Apr 20 09:35:33 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Microwave Processing Message-ID: Hi Terry & HistoNetters Yes, in the Leong method for biopsies, the specimens are heat stabilized in the microwave in saline, and not really chemically fixed. They get fixed in the ethanol in a traditional processor (Leong used ethanol, chloroform and paraffin- no xylene). I haven't seen the resulting morphology myself. As for breasts, the Milestone unit has a module, the DDG, which is 280mm x 250mm, for stabilization of breasts and other large organs. One advantage with breasts is that the nodes just light up bright white. Parenthetically, I don't believe that the breasts in Yorkshire could possibly be bigger than in the American South... best regards, Steven >Thanks Steven. It was the Leong method to which I referred. >I am totally befuddled over fixation in the microwave. Surely in the ... >...snip... >... heat fixation? > >Firming up breasts in the microwave seems fun - but the breasts in >Yorkshire are bigger than the microwaves:-) From jkiernan <@t> uwo.ca Tue Apr 20 09:51:54 2004 From: jkiernan <@t> uwo.ca (J. A. Kiernan) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Mesentery tissue References: Message-ID: <4085390A.7BBB2CA4@uwo.ca> The way I do mesentery is as follows. The preparations are very well suited to lab teaching. 1. Remove a rat's small intestine with the mesentery attached. Rinse in saline to get rid of blood etc. 2. Put a loop of gut over one end of a microscope slide, and pull on the mesentery so that it's stretched over the slide. 3. Trim round the edges of the glass with a sharp scalpel or razor blade, and scrape off any thick bits, such as larger blood vessels, from the middle of the slide. 4. Before the preparation dries out completely, immerse the slide in a coagulant fixative (Carnoy's is good) for a few minutes. Store the slides in 70% alcohol until you're ready to stain them. Many methods work really well on such preparations. Dilute toluidine blue will provide spectacular metachromatic staining of mast cells, for example. If you miss out Step 4 above, you can expose the preparation to hot formaldehyde vapour and see the sympathetic nerve fibres by fluorescence microscopy (a histochemical test for noradrenaline). You can also use staining methods for different types of connective tissue fibres. -- ------------------------------- 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 _______________________________ Donna Kumiski wrote: > > I am looking for a protocol to stain and mount mesentery. > In a past discussion I asked about mounting the tissue but I need > more details about the staining. I would like to do an H&E on the > tissue then mount it on slides. These slides will be used in a > Histology > course for first year medical students > Donna > > ---------------------------------------------- > Donna Kumiski HT (ASCP) > Medical College of Georgia > Dept. of Cellular Biology and Anatomy > CB 1202 > Augusta, Georgia 30912-2000 > (706) 721-6278 > dkumiski@mail.mcg.edu > ----------------------------------------------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Don.Birgerson <@t> leica-microsystems.com Tue Apr 20 10:04:05 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Reichert-Jung 2030 Message-ID: Hi Sue, Reichert-Jung microtomes are still made by the same people but we have changed our name to Leica-Microsystems. Give me a call and I am sure we can solve your problem. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Kapoor, Sue" To: "'histonet@lists.utsouthwestern.edu'" Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Reichert-Jung 2030 western.edu 04/20/2004 08:18 AM Can anyone in histoland help me? I have a Reichert-Jung 2030 Biocut microtome that suddenly stoped advancing, I think something inside slipped, does anyone know how to open this microtome? Thank you in advance! Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Apr 20 10:03:57 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Blade Mouse Brain AND Dissection In-Reply-To: <00ec01c42676$5eb197f0$87997f81@LaptopRowani> References: Message-ID: <3.0.6.32.20040420090357.00bc2778@gemini.msu.montana.edu> Ultrastructure studies usually means electron microscopy, is that what you are doing? How thin are you sectioning? How are you processing your tissues? Into paraffin? Epoxy resins for EM? IfAt 10:55 AM 4/20/2004 +0930, you wrote: >I like to cut small specific areas of mouse brain (fixed or fresh) for >various purposes including ultrastructure studies. Could anyone suggest the >best blade that will result in minimum tissue distortion. > >Rowani > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From ALAbeyta <@t> salud.unm.edu Tue Apr 20 10:07:15 2004 From: ALAbeyta <@t> salud.unm.edu (Antonia Abeyta) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] more ?? on rat turbs... Message-ID: Hi again, I am still struggling with these rat turbinates. For those who are experienced in cutting frozen rat turbinates, what would you suggest for their storage both before and after cutting? Are they better off in the -80 or -20? Has anyone ever had trouble with them drying out, and if so, did you find a solution? We are cutting these sections thinly on a cryostat and mounting them on probe plus slides. Because we are having trouble keeping them on, we are also thinking about using coated slides. Any suugestions on a slilde coating media we should use? A bunch of question, I know, I appreciate everyone's help. Antonia Antonia L. Abeyta Health Sciences Tech. III Community Environmental Health Program University of New Mexico Surge Bldg. Room 140 Albuquerque, NM 87131 (505) 272-4028 From siksik03 <@t> comcast.net Tue Apr 20 10:16:21 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] re antigen retrieval In-Reply-To: <001e01c4264c$8637f280$58a79951@home> References: <001e01c4264c$8637f280$58a79951@home> Message-ID: Hi Carl & HistoNetters Good point, Carl- makes you wonder, doesn't it? According to Dr. Shi, the factors that really matter are time, temperature and pH. Some antigens turn out to be "easy", others "hard". It doesn't seem to matter too much what solution is used so much, as you say, so long as the pH is right for the particular Ab, and the time and temperature are accurately controlled. That being said, I have usually found that high pH solutions damage morphology, and prefer to use pH 6.0 citrate buffer whenever I can get away with it, and raise the temperature or lengthen the time rather than raise the pH to get the "hard" aBs to work. best regards, Steven At 9:25 PM +0100 4/19/04, Carl wrote: >Blew the cobwebs there, John K. I remember reeling with horror at >the thought of using it ( I did....once). Was I justified in my >fears? It didn't do anything for my MIB1s/p53s , I have to >disclose.Neither did Urea........ >NB Must admit, Urea never did anything for me with other Ags( except >strip my sections off). >Also,how come so many of us have best results with different high >temp AR solutions, yet with same( it seems) Ab reagents? From carl.hobbs <@t> kcl.ac.uk Tue Apr 20 10:14:48 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] re PGP cross reactivity Message-ID: <008601c426ea$38c4c980$e8345c9f@Carlos> Ultraclone's give's me a very similar patterning in mouse as in human. Of course, I cannot guarantee that the positivity you see in mouse/chick is definitely PGP9.5. Please check with Ultraclone From gcallis <@t> montana.edu Tue Apr 20 10:28:11 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Re: Decalcification of Rat Hind paws In-Reply-To: Message-ID: <3.0.6.32.20040420092811.00bdf320@gemini.msu.montana.edu> We are doing a similar study with mouse paws, difference being larger rat paws, yours will need longer fixation, and you can speed up decalcification and not damage the cartilage by using either of the two following decalcifiers. Make them up in house. Both decalcifiers do not damage proteoglycans, and work beautifully with SafO/FG staining, also T blue. Fast: 8% formic acid/4% hydrochloric acid, change daily, do endpoint determinations as pure estimation of time CAN get you in trouble with under- and over removal of calcium. There is a way to do it without going to a lot of work, OR use xray methods with Faxitron for the most sensitive detection. You can also do chemical testing, but we are finding the weight loss/weight gain method works fine. You will probably have paws done in a couple of days this decalcifier, but I wouldn't bet on it via estimation. Paws take longer than you think - the arrangement of tiny bones so close to each other tends to slow down fixation AND decalcification. Slit open open skin on back of paws to help out too if you can get this done. Faster than your buffered formic acid: 15% formic acid, do endpoint determinations and change daily but decalcification will take longer than the above combo acid decalcifying solution. Once you do the endpoint determination on a few of the paws (they are all about the same size, age, etc) you have an estimate of time for the rest. I will be happy to send the quick method for doing this. I have found paws need to have extended processing due to the same tissue density/boney problems. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From carl.hobbs <@t> kcl.ac.uk Tue Apr 20 10:34:06 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] re Ag retrieval Message-ID: <009d01c426ec$eafb1f80$e8345c9f@Carlos> Thanks for that , Steve. I must admit to the opposite lol. TRIS at pH 10 gives me better morphological preservation than pH9 ( sections more inclined to fall off, and much better/consistent preservation than at pH6..............(Nuclei lose quite a bit of Haematein- reactivity equally) Also, I have just seen some pictures of Chris van der Loos, clearly demonstrating greatly increased disclosure of endog biotin at the higher pH values. Again, the opposite to my experiences. Ok, ok...someone out there must know what's going on? From kappeler <@t> patho.unibe.ch Tue Apr 20 10:37:07 2004 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] RE: Urea disclosure of endog biotin References: <2a52ef2a6498.2a64982a52ef@amc.uva.nl> Message-ID: <00af01c426ed$56a0a610$27955c82@patho.unibe.ch> Hi Chris & Carl Similar results here. I don't worry too much about endogenous biotin after citrate 6.0, however we fear EDTA 9.0 and even more Tris-Urea 9.5 ... Sometimes I feel like it should be called immunohistoalchemy rather than immunohistochemistry ... (never revealed any gold, though) Andi Kappeler Institute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "C.M. van der Loos" To: Cc: Sent: Tuesday, April 20, 2004 1:39 PM Subject: [Histonet] RE: Urea disclosure of endog biotin > Hi Carl, > In my hands on human kidney FFPE sections it's just the other way around: after citrate 6.0 retrieval there is hardly any endog. biotin (like with untreated sections) but after EDTA 9.0 there is lots of endog. biotin around again! > Who ever said that IHC is a simple and straightforward technique??? > > Chris van der Loos, PhD > Dept. of Pathology > Academic Medical Center > Amsterdam - The Netherlands > > ----- Original Message ----- > >From Carl > Date Mon, 19 Apr 2004 21:06:04 +0100 > To histonet@lists.utsouthwestern.edu > Subject [Histonet] re Urea disclosure of endog biotin > Mmmmm...I have to "disclose" that citric acid M/Wing at pH6 does lead to endog biotin disclosure yet, when I do same with TRIS adjusted to pH10, I lose practically all endog biotin. (chick tissue being a perfect example) > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Nancy.Walker <@t> sanofi-synthelabo.com Tue Apr 20 09:52:51 2004 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] fat cryostat sections: cutting pb, IHC? Message-ID: Any pointers for cutting adipose tissue with a cryostat? I have mouse epididyme adipose tissue that is unfixed and embedded in tissue tek. At - 30?C I can cut without it melting but most of the specimen stays on the knife edge leaving a nicely cut section of tissue tek, a big hole and a little fat tissue around the edges. And this at 45 ?!!! Any thinner and I get just the hole. I have a crostat which I can adjust the block and chamber temp separately. Haven't fiddled with the knife angle yet... WHAT TO DO??? any special tratments to get IHC reagents to penetrate this hydrophobic stuff? thank you for you advice, Nancy Walker Molecular Pharmacology Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 From Nancy.Walker <@t> sanofi-synthelabo.com Tue Apr 20 09:13:37 2004 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:50 2005 Subject: =?iso-8859-1?Q?R=E9f=2E_=3A_[Histonet]_tissue_slicer?= Message-ID: Try the McElwain tissue chopper, it's very fast. I do 400? brain slices. Maximum is 1 mm, minimum probably depends on the type of tissue, it has graduations of 1?. See website : http://www.campden-inst.com good luck Nancy Walker Molecular Pharmacology Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 From gcallis <@t> montana.edu Tue Apr 20 11:37:09 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Weight loss weight gain decalcification endpoint test explanation In-Reply-To: Message-ID: <3.0.6.32.20040420103709.00bc0c70@gemini.msu.montana.edu> Mark, As calcium is ionized and leaves bone collagen matrix, the bone loses weight. At the end of this process when calcium is gone, the bone begins to gain weight again, back to its original weight. This may be once calcium is removed, the weight gain is attributed to water now in those space occupied by calcium. Ones needs to use a balance that weighs in milligrams for greater accuracy. At any rate, the test is quick, saves time and does work. It was originally published many years ago as an nitric acid endpoint test (pretty harsh, rapid inorganic acid decalcifier). In the original publication, and at the end, the decalcifying fluid was chemically endpoint tested for presence of calcium. We have never done the latter and if one does a chemical endpoint test, the decalcifying fluid would have to be replaced daily near end of decalcification. You can't leave just multiple bones in decalcifier for accurate chemical testing - that is better done on one sample at a time for accuracy and very time consuming when 50 mouse paws are in the experiment. One joy of this method - it works with EDTA decalcification which is difficult to do chemical endpoint test. Cathy Mayton has a publication using EDTA with this endpoint test so it is useful for both acid and chelation decalcification. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jcline <@t> wchsys.org Tue Apr 20 11:49:22 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] KEVIN WILLIAMS/DERM SHAVES Message-ID: <007f01c426f7$6efa6de0$1d2a14ac@wchsys.org> You might want to talk to Steven Slap from Micromed. He helped develope the microwave processor that I have. The T/T microwave I have only takes about a total of 40 minutes to process biopsies. It is a two stage procedure with a processing solution and paraffin. I run this two times a day for all our GI and cervical biopsies. The biopsies are not crispy, like with the normal processing run. ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. 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From Sue.Kapoor <@t> uhsi.org Tue Apr 20 11:55:37 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Reichert-Jung 2030 Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F55A7@khmcexch.uhsi.org> Thanks to everyone, we've fix the problem and am back up and crankin' them out :) Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 -----Original Message----- From: Don.Birgerson@leica-microsystems.com [mailto:Don.Birgerson@leica-microsystems.com] Sent: Tuesday, April 20, 2004 10:04 AM To: Kapoor, Sue Cc: 'histonet@lists.utsouthwestern.edu'; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Reichert-Jung 2030 Hi Sue, Reichert-Jung microtomes are still made by the same people but we have changed our name to Leica-Microsystems. Give me a call and I am sure we can solve your problem. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Kapoor, Sue" To: "'histonet@lists.utsouthwestern.edu'" Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Reichert-Jung 2030 western.edu 04/20/2004 08:18 AM Can anyone in histoland help me? I have a Reichert-Jung 2030 Biocut microtome that suddenly stoped advancing, I think something inside slipped, does anyone know how to open this microtome? Thank you in advance! Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rocan <@t> mac.com Tue Apr 20 12:27:05 2004 From: rocan <@t> mac.com (rocan@mac.com) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] fat cryostat sections: cutting pb, IHC? In-Reply-To: References: Message-ID: Fat is notoriously difficult to cut on a cryostat. At one time we "delipidated" the sample by leaving small pieces in acetone at 4?C; then we would freeze in OCT and cut. More recently we embed in Cryogel from www.instrumedics.com and we do not need the acetone incubation anymore. Cryogel is a lot easier to cut than OCT. For mouse skin with fat this Cryogel make an enormous difference. A warmer temperature may be very helpful. Think of it as if you were slicing frozen butter! Also, we now also use instrumedics tape transfer system called CryoJane for fat and it works particularly well but it is expensive and has a good learning curve before you get the best results. I only would buy this system if you are going to do fat sectioning on a regular basis. I hope this helps. Rocio M. Rocio Sierra-Honigmann Director Engineered Wound Repair Laboratory Cedars Sinai Research Institute Tel.310-423-1882 Fax 310-423-2325 Honigmannr@cshs.org On Apr 20, 2004, at 7:52 AM, Nancy.Walker@sanofi-synthelabo.com wrote: > Any pointers for cutting adipose tissue with a cryostat? > > I have mouse epididyme adipose tissue that is unfixed and embedded > in > tissue tek. At - 30?C I can cut without it melting but most of the > specimen stays on the knife edge leaving a nicely cut section of tissue > tek, a big hole and a little fat tissue around the edges. And this at > 45 > ?!!! Any thinner and I get just the hole. > > I have a crostat which I can adjust the block and chamber temp > separately. > Haven't fiddled with the knife angle yet... > > WHAT TO DO??? > > any special tratments to get IHC reagents to penetrate this hydrophobic > stuff? > > thank you for you advice, > > Nancy Walker > Molecular Pharmacology > > Sanofi-Synthelbo Research > B.P. 37 Lab?ge Innopole > 31676 LABEGE CEDEX FRANCE > > nancy.walker@sanofi-synthelabo.com > tel : (33)561004179? fax :(33)561004001 > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Patricia.Patterson <@t> uhhs.com Tue Apr 20 13:08:03 2004 From: Patricia.Patterson <@t> uhhs.com (Patterson, Patricia) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Coverslipper Agony Message-ID: <11B1D28B53C82B4FA90552E5F5E938C701F8BB7C@uhmailbox1.uhhs.com> Hi All! I'd like some suggestions from you - we have a Hacker Coverslipper - using glass coverslips - and we get air bubbles. Not all the time, but enough to drive us crazy. We try to keep the dispensing needle clean and the amount of xylene in the rack holder consistent. What is everyone else doing?? Thanks Pat Patterson University Hospitals of Cleveland. The enclosed information is STRICTLY CONFIDENTIAL and is intended for the use of the addressee only. University Hospitals Health System and its affiliates disclaim any responsibility for unauthorized disclosure of this information to anyone other than the addressee. Federal and Ohio law protect patient medical information disclosed in this email, including psychiatric disorders, (HIV) test results, AIDs-related conditions, alcohol, and/or drug dependence or abuse. Federal regulation (42 CFR Part 2) and Ohio Revised Code section 5122.31 and 3701.243 prohibit disclosure of this information without the specific written consent of the person to whom it pertains, or as otherwise permitted by law. From carrie <@t> ethossearch.com Tue Apr 20 15:15:19 2004 From: carrie <@t> ethossearch.com (Carrie Roy) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Job Opportunity - Southern California Message-ID: Ethos Search Consultants specializes in placing sales, marketing and technical applications candidates, who are scientifically trained, into positions in the life and analytical sciences and we are working on an Instrument Sales Representative position in the Southern California/Las Vegas area. I am seeking a histology professional with a strong technical background to cover a sales territory in the Southern California/Las Vegas area. This position will allow someone to utilize their scientific expertise to assist others by representing products into the diagnostic lab environment. This is an opportunity to join the worldwide leader in reagent, consumable, and equipment sales to the anatomical pathology market at a time when the company is growing and expanding its product offerings. First year income is expected at $65-$75k, plus a company car If you or anyone you know is interested in this, or any other biotechnology/scientific related sales, marketing, or technical applications position, please feel free to contact me at: Carrie Roy Ethos Search Consultants 310-791-4396 carrie@ethossearch.com From convmcm <@t> cc.usu.edu Tue Apr 20 14:52:11 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Microwave Processing In-Reply-To: Message-ID: <000001c42710$f87fedb0$4a737b81@Cygnus> Steve & Terry, Thanks, boys, for keeping us all abreast of these developments *g* (couldn't resist) Connie McManus >Parenthetically, I don't believe that the breasts in Yorkshire could >possibly be bigger than in the American South... >Firming up breasts in the microwave seems fun - but the breasts in >Yorkshire are bigger than the microwaves:-) From histo <@t> compbio.com Tue Apr 20 16:21:23 2004 From: histo <@t> compbio.com (Histology) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Job Posting!!! Message-ID: <000f01c4271d$705d15e0$fda55742@ca.sprintbbd.net> Comparative Biosciences, Inc. Looking for Histology Teachnician and Laboratory Assistant. Please contact Nancy Pinard at 650-966-0330 x114 for further information. From histo <@t> compbio.com Tue Apr 20 16:16:32 2004 From: histo <@t> compbio.com (Histology) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Job Posting!!! Message-ID: <000e01c4271d$6fe797c0$fda55742@ca.sprintbbd.net> Comparative Biosciences, Inc. Looking for a Histology Technician and Laboratory Assistant. Please contact Nancy Pinard at 650-966-0330 x105 for further information. From Stanley.Stylli <@t> mh.org.au Tue Apr 20 17:24:18 2004 From: Stanley.Stylli <@t> mh.org.au (Stylli, Stanley) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] antibodies for FFPE human tissue Message-ID: <00519F9F41D2844D9C75BEE1F6AC09AFC89F6D@rmhmail1.ssg.org.au> Can someone please recommend a source for the following antibodies that they have used successfully (with or without antigen retrieval) on human tissue (preferably brain) p53 bax bid bcl-2 XIAP bcl-XL bcl-XS caspase 3 Thanks Stan From balajimr <@t> drreddys.com Tue Apr 20 22:12:27 2004 From: balajimr <@t> drreddys.com (balajimr@drreddys.com) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Proteoglycan - decalcification In-Reply-To: <3.0.6.32.20040420092811.00bdf320@gemini.msu.montana.edu> Message-ID: Dear Callis, Thank you very much for your replies to my earlier mails. This is with regard to Saf/FG staining of cartilage. I want to demonstrate proteoglycans in cartilage of femur joint, tarsal joint and ankle joint. I am in the process of standardization of the same. I have taken a trial tissue of joints from Rat and I have used 20% EDTA for decalcification. I read somewhere that it affects the proteoglycan staining. Is it true? If so could you please suggest a good decalcifier for the same.Also let me know how long I can fix these joints for good fixation in NBF. Thanks in advance. Dr. Balaji Dept. Pre clinical safety evaluation, Discovery research, Dr. Reddys Laboratories ltd. Bollaram Road, Miyapur, Hyderabad, 500 049 Andhra Pradseh, INDIA Tel: 23045439 - Ext.432. From jkiernan <@t> uwo.ca Tue Apr 20 23:22:32 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Re: Proteoglycan - decalcification - Why safranine? References: Message-ID: <4085F708.BA069836@uwo.ca> Dear Dr Balaji, Adequate fixation is important before you decalcify anything. That means at least 24 hours (a week is probably better) in a neutral buffered (or any other) formaldehyde solution. Is your "Saf/FG" method safranine O followed by fast green FCF? There are many such methods. One group is, to plant anatomists, what H&E is to animal pathologists. (I apologize for using nouns as adjectives, implying that anatomists are plants whereas pathologists are animals.) The other lot of safranine-fast green methods are the stuff of traditional light microscope cytology, for showing chromosomes and cytoplasmic organelles in truly thin paraffin sections of tiny bits of tissues fixed in solutions containing osmium tetroxide and potassium dichromate. I have used both groups of methods. The plant anatomy ones are straightforward if you follow the instructions, which include checking with a microscope before making the preparation a permanent one - just like H&E(!). (My limited trials of safranine and fast green FCF for traditional cytology were all dismal failures; the colours initially seen were always largely lost in the dehydrating alcohol.) If you need to stain proteoglycans in cartilage, why not use an easy staining method that does not involve critical differentiations? There are many such methods. In cartilage, they are very clean and selective. Sulphate ions (in the chondroitin sulphate glycosaminoglycans of the proteoglycans) bind cationic dyes at pH 1.0 or even lower. Alcian blue at pH 1 can be followed by any counterstain because alcian blue changes into a permanently insoluble pigment while the unbound dye is being washed out of the sections. Carbohydrate histochemistry is a large subject. It includes many inexpensive but discriminating techniques that are neglected by well funded researchers and clinical pathologists. John A. Kiernan London, Canada. _______________________________________ balajimr@drreddys.com wrote: > Thank you very much for your replies to my earlier mails. This is with > regard to Saf/FG staining of cartilage. I want to demonstrate > proteoglycans in cartilage of femur joint, tarsal joint and ankle joint. I > am in the process of standardization of the same. I have taken a trial > tissue of joints from Rat and I have used 20% EDTA for decalcification. I > read somewhere that it affects the proteoglycan staining. Is it true? If > so could you please suggest a good decalcifier for the same.Also let me > know how long I can fix these joints for good fixation in NBF. Thanks in > advance. > > Dr. Balaji > Dept. Pre clinical safety evaluation, > Discovery research, > Dr. Reddys Laboratories ltd. Bollaram Road, > Miyapur, Hyderabad, 500 049 > Andhra Pradseh, INDIA -------------------------------------------- From Inga.Hansson <@t> neuro.uu.se Wed Apr 21 00:52:54 2004 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Apostain Message-ID: Hi all! Anyone ever tried the APOSTAIN antibody to ssDNA from Alexis that is supposed to stain apoptotic cells??? I followed the protocol in detail but I see absolutely NOTHING! Inga From jim.manavis <@t> imvs.sa.gov.au Wed Apr 21 01:47:55 2004 From: jim.manavis <@t> imvs.sa.gov.au (jim) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Substance P receptor (NK1) Message-ID: <00a701c4276c$9359bea0$7a6c140a@imvs.sa.gov.au> Does anyone know of a good substance P receptor (NK1) antibody for FFPE human tissue, and/or rat and mouse. Thank you Jim Manavis Laboratory Manager Hanson Institute Centre for Neurological Diseases Adelaide, SA, 5000 Australia Phone: 61-08-8222-3668 FAX: 61-08-8222 3204 email: jim.manavis@imvs.sa.gov.au Disclaimer: Not this little black duck! From jim.manavis <@t> imvs.sa.gov.au Wed Apr 21 01:48:45 2004 From: jim.manavis <@t> imvs.sa.gov.au (jim) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] CGRP Message-ID: <00ac01c4276c$b16739e0$7a6c140a@imvs.sa.gov.au> Does anyone know of a good CGRP antibody for FFPE human tissue, and/or rat and mouse. Jim Manavis Laboratory Manager Hanson Institute Centre for Neurological Diseases Adelaide, SA, 5000 Australia Phone: 61-08-8222-3668 FAX: 61-08-8222 3204 email: jim.manavis@imvs.sa.gov.au Disclaimer: Not this little black duck! From chris.goodall <@t> bristol.ac.uk Wed Apr 21 04:50:12 2004 From: chris.goodall <@t> bristol.ac.uk (anclg) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Macneals Tetrachrome Message-ID: <000101c42786$0a52dc80$8f6ede89@hist143> Hi everyone, I`m ordering the stains to make up Macneals Tetrachrome, I really like the sound of this, but Sigma sell two methylene violets one with C.I. 52041 and one with C.I. 50206, can anyone tell me which one to order? Bye the way , I am enjoying the saline in the microwave saga.:-) Many thanks. Chris Goodall From francesca.wannenes <@t> uniroma2.it Wed Apr 21 05:54:53 2004 From: francesca.wannenes <@t> uniroma2.it (Francesca Wannenes) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] IHC for PECAM-1 Message-ID: <001c01c4278f$1514ae60$c13450a0@Silvia> Hallo! I want to test the PECAM-1 expression in PC3 xenograft tumors. I have the primary antibody PECAM-1 (M20) and the ABC staining system of Santa Cruz Biotechnology. My samples are paraffin-formalin embedded. Have you a method to use these antibodies for the immunohistochemistry? If not, have you similar methods for immunohistochemistry for to test CD31 (PECAM-1) expression? Thank you to everybody Francesca From angela.mcnabola.b <@t> bayer.com Wed Apr 21 05:58:17 2004 From: angela.mcnabola.b <@t> bayer.com (Angela McNabola) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] IHC for PECAM-1 Message-ID: Hi Francesca, We have used this antibody on several xenographs (including PC3) with nice results. Incidentally, this is the only PECAM-1 (M20) from SC that we have gotten to work in FFPE tissues. We do it on an automated stainer, but it can be done offline as well. Here is our protocol: 1. 5um paraffin sections were deparaffinized in xylene and brought to water through a graded series of alcohols. 2. Antigen retrieval was performed using Dako Target Retrieval Solution (DakoCytomation, S1699, Carpinteria,CA). Buffer was prepared according to package insert. Slides were placed in heated buffer and steamed (in a pre-heated Black and Decker steamer) for 30 minutes, then cooled for 20 minutes at room temperature. 3. Slides were rinsed in Dako wash buffer (S3006, DakoCytomation, Carpinteria, CA) and then placed in 3% hydrogen peroxide (in water) for 5 minutes to block endogenous peroxides. 4. Slides were rinsed with Dako wash buffer and an avidin /biotin block was performed using an Avidin/Biotin Blocking kit (Vector Laboratories, Inc., Burlingame CA, cat# SP-2001). Rabbit serum was used as the protein block (30 minute incubation), and the Avidin and Biotin were incubated for 10 minutes each. 5. Immunohistochemistry was performed using a goat Vectastain ABC elite kit (Vector Laboratories, cat # PK-6105) following package insert beginning at the primary antibody step with the following modifications: two thirty minute incubations were performed with primary antibody and following the incubation of the secondary antibody slides were rinsed in buffer, distilled water, and buffer again. Primary antibody used with the kit was CD31 antibody (PECAM-1 (M-20) SC-1506 goat polyclonal, 200 ug/ml, Santa Cruz Biotechnology) at 1:750 (using Dako Antibody Diluent ? cat# S0809, DakoCytomation, Carpinteria, CA). Goat IgG (1:750) ( code# 005-000-003, Jackson Immuno Research Laboratories, INC., West Grove, PA) and rabbit serum (protein block) were used as negative controls. DAB was the chromagen used (DakoCytomation., Carpinteria Ca.). 6. Slides were counterstained with Hematoxylin (Automation Hematoxylin, S3301, DakoCytomation, Carpinteria,CA). "Francesca Wannenes" t> cc: Sent by: Subject: [Histonet] IHC for PECAM-1 histonet-bounces@lists.utsouth western.edu 04/21/2004 06:54 AM Hallo! I want to test the PECAM-1 expression in PC3 xenograft tumors. I have the primary antibody PECAM-1 (M20) and the ABC staining system of Santa Cruz Biotechnology. My samples are paraffin-formalin embedded. Have you a method to use these antibodies for the immunohistochemistry? If not, have you similar methods for immunohistochemistry for to test CD31 (PECAM-1) expression? Thank you to everybody Francesca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Wed Apr 21 06:36:39 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:50 2005 Subject: FW: [Histonet] Microwave Processing Message-ID: 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 20 April 2004 14:55 To: Marshall Terry Dr, Consultant Histopathologist Subject: RE: [Histonet] Microwave Processing Hi Terry & HistoNetters Yes, in the Leong method for biopsies, the specimens are heat stabilized in the microwave in saline, and not really chemically fixed. They get fixed in the ethanol in a traditional processor (Leong used ethanol, chloroform and paraffin- no xylene). I haven't seen the resulting morphology myself. As for breasts, the Milestone unit has a module, the DDG, which is 280mm x 250mm, for stabilization of breasts and other large organs. One advantage with breasts is that the nodes just light up bright white. Parenthetically, I don't believe that the breasts in Yorkshire could possibly be bigger than in the American South... best regards, Steven >Thanks Steven. It was the Leong method to which I referred. >I am totally befuddled over fixation in the microwave. Surely in the >Leong method (for blocks), the saline is irrelevent other than as a >carrier and buffer medium, and it is heat fixation? > >Firming up breasts in the microwave seems fun - but the breasts in >Yorkshire are bigger than the microwaves:-) From abright <@t> brightinstruments.com Wed Apr 21 06:48:54 2004 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] fat cryostat sections: cutting pb, IHC? Message-ID: Nancy, The sample you are trying to section should not be a problem, it looks to me that your knife temperature and anti-roll plate are not cold enough, you omitted this temperature in your email perhaps you could inform me. Also you may need to have the specimen slightly warmer. I hope this assists you. 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: Nancy.Walker@sanofi-synthelabo.com [mailto:Nancy.Walker@sanofi-synthelabo.com] Sent: 20 April 2004 15:53 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fat cryostat sections: cutting pb, IHC? Any pointers for cutting adipose tissue with a cryostat? I have mouse epididyme adipose tissue that is unfixed and embedded in tissue tek. At - 30?C I can cut without it melting but most of the specimen stays on the knife edge leaving a nicely cut section of tissue tek, a big hole and a little fat tissue around the edges. And this at 45 ?!!! Any thinner and I get just the hole. I have a crostat which I can adjust the block and chamber temp separately. Haven't fiddled with the knife angle yet... WHAT TO DO??? any special tratments to get IHC reagents to penetrate this hydrophobic stuff? thank you for you advice, Nancy Walker Molecular Pharmacology Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Wed Apr 21 06:48:12 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Microwave Processing Message-ID: My confusion has not been dispelled. Steven writes: "The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- ..." In the next post, we get: "Yes, in the Leong method for biopsies, the specimens are heat stabilized in the microwave in saline, and not really chemically fixed. They get fixed in the ethanol in a traditional processor ..." These statements are mutually incompatible. Moreover, what is the difference between "stabilisation" and "fixation". A further muddle is introduced by: "not really chemically fixed." Well, hell, are they fixed or no? Heat fixation is fixation - who cares whether chemically fixed? Of course, the major problem is that mechanisms of fixation are varied (with the fixative) and not well understood. (Speak for yourself do I hear someone say?) 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 17 April 2004 22:54 To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hi Terry & HistoNetters I have done lots of successful large organ stabilization in saline at 65?C for 20-35 minutes (depending on the size of the organ. However, it is important to emphasize that the purpose of this step usually is to firm up the tissue so that it can be subsequently cut up into thinner pieces for processing, which must include a fixation step, either in the microwave or on a conventional processor, in formalin or some non-formalin fixative. The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- a procedure I have not tried myself. best regards, Steven Slap Microwave Consultant At 3:58 PM +0100 4/16/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Sorry to but in - but on a related question - >does anyone do microwave fixation in saline as >described by for instance the Melbourne group, >which involves bringing up to 65C in saline? >Have tried it this week and the sections are even crummier than usual. >When I was in Tasmania, a local private lab. did >it and their sections were fine. From MAUGER <@t> email.chop.edu Wed Apr 21 06:51:34 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Coverslipper Agony Message-ID: Pat, We had crazy problems with our Hacker too, until I realized we were using a toluene based mounting medium(which was recommended by the salesperson!) but we were mounting from xylene. The toluene dries much quicker, and not only did we get bubbles, after a while the coverslips were popping off! Switch to a xylene based mountant like Protocol from Fisher and hopefully your problem will be solved. Good luck, Jo >>> "Patterson, Patricia" 04/20/04 02:08PM >>> Hi All! I'd like some suggestions from you - we have a Hacker Coverslipper - using glass coverslips - and we get air bubbles. Not all the time, but enough to drive us crazy. We try to keep the dispensing needle clean and the amount of xylene in the rack holder consistent. What is everyone else doing?? Thanks Pat Patterson University Hospitals of Cleveland. The enclosed information is STRICTLY CONFIDENTIAL and is intended for the use of the addressee only. University Hospitals Health System and its affiliates disclaim any responsibility for unauthorized disclosure of this information to anyone other than the addressee. Federal and Ohio law protect patient medical information disclosed in this email, including psychiatric disorders, (HIV) test results, AIDs-related conditions, alcohol, and/or drug dependence or abuse. Federal regulation (42 CFR Part 2) and Ohio Revised Code section 5122.31 and 3701.243 prohibit disclosure of this information without the specific written consent of the person to whom it pertains, or as otherwise permitted by law. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ian.montgomery <@t> bio.gla.ac.uk Wed Apr 21 06:52:24 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:22:50 2005 Subject: Fwd: [Histonet] Mesentery tissue Message-ID: <6.0.1.1.2.20040421122801.02d67160@udcf.gla.ac.uk> Donna, For smaller animals I dissect the small intestine from stomach to rectum as one piece. Then cut lengths of the gut, with mesentery attached, and pin onto cork mats keeping the mesentery stretched. Fix in your fixative of choice. After an appropriate fixing time, remove from the cork mat and trim the mesentery away from the intestine. You'll now have sheets of fixed mesentery that you can stain as appropriate. For ease in handling I normally stain the entire sheet and cut into smaller pieces just before mounting. H&E staining time is just as normal but remember, there are large blood vessels and depending on animal quite a lot of fat in the tissue, so increase the dehydration time accordingly. In fact, I trim away the large vessels and fat during dehydration. If you want to study the blood vessels in TS then dissect out the mesentery, spread it out then roll up like a Swiss roll. Tie both ends and stretch lightly over a cork mat. Fix using your fixative of choice. A slot in the cork mat gives the fixative better access to the tissue. Dehydrate as one long piece then cut into block size pieces just before embedding. Cut and stain as normal. It can be a wee bit fiddly at first, but you'll soon master the technique. Ian. >X-Mailer: Novell GroupWise Internet Agent 6.5.2 Beta >Date: Tue, 20 Apr 2004 09:40:37 -0400 >From: "Donna Kumiski" >To: >X-Scan-Signature: ab3f4c55490e4d672805895add812bc5 >Cc: >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.3 >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: 7bf0d7e947a05afa907d3090d45e1f8d >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] Mesentery tissue >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=6.5 tests=none autolearn=no version=2.63 >X-SA-Exim-Version: 3.1 (built Tue Oct 14 16:21:02 CDT 2003) >X-SA-Exim-Scanned: Yes >X-GLA-Spam-Scan: R >X-GLA-Spam-Score: 0.0 (/) >X-GLA-Spam-Report: > > >I am looking for a protocol to stain and mount mesentery. >In a past discussion I asked about mounting the tissue but I need > more details about the staining. I would like to do an H&E on the >tissue then mount it on slides. These slides will be used in a >Histology >course for first year medical students > Donna > > > >---------------------------------------------- >Donna Kumiski HT (ASCP) >Medical College of Georgia >Dept. of Cellular Biology and Anatomy >CB 1202 >Augusta, Georgia 30912-2000 >(706) 721-6278 > dkumiski@mail.mcg.edu >----------------------------------------------- > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk From DKUMISKI <@t> mail.mcg.edu Wed Apr 21 07:04:58 2004 From: DKUMISKI <@t> mail.mcg.edu (Donna Kumiski) Date: Fri Sep 16 15:22:50 2005 Subject: Fwd: [Histonet] Mesentery tissue Message-ID: Ian, Should I mount the smaller pieces of the sheet with an aqueous mounting media? If so, what would you recommend? Or should I clear after dehydration and mount with a permenent mounting media. Cheers, Donna ---------------------------------------------- Donna Kumiski HT (ASCP) Medical College of Georgia Dept. of Cellular Biology and Anatomy CB 1202 Augusta, Georgia 30912-2000 (706) 721-6278 dkumiski@mail.mcg.edu ----------------------------------------------- From kmerriam2003 <@t> yahoo.com Wed Apr 21 07:41:18 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Re: Proteoglycan - decalcification - Why safranine? In-Reply-To: <4085F708.BA069836@uwo.ca> Message-ID: <20040421124118.77048.qmail@web10003.mail.yahoo.com> At my last job, we were looking for retention of glycosaminoglycans, in both soft tissue and bone. We would fix the tissue in methacarn (methyl carnoys) and decal in 10% formic acid containing ion exchange resin. EDTA will cause a severe decrease GAG retention in the bone (we tested this decal method for GAG retention, and it was amazing how different it was than with the formic acid decal method). Good luck, Kim Merriam Novartis Cambridge, MA John Kiernan wrote: Dear Dr Balaji, Adequate fixation is important before you decalcify anything. That means at least 24 hours (a week is probably better) in a neutral buffered (or any other) formaldehyde solution. Is your "Saf/FG" method safranine O followed by fast green FCF? There are many such methods. One group is, to plant anatomists, what H&E is to animal pathologists. (I apologize for using nouns as adjectives, implying that anatomists are plants whereas pathologists are animals.) The other lot of safranine-fast green methods are the stuff of traditional light microscope cytology, for showing chromosomes and cytoplasmic organelles in truly thin paraffin sections of tiny bits of tissues fixed in solutions containing osmium tetroxide and potassium dichromate. I have used both groups of methods. The plant anatomy ones are straightforward if you follow the instructions, which include checking with a microscope before making the preparation a permanent one - just like H&E(!). (My limited trials of safranine and fast green FCF for traditional cytology were all dismal failures; the colours initially seen were always largely lost in the dehydrating alcohol.) If you need to stain proteoglycans in cartilage, why not use an easy staining method that does not involve critical differentiations? There are many such methods. In cartilage, they are very clean and selective. Sulphate ions (in the chondroitin sulphate glycosaminoglycans of the proteoglycans) bind cationic dyes at pH 1.0 or even lower. Alcian blue at pH 1 can be followed by any counterstain because alcian blue changes into a permanently insoluble pigment while the unbound dye is being washed out of the sections. Carbohydrate histochemistry is a large subject. It includes many inexpensive but discriminating techniques that are neglected by well funded researchers and clinical pathologists. John A. Kiernan London, Canada. _______________________________________ balajimr@drreddys.com wrote: > Thank you very much for your replies to my earlier mails. This is with > regard to Saf/FG staining of cartilage. I want to demonstrate > proteoglycans in cartilage of femur joint, tarsal joint and ankle joint. I > am in the process of standardization of the same. I have taken a trial > tissue of joints from Rat and I have used 20% EDTA for decalcification. I > read somewhere that it affects the proteoglycan staining. Is it true? If > so could you please suggest a good decalcifier for the same.Also let me > know how long I can fix these joints for good fixation in NBF. Thanks in > advance. > > Dr. Balaji > Dept. Pre clinical safety evaluation, > Discovery research, > Dr. Reddys Laboratories ltd. Bollaram Road, > Miyapur, Hyderabad, 500 049 > Andhra Pradseh, INDIA -------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Photos: High-quality 4x6 digital prints for 25¢ From Barry.R.Rittman <@t> uth.tmc.edu Wed Apr 21 08:20:26 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Microwave Processing Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635A24@UTHEVS3.mail.uthouston.edu> The term fixation is a generic one that is a greatly misused one in the literature. We use the term fixation to include the following three steps. 1. When a fixative reaches tissue there is an in initial "killing" of the cells. 2. This is followed by a "stabilization" of components. The bonds that are formed are often temporary bonds that can be broken. (An example is the use of formalin for a short time of 1-2 hours, wehen the "temporary" bonds that have been formed can usually be broken by washing in running tap water.). This has been used to advantage in histochemistry when enzymes are prevented from diffusing but can still be demonstrated. 3. If tissue is left for a sufficient length of time then "fixation" occurs. The bonds that are formed are more permanent, more stable and often require the use of procedures such as antigen retrieval to break these bonds. This last step continues sometimes for years with more and more permanent bonds formed. This results in the increasing difficulty of staining tissues that have been fixed for a long period of time. It is important therefore when discussing fixation to note the duration of "fixation" and the specific conditions under which the fixative is applied. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Wednesday, April 21, 2004 6:48 AM To: Steven E. Slap; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing My confusion has not been dispelled. Steven writes: "The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- ..." In the next post, we get: "Yes, in the Leong method for biopsies, the specimens are heat stabilized in the microwave in saline, and not really chemically fixed. They get fixed in the ethanol in a traditional processor ..." These statements are mutually incompatible. Moreover, what is the difference between "stabilisation" and "fixation". A further muddle is introduced by: "not really chemically fixed." Well, hell, are they fixed or no? Heat fixation is fixation - who cares whether chemically fixed? Of course, the major problem is that mechanisms of fixation are varied (with the fixative) and not well understood. (Speak for yourself do I hear someone say?) 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 17 April 2004 22:54 To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hi Terry & HistoNetters I have done lots of successful large organ stabilization in saline at 65?C for 20-35 minutes (depending on the size of the organ. However, it is important to emphasize that the purpose of this step usually is to firm up the tissue so that it can be subsequently cut up into thinner pieces for processing, which must include a fixation step, either in the microwave or on a conventional processor, in formalin or some non-formalin fixative. The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- a procedure I have not tried myself. best regards, Steven Slap Microwave Consultant At 3:58 PM +0100 4/16/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Sorry to but in - but on a related question - >does anyone do microwave fixation in saline as >described by for instance the Melbourne group, >which involves bringing up to 65C in saline? >Have tried it this week and the sections are even crummier than usual. >When I was in Tasmania, a local private lab. did >it and their sections were fine. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ljb <@t> medicine.wisc.edu Wed Apr 21 08:49:49 2004 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] MacNeals Tetrachrome Message-ID: Good Morning Gail and Histonet! I've gone back through some past postings regarding MacNeal's tetrachrome but, have been unable to find the actual protocol. Would you be willing to share it? Thanks, Cindy LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 From mab70 <@t> medschl.cam.ac.uk Wed Apr 21 08:54:31 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Attention Gayle Callis Message-ID: <000601c427a8$2bf80500$42536f83@medlan.cam.ac.uk> Hi, Gayle, While searching the histonet archive for information about processing some rodent tissues, I came across a comment by yourself that it was planned to produce a book on this subject listing the best techniques. Is this book out yet and how can I get access to it? Incidentally the search facility on the histonet website is brilliant. Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR From kemlo <@t> tiscali.co.uk Wed Apr 21 09:16:10 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:50 2005 Subject: [Histonet] Microwave Processing In-Reply-To: Message-ID: <001601c427ab$378b1850$56c4e150@KEMLOS> Um....... A boiled egg is stabilized but not fixed; does that help? A fixative is designed not only to stabilize proteins by coagulation, but in most cases it links chemically to the proteins. Its job is also that of preventing putrefaction both by endogenous and exogenous means (lysosomes and bugs). A boiled egg will rot by putrefaction, but if you fixed it, then it wouldn't; would it? Heat stabilizes by coagulating protein doesn't it, formalin forms links between reactive sites on the proteins, doesn't it? Am I talking a load of........... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 21 April 2004 12:48 To: Steven E. Slap; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing My confusion has not been dispelled. Steven writes: "The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- ..." In the next post, we get: "Yes, in the Leong method for biopsies, the specimens are heat stabilized in the microwave in saline, and not really chemically fixed. They get fixed in the ethanol in a traditional processor ..." These statements are mutually incompatible. Moreover, what is the difference between "stabilisation" and "fixation". A further muddle is introduced by: "not really chemically fixed." Well, hell, are they fixed or no? Heat fixation is fixation - who cares whether chemically fixed? Of course, the major problem is that mechanisms of fixation are varied (with the fixative) and not well understood. (Speak for yourself do I hear someone say?) 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 17 April 2004 22:54 To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hi Terry & HistoNetters I have done lots of successful large organ stabilization in saline at 65?C for 20-35 minutes (depending on the size of the organ. However, it is important to emphasize that the purpose of this step usually is to firm up the tissue so that it can be subsequently cut up into thinner pieces for processing, which must include a fixation step, either in the microwave or on a conventional processor, in formalin or some non-formalin fixative. The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- a procedure I have not tried myself. best regards, Steven Slap Microwave Consultant At 3:58 PM +0100 4/16/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Sorry to but in - but on a related question - >does anyone do microwave fixation in saline as >described by for instance the Melbourne group, >which involves bringing up to 65C in saline? >Have tried it this week and the sections are even crummier than usual. >When I was in Tasmania, a local private lab. did >it and their sections were fine. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Wed Apr 21 09:00:23 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] ISH - A Review Article in Journal of Histotechnology Message-ID: For those of you out there doing ISH or wanting to learn more about it, there is a fantastic article in the March 2004 issue of The Journal of Histotechnology written by Xiang Qian, Long Jin, and Ricardo V. Lloyd. A must read! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From Terry.Marshall <@t> rothgen.nhs.uk Wed Apr 21 09:51:02 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing Message-ID: Kemlo asks: "Am I talking a load of..........." Inevitably:-) Kemlo, You talk as if formalin was the only fixative. What about the group called, oddly enough, coagulative fixatives? Now .... I wonder how they work. Let me see ...... Furthermore, you flit between the several different subjects in an unstructured way. When you say : "Its job is also that of preventing putrefaction both by endogenous and exogenous means (lysosomes and bugs)." You are wrong on 2 counts. Its effect may be to prevent putrefaction, but hardly its job. (I realise that some books list this, but a dehydrated specimen in wax is more at risk from mice than bacteria). Furthermore, the action you refer to, attributed to lysosomes, is autolysis, not putrefaction. Furtherfurthermore, a fixed egg will only elude putrefaction by virtue of the retained/residual fixative fixing any bacteria with ungodly thoughts toward the egg, wouldn't it? That is to say, "resistance to putrefaction" will be a property that resides in the fixative not in the fixed egg. Yours lovingly, Confused from Rotherham 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 21 April 2004 15:16 To: Marshall Terry Dr, Consultant Histopathologist; 'Steven E. Slap'; 'kevin williams'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Um....... A boiled egg is stabilized but not fixed; does that help? A fixative is designed not only to stabilize proteins by coagulation, but in most cases it links chemically to the proteins. Its job is also that of preventing putrefaction both by endogenous and exogenous means (lysosomes and bugs). A boiled egg will rot by putrefaction, but if you fixed it, then it wouldn't; would it? Heat stabilizes by coagulating protein doesn't it, formalin forms links between reactive sites on the proteins, doesn't it? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 21 April 2004 12:48 To: Steven E. Slap; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing My confusion has not been dispelled. Steven writes: "The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- ..." In the next post, we get: "Yes, in the Leong method for biopsies, the specimens are heat stabilized in the microwave in saline, and not really chemically fixed. They get fixed in the ethanol in a traditional processor ..." These statements are mutually incompatible. Moreover, what is the difference between "stabilisation" and "fixation". A further muddle is introduced by: "not really chemically fixed." Well, hell, are they fixed or no? Heat fixation is fixation - who cares whether chemically fixed? Of course, the major problem is that mechanisms of fixation are varied (with the fixative) and not well understood. (Speak for yourself do I hear someone say?) 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 17 April 2004 22:54 To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hi Terry & HistoNetters I have done lots of successful large organ stabilization in saline at 65?C for 20-35 minutes (depending on the size of the organ. However, it is important to emphasize that the purpose of this step usually is to firm up the tissue so that it can be subsequently cut up into thinner pieces for processing, which must include a fixation step, either in the microwave or on a conventional processor, in formalin or some non-formalin fixative. The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- a procedure I have not tried myself. best regards, Steven Slap Microwave Consultant At 3:58 PM +0100 4/16/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Sorry to but in - but on a related question - >does anyone do microwave fixation in saline as >described by for instance the Melbourne group, >which involves bringing up to 65C in saline? >Have tried it this week and the sections are even crummier than usual. >When I was in Tasmania, a local private lab. did >it and their sections were fine. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Wed Apr 21 10:37:42 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Chemokine staining Message-ID: We have a colleague that is somewhat new to IHC staining who is being asked to stain for a number of antigens in the chemokine class. The antibodies he was given to use have not been tried on FFPE tissues. We're not familiar with staining of this class of antigens and are wondering if anyone out in Histonet land has experience in this area. I'm thinking, for example, of having to permeabilize the cell nucleus for staining of cytokines. Any suggestions would be helpful as right now he's getting no staining at all. Thanks again, 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 From cfranci <@t> rigel.com Wed Apr 21 10:42:53 2004 From: cfranci <@t> rigel.com (Christian Franci) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 33 In-Reply-To: <20040421144046.81271168008@relay.rigel.com> Message-ID: Ciao! You use Santa Cruz Ab's? I've had problems with that Ab. Try a company called BioCare Medical. (www.biocare.net) I got their mAb CD31 to work great on Paraffin sections. They offer a solution for a one step HIER and deparaffinizing. Lastly, I've moved away from HRP development and now use Biocare's Vulcan Alkaline Phosphatase. I get beautiful, clean staining of my xenograft vessels for bright field and, as a bonus, the stain fluoresces hot pink under a Texas red filter... You get really cool pictures. Hope this helps! Arrivederci! C From kemlo <@t> tiscali.co.uk Wed Apr 21 10:51:33 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing In-Reply-To: Message-ID: <000001c427b8$8931e230$54332850@KEMLOS> Geez, tetchy begger aren't you. I'll answer when I've thunk about it and give you an answer that is both lucid and structured. this used to be fun; why are some people so...... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 21 April 2004 15:51 To: Kemlo Rogerson; Steven E. Slap; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Kemlo asks: "Am I talking a load of..........." Inevitably:-) Kemlo, You talk as if formalin was the only fixative. What about the group called, oddly enough, coagulative fixatives? Now .... I wonder how they work. Let me see ...... Furthermore, you flit between the several different subjects in an unstructured way. When you say : "Its job is also that of preventing putrefaction both by endogenous and exogenous means (lysosomes and bugs)." You are wrong on 2 counts. Its effect may be to prevent putrefaction, but hardly its job. (I realise that some books list this, but a dehydrated specimen in wax is more at risk from mice than bacteria). Furthermore, the action you refer to, attributed to lysosomes, is autolysis, not putrefaction. Furtherfurthermore, a fixed egg will only elude putrefaction by virtue of the retained/residual fixative fixing any bacteria with ungodly thoughts toward the egg, wouldn't it? That is to say, "resistance to putrefaction" will be a property that resides in the fixative not in the fixed egg. Yours lovingly, Confused from Rotherham 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 21 April 2004 15:16 To: Marshall Terry Dr, Consultant Histopathologist; 'Steven E. Slap'; 'kevin williams'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Um....... A boiled egg is stabilized but not fixed; does that help? A fixative is designed not only to stabilize proteins by coagulation, but in most cases it links chemically to the proteins. Its job is also that of preventing putrefaction both by endogenous and exogenous means (lysosomes and bugs). A boiled egg will rot by putrefaction, but if you fixed it, then it wouldn't; would it? Heat stabilizes by coagulating protein doesn't it, formalin forms links between reactive sites on the proteins, doesn't it? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 21 April 2004 12:48 To: Steven E. Slap; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing My confusion has not been dispelled. Steven writes: "The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- ..." In the next post, we get: "Yes, in the Leong method for biopsies, the specimens are heat stabilized in the microwave in saline, and not really chemically fixed. They get fixed in the ethanol in a traditional processor ..." These statements are mutually incompatible. Moreover, what is the difference between "stabilisation" and "fixation". A further muddle is introduced by: "not really chemically fixed." Well, hell, are they fixed or no? Heat fixation is fixation - who cares whether chemically fixed? Of course, the major problem is that mechanisms of fixation are varied (with the fixative) and not well understood. (Speak for yourself do I hear someone say?) 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 17 April 2004 22:54 To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hi Terry & HistoNetters I have done lots of successful large organ stabilization in saline at 65?C for 20-35 minutes (depending on the size of the organ. However, it is important to emphasize that the purpose of this step usually is to firm up the tissue so that it can be subsequently cut up into thinner pieces for processing, which must include a fixation step, either in the microwave or on a conventional processor, in formalin or some non-formalin fixative. The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- a procedure I have not tried myself. best regards, Steven Slap Microwave Consultant At 3:58 PM +0100 4/16/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Sorry to but in - but on a related question - >does anyone do microwave fixation in saline as >described by for instance the Melbourne group, >which involves bringing up to 65C in saline? >Have tried it this week and the sections are even crummier than usual. >When I was in Tasmania, a local private lab. did >it and their sections were fine. _______________________________________________ Histonet mailing list Histonet@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 Terry.Marshall <@t> rothgen.nhs.uk Wed Apr 21 11:06:14 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing Message-ID: Kemlo feebly blurts, for want of anything intelligent to say: "Geez, tetchy begger aren't you. I'll answer when I've thunk about it and give you an answer that is both lucid and structured." You can't see the twinkle in my eyes? 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 21 April 2004 16:52 To: Marshall Terry Dr, Consultant Histopathologist; 'Steven E. Slap'; 'kevin williams'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing this used to be fun; why are some people so...... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 21 April 2004 15:51 To: Kemlo Rogerson; Steven E. Slap; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Kemlo asks: "Am I talking a load of..........." Inevitably:-) Kemlo, You talk as if formalin was the only fixative. What about the group called, oddly enough, coagulative fixatives? Now .... I wonder how they work. Let me see ...... Furthermore, you flit between the several different subjects in an unstructured way. When you say : "Its job is also that of preventing putrefaction both by endogenous and exogenous means (lysosomes and bugs)." You are wrong on 2 counts. Its effect may be to prevent putrefaction, but hardly its job. (I realise that some books list this, but a dehydrated specimen in wax is more at risk from mice than bacteria). Furthermore, the action you refer to, attributed to lysosomes, is autolysis, not putrefaction. Furtherfurthermore, a fixed egg will only elude putrefaction by virtue of the retained/residual fixative fixing any bacteria with ungodly thoughts toward the egg, wouldn't it? That is to say, "resistance to putrefaction" will be a property that resides in the fixative not in the fixed egg. Yours lovingly, Confused from Rotherham 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 21 April 2004 15:16 To: Marshall Terry Dr, Consultant Histopathologist; 'Steven E. Slap'; 'kevin williams'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Um....... A boiled egg is stabilized but not fixed; does that help? A fixative is designed not only to stabilize proteins by coagulation, but in most cases it links chemically to the proteins. Its job is also that of preventing putrefaction both by endogenous and exogenous means (lysosomes and bugs). A boiled egg will rot by putrefaction, but if you fixed it, then it wouldn't; would it? Heat stabilizes by coagulating protein doesn't it, formalin forms links between reactive sites on the proteins, doesn't it? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 21 April 2004 12:48 To: Steven E. Slap; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing My confusion has not been dispelled. Steven writes: "The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- ..." In the next post, we get: "Yes, in the Leong method for biopsies, the specimens are heat stabilized in the microwave in saline, and not really chemically fixed. They get fixed in the ethanol in a traditional processor ..." These statements are mutually incompatible. Moreover, what is the difference between "stabilisation" and "fixation". A further muddle is introduced by: "not really chemically fixed." Well, hell, are they fixed or no? Heat fixation is fixation - who cares whether chemically fixed? Of course, the major problem is that mechanisms of fixation are varied (with the fixative) and not well understood. (Speak for yourself do I hear someone say?) 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: Steven E. Slap [mailto:siksik03@comcast.net] Sent: 17 April 2004 22:54 To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hi Terry & HistoNetters I have done lots of successful large organ stabilization in saline at 65?C for 20-35 minutes (depending on the size of the organ. However, it is important to emphasize that the purpose of this step usually is to firm up the tissue so that it can be subsequently cut up into thinner pieces for processing, which must include a fixation step, either in the microwave or on a conventional processor, in formalin or some non-formalin fixative. The saline will not work alone as a fixation step, except in the case of very small biopsies (as written up in an article by Tony Leong)- a procedure I have not tried myself. best regards, Steven Slap Microwave Consultant At 3:58 PM +0100 4/16/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Sorry to but in - but on a related question - >does anyone do microwave fixation in saline as >described by for instance the Melbourne group, >which involves bringing up to 65C in saline? >Have tried it this week and the sections are even crummier than usual. >When I was in Tasmania, a local private lab. did >it and their sections were fine. _______________________________________________ Histonet mailing list Histonet@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 bryand <@t> netbistro.com Wed Apr 21 11:12:52 2004 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Macneals Tetrachrome In-Reply-To: <000101c42786$0a52dc80$8f6ede89@hist143> References: <000101c42786$0a52dc80$8f6ede89@hist143> Message-ID: <40869D84.1080703@netbistro.com> Conn specifically comments that Bernthen's methylene violet is used in MacNeal's stain. That is CI #52041 according to Aldrich. Bryan Llewellyn anclg wrote: > Hi everyone, > I`m ordering the stains to make up Macneals Tetrachrome, I really > like the sound of this, but Sigma sell two methylene violets one with > C.I. 52041 and one with C.I. 50206, can anyone tell me which one to > order? Bye the way , I am enjoying the saline in the microwave saga.:-) > Many thanks. Chris Goodall > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From limberis <@t> mail.med.upenn.edu Wed Apr 21 11:34:19 2004 From: limberis <@t> mail.med.upenn.edu (Maria Limberis) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing In-Reply-To: Message-ID: <5.1.3.2.2.20040421122338.00b6fd38@mail.med.upenn.edu> Hi everyone- I have instilled a vector expressing human placental Alkaline Phosphatase in the lung of rhesus monkeys. However, when I use a protocol routinely used in our lab to prevent background in mouse lung (which works very well) the background is very high preventing assessment of gene transfer. I am wondering if anyone has any suggestions or ideas! Much appreciated Maria ------------------------------------------------------------------------- Maria Limberis, PhD Gene Therapy Program University of Pennsylvania School of Medicine Department of Medicine Division of Medical Genetics The Wistar Institute, Room 201 3601 Spruce Street Philadelphia, PA 19104 Tel: 215 8987166 Fax: 215 5737414 Email: limberis@mail.med.upenn.edu From kspencer <@t> utmem.edu Wed Apr 21 11:43:29 2004 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing In-Reply-To: <000001c427b8$8931e230$54332850@KEMLOS> Message-ID: <04B8DCB2-93B3-11D8-86DE-000393967904@utmem.edu> Do you guys know each other? I love this battle of the minds, but who am I really to believe? You have truly brightened my day and made me laugh when I was blue. -Kathleen On Wednesday, April 21, 2004, at 10:51 AM, Kemlo Rogerson wrote: > Geez, tetchy begger aren't you. I'll answer when I've thunk about it and > give you an answer that is both lucid and structured. > > this used to be fun; why are some people so...... > > Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS > Tel: 0208 970 8414 > Mob: 07830 196072 > Mobile E-Mail kemlorogerson@3mail.com > FAX & Answer Phone 0871 242 8094 > E-mail Accounts: > kemlo@tiscali.co.uk or > kemlo1@btinternet.com > Disclaimer: The information contained in this message and/or any > attachments(s) may be of a private and confidential nature, and is > intended solely for the attention of the addressee. If you have received > this message in error or feel you should not have been the intended > recipient, please return it and any attachments to the sender > immediately. All messages relating to this communication should then be > deleted from your system. Unauthorised usage, copying, disclosure or > alteration of this message and/or attachment(s) is strictly prohibited. > Barking, Havering and Redbridge Hospitals NHS Trust will not be held > responsible for any direct or indirect damages which may arise from > alteration of this message or any attachment(s), by a third party or > resulting from the transmission of a virus. > > > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall > Terry Dr,Consultant Histopathologist > Sent: 21 April 2004 15:51 > To: Kemlo Rogerson; Steven E. Slap; kevin williams; > histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > Kemlo asks: > > "Am I talking a load of..........." > > Inevitably:-) > > Kemlo, > > You talk as if formalin was the only fixative. What about the group > called, oddly enough, coagulative fixatives? > Now .... I wonder how they work. Let me see ...... > Furthermore, you flit between the several different subjects in an > unstructured way. > > When you say : > "Its job is also that of preventing putrefaction both by endogenous and > exogenous means > (lysosomes and bugs)." > > You are wrong on 2 counts. > Its effect may be to prevent putrefaction, but hardly its job. (I > realise that some books list this, but a dehydrated specimen in wax is > more at risk from mice than bacteria). > Furthermore, the action you refer to, attributed to lysosomes, is > autolysis, not putrefaction. > > Furtherfurthermore, a fixed egg will only elude putrefaction by virtue > of the retained/residual fixative fixing any bacteria with ungodly > thoughts toward the egg, wouldn't it? > That is to say, "resistance to putrefaction" will be a property that > resides in the fixative not in the fixed egg. > > Yours lovingly, > > Confused from Rotherham > > 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] > Sent: 21 April 2004 15:16 > To: Marshall Terry Dr, Consultant Histopathologist; 'Steven E. Slap'; > 'kevin williams'; histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > > Um....... A boiled egg is stabilized but not fixed; does that help? A > fixative is designed not only to stabilize proteins by coagulation, but > in most cases it links chemically to the proteins. Its job is also that > of preventing putrefaction both by endogenous and exogenous means > (lysosomes and bugs). A boiled egg will rot by putrefaction, but if you > fixed it, then it wouldn't; would it? Heat stabilizes by coagulating > protein doesn't it, formalin forms links between reactive sites on the > proteins, doesn't it? > > > > Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS > Tel: 0208 970 8414 > Mob: 07830 196072 > Mobile E-Mail kemlorogerson@3mail.com > FAX & Answer Phone 0871 242 8094 > E-mail Accounts: > kemlo@tiscali.co.uk or > kemlo1@btinternet.com > Disclaimer: The information contained in this message and/or any > attachments(s) may be of a private and confidential nature, and is > intended solely for the attention of the addressee. If you have received > this message in error or feel you should not have been the intended > recipient, please return it and any attachments to the sender > immediately. All messages relating to this communication should then be > deleted from your system. Unauthorised usage, copying, disclosure or > alteration of this message and/or attachment(s) is strictly prohibited. > Barking, Havering and Redbridge Hospitals NHS Trust will not be held > responsible for any direct or indirect damages which may arise from > alteration of this message or any attachment(s), by a third party or > resulting from the transmission of a virus. > > > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall > Terry Dr,Consultant Histopathologist > Sent: 21 April 2004 12:48 > To: Steven E. Slap; kevin williams; histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > My confusion has not been dispelled. > > Steven writes: > > "The saline will not work > alone as a fixation step, except in the case of > very small biopsies (as written up in an article > by Tony Leong)- ..." > > In the next post, we get: > > "Yes, in the Leong method for biopsies, the specimens are heat > stabilized in the microwave in saline, and not really chemically > fixed. They get fixed in the ethanol in a traditional processor ..." > > These statements are mutually incompatible. > > Moreover, what is the difference between "stabilisation" and "fixation". > > A further muddle is introduced by: > > "not really chemically fixed." > > Well, hell, are they fixed or no? Heat fixation is fixation - who cares > whether chemically fixed? > > Of course, the major problem is that mechanisms of fixation are varied > (with the fixative) and not well understood. > (Speak for yourself do I hear someone say?) > > 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: Steven E. Slap [mailto:siksik03@comcast.net] > Sent: 17 April 2004 22:54 > To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; > histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > > Hi Terry & HistoNetters > > I have done lots of successful large organ > stabilization in saline at 65?C for 20-35 minutes > (depending on the size of the organ. However, it > is important to emphasize that the purpose of > this step usually is to firm up the tissue so > that it can be subsequently cut up into thinner > pieces for processing, which must include a > fixation step, either in the microwave or on a > conventional processor, in formalin or some > non-formalin fixative. The saline will not work > alone as a fixation step, except in the case of > very small biopsies (as written up in an article > by Tony Leong)- a procedure I have not tried > myself. > > best regards, > Steven Slap > Microwave Consultant > > > At 3:58 PM +0100 4/16/04, Marshall Terry Dr, > Consultant Histopathologist wrote: >> Sorry to but in - but on a related question - >> does anyone do microwave fixation in saline as >> described by for instance the Melbourne group, >> which involves bringing up to 65C in saline? >> Have tried it this week and the sections are even crummier than usual. >> When I was in Tasmania, a local private lab. did >> it and their sections were fine. > > > _______________________________________________ > Histonet mailing list > Histonet@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 akwilliams75 <@t> hotmail.com Wed Apr 21 12:01:56 2004 From: akwilliams75 <@t> hotmail.com (kevin williams) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] OSHA compliance, Procedures and "Assessments of Potential Hazards" Message-ID: Dear all: Can I use my procedure manual as my compliance with the OSHA regulation requireing me to do a hazard check on all my work practices? A procedure manual is set up to tell people how to perform a specific task in a safe set of steps with a section on the safty precautions laid out. Am I reinventing the wheel? Thanks, A. Kevin Williams Vermont Dermatopathology _________________________________________________________________ [1]Check out MSN PC Safety & Security to help ensure your PC is protected and safe. References 1. http://g.msn.com/8HMAENUS/2749??PS= From gcallis <@t> montana.edu Wed Apr 21 12:14:38 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Macneals Tetrachrome In-Reply-To: <000101c42786$0a52dc80$8f6ede89@hist143> Message-ID: <3.0.6.32.20040421111438.00bc58f8@gemini.msu.montana.edu> Chris, The CI 52041 is the correct methylene violet. MacNeals used this, and it also has a common name of Bernthsen. At 10:50 AM 4/21/2004 +0100, you wrote: >Hi everyone, > I`m ordering the stains to make up Macneals Tetrachrome, I really >like the sound of this, but Sigma sell two methylene violets one with >C.I. 52041 and one with C.I. 50206, can anyone tell me which one to >order? Bye the way , I am enjoying the saline in the microwave saga.:-) >Many thanks. Chris Goodall > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed Apr 21 12:24:39 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] MacNeals Tetrachrome stain solution recipe In-Reply-To: Message-ID: <3.0.6.32.20040421112439.00bc58f8@gemini.msu.montana.edu> Dear LaCinda, Here is the recipe you are looking for. I have stained PMMA embedded bone sections, thicker/ground with this, but I did use 1% formic acid etching for 30 seconds in ultrasonicator, a very good water rinse after this, then dry section and immerse. This stain can also be combined with Toluidine Blue, Sterchi method for even more brilliant results particularly if you want to demonstrate ostoid! MACNEAL'S TETRACHROME STOCK SOLUTION 0.5g methylene blue 0.8g azure A eosinate 0.1g methylene violet CI# 52041 250 ml methanol 250 glycerin Stir, leave at 50C for 12 hours, then 37C for 3 days. Filter, store in dark. It is good for up to years! Older solutions stain very well, if not better. WORKING STAINING SOLUTION 5 mls stock stain solution, 95mls distilled water Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed Apr 21 12:30:09 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Animal Tissue Processing Manual In-Reply-To: <000601c427a8$2bf80500$42536f83@medlan.cam.ac.uk> Message-ID: <3.0.6.32.20040421113009.00bc52b0@gemini.msu.montana.edu> Margaret, Diane Sterchi and I were editors on the National Society for Histotechnology, Veterinary, Industry and Research Committee (VIR) Animal Tissue Processing Manual. It has many processing schedules for different species, also tissues ie. eyes, bone, etc. Booklet comes with a CD. Contact NSH at Histo@NSH.org or go onto their website on how to order. You will find this manual listed under education in their webpage for ordering instructions. At 02:54 PM 4/21/2004 +0100, you wrote: >Hi, Gayle, > >While searching the histonet archive for information about processing some >rodent tissues, I came across a comment by yourself that it was planned to >produce a book on this subject listing the best techniques. Is this book out >yet and how can I get access to it? Incidentally the search facility on the >histonet website is brilliant. > >Margaret > >Margaret Blount >Chief Technician >Clinical Biochemistry >University of Cambridge >Addenbrooke's Hospital >Hills Road >Cambridge >CB2 2QR > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From sharon.willman <@t> bms.com Wed Apr 21 13:11:53 2004 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] S-100 Protein Message-ID: <4086B969.37F697BB@bms.com> I was wondering if anyone has a method for S-100 protein staining method. Any help would be most appreciated. Thanks in advance, Sharon Willman From siksik03 <@t> comcast.net Wed Apr 21 13:17:45 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing In-Reply-To: References: Message-ID: Hi Terry & HistoNetters Let me take a stab at this, although I am not a chemist (Dick Dapson, where are you when I really need you?)... Microwave stabilization uses heat to harden the tissue through a process of coagulation of the proteins in the tissue. Kok & Boon liken this to poaching an egg. Alcoholic "fixation" also hardens the tissue by coagulation of the proteins, but also removes the water (the higher the percentage of alcohol, the more the hardening effect). The use of alcohol as the primary fixative results in a characteristic shrinkage within the tissue. Aldehydes "fix" the tissue through cross-linking of the proteins. When I talk about microwave stabilization not chemically "fixing" tissue, I mean to say that the proteins are not cross-linked. In Leong's work, the biopsies were microwave-stabilized in saline (hardened), then further hardened in graded ethanols on a conventional processor (normally referred to as "alcoholic fixation"), but still not cross-linked. With larger tissues, I would worry about shrinkage with this method, but Leong seemed to have success with it on biopsies. I still don't think they would look quite the same as formalin-fixed specimens. I'm sorry if I didn't make myself clearer. To muddy the waters further, some labs carry out microwave stabilization of gross tissue specimens in formalin at 50?C, rather than in saline at 65?C, but this still isn't fixation, because the formalin doesn't penetrate the tissue sufficiently to cross-link. Whew. best regards, Steven Slap Microwave Consultant At 12:48 PM +0100 4/21/04, Marshall Terry Dr, Consultant Histopathologist wrote: >My confusion has not been dispelled. > >Steven writes: > >"The saline will not work >alone as a fixation step, except in the case of >very small biopsies (as written up in an article >by Tony Leong)- ..." > >In the next post, we get: > >"Yes, in the Leong method for biopsies, the specimens are heat >stabilized in the microwave in saline, and not really chemically >fixed. They get fixed in the ethanol in a traditional processor ..." > >These statements are mutually incompatible. > >Moreover, what is the difference between "stabilisation" and "fixation". > >A further muddle is introduced by: > >"not really chemically fixed." > >Well, hell, are they fixed or no? Heat fixation >is fixation - who cares whether chemically fixed? > >Of course, the major problem is that mechanisms >of fixation are varied (with the fixative) and >not well understood. >(Speak for yourself do I hear someone say?) From gcallis <@t> montana.edu Wed Apr 21 13:35:04 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Animal Tissue Processing Manual from NSH Message-ID: <3.0.6.32.20040421123504.00bcd780@gemini.msu.montana.edu> It has been brought to my attention that ordering the Animal Tissue Processing Manual or rather finding it in the NSH website is a tidge difficult. So here it is: http://www.nsh.org/education/materials.html Booklet was placed in materials section under Education committee. It will be there at bottom of the webpage after all the self-assessment booklets. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From ROENN <@t> surgery.wisc.edu Wed Apr 21 13:46:26 2004 From: ROENN <@t> surgery.wisc.edu (Drew Allan Roenneburg) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Chemokine antibodies anti human MIG and I-TAC Message-ID: Hi everyone, I was wondering if anyone knows of a company who carries the two antibodies anti human MIG and I-TAC? And if so, has anyone ever used them successfully on FFPE tissue? Any information would be greatly appreciated. Thanks Drew Roenneburg UW Medical School Madison, WI From siksik03 <@t> comcast.net Wed Apr 21 14:09:26 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing In-Reply-To: References: Message-ID: Hello again HistoNetters Yes, there are these hybrid mixtures called coagulative fixatives. They are usually combinations of ethyl alcohol and polyethylene glycol, and, occasionally, acetic acid. They do not cross-link proteins. The PEG helps prevent the shrinkage associated with the use of ethanol as a fixative. The advantage of all these mixtures is that, no cross-linking having taken place, no subsequent antigen retrieval (or "unmasking") of the proteins would be necessary for IHC. Dr. Boon has a good chapter on them in support of her "Boon-Fix". To muddy the waters further, there are still other non-formalin fixatives, which are neither coagulative fixatives, nor cross-linking fixatives, based on glyoxal, such as Anatech's "Prefer", but which still are said to chemically "fix" the tissue. I'll leave the mechanism of their chemistry to the experts. best regards, Steven Slap At 3:51 PM +0100 4/21/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Kemlo asks: > >"Am I talking a load of..........." > >Inevitably:-) > >Kemlo, > >You talk as if formalin was the only fixative. What about the group >called, oddly enough, coagulative fixatives? >Now .... I wonder how they work. Let me see ...... >Furthermore, you flit between the several different subjects in an >unstructured way. From GDawson <@t> Milw.Dynacare.com Wed Apr 21 14:45:57 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] 88342 regulation comiserations Message-ID: All, I'm not looking for a quote on the rules and regulations on how and why only one 88342 can be billed on multiple sub-numbers from the same specimen. I am, however, going to share an example of how this rule can harm today's IHC lab. I just received a request for a 3 antibody cocktail to be performed on blocks A1 thru A10 from a given surgical case. While doing this work, it really ate at me that, due to the current IHC billing policies, my lab will be reimbursed one 88342 for what amounts to 10 88342's, not to mention that there is triple the amount of usual antibody in this cocktail. That's nine 88342's and 27 antibody applications that my lab gets to eat. When I think of all the sentinel node protocols that I perform which inevitably contain multiple blocks from the same specimen it starts to worry me. If I get one more pathologist who is gun-shy about possible liabilities that may arise if they don't cover all the blocks in a given specimen or flat out refuses to pick a representative block based on the morphology staring them in the face on the H&E I think I'll have to declare bankrupcy. I know we all have cases like this to talk about but I just had to vent. I am a firm believer that a lab should be able to charge for X number of IHC's when X number of IHC's are performed. If the pathologist is worried about professional fee's associated with it, I think there should be a method to charge for just the additional technical work done. I do have one question related to this rambling babble. What is the largest number of IHC's that you were unable to bill for due to this rule. Mine: a dermatopatholgist ordered a cytokeratin on blocks A1 thru A16 on me. I'll shut up now, Glen Dawson BS, HT & QIHC Lead IHC Technologist Milwaukee, WI From shive003 <@t> umn.edu Wed Apr 21 15:17:59 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] CGRP References: <00ac01c4276c$b16739e0$7a6c140a@imvs.sa.gov.au> Message-ID: <00fa01c427dd$bd5e8340$78065486@vdl220FAC> I use xCGRP from Serotec, at 1:1000 with no pretreatment necessary. The data sheet says it reacts with human and rat tissue. I use it on dog tissue with strong positive staining results. Jan Shivers Univ. of Minnesota Veterinary Diagnostic Lab St. Paul, MN USA ----- Original Message ----- From: "jim" To: "Histonet" Sent: Wednesday, April 21, 2004 1:48 AM Subject: [Histonet] CGRP > Does anyone know of a good CGRP antibody for FFPE human tissue, and/or rat > and mouse. > > Jim Manavis > Laboratory Manager > Hanson Institute > Centre for Neurological Diseases > Adelaide, SA, 5000 > Australia > Phone: 61-08-8222-3668 > FAX: 61-08-8222 3204 > email: jim.manavis@imvs.sa.gov.au > Disclaimer: Not this little black duck! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From convmcm <@t> cc.usu.edu Wed Apr 21 15:52:08 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Animal Tissue Processing Manual from NSH In-Reply-To: <3.0.6.32.20040421123504.00bcd780@gemini.msu.montana.edu> Message-ID: <000201c427e2$82e1e130$4a737b81@Cygnus> Thank you, Gayle, for the link. This will come in handy for me as one of my tasks for this year is to build up our lab library. All of our books are ancient. I'm in need of good books on IHC & general histology (Freida Carson's book comes to mind). If anyone would care to recommend a good book on IHC, feel free to let me know. *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Wednesday, April 21, 2004 11:35 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Animal Tissue Processing Manual from NSH It has been brought to my attention that ordering the Animal Tissue Processing Manual or rather finding it in the NSH website is a tidge difficult. So here it is: http://www.nsh.org/education/materials.html Booklet was placed in materials section under Education committee. It will be there at bottom of the webpage after all the self-assessment booklets. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SCheasty <@t> ahs.llumc.edu Wed Apr 21 16:41:19 2004 From: SCheasty <@t> ahs.llumc.edu (Cheasty, Sandra) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Drain Hose for Leica Stainer Message-ID: <2E50F33F91EEDA46A77BC3B2575BB0910587AD@mars.llumc.edu> Can I just go to Home Depot and get a new drain hose for our Leica stainer? Ours is cracked, our clinical/facilities engineering folks won't touch it because we have a maintenance agreement, and we're not due for a maintenance call for several months... It doesn't look like rocket-science... can't I just go get a length of solvent resistant hose, a new hose clamp and be done with it? Thanks, Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. From Rcartun <@t> harthosp.org Wed Apr 21 17:13:11 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] 88342 regulation comiserations Message-ID: Is this for metastatic melanoma? If so, what is in your cocktail? Richard Cartun >>> "Dawson, Glen" 04/21/04 03:45PM >>> All, I'm not looking for a quote on the rules and regulations on how and why only one 88342 can be billed on multiple sub-numbers from the same specimen. I am, however, going to share an example of how this rule can harm today's IHC lab. I just received a request for a 3 antibody cocktail to be performed on blocks A1 thru A10 from a given surgical case. While doing this work, it really ate at me that, due to the current IHC billing policies, my lab will be reimbursed one 88342 for what amounts to 10 88342's, not to mention that there is triple the amount of usual antibody in this cocktail. That's nine 88342's and 27 antibody applications that my lab gets to eat. When I think of all the sentinel node protocols that I perform which inevitably contain multiple blocks from the same specimen it starts to worry me. If I get one more pathologist who is gun-shy about possible liabilities that may arise if they don't cover all the blocks in a given specimen or flat out refuses to pick a representative block based on the morphology staring them in the face on the H&E I think I'll have to declare bankrupcy. I know we all have cases like this to talk about but I just had to vent. I am a firm believer that a lab should be able to charge for X number of IHC's when X number of IHC's are performed. If the pathologist is worried about professional fee's associated with it, I think there should be a method to charge for just the additional technical work done. I do have one question related to this rambling babble. What is the largest number of IHC's that you were unable to bill for due to this rule. Mine: a dermatopatholgist ordered a cytokeratin on blocks A1 thru A16 on me. I'll shut up now, Glen Dawson BS, HT & QIHC Lead IHC Technologist Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cuttingedgehistology <@t> yahoo.com Wed Apr 21 17:23:19 2004 From: cuttingedgehistology <@t> yahoo.com (Brandon Stokes) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Oven temp and time.... Message-ID: <20040421222319.88092.qmail@web41401.mail.yahoo.com> Hello everyone.....I was just wondering if a few people out there will tell me what temp. they keep their slide dryer at.....and also how long? Thanks, Brandon Stokes, HT(ASCP) --------------------------------- Do you Yahoo!? Yahoo! Photos: High-quality 4x6 digital prints for 25¢ From caroline.stott <@t> anatomy.otago.ac.nz Wed Apr 21 18:00:52 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Oven temp and time.... Message-ID: <5.2.1.1.0.20040422105951.00a086c0@anatomy.otago.ac.nz> 37 degrees overnight. Or if in a real hurry, 60 degrees for 20minutes. Caroline Caroline Stott Histology Service Unit Medical School University of Otago Dunedin (03) 479 7152 From kwuny <@t> email.cs.nsw.gov.au Wed Apr 21 18:55:37 2004 From: kwuny <@t> email.cs.nsw.gov.au (Young Kwun) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] CGRP In-Reply-To: <00ac01c4276c$b16739e0$7a6c140a@imvs.sa.gov.au> Message-ID: <200404220953984.SM01292@crgcsls814> Hi Jim, I used polyclonal rat CGRP and synthetic SP (porcine VIP,NPY as well) from Amersham International, Buckinghamshire. It worked well on rat and ovine tissues and it should work on human tissue. Young Kwun Senior Hospital Scientist Dept of Anatomical Pathology Concord Hospital Concord NSW 2139 Australia Phone:61-2-9767 6075 Fax:61-2-9767 8427 Email:kwuny@email.cs.nsw.gov.au -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of jim Sent: Wednesday, 21 April 2004 4:49 PM To: Histonet Subject: [Histonet] CGRP Does anyone know of a good CGRP antibody for FFPE human tissue, and/or rat and mouse. Jim Manavis Laboratory Manager Hanson Institute Centre for Neurological Diseases Adelaide, SA, 5000 Australia Phone: 61-08-8222-3668 FAX: 61-08-8222 3204 email: jim.manavis@imvs.sa.gov.au Disclaimer: Not this little black duck! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please destroy it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of the Central Sydney Area Health Service." From MajorFocus <@t> aol.com Wed Apr 21 19:32:12 2004 From: MajorFocus <@t> aol.com (MajorFocus@aol.com) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Re: Sheets of Slide Labels that work in a LaserJet Message-ID: <407C083C.714CD376.0BD82245@aol.com> Major Focus carries laser sheet labels that work with laserjet or inkjet printers. The sheets are standard 8.5 x 11" sheets with eight labels across and 11 labels down. You can check out the catalog and pricing information on their website at www.majorfocus.com Greg Good, HT(ASCP) From KAELPERS <@t> aol.com Wed Apr 21 20:25:42 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Coverslipper Agony Message-ID: <7e.4c527e93.2db87916@aol.com> We also have a hacker coverslipper and experience these issues. We have used permount from Fisher and we are now using Richard Allans mountant. I have always noticed on the first couple set of coverslips that there is a problem and then sporadic from there. A couple things I do is to #1 dispense quite a bit of mountant from the needle before beginning and #2 the springs that are on your plunges for picking up the coverslip and laying the coverslip onto the slide can also play a huge part. You want to make sure that the coverslip is placed at the bottom edge of the slide first and then slowly pressed onto the slide by the other. Adjusting these springs can greatly increase or decrease the pressure applied. If you have it adjusted nicely they will actually press the air bubbles away while applying the coverslip. It has always been trial and error and something you can't just walk away from. We exam every slide and wipe the backs after each rack is complete and then there is no issues after drying of the slide. We have the person staining (hacker too) also operating the coverslipper and it has worked out nicely. lge From jkiernan <@t> uwo.ca Wed Apr 21 22:23:48 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing (Egg) References: <001601c427ab$378b1850$56c4e150@KEMLOS> Message-ID: <40873AC4.FF00894B@uwo.ca> No! The egg is stable for weeks, especially if refrigerated. Boiling an egg solidifies the proteins, especially the albumen (white). This makes the egg easier to eat than the liquid contents of a broken raw egg. Cooking has enabled us humans to collect all the fat and protein from the eggs we take from birds. Fixation means preserving structural detail for microscopy. Heat alone will not do that. John Kiernan London, Canada. _____________________ Kemlo Rogerson wrote: > > Um....... A boiled egg is stabilized but not fixed; does that help? A > fixative is designed not only to stabilize proteins by coagulation, but > in most cases it links chemically to the proteins. Its job is also that > of preventing putrefaction both by endogenous and exogenous means > (lysosomes and bugs). A boiled egg will rot by putrefaction, but if you > fixed it, then it wouldn't; would it? Heat stabilizes by coagulating > protein doesn't it, formalin forms links between reactive sites on the > proteins, doesn't it? > > Am I talking a load of........... > > Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS > Tel: 0208 970 8414 > Mob: 07830 196072 > Mobile E-Mail kemlorogerson@3mail.com > FAX & Answer Phone 0871 242 8094 > E-mail Accounts: > kemlo@tiscali.co.uk or > kemlo1@btinternet.com > Disclaimer: The information contained in this message and/or any > attachments(s) may be of a private and confidential nature, and is > intended solely for the attention of the addressee. If you have received > this message in error or feel you should not have been the intended > recipient, please return it and any attachments to the sender > immediately. All messages relating to this communication should then be > deleted from your system. Unauthorised usage, copying, disclosure or > alteration of this message and/or attachment(s) is strictly prohibited. > Barking, Havering and Redbridge Hospitals NHS Trust will not be held > responsible for any direct or indirect damages which may arise from > alteration of this message or any attachment(s), by a third party or > resulting from the transmission of a virus. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall > Terry Dr,Consultant Histopathologist > Sent: 21 April 2004 12:48 > To: Steven E. Slap; kevin williams; histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > My confusion has not been dispelled. > > Steven writes: > > "The saline will not work > alone as a fixation step, except in the case of > very small biopsies (as written up in an article > by Tony Leong)- ..." > > In the next post, we get: > > "Yes, in the Leong method for biopsies, the specimens are heat > stabilized in the microwave in saline, and not really chemically > fixed. They get fixed in the ethanol in a traditional processor ..." > > These statements are mutually incompatible. > > Moreover, what is the difference between "stabilisation" and "fixation". > > A further muddle is introduced by: > > "not really chemically fixed." > > Well, hell, are they fixed or no? Heat fixation is fixation - who cares > whether chemically fixed? > > Of course, the major problem is that mechanisms of fixation are varied > (with the fixative) and not well understood. > (Speak for yourself do I hear someone say?) > > 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: Steven E. Slap [mailto:siksik03@comcast.net] > Sent: 17 April 2004 22:54 > To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; > histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > Hi Terry & HistoNetters > > I have done lots of successful large organ > stabilization in saline at 65?C for 20-35 minutes > (depending on the size of the organ. However, it > is important to emphasize that the purpose of > this step usually is to firm up the tissue so > that it can be subsequently cut up into thinner > pieces for processing, which must include a > fixation step, either in the microwave or on a > conventional processor, in formalin or some > non-formalin fixative. The saline will not work > alone as a fixation step, except in the case of > very small biopsies (as written up in an article > by Tony Leong)- a procedure I have not tried > myself. > > best regards, > Steven Slap > Microwave Consultant > > At 3:58 PM +0100 4/16/04, Marshall Terry Dr, > Consultant Histopathologist wrote: > >Sorry to but in - but on a related question - > >does anyone do microwave fixation in saline as > >described by for instance the Melbourne group, > >which involves bringing up to 65C in saline? > >Have tried it this week and the sections are even crummier than usual. > >When I was in Tasmania, a local private lab. did > >it and their sections were fine. > > _______________________________________________ > Histonet mailing list > Histonet@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 <@t> tiscali.co.uk Thu Apr 22 02:04:31 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing (Egg) In-Reply-To: <40873AC4.FF00894B@uwo.ca> Message-ID: <000901c42838$13338680$3d362850@KEMLOS> We put our boiled eggs into vinegar to preserve them. Nothing better than pickled eggs with a nice pint of Marstons Pedigree. Anyone cut sections on a pickled egg? Is cellular detail preserved by the acetic acid or do the egg cells swell? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 22 April 2004 04:24 To: Kemlo Rogerson Cc: 'Steven E. Slap'; histonet@pathology.swmed.edu; 'Marshall Terry Dr,Consultant Histopathologist' Subject: Re: [Histonet] Microwave Processing (Egg) No! The egg is stable for weeks, especially if refrigerated. Boiling an egg solidifies the proteins, especially the albumen (white). This makes the egg easier to eat than the liquid contents of a broken raw egg. Cooking has enabled us humans to collect all the fat and protein from the eggs we take from birds. Fixation means preserving structural detail for microscopy. Heat alone will not do that. John Kiernan London, Canada. _____________________ Kemlo Rogerson wrote: > > Um....... A boiled egg is stabilized but not fixed; does that help? A > fixative is designed not only to stabilize proteins by coagulation, but > in most cases it links chemically to the proteins. Its job is also that > of preventing putrefaction both by endogenous and exogenous means > (lysosomes and bugs). A boiled egg will rot by putrefaction, but if you > fixed it, then it wouldn't; would it? Heat stabilizes by coagulating > protein doesn't it, formalin forms links between reactive sites on the > proteins, doesn't it? > > Am I talking a load of........... > > Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS > Tel: 0208 970 8414 > Mob: 07830 196072 > Mobile E-Mail kemlorogerson@3mail.com > FAX & Answer Phone 0871 242 8094 > E-mail Accounts: > kemlo@tiscali.co.uk or > kemlo1@btinternet.com > Disclaimer: The information contained in this message and/or any > attachments(s) may be of a private and confidential nature, and is > intended solely for the attention of the addressee. If you have received > this message in error or feel you should not have been the intended > recipient, please return it and any attachments to the sender > immediately. All messages relating to this communication should then be > deleted from your system. Unauthorised usage, copying, disclosure or > alteration of this message and/or attachment(s) is strictly prohibited. > Barking, Havering and Redbridge Hospitals NHS Trust will not be held > responsible for any direct or indirect damages which may arise from > alteration of this message or any attachment(s), by a third party or > resulting from the transmission of a virus. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall > Terry Dr,Consultant Histopathologist > Sent: 21 April 2004 12:48 > To: Steven E. Slap; kevin williams; histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > My confusion has not been dispelled. > > Steven writes: > > "The saline will not work > alone as a fixation step, except in the case of > very small biopsies (as written up in an article > by Tony Leong)- ..." > > In the next post, we get: > > "Yes, in the Leong method for biopsies, the specimens are heat > stabilized in the microwave in saline, and not really chemically > fixed. They get fixed in the ethanol in a traditional processor ..." > > These statements are mutually incompatible. > > Moreover, what is the difference between "stabilisation" and "fixation". > > A further muddle is introduced by: > > "not really chemically fixed." > > Well, hell, are they fixed or no? Heat fixation is fixation - who cares > whether chemically fixed? > > Of course, the major problem is that mechanisms of fixation are varied > (with the fixative) and not well understood. > (Speak for yourself do I hear someone say?) > > 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: Steven E. Slap [mailto:siksik03@comcast.net] > Sent: 17 April 2004 22:54 > To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; > histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > Hi Terry & HistoNetters > > I have done lots of successful large organ > stabilization in saline at 65?C for 20-35 minutes > (depending on the size of the organ. However, it > is important to emphasize that the purpose of > this step usually is to firm up the tissue so > that it can be subsequently cut up into thinner > pieces for processing, which must include a > fixation step, either in the microwave or on a > conventional processor, in formalin or some > non-formalin fixative. The saline will not work > alone as a fixation step, except in the case of > very small biopsies (as written up in an article > by Tony Leong)- a procedure I have not tried > myself. > > best regards, > Steven Slap > Microwave Consultant > > At 3:58 PM +0100 4/16/04, Marshall Terry Dr, > Consultant Histopathologist wrote: > >Sorry to but in - but on a related question - > >does anyone do microwave fixation in saline as > >described by for instance the Melbourne group, > >which involves bringing up to 65C in saline? > >Have tried it this week and the sections are even crummier than usual. > >When I was in Tasmania, a local private lab. did > >it and their sections were fine. > > _______________________________________________ > Histonet mailing list > Histonet@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 <@t> tiscali.co.uk Thu Apr 22 02:48:22 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Pathology modernisation In-Reply-To: Message-ID: <000001c4283e$3032b750$3d362850@KEMLOS> Leaving behind Terry's fixation on fixation I would like, for once, to ask a serious question. Pathology modernisation in London appears to be 'ahead of the game', I assume because in London Pathology modernisation was already happening before it became a National Sport. The advent of LBC means that, to justfy a Processor, then Labs may have to merge or employ a 'hub and spoke' method of delivering Cervical Cytology. I don't wish to get into a LBC debate as I have had enough of that to last me a lifetime. I am assuming that what I have just said is the future; collaboration or merging to produce a workload that can use the capacity of T3000, multiple T2000 or a SurePath processor. But where does Non Gynae stand in all this? Traditionally, it has been the remit of the Cytology Lab to service Pathology's Non-Gynae commitment, but is that logical? I assume Non-Gynae will be served by either a T2000 or something SurePath can come up with; it may be we stay with traditional methods. My question is, given that many histological techniques are carried out on Non-Gynae specimens, could Non-Gynae sit within Histology. It would divorce it from the 'Pap Factory' scenario and could place it fairly and squarely where, in my opinion, it belongs. Could Non-Gynae Cytology actually find its 'place in the sun' if it was within the Histology Department of Cell Path? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Sho Dan BSK Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven E. Slap Sent: 21 April 2004 20:09 To: Marshall Terry Dr,Consultant Histopathologist; Kemlo Rogerson; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hello again HistoNetters Yes, there are these hybrid mixtures called coagulative fixatives. They are usually combinations of ethyl alcohol and polyethylene glycol, and, occasionally, acetic acid. They do not cross-link proteins. The PEG helps prevent the shrinkage associated with the use of ethanol as a fixative. The advantage of all these mixtures is that, no cross-linking having taken place, no subsequent antigen retrieval (or "unmasking") of the proteins would be necessary for IHC. Dr. Boon has a good chapter on them in support of her "Boon-Fix". To muddy the waters further, there are still other non-formalin fixatives, which are neither coagulative fixatives, nor cross-linking fixatives, based on glyoxal, such as Anatech's "Prefer", but which still are said to chemically "fix" the tissue. I'll leave the mechanism of their chemistry to the experts. best regards, Steven Slap At 3:51 PM +0100 4/21/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Kemlo asks: > >"Am I talking a load of..........." > >Inevitably:-) > >Kemlo, > >You talk as if formalin was the only fixative. What about the group >called, oddly enough, coagulative fixatives? Now .... I wonder how they >work. Let me see ...... Furthermore, you flit between the several >different subjects in an unstructured way. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ Upgrade Your Email - Click here! From nick.kirk3 <@t> btopenworld.com Thu Apr 22 03:30:35 2004 From: nick.kirk3 <@t> btopenworld.com (=?iso-8859-1?q?NICK=20KIRK?=) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Pathology modernisation In-Reply-To: <000001c4283e$3032b750$3d362850@KEMLOS> Message-ID: <20040422083035.41484.qmail@web86307.mail.ukl.yahoo.com> Kemlo We are developing the sort of centralised Gynae cytology network you describe, here in sunny Cambridgeshire with a centralised Gynae preparation/staining/screening laboratory based in a new building in Newmarket. Currently this serves all of Cambridge and West Suffolk and West Cambridgeshire (us here in Huntingdon) will be joining in at a later date. The Non-gynae however, has remained where it has always been at the original labs and the prep work etc is now being done by Histology staff instead of Cytology staff. I think the idea is to keep it close to the Pathologists who are mainly based at the original hospitals (Addenbrooke's and West Suffolk). Changing from conventional smears to LBC in Gynae is going to be hard enough as it is without involving Non-gynae, at least in the first instance, although there are obvious merits in including Non-Gynae specimens as well. >From what I understand, you can already do a certain amount of Non-Gynae specimens such as body cavity fluids etc on the conventional LBC machines. Interesting times I think. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England Kemlo Rogerson wrote: Leaving behind Terry's fixation on fixation I would like, for once, to ask a serious question. Pathology modernisation in London appears to be 'ahead of the game', I assume because in London Pathology modernisation was already happening before it became a National Sport. The advent of LBC means that, to justfy a Processor, then Labs may have to merge or employ a 'hub and spoke' method of delivering Cervical Cytology. I don't wish to get into a LBC debate as I have had enough of that to last me a lifetime. I am assuming that what I have just said is the future; collaboration or merging to produce a workload that can use the capacity of T3000, multiple T2000 or a SurePath processor. But where does Non Gynae stand in all this? Traditionally, it has been the remit of the Cytology Lab to service Pathology's Non-Gynae commitment, but is that logical? I assume Non-Gynae will be served by either a T2000 or something SurePath can come up with; it may be we stay with traditional methods. My question is, given that many histological techniques are carried out on Non-Gynae specimens, could Non-Gynae sit within Histology. It would divorce it from the 'Pap Factory' scenario and could place it fairly and squarely where, in my opinion, it belongs. Could Non-Gynae Cytology actually find its 'place in the sun' if it was within the Histology Department of Cell Path? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Sho Dan BSK Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven E. Slap Sent: 21 April 2004 20:09 To: Marshall Terry Dr,Consultant Histopathologist; Kemlo Rogerson; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hello again HistoNetters Yes, there are these hybrid mixtures called coagulative fixatives. They are usually combinations of ethyl alcohol and polyethylene glycol, and, occasionally, acetic acid. They do not cross-link proteins. The PEG helps prevent the shrinkage associated with the use of ethanol as a fixative. The advantage of all these mixtures is that, no cross-linking having taken place, no subsequent antigen retrieval (or "unmasking") of the proteins would be necessary for IHC. Dr. Boon has a good chapter on them in support of her "Boon-Fix". To muddy the waters further, there are still other non-formalin fixatives, which are neither coagulative fixatives, nor cross-linking fixatives, based on glyoxal, such as Anatech's "Prefer", but which still are said to chemically "fix" the tissue. I'll leave the mechanism of their chemistry to the experts. best regards, Steven Slap At 3:51 PM +0100 4/21/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Kemlo asks: > >"Am I talking a load of..........." > >Inevitably:-) > >Kemlo, > >You talk as if formalin was the only fixative. What about the group >called, oddly enough, coagulative fixatives? Now .... I wonder how they >work. Let me see ...... Furthermore, you flit between the several >different subjects in an unstructured way. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ =48017&partner=hotbar> Upgrade Your Email - Click here! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo <@t> tiscali.co.uk Thu Apr 22 04:32:44 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Pathology modernisation In-Reply-To: <20040422083035.41484.qmail@web86307.mail.ukl.yahoo.com> Message-ID: <000301c4284c$c7f31a40$66362850@KEMLOS> Thanks for that, I wouldn't mind seeing the outcome of your modernisation if I may. The major problem I will be having is the threat the Staff see from their jobs being eroded and the loss of Non-Gynae. I see their point but my feeling is that Non-Gynae certainly has been the Cinderella of Cytology, maybe it will blossom under the control of Histology. I say this as primarily a 'Cytologist' who also has an interest in Histology; my job entails not only managing the Department but Primary Screening, Checking and carrying out the 'cut up'. I also work in the Evenings in Central London screening LBC; I hope I have balanced view deviod of partisan bigotry. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: NICK KIRK [mailto:nick.kirk3@btopenworld.com] Sent: 22 April 2004 09:31 To: Kemlo Rogerson; histonet@pathology.swmed.edu Subject: Re: [Histonet] Pathology modernisation Kemlo We are developing the sort of centralised Gynae cytology network you describe, here in sunny Cambridgeshire with a centralised Gynae preparation/staining/screening laboratory based in a new building in Newmarket. Currently this serves all of Cambridge and West Suffolk and West Cambridgeshire (us here in Huntingdon) will be joining in at a later date. The Non-gynae however, has remained where it has always been at the original labs and the prep work etc is now being done by Histology staff instead of Cytology staff. I think the idea is to keep it close to the Pathologists who are mainly based at the original hospitals (Addenbrooke's and West Suffolk). Changing from conventional smears to LBC in Gynae is going to be hard enough as it is without involving Non-gynae, at least in the first instance, although there are obvious merits in including Non-Gynae specimens as well. >From what I understand, you can already do a certain amount of Non-Gynae specimens such as body cavity fluids etc on the conventional LBC machines. Interesting times I think. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England Kemlo Rogerson wrote: Leaving behind Terry's fixation on fixation I would like, for once, to ask a serious question. Pathology modernisation in London appears to be 'ahead of the game', I assume because in London Pathology modernisation was already happening before it became a National Sport. The advent of LBC means that, to justfy a Processor, then Labs may have to merge or employ a 'hub and spoke' method of delivering Cervical Cytology. I don't wish to get into a LBC debate as I have had enough of that to last me a lifetime. I am assuming that what I have just said is the future; collaboration or merging to produce a workload that can use the capacity of T3000, multiple T2000 or a SurePath processor. But where does Non Gynae stand in all this? Traditionally, it has been the remit of the Cytology Lab to service Pathology's Non-Gynae commitment, but is that logical? I assume Non-Gynae will be served by either a T2000 or something SurePath can come up with; it may be we stay with traditional methods. My question is, given that many histological techniques are carried out on Non-Gynae specimens, could Non-Gynae sit within Histology. It would divorce it from the 'Pap Factory' scenario and could place it fairly and squarely where, in my opinion, it belongs. Could Non-Gynae Cytology actually find its 'place in the sun' if it was within the Histology Department of Cell Path? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Sho Dan BSK Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven E. Slap Sent: 21 April 2004 20:09 To: Marshall Terry Dr,Consultant Histopathologist; Kemlo Rogerson; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hello again HistoNetters Yes, there are these hybrid mixtures called coagulative fixatives. They are usually combinations of ethyl alcohol and polyethylene glycol, and, occasionally, acetic acid. They do not cross-link proteins. The PEG helps prevent the shrinkage associated with the use of ethanol as a fixative. The advantage of all these mixtures is that, no cross-linking having taken place, no subsequent antigen retrieval (or "unmasking") of the proteins would be necessary for IHC. Dr. Boon has a good chapter on them in support of her "Boon-Fix". To muddy the waters further, there are still other non-formalin fixatives, which are neither coagulative fixatives, nor cross-linking fixatives, based on glyoxal, such as Anatech's "Prefer", but which still are said to chemically "fix" the tissue. I'll leave the mechanism of their chemistry to the experts. best regards, Steven Slap At 3:51 PM +0100 4/21/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Kemlo asks: > >"Am I talking a load of..........." > >Inevitably:-) > >Kemlo, > >You talk as if formalin was the only fixative. What about the group >called, oddly enough, coagulative fixatives? Now .... I wonder how they >work. Let me see ...... Furthermore, you flit between the several >different subjects in an unstructured way. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ =48017&partner=hotbar> Upgrade Your Email - Click here! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mab70 <@t> medschl.cam.ac.uk Thu Apr 22 04:48:31 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing In-Reply-To: <5.1.3.2.2.20040421122338.00b6fd38@mail.med.upenn.edu> Message-ID: <000201c4284e$f99c0b90$42536f83@medlan.cam.ac.uk> Just a quick suggestion which maybe you have already tried. Most alkaline phosphatase commercial substrates use Levamisole as an inhibitor of endogenous alkaline phosphatase. This does not inhibit the intestinal enzyme as used for labelling antibodies, so maybe that is what you need. The method was published by Ponder and Wilkinson, but I don't have my notes to hand so can't give you the exact reference, however if you look in any reputable textbook on immunohistochemistry you should find the details there. margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Maria Limberis Sent: Wednesday, April 21, 2004 5:34 PM To: Marshall Terry Dr,Consultant Histopathologist; Kemlo Rogerson; Steven E. Slap; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hi everyone- I have instilled a vector expressing human placental Alkaline Phosphatase in the lung of rhesus monkeys. However, when I use a protocol routinely used in our lab to prevent background in mouse lung (which works very well) the background is very high preventing assessment of gene transfer. I am wondering if anyone has any suggestions or ideas! Much appreciated Maria ------------------------------------------------------------------------- Maria Limberis, PhD Gene Therapy Program University of Pennsylvania School of Medicine Department of Medicine Division of Medical Genetics The Wistar Institute, Room 201 3601 Spruce Street Philadelphia, PA 19104 Tel: 215 8987166 Fax: 215 5737414 Email: limberis@mail.med.upenn.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gudrun.lang <@t> aon.at Thu Apr 22 05:41:52 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Oven temp and time.... References: <20040421222319.88092.qmail@web41401.mail.yahoo.com> Message-ID: <004d01c42856$6cf72050$eeeea8c0@SERVER> In routine histology we dry the cuts at 60 degrees for 30 min. (Umluftbrutschrank = airmoving oven?) For Immunhisto they let the slides over night at 37 degrees, then at 60 degrees for a short time. Gudrun Lang ----- Original Message ----- From: "Brandon Stokes" To: "HistologyList" Sent: Thursday, April 22, 2004 12:23 AM Subject: [Histonet] Oven temp and time.... > Hello everyone.....I was just wondering if a few people out there will tell me what temp. they keep their slide dryer at.....and also how long? > > Thanks, > Brandon Stokes, HT(ASCP) > > > > --------------------------------- > Do you Yahoo!? > Yahoo! Photos: High-quality 4x6 digital prints for 25? > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Terry.Marshall <@t> rothgen.nhs.uk Thu Apr 22 06:40:53 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Microwave Processing Message-ID: Of course we know each other. I used to work for him. 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: Kathleen Spencer [mailto:kspencer@utmem.edu] Sent: 21 April 2004 17:43 To: Kemlo Rogerson Cc: Marshall Terry Dr, Consultant Histopathologist; 'Steven E. Slap'; 'kevin williams'; histonet@pathology.swmed.edu Subject: Re: [Histonet] Microwave Processing Do you guys know each other? I love this battle of the minds, but who am I really to believe? You have truly brightened my day and made me laugh when I was blue. -Kathleen On Wednesday, April 21, 2004, at 10:51 AM, Kemlo Rogerson wrote: > Geez, tetchy begger aren't you. I'll answer when I've thunk about it and > give you an answer that is both lucid and structured. > > this used to be fun; why are some people so...... > > Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS > Tel: 0208 970 8414 > Mob: 07830 196072 > Mobile E-Mail kemlorogerson@3mail.com > FAX & Answer Phone 0871 242 8094 > E-mail Accounts: > kemlo@tiscali.co.uk or > kemlo1@btinternet.com > Disclaimer: The information contained in this message and/or any > attachments(s) may be of a private and confidential nature, and is > intended solely for the attention of the addressee. If you have received > this message in error or feel you should not have been the intended > recipient, please return it and any attachments to the sender > immediately. All messages relating to this communication should then be > deleted from your system. Unauthorised usage, copying, disclosure or > alteration of this message and/or attachment(s) is strictly prohibited. > Barking, Havering and Redbridge Hospitals NHS Trust will not be held > responsible for any direct or indirect damages which may arise from > alteration of this message or any attachment(s), by a third party or > resulting from the transmission of a virus. > > > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall > Terry Dr,Consultant Histopathologist > Sent: 21 April 2004 15:51 > To: Kemlo Rogerson; Steven E. Slap; kevin williams; > histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > Kemlo asks: > > "Am I talking a load of..........." > > Inevitably:-) > > Kemlo, > > You talk as if formalin was the only fixative. What about the group > called, oddly enough, coagulative fixatives? > Now .... I wonder how they work. Let me see ...... > Furthermore, you flit between the several different subjects in an > unstructured way. > > When you say : > "Its job is also that of preventing putrefaction both by endogenous and > exogenous means > (lysosomes and bugs)." > > You are wrong on 2 counts. > Its effect may be to prevent putrefaction, but hardly its job. (I > realise that some books list this, but a dehydrated specimen in wax is > more at risk from mice than bacteria). > Furthermore, the action you refer to, attributed to lysosomes, is > autolysis, not putrefaction. > > Furtherfurthermore, a fixed egg will only elude putrefaction by virtue > of the retained/residual fixative fixing any bacteria with ungodly > thoughts toward the egg, wouldn't it? > That is to say, "resistance to putrefaction" will be a property that > resides in the fixative not in the fixed egg. > > Yours lovingly, > > Confused from Rotherham > > 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] > Sent: 21 April 2004 15:16 > To: Marshall Terry Dr, Consultant Histopathologist; 'Steven E. Slap'; > 'kevin williams'; histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > > Um....... A boiled egg is stabilized but not fixed; does that help? A > fixative is designed not only to stabilize proteins by coagulation, but > in most cases it links chemically to the proteins. Its job is also that > of preventing putrefaction both by endogenous and exogenous means > (lysosomes and bugs). A boiled egg will rot by putrefaction, but if you > fixed it, then it wouldn't; would it? Heat stabilizes by coagulating > protein doesn't it, formalin forms links between reactive sites on the > proteins, doesn't it? > > > > Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS > Tel: 0208 970 8414 > Mob: 07830 196072 > Mobile E-Mail kemlorogerson@3mail.com > FAX & Answer Phone 0871 242 8094 > E-mail Accounts: > kemlo@tiscali.co.uk or > kemlo1@btinternet.com > Disclaimer: The information contained in this message and/or any > attachments(s) may be of a private and confidential nature, and is > intended solely for the attention of the addressee. If you have received > this message in error or feel you should not have been the intended > recipient, please return it and any attachments to the sender > immediately. All messages relating to this communication should then be > deleted from your system. Unauthorised usage, copying, disclosure or > alteration of this message and/or attachment(s) is strictly prohibited. > Barking, Havering and Redbridge Hospitals NHS Trust will not be held > responsible for any direct or indirect damages which may arise from > alteration of this message or any attachment(s), by a third party or > resulting from the transmission of a virus. > > > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall > Terry Dr,Consultant Histopathologist > Sent: 21 April 2004 12:48 > To: Steven E. Slap; kevin williams; histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > My confusion has not been dispelled. > > Steven writes: > > "The saline will not work > alone as a fixation step, except in the case of > very small biopsies (as written up in an article > by Tony Leong)- ..." > > In the next post, we get: > > "Yes, in the Leong method for biopsies, the specimens are heat > stabilized in the microwave in saline, and not really chemically > fixed. They get fixed in the ethanol in a traditional processor ..." > > These statements are mutually incompatible. > > Moreover, what is the difference between "stabilisation" and "fixation". > > A further muddle is introduced by: > > "not really chemically fixed." > > Well, hell, are they fixed or no? Heat fixation is fixation - who cares > whether chemically fixed? > > Of course, the major problem is that mechanisms of fixation are varied > (with the fixative) and not well understood. > (Speak for yourself do I hear someone say?) > > 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: Steven E. Slap [mailto:siksik03@comcast.net] > Sent: 17 April 2004 22:54 > To: Marshall Terry Dr, Consultant Histopathologist; kevin williams; > histonet@pathology.swmed.edu > Subject: RE: [Histonet] Microwave Processing > > > Hi Terry & HistoNetters > > I have done lots of successful large organ > stabilization in saline at 65?C for 20-35 minutes > (depending on the size of the organ. However, it > is important to emphasize that the purpose of > this step usually is to firm up the tissue so > that it can be subsequently cut up into thinner > pieces for processing, which must include a > fixation step, either in the microwave or on a > conventional processor, in formalin or some > non-formalin fixative. The saline will not work > alone as a fixation step, except in the case of > very small biopsies (as written up in an article > by Tony Leong)- a procedure I have not tried > myself. > > best regards, > Steven Slap > Microwave Consultant > > > At 3:58 PM +0100 4/16/04, Marshall Terry Dr, > Consultant Histopathologist wrote: >> Sorry to but in - but on a related question - >> does anyone do microwave fixation in saline as >> described by for instance the Melbourne group, >> which involves bringing up to 65C in saline? >> Have tried it this week and the sections are even crummier than usual. >> When I was in Tasmania, a local private lab. did >> it and their sections were fine. > > > _______________________________________________ > Histonet mailing list > Histonet@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 Myri37 <@t> aol.com Thu Apr 22 06:42:16 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Villanueva bone stain Message-ID: <1EB1DB4A.1EBE446C.0005167B@aol.com> Hi everyone I would like to use villanueva bone stain in staining MMA sections of bone , but i didn't stain bone before embedding, my sections are beween 50 -100 um thick do you have any protocol or advice Thank you very much Myriam baali Natural Implant From MDiCarlo <@t> KaleidaHealth.Org Thu Apr 22 07:06:22 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] mounting medium Message-ID: Hello Histonetters My 5 x 7 inch slides are air dried flat for at least a month before I file them upright. The problem I'm having is that once the slides are filed some of the toluene based mounting medium oozes out. After I clean the slides off there is usually a big air bubble where the tissue is and I don't like to re-coverslip for fear of the tissue falling off since it has happened. I spoke to a sales rep and he is sending me different mountants to try ( I assume all xylene based since I coverslip from xylene) but I was wondering what everyone else uses to prevent this from happening. Any suggestions to eliminate this problem? Thanks for your help. Peggy DiCarlo HT (ASCP) 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 Marion.Hiles <@t> north-bristol.swest.nhs.uk Thu Apr 22 08:18:23 2004 From: Marion.Hiles <@t> north-bristol.swest.nhs.uk (Marion Hiles) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] S-100 Protein Message-ID: <2EE924DF60902943AC6E2EF35155451F19CD07@nbfexch03.north-bristol.nhs> What sort of sections will you be using? Frozen or paraffin? Am I right to assume you have no immuno experience or are you specifically asking about the antibody. Bob Quilty Neuropathology Frenchay Hospital Bristol UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sharon E Willman Sent: 21 April 2004 19:12 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] S-100 Protein I was wondering if anyone has a method for S-100 protein staining method. Any help would be most appreciated. Thanks in advance, Sharon Willman _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: The information in this message is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorised. If you are not the intended recipient, any disclosure, copying, or distribution of the message, or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. Please immediately contact the sender if you have received this message in error. Thank you. From gsp26 <@t> drexel.edu Thu Apr 22 08:49:32 2004 From: gsp26 <@t> drexel.edu (gsp26@drexel.edu) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Antibody for Tissue factor for IHC on rabbit FFPE Message-ID: <4e1d324e05f9.4e05f94e1d32@drexel.edu> I need advice on antibody for tissue factor for IHC staining on rabbit FFPE slides. Please advice on antigen retrieval . Thanks in advance!! Gurusher Panjrath Research Fellow, Cardiovascular Nuclear Imaging Laboratories Drexel University College of Medicine 245 N 15 Street, MS 470, Philadelphia, PA,19102 Off-215-762-4218 Fax-215-762-1525 From BMolinari <@t> heart.thi.tmc.edu Thu Apr 22 09:16:16 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Ayoub-Shklar stain Message-ID: Hi, Does anyone have a copy of the Ayoub-Shklar stain for fibrin? I Thanks, Betsy Molinari HT (ASCP) Texas Heart Institute 832-355-8524 832-355-6812 (fax) From fcs <@t> uevora.pt Thu Apr 22 08:45:01 2004 From: fcs <@t> uevora.pt (Fernando Capela e Silva) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] S-100 Protein References: <2EE924DF60902943AC6E2EF35155451F19CD07@nbfexch03.north-bristol.nhs> Message-ID: <00ca01c42870$05f35da0$92d988c1@uevora.pt> I used S-100 from LabVision in chicken growth plate(Formalin/Paraffin) and worked fine http://www.labvision.com/ ----- Original Message ----- From: "Marion Hiles" To: "'Sharon E Willman'" Cc: "Histonet (E-mail)" Sent: Thursday, April 22, 2004 2:18 PM Subject: RE: [Histonet] S-100 Protein > What sort of sections will you be using? Frozen or Am I right to > assume you have no immuno experience or are you specifically asking about > the antibody. > > Bob Quilty > Neuropathology > Frenchay Hospital > Bristol UK > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sharon E > Willman > Sent: 21 April 2004 19:12 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] S-100 Protein > > > I was wondering if anyone has a method for S-100 protein staining > method. Any help would be most appreciated. > > Thanks in advance, > > Sharon Willman > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > DISCLAIMER: The information in this message is confidential and may be > legally privileged. It is intended solely for the addressee. Access to this > message by anyone else is unauthorised. If you are not the intended > recipient, any disclosure, copying, or distribution of the message, or any > action or omission taken by you in reliance on it, is prohibited and may be > unlawful. Please immediately contact the sender if you have received this > message in error. Thank you. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Terry.Marshall <@t> rothgen.nhs.uk Thu Apr 22 09:46:51 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Ayoub-Shklar stain Message-ID: You mean the deja vu stain? http://www.histosearch.com/histonet/Sep/Re.Ayoub-Shklarmethod.html 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: Molinari, Betsy [mailto:BMolinari@heart.thi.tmc.edu] Sent: 22 April 2004 15:16 To: histonet@pathology.swmed.edu Subject: [Histonet] Ayoub-Shklar stain Hi, Does anyone have a copy of the Ayoub-Shklar stain for fibrin? I Thanks, Betsy Molinari HT (ASCP) Texas Heart Institute 832-355-8524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mab70 <@t> medschl.cam.ac.uk Thu Apr 22 10:04:27 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] RE: Animal Tissue Processing Manual In-Reply-To: <3.0.6.32.20040421113009.00bc52b0@gemini.msu.montana.edu> Message-ID: <000d01c4287b$1c437d50$36536f83@medlan.cam.ac.uk> Dear Gayle, Thank you for your speedy reply to my enquiry, I will follow this up. All the best Margaret -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Wednesday, April 21, 2004 6:30 PM To: Margaret Blount; Histonet@lists.utsouthwestern.edu Subject: Animal Tissue Processing Manual Margaret, Diane Sterchi and I were editors on the National Society for Histotechnology, Veterinary, Industry and Research Committee (VIR) Animal Tissue Processing Manual. It has many processing schedules for different species, also tissues ie. eyes, bone, etc. Booklet comes with a CD. Contact NSH at Histo@NSH.org or go onto their website on how to order. You will find this manual listed under education in their webpage for ordering instructions. At 02:54 PM 4/21/2004 +0100, you wrote: >Hi, Gayle, > >While searching the histonet archive for information about processing some >rodent tissues, I came across a comment by yourself that it was planned to >produce a book on this subject listing the best techniques. Is this book out >yet and how can I get access to it? Incidentally the search facility on the >histonet website is brilliant. > >Margaret > >Margaret Blount >Chief Technician >Clinical Biochemistry >University of Cambridge >Addenbrooke's Hospital >Hills Road >Cambridge >CB2 2QR > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From MTitford <@t> aol.com Thu Apr 22 10:24:12 2004 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Is Cytology in for big changes? Message-ID: <6BF2BA24.488D033E.00762DB1@aol.com> I have been following Mr Rogersons discussions of changes in pathology services in the United Kingdom. I wonder if overall, cytology sections are in for the biggest changes, not histology sections, as yet. Automated cytology screening instruments are becoming popular, (meaning less cytotechnologists required) and the advent of virus testing (HPV is it?) may reduce the need or eliminate GYN screening. (I hope I don't start a big argument!) Mike Titford USA Pathology Mobile AL USA From Marjorie.Lehman <@t> unilever.com Thu Apr 22 10:19:38 2004 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Ayoub-Shklar stain Message-ID: Isn't A-S for keratin & prekeratin? I have used it for that and it is dandy, but I don't think it deliniates fibrin. What about the Weigert's and Carstairs' methods for fibrin in Sheehan and Hrapchak? Marjorie -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [SMTP:Terry.Marshall@rothgen.nhs.uk] Sent: Thursday, April 22, 2004 10:47 AM To: Molinari, Betsy; histonet@pathology.swmed.edu Subject: RE: [Histonet] Ayoub-Shklar stain You mean the deja vu stain? http://www.histosearch.com/histonet/Sep/Re.Ayoub-Shklarmethod.html 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: Molinari, Betsy [mailto:BMolinari@heart.thi.tmc.edu] Sent: 22 April 2004 15:16 To: histonet@pathology.swmed.edu Subject: [Histonet] Ayoub-Shklar stain Hi, Does anyone have a copy of the Ayoub-Shklar stain for fibrin? I Thanks, Betsy Molinari HT (ASCP) Texas Heart Institute 832-355-8524 832-355-6812 (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 BMolinari <@t> heart.thi.tmc.edu Thu Apr 22 10:33:26 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Ayoub-Shklar stain Message-ID: Thanks Marjorie , you are right. I am using it for keratin. More coffee please. Have a good day! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of marjorie lehman Sent: Thursday, April 22, 2004 10:20 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] Ayoub-Shklar stain Isn't A-S for keratin & prekeratin? I have used it for that and it is dandy, but I don't think it deliniates fibrin. What about the Weigert's and Carstairs' methods for fibrin in Sheehan and Hrapchak? Marjorie -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [SMTP:Terry.Marshall@rothgen.nhs.uk] Sent: Thursday, April 22, 2004 10:47 AM To: Molinari, Betsy; histonet@pathology.swmed.edu Subject: RE: [Histonet] Ayoub-Shklar stain You mean the deja vu stain? http://www.histosearch.com/histonet/Sep/Re.Ayoub-Shklarmethod.html 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: Molinari, Betsy [mailto:BMolinari@heart.thi.tmc.edu] Sent: 22 April 2004 15:16 To: histonet@pathology.swmed.edu Subject: [Histonet] Ayoub-Shklar stain Hi, Does anyone have a copy of the Ayoub-Shklar stain for fibrin? I Thanks, Betsy Molinari HT (ASCP) Texas Heart Institute 832-355-8524 832-355-6812 (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 From gcallis <@t> montana.edu Thu Apr 22 10:42:41 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Oozing mounting medium In-Reply-To: Message-ID: <3.0.6.32.20040422094241.00bc5558@gemini.msu.montana.edu> Peggy, I have had this happen and one needs to do longer drying, it happened with both xylene and toluene based mounting media. Been there and experienced your problem with gigantic sheep tibia and human femoral bone sections. The problem is with that large section in fact, everything is BIGGER! Even though you think section is truly flat, it often had thick thin areas, more so than routine, small paraffin section. Don't try to overclean these slides, this removes enough media to cause an air bubble particularly if you use dipping into xylene, then wiping to clean. This dissolves enough media away to create air bubbles. It also sounds as though longer is needed, these are huge sections with a lot of goo under that coverslip. If the bubble connects to outer edge of coverslip, use a very fine pipette to add media at bubbble edge and let it flow into bubble space, redry. We have repaired bubbles this way before. A #2 thickness coverslip is often nicer. One could also try a weight in middle of section to keep large coverslips flat. At 08:06 AM 4/22/2004 -0400, you wrote: >Hello Histonetters > >My 5 x 7 inch slides are air dried flat for at least a month before I file >them upright. The problem I'm having is that once the slides are filed some >of the toluene based mounting medium oozes out. After I clean the slides >off there is usually a big air bubble where the tissue is and I don't like >to re-coverslip for fear of the tissue falling off since it has happened. I >spoke to a sales rep and he is sending me different mountants to try ( I >assume all xylene based since I coverslip from xylene) but I was wondering >what everyone else uses to prevent this from happening. Any suggestions to >eliminate this problem? > >Thanks for your help. >Peggy DiCarlo HT (ASCP) > >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 MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Terry.Marshall <@t> rothgen.nhs.uk Thu Apr 22 11:05:30 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Pathology modernisation Message-ID: You are right in your thoughts about non-gynae - it should/could be part of histology. I have always regarded cytology as a rather challenging way of making a tissue diagnosis. So is Gynae-cytology, but it is so vast and so damned boring that it should be kept apart. That said, non-gynae cytology has suffered from the onslaught of trucuts, endoscopy and to some extent, radiology. (I *do* wish radiologist wouldn't make histologic diagnoses). For these reasons, I think it's dying on its feet. I have written before about how good ThinPrep is for non-gynae stuff, and it looks as if the likely centralisation of LBC machines will mean that non-gynae will continue to be pursued amidst piles of overlapping cells and blood, just when we had this opportunity for it to be otherwise. (BTW, and for Kemlo, I do mean ThinPrep, as that is the only system of which I have had experience). Also BTW, for the majority who do not know about pathology modernisation in the UK, its political speak for centralisation and cost cutting - nothing to do with modernisation at all. 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 22 April 2004 08:48 To: histonet@pathology.swmed.edu Subject: [Histonet] Pathology modernisation Leaving behind Terry's fixation on fixation I would like, for once, to ask a serious question. Pathology modernisation in London appears to be 'ahead of the game', I assume because in London Pathology modernisation was already happening before it became a National Sport. The advent of LBC means that, to justfy a Processor, then Labs may have to merge or employ a 'hub and spoke' method of delivering Cervical Cytology. I don't wish to get into a LBC debate as I have had enough of that to last me a lifetime. I am assuming that what I have just said is the future; collaboration or merging to produce a workload that can use the capacity of T3000, multiple T2000 or a SurePath processor. But where does Non Gynae stand in all this? Traditionally, it has been the remit of the Cytology Lab to service Pathology's Non-Gynae commitment, but is that logical? I assume Non-Gynae will be served by either a T2000 or something SurePath can come up with; it may be we stay with traditional methods. My question is, given that many histological techniques are carried out on Non-Gynae specimens, could Non-Gynae sit within Histology. It would divorce it from the 'Pap Factory' scenario and could place it fairly and squarely where, in my opinion, it belongs. Could Non-Gynae Cytology actually find its 'place in the sun' if it was within the Histology Department of Cell Path? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Sho Dan BSK Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven E. Slap Sent: 21 April 2004 20:09 To: Marshall Terry Dr,Consultant Histopathologist; Kemlo Rogerson; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hello again HistoNetters Yes, there are these hybrid mixtures called coagulative fixatives. They are usually combinations of ethyl alcohol and polyethylene glycol, and, occasionally, acetic acid. They do not cross-link proteins. The PEG helps prevent the shrinkage associated with the use of ethanol as a fixative. The advantage of all these mixtures is that, no cross-linking having taken place, no subsequent antigen retrieval (or "unmasking") of the proteins would be necessary for IHC. Dr. Boon has a good chapter on them in support of her "Boon-Fix". To muddy the waters further, there are still other non-formalin fixatives, which are neither coagulative fixatives, nor cross-linking fixatives, based on glyoxal, such as Anatech's "Prefer", but which still are said to chemically "fix" the tissue. I'll leave the mechanism of their chemistry to the experts. best regards, Steven Slap At 3:51 PM +0100 4/21/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Kemlo asks: > >"Am I talking a load of..........." > >Inevitably:-) > >Kemlo, > >You talk as if formalin was the only fixative. What about the group >called, oddly enough, coagulative fixatives? Now .... I wonder how they >work. Let me see ...... Furthermore, you flit between the several >different subjects in an unstructured way. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ Upgrade Your Email - Click here! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From k.whalley <@t> ich.ucl.ac.uk Thu Apr 22 11:13:34 2004 From: k.whalley <@t> ich.ucl.ac.uk (Katy Whalley) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] methanol and wax Message-ID: <4750.194.82.240.48.1082650414.squirrel@jenner.ich.ucl.ac.uk> Hi, A colleague of mine hs been having some trouble with wax embedding/ sectioning her samples. They have been dehydrated through a series of methanol, rather than ethanol steps, follwed by histoclear and wax as usual. The samples did not section very well, either not cutting at all (i.e. the wax cuts leaving a hole) or giving a cracked appearance. She has been advised that the problem was probably the methanol. Has anyone ever had this problem, and do you know if there is some way her samples can be rescued as they are very precious! thanks Katy Whalley, UCL From kemlo <@t> tiscali.co.uk Thu Apr 22 11:18:03 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Pathology modernisation In-Reply-To: Message-ID: <000101c42885$63cb5d50$79cce150@KEMLOS> You are right in your thoughts about non-gynae - it should/could be part of histology. I have always regarded cytology as a rather challenging way of making a tissue diagnosis. So is Gynae-cytology, but it is so vast and so damned boring that it should be kept apart. That said, non-gynae cytology has suffered from the onslaught of trucuts, endoscopy and to some extent, radiology. (I *do* wish radiologist wouldn't make histologic diagnoses). For these reasons, I think it's dying on its feet. AGREED. I have written before about how good ThinPrep is for non-gynae stuff, and it looks as if the likely centralisation of LBC machines will mean that non-gynae will continue to be pursued amidst piles of overlapping cells and blood, just when we had this opportunity for it to be otherwise. (BTW, and for Kemlo, I do mean ThinPrep, as that is the only system of which I have had experience). I accept your limited experience, but your point remains valid Also BTW, for the majority who do not know about pathology modernisation in the UK, its political speak for centralisation and cost cutting - nothing to do with modernisation at all. Or maybe it could limit the power of the Consultant. Introduce evidence based testing, demand management and effective clinical pathways? Nah, you are right, aint gonna happen! 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: Kemlo Rogerson [mailto:kemlo@tiscali.co.uk] Sent: 22 April 2004 08:48 To: histonet@pathology.swmed.edu Subject: [Histonet] Pathology modernisation Leaving behind Terry's fixation on fixation I would like, for once, to ask a serious question. Pathology modernisation in London appears to be 'ahead of the game', I assume because in London Pathology modernisation was already happening before it became a National Sport. The advent of LBC means that, to justfy a Processor, then Labs may have to merge or employ a 'hub and spoke' method of delivering Cervical Cytology. I don't wish to get into a LBC debate as I have had enough of that to last me a lifetime. I am assuming that what I have just said is the future; collaboration or merging to produce a workload that can use the capacity of T3000, multiple T2000 or a SurePath processor. But where does Non Gynae stand in all this? Traditionally, it has been the remit of the Cytology Lab to service Pathology's Non-Gynae commitment, but is that logical? I assume Non-Gynae will be served by either a T2000 or something SurePath can come up with; it may be we stay with traditional methods. My question is, given that many histological techniques are carried out on Non-Gynae specimens, could Non-Gynae sit within Histology. It would divorce it from the 'Pap Factory' scenario and could place it fairly and squarely where, in my opinion, it belongs. Could Non-Gynae Cytology actually find its 'place in the sun' if it was within the Histology Department of Cell Path? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Sho Dan BSK Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven E. Slap Sent: 21 April 2004 20:09 To: Marshall Terry Dr,Consultant Histopathologist; Kemlo Rogerson; kevin williams; histonet@pathology.swmed.edu Subject: RE: [Histonet] Microwave Processing Hello again HistoNetters Yes, there are these hybrid mixtures called coagulative fixatives. They are usually combinations of ethyl alcohol and polyethylene glycol, and, occasionally, acetic acid. They do not cross-link proteins. The PEG helps prevent the shrinkage associated with the use of ethanol as a fixative. The advantage of all these mixtures is that, no cross-linking having taken place, no subsequent antigen retrieval (or "unmasking") of the proteins would be necessary for IHC. Dr. Boon has a good chapter on them in support of her "Boon-Fix". To muddy the waters further, there are still other non-formalin fixatives, which are neither coagulative fixatives, nor cross-linking fixatives, based on glyoxal, such as Anatech's "Prefer", but which still are said to chemically "fix" the tissue. I'll leave the mechanism of their chemistry to the experts. best regards, Steven Slap At 3:51 PM +0100 4/21/04, Marshall Terry Dr, Consultant Histopathologist wrote: >Kemlo asks: > >"Am I talking a load of..........." > >Inevitably:-) > >Kemlo, > >You talk as if formalin was the only fixative. What about the group >called, oddly enough, coagulative fixatives? Now .... I wonder how they >work. Let me see ...... Furthermore, you flit between the several >different subjects in an unstructured way. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ &El=em%3b&SG=&RAND =48017&partner=hotbar> Upgrade Your Email - Click here! _______________________________________________ Histonet mailing list Histonet@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 cindyk <@t> centrexlabs.com Thu Apr 22 12:31:58 2004 From: cindyk <@t> centrexlabs.com (Cindy Kosuda) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] (no subject) Message-ID: <000101c4288f$b87694f0$f4fca8c0@gm55r01> Centrex Clinical Laboratories, Inc. Working to give you results! Bring your valuable clinical laboratory skills to Centrex Labs and you?ll make a difference in healthcare. Our mission is to provide a full spectrum of diagnostic and patient status information for physicians, hospitals and other qualified individuals benefiting patients and improving healthcare in the communities we serve. Centrex offers competitive pay and an excellent benefits package. Sign on bonuses are available. The following opportunities are available: Histotechnician - 1st and 2nd shift positions available Apply to Human Resources, Centrex Labs, 28 Campion Road, New Hartford, New York 13413 or fax to 315/797-1849. You can also visit us on the web at www.centrexlabs.com and email heatherf@centrexlabs.com . From carl.hobbs <@t> kcl.ac.uk Thu Apr 22 15:25:16 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] re fixation v preservation Message-ID: <001401c428a7$ecef1f00$38292bd9@home> Would that THE man HCook would engage in this......many of my essays turned on my definitions of the above...still not sure. Is there a difference between fixation and preservation? Yes, I know well the textbooks..... From JColCLEFA <@t> aol.com Thu Apr 22 16:40:57 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] slide drying, cyto/hist field changes/ non formalin fix Message-ID: If this is for drying after cutting & before staining, we are all ( routine and IHC) m-wave and nuke em for 3 minutes. We use 130 different primary antibodies and have never had an issue with staining compared to oven dried. Ovens we kept at 60C and dried for 30 min. For Brains when we stain Bielschowsky(sp?) we nuke and then hold at 40C overnight. If you mean drying after coverslipping, 40c for two-three days for permount and not at all for auto celluloid machines. Cyto changes- in addition to the popularity of mechanized cyto readers the development of an HPV vaccine is on the horizon further squeezing the future utility of pap smears (HPV vaccine=less HPV = very little cx ca=less PAP smear relevance) The looming change for Histo is the 2005 AS degree requirement for HT's, adding an additional roadblock for some prospective techs. (I believe in the change though- we need more respect!) As most IHC procedures are optimized for formalin, I would really hate the headache of retitring and reworking all my antibodies to accomodate these "alternative" fixatives From convmcm <@t> cc.usu.edu Thu Apr 22 16:52:50 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Drain Hose for Leica Stainer In-Reply-To: <2E50F33F91EEDA46A77BC3B2575BB0910587AD@mars.llumc.edu> Message-ID: <000e01c428b4$287a8940$4a737b81@Cygnus> This happened to my Leica stainer about 2 years ago. I went to Lowe's and got some tubing that has actually served us better than the hose that comes with the stainer. I did NOT get that crinkly wrinkled up hose, but some clear, smooth tubing. There is no more nasty fungus and bacteria growth in the tubing like the wrinkled up hose used to get. Be sure to get a hose clamp, too. I found that the clamp that came with our stainer had rusted through. It's not a bad idea to change that when you change the hose. >From the FWIW Dept.... Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cheasty, Sandra Sent: Wednesday, April 21, 2004 2:41 PM To: HistoNet (E-mail) Subject: [Histonet] Drain Hose for Leica Stainer Can I just go to Home Depot and get a new drain hose for our Leica stainer? Ours is cracked, our clinical/facilities engineering folks won't touch it because we have a maintenance agreement, and we're not due for a maintenance call for several months... It doesn't look like rocket-science... can't I just go get a length of solvent resistant hose, a new hose clamp and be done with it? Thanks, Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Thu Apr 22 17:49:07 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Pathology modernisation Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E19C@simba.kids> Kemlo, Some comments below: " My question is, given that many histological techniques are carried out on Non-Gynae specimens, could Non-Gynae sit within Histology. It would divorce it from the 'Pap Factory' scenario and could place it fairly and squarely where, in my opinion, it belongs. Could Non-Gynae Cytology actually find its 'place in the sun' if it was within the Histology Department of Cell Path? " Having worked in a Private Gyne Cytology Lab as well as Hospital Cytology Labs, I would miss screening the occasional General cytology smear or FNA. It breaks up the monotony of many negative PAP smears. (Don't get me wrong, I look forward to the day when there are no more CINs or CIS or other abnormalities). Regards Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From KAELPERS <@t> aol.com Thu Apr 22 18:51:50 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Drain Hose for Leica Stainer Message-ID: <114.31c33fce.2db9b496@aol.com> We use this on our stainer too! Definitely right in that there is no build up, no grooves for things to settle in and grow. Lowes knows! lge From histo20 <@t> hotmail.com Thu Apr 22 19:13:15 2004 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Cytology cost per test Message-ID: We are being asked to calculate cost per test in pathology and cytology. Does anyone have any experience with this? Calculations, formulas, etc., especially in cytology. Would someone point us in the right direction? Any help would be most appreciated!!! Paula Wilder St. Joseph Medical Center Towson, MD 21204 _________________________________________________________________ Test your ‘Travel Quotient’ and get the chance to win your dream trip! http://travel.msn.com From Mplhisto <@t> aol.com Thu Apr 22 19:53:41 2004 From: Mplhisto <@t> aol.com (Mplhisto@aol.com) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] MSI testing Message-ID: <121.2dea8841.2db9c315@aol.com> We are about to start MSI testing at our lab MLH 1, MSH@ and 6, possibly EGFR, does anyone have any advice they would like to share, are you performing tests manually or automated? Any advice would be appreciated . Thanks, Meredith Hale HT(ASCP) From jim.manavis <@t> imvs.sa.gov.au Thu Apr 22 20:31:02 2004 From: jim.manavis <@t> imvs.sa.gov.au (jim) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Slide & Cassette writers Message-ID: <013401c428d2$a3795ea0$7a6c140a@imvs.sa.gov.au> We are in the process of purchasing both a slide and cassette writer. What comments has anyone on the Leica IPC cassette printer and Leica IPS slide printer or any other manufacturers models they might be using. Thanks Jim Manavis Laboratory Manager Hanson Institute Centre for Neurological Diseases Adelaide, SA, 5000 Australia Phone: 61-08-8222-3668 FAX: 61-08-8222 3204 email: jim.manavis@imvs.sa.gov.au Disclaimer: Not this little black duck! From jim.manavis <@t> imvs.sa.gov.au Thu Apr 22 21:02:27 2004 From: jim.manavis <@t> imvs.sa.gov.au (jim) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] MSI testing In-Reply-To: <121.2dea8841.2db9c315@aol.com> Message-ID: <013c01c428d7$06fadcc0$7a6c140a@imvs.sa.gov.au> Meredith Please see recent publication in Applied Immunohistochemistry & Molecular Morphology Vol 11(1): p73-77, 2003. Cheers Jim Manavis Laboratory Manager Hanson Institute Centre for Neurological Diseases Adelaide, SA, 5000 Australia Phone: 61-08-8222-3668 FAX: 61-08-8222 3204 email: jim.manavis@imvs.sa.gov.au Disclaimer: Not this little black duck! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mplhisto@aol.com Sent: Friday, 23 April 2004 10:24 AM To: histonet Subject: [Histonet] MSI testing We are about to start MSI testing at our lab MLH 1, MSH@ and 6, possibly EGFR, does anyone have any advice they would like to share, are you performing tests manually or automated? Any advice would be appreciated . Thanks, Meredith Hale HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jim.manavis <@t> imvs.sa.gov.au Thu Apr 22 21:03:50 2004 From: jim.manavis <@t> imvs.sa.gov.au (jim) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Antibody for Tissue factor for IHC on rabbit FFPE In-Reply-To: <4e1d324e05f9.4e05f94e1d32@drexel.edu> Message-ID: <013d01c428d7$388c1240$7a6c140a@imvs.sa.gov.au> Gurusher I have used American Diagnostics Inc Tissue Factor antibody (product no. 4511) on rabbit FFPE material using citrate buffer, as well as in frozen tissue. Cheers Jim Manavis Laboratory Manager Hanson Institute Centre for Neurological Diseases Adelaide, SA, 5000 Australia Phone: 61-08-8222-3668 FAX: 61-08-8222 3204 email: jim.manavis@imvs.sa.gov.au Disclaimer: Not this little black duck! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of gsp26@drexel.edu Sent: Thursday, 22 April 2004 11:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Antibody for Tissue factor for IHC on rabbit FFPE I need advice on antibody for tissue factor for IHC staining on rabbit FFPE slides. Please advice on antigen retrieval . Thanks in advance!! Gurusher Panjrath Research Fellow, Cardiovascular Nuclear Imaging Laboratories Drexel University College of Medicine 245 N 15 Street, MS 470, Philadelphia, PA,19102 Off-215-762-4218 Fax-215-762-1525 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From michael_lafriniere <@t> memorial.org Thu Apr 22 22:20:23 2004 From: michael_lafriniere <@t> memorial.org (LaFriniere, Mike) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] REGION III MEETING BIRMINGHAM ALABAMA Message-ID: Final notice on the Histonet for the Region III Annual Meeting beginning Thursday evening May 20th and ending at noon Sunday May 23 at the Wynfrey Hotel in Birmingham Alabama, fun and education in the Magic City! 14 great workshops are in the program from basic to advanced, 28 vendors, great food with registration and a DJ sponsored by Thermo Electron at the banquet! Most vendors will have great door prizes from CASH to great gifts! In addition, shopping till you drop at the Galleria Mall attached to the Wynfrey during your free time! We have extended the special rates on the rooms till May 1st . Hurry and get your reservations at the hotel by calling the Wynfrey 205-444-5784 and state you are with the Region III/Alabama Histology Society to get the special rate! Workshop and additional information can be obtained on line www.timjday.com and click on Region III, workshop registration is welcome through the website. NSH Region III continues to demonstrate education as our top priority and is offering financial assistance for those of you who have had cut backs with educational funds from your employer. If you find yourself wanting to come to this meeting but do not have all of the funds available, feel free to contact me directly prior to May 1st to discuss your need, do not let a few dollars interfere with your continued educational needs at this outstanding histology educational meeting! Sincerely, Michael LaFriniere PA, HT(ASCP) NSH Region III Director 423-495-6117 w 423-902-9340 c From kemlo <@t> tiscali.co.uk Fri Apr 23 02:25:17 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Pathology modernisation In-Reply-To: <1CF2E2E5BB36D5119E7A0008C791F3740800E19C@simba.kids> Message-ID: <001201c42904$243222d0$5c062850@KEMLOS> But I think that is my point Tony; Non-Gynae is seen as a bit of light relief to make a 'Gynae Day' more palitable Cytology Labs are mainly funded for their Gynae commitment and, it's my own belief, Non-Gynae takes second place. You don't need to enter EQA to do Non-Gynae, do you? You don't actually need the 'Screening Certificate' do you? In my day the 'Cert of Competence for Gynae and Systemic Cytology' had both elements; am I right that the present one is only for Gynae? I know the IBMS does an exam but I'm sure it is not needed to 'check' Non-Gynae. I wait to be corrected by those that participate and those that lurk. Non-Gynae needs to sit somewhere and if it's the Histologists that set a better table, then so be it. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tony Henwood Sent: 22 April 2004 23:49 To: 'Kemlo Rogerson'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Pathology modernisation Kemlo, Some comments below: " My question is, given that many histological techniques are carried out on Non-Gynae specimens, could Non-Gynae sit within Histology. It would divorce it from the 'Pap Factory' scenario and could place it fairly and squarely where, in my opinion, it belongs. Could Non-Gynae Cytology actually find its 'place in the sun' if it was within the Histology Department of Cell Path? " Having worked in a Private Gyne Cytology Lab as well as Hospital Cytology Labs, I would miss screening the occasional General cytology smear or FNA. It breaks up the monotony of many negative PAP smears. (Don't get me wrong, I look forward to the day when there are no more CINs or CIS or other abnormalities). Regards Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.Eyres <@t> sanofi-synthelabo.com Fri Apr 23 02:35:53 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] re fixation v preservation Message-ID: Hi Carl, Are you referring to THE Harry Cook that lectured at Paddington? Cheers Steve "Carl" Sent by: To: histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] re fixation v preservation 22/04/2004 21:25 Would that THE man HCook would engage in this......many of my essays turned on my definitions of the above...still not sure. Is there a difference between fixation and preservation? Yes, I know well the textbooks..... _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Myri37 <@t> aol.com Fri Apr 23 03:07:56 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] MacNeal's tetrachrome Message-ID: <05528F44.695581DE.0005167B@aol.com> hi i searched on histonet archives a protocol of Macneal's tetrachrome, but i didn't find please could someone give me a recipe ? thank you very much Myriam baali natural implant From shaumik.adhya <@t> ucl.ac.uk Fri Apr 23 04:46:59 2004 From: shaumik.adhya <@t> ucl.ac.uk (Shaumik Adhya) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] beta galactosidase nuclear counterstain Message-ID: <1082713619.4088e613a2bb6@www.webmail.ucl.ac.uk> Hi Histonetters, I'm trying to stain myocardium for beta galactosidase (resulting in a blue precipitate), and have been trying to use Nuclear Fast Red as a nuclear counterstain - whilst this gives me reasonable nuclear staining, the process of dehydrating and mounting makes me lose all the beta gal. If I use an aqueous mount such as gycergel, the Nuclear Fast Red dissolves out over the course of a week, which is less than ideal. My protocol for dehydrating and mounting is: 10 quick dips in 70% ethanol 2 min in 100% ethanol 5 min in 100% ethanol 5 min in Histoclear 2 min in Histoclear DPX to coverslip with. I'm wondering what people use as a nuclear counterstain for beta-gal, and if you had any tips or advice. Thanks Shaumik From HETZER <@t> surgery.wisc.edu Fri Apr 23 08:51:07 2004 From: HETZER <@t> surgery.wisc.edu (Michael Hetzer) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Mohs Surgery Tech Opening Message-ID: We have a full time day opening for a frozen section histologist at the birthplace of Mohs surgery, The Mohs Surgery Clinic of the University of Wisconsin, Madsion. Experience cutting Mohs sections is not necessary but we would prefer some experience in histology. Contact Julie Schrader at UW hospital human resources at 608-264-9136 and refer to position #017081. From Ian.Bernard <@t> LACKLAND.AF.MIL Fri Apr 23 09:21:44 2004 From: Ian.Bernard <@t> LACKLAND.AF.MIL (Bernard Ian R SSgt 59 CRES/MSROP) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] REGION III MEETING BIRMINGHAM ALABAMA Message-ID: You all I miss Alabama -----Original Message----- From: LaFriniere, Mike [mailto:michael_lafriniere@memorial.org] Sent: Thursday, April 22, 2004 10:20 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] REGION III MEETING BIRMINGHAM ALABAMA Final notice on the Histonet for the Region III Annual Meeting beginning Thursday evening May 20th and ending at noon Sunday May 23 at the Wynfrey Hotel in Birmingham Alabama, fun and education in the Magic City! 14 great workshops are in the program from basic to advanced, 28 vendors, great food with registration and a DJ sponsored by Thermo Electron at the banquet! Most vendors will have great door prizes from CASH to great gifts! In addition, shopping till you drop at the Galleria Mall attached to the Wynfrey during your free time! We have extended the special rates on the rooms till May 1st . Hurry and get your reservations at the hotel by calling the Wynfrey 205-444-5784 and state you are with the Region III/Alabama Histology Society to get the special rate! Workshop and additional information can be obtained on line www.timjday.com and click on Region III, workshop registration is welcome through the website. NSH Region III continues to demonstrate education as our top priority and is offering financial assistance for those of you who have had cut backs with educational funds from your employer. If you find yourself wanting to come to this meeting but do not have all of the funds available, feel free to contact me directly prior to May 1st to discuss your need, do not let a few dollars interfere with your continued educational needs at this outstanding histology educational meeting! Sincerely, Michael LaFriniere PA, HT(ASCP) NSH Region III Director 423-495-6117 w 423-902-9340 c _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Fri Apr 23 09:51:58 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Cytology cost per test Message-ID: See Model for Determining Manual Screening Pap Smear Costs: http://www.cap.org/apps/docs/medicare/resources/papcost.html Gary Gill -----Original Message----- From: Paula Wilder [mailto:histo20@hotmail.com] Sent: Thursday, April 22, 2004 7:13 PM To: histonet@pathology.swmed.edu Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cytology cost per test We are being asked to calculate cost per test in pathology and cytology. Does anyone have any experience with this? Calculations, formulas, etc., especially in cytology. Would someone point us in the right direction? Any help would be most appreciated!!! Paula Wilder St. Joseph Medical Center Towson, MD 21204 _________________________________________________________________ Test your 'Travel Quotient' and get the chance to win your dream trip! http://travel.msn.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Fri Apr 23 09:51:58 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Cytology cost per test Message-ID: See Model for Determining Manual Screening Pap Smear Costs: http://www.cap.org/apps/docs/medicare/resources/papcost.html Gary Gill -----Original Message----- From: Paula Wilder [mailto:histo20@hotmail.com] Sent: Thursday, April 22, 2004 7:13 PM To: histonet@pathology.swmed.edu Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cytology cost per test We are being asked to calculate cost per test in pathology and cytology. Does anyone have any experience with this? Calculations, formulas, etc., especially in cytology. Would someone point us in the right direction? Any help would be most appreciated!!! Paula Wilder St. Joseph Medical Center Towson, MD 21204 _________________________________________________________________ Test your 'Travel Quotient' and get the chance to win your dream trip! http://travel.msn.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From HETZER <@t> surgery.wisc.edu Fri Apr 23 11:10:10 2004 From: HETZER <@t> surgery.wisc.edu (Michael Hetzer) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Apply online Message-ID: We have a full time day opening for a frozen section histologist at the birthplace of Mohs surgery, The Mohs Surgery Clinic of the University of Wisconsin, Madsion. Experience cutting Mohs sections is not necessary but we would prefer some experience in histology. Contact Julie Schrader at UW hospital human resources at 608-264-9136 and refer to position #017081. You can also apply online at www.hosp.wisc.edu/HR/vacancy/index.asp From TJJ <@t> Stowers-Institute.org Fri Apr 23 12:47:57 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Re: Beta galactosidase nuclear counterstain Message-ID: It is probably not necessary to leave the slides for those times in the alcohol and xylene. However, we don't have the problem of losing the x-gal staining on tissue sections or even in the processor for stained whole mounts for paraffin embedding. We use the x-gal staining protocol recommended by Lobe, et al (Developmental Biology 208; 281-292) and counterstain in nuclear fast red, rinse in water, and dehydrate, clear and mount as usual with no loss of staining. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From JPCOLEMA <@t> sentara.com Fri Apr 23 13:42:57 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Cost per test Message-ID: The way I go about a cost per test analysis is by taking the cost of each item used in a procedure (including FTE per hour salaries), dividing by the number of tests each unit handles, and adding the quotients. Example: IHC kit costs $300 and stains 50 slides, $300 / 50 slds x 3 slds per test = $18. CD45 antibody is diluted 1:400, uses .3ml per slide, costs $250. per ml-$250/ 400ml x .3ml x 2 sldper test= $.38 worth of CD45 antibody per test. Slides are $50 per gross (144 slds)- $50/144 slds x 3 slides per test = 1.04 per test Salary $10 bucks an hour, 3 hours to perform a set of 25 slides= $10 per H x 3 h / 25 slides per run x 3 slides per test = $3.6 per test. Total- 18 + .38 + 1.04 + 3.6 = 23.02 total cost per test assumption: 1 patient slide and 1 pos control and 1 neg control= 1 test Note- all prices are fictitious and used here for example only. Also, more items per test are considered in the real world- I tried to keep this simple. Also if my math is wrong feel free to correct it for yourself without Emailing me about how shabby my math is. JC(QIHC) questions? email me directly jcolclefa@aol.com. From RCazares <@t> schosp.org Fri Apr 23 14:30:49 2004 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] PTAH Procedure Message-ID: <229A3566B9F0D311826E00D0B7441D79061A44E1@swedish_nt1.schosp.org> Hello again Histonetters, I was wondering if anyone out there knows if there is a procedure for PTAH that does not require the tissue to be treated in Zenker's? Any and all information will be greatly appreciated. Thanks. Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ************************* From jkiernan <@t> uwo.ca Fri Apr 23 16:02:08 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] beta galactosidase nuclear counterstain References: <1082713619.4088e613a2bb6@www.webmail.ucl.ac.uk> Message-ID: <40898450.22DC034E@uwo.ca> The blue product of the galactosidase reaction is an indigoid dye and it's insoluble in alcohols and xylene. I think you must be doing something wrong here! Nuclear fast red isn't always the dye that it should be. (The name has been applied to different compounds; the one to use for nuclei is CI 60760, made up usually as a 0.1% solution in 5% alum. Boil; cool overnight; filter to remove crud.) John Kiernan London, Canada _____________________ Shaumik Adhya wrote: > > Hi Histonetters, > > I'm trying to stain myocardium for beta galactosidase (resulting in a blue > precipitate), and have been trying to use Nuclear Fast Red as a nuclear > counterstain - whilst this gives me reasonable nuclear staining, the process > of dehydrating and mounting makes me lose all the beta gal. If I use an > aqueous mount such as gycergel, the Nuclear Fast Red dissolves out over the > course of a week, which is less than ideal. > > My protocol for dehydrating and mounting is: > > 10 quick dips in 70% ethanol > 2 min in 100% ethanol > 5 min in 100% ethanol > 5 min in Histoclear > 2 min in Histoclear > DPX to coverslip with. > > I'm wondering what people use as a nuclear counterstain for beta-gal, and if > you had any tips or advice. Thanks > > Shaumik > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SUSANLMCCOY <@t> aol.com Fri Apr 23 16:35:03 2004 From: SUSANLMCCOY <@t> aol.com (SUSANLMCCOY@aol.com) Date: Fri Sep 16 15:22:51 2005 Subject: [Histonet] Awards / Scholarships Message-ID: <314725EB.033E2430.3C887B41@aol.com> Dear NSH Memebers, It is not too late!!! Please take advantage of the generous benefits the NSH Awards / Scholarship program has to offer you as members, just remember you do not have to be nominated, "you" can just simply apply. Send your applications this next week, just submit the nomination and follow up with paperwork during the first 3 weeks in May. Thank you, Susan McCoy Awards Chairman From cwscouten <@t> myneurolab.com Sat Apr 24 10:30:52 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] tissue slicer Message-ID: I have been away at Experimental Biology. Is it too late to get in on this one? I want to correct a missimpresion. The Vibratome(TM) requires no embedding, is routinely used with living tissue to create sections for brain slice recording. Most brain slice people regard the Vibratome as necessary for gentlest handling of the tissue. Simple block the tissue with a razor blade, then use instant cyanoacrylate glue to hold the tissue in place on the pedestal. Cut any thickness you want down to as low as 30 microns. The egg slicer will crush the tissue, unless it is quite firm like a boiled egg, for the same reason you can walk on a bed of nails if there are lots of them, the weight is distributed to avoid penetration. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: Alan Bright [mailto:abright@brightinstruments.com] Sent: Tuesday, April 20, 2004 10:33 AM To: Doug Martin; Charles Scouten; Jim G. Unnerstall Subject: FW: [Histonet] tissue slicer And this Best regards Alan Bright Bright Instrument Co. Ltd England -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: 20 April 2004 18:05 To: k.whalley@ich.ucl.ac.uk Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] tissue slicer Almost forgot ........... There is a device called a "Stadie-Riggs tissue slicer" that will slice fresh tissue fairly thin, a few hundred microns. Thomas Scientific sells them, I don't know if anyone else does. A few hundred dollars in US currency, I think. Geoff Katy Whalley wrote: >Hi, > >Thanks to everyone for your advice and suggestions. It seems I might >have to re-think the thickness of the slices required - in retrospect, >I was probably aiming too low anyway. Basically, the reason we want to >cut them quickly is in order to culture the slices later, so I'm not >sure a vibratome or cryostat would be appropriate as I think the tissue >would have to be embedded/ frozen first. Overall the 'egg-slicer' or >matrix type of device seem like they may be the best option, but I may >try to make my own, as described by Geoff, to cut costs a bit. > >Katy > > Bad vibes, man. > > >>But, Geoff, your device would work, but as you note, not for less than >>1000 micron slices, definitely not 30 - 300 micron, and there'd be >>more tissue damage from compression. A vibratome is pretty much the >>only choice, there is no "all slices at once" instrument that I've >>seen. >> >>Phil >> >> >> >>>Always wondered what a vibratome was. >>>I've got a wifatome. >>> >>>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: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] >>>Sent: 19 April 2004 19:08 >>>To: k.whalley@ich.ucl.ac.uk >>>Cc: histonet@lists.utsouthwestern.edu >>>Subject: Re: [Histonet] tissue slicer >>> >>> >>>Hi Katy: >>> >>> You could buy a Vibratome, a device with a vibrating blade that >>>will cut fixed or unfixed tissue at a thickness you select. I think >>>there are several models and vendors. >>> Or, you could make an inexpensive device for little more than >>>pocket change. Buy some high-quality double-edge razor blades and >>>some material to use for spacing the blades. For 1 mm or more use >>>square aluminum rod, for 0.5 mm or less use "shim stock". A >>>well-stocked hardware store or maching shop will have these items. >>>Use "super-glue" to glue up a "blade-spacer-blade-spacer-blade ..." >>>tool with as many blades as your project demands. One 'application' >>>of the tool to the sample will give you uniform and reproducable >>>slices. Be sure to cut off or mask the edge of the blade not in use >>>so you won't cut yourself. >>> >>>Geoff >>> >>>Katy Whalley wrote: >>> >>> >>> >>>>Hi, >>>> >>>>We are looking for a device which can be used to cut tissue quickly >>>>into slices of an even thickness. We're not sure yet exactly how >>>>thick these will be but something in the range 30-300 microns is >>>>likely. My supervisor has in mind something in which several blades >>>>are attached to a holder that keeps them the correct distance apart, >>>>so that all the slices are cut >>>>at once. Has anyone ever used/ seen this kind of thing, or anything else >>>>which would do the job? >>>> >>>>thanks, >>>>Katy, UCL >>>> >>>> >>-- >>Philip Oshel >>Supervisor, BBPIC microscopy facility >>Department of Animal Sciences >>University of Wisconsin >>1675 Observatory Drive >>Madison, WI 53706 - 1284 >>voice: (608) 263-4162 >>fax: (608) 262-5157 (dept. 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 > > > -- -- ********************************************** 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 cwscouten <@t> myneurolab.com Sat Apr 24 10:53:11 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] vibratome or frozen sections for immuno Message-ID: Geoff, if I sent 30 or 40 refereces that used both frozen cryostat sections and Vibratome sections, would you have time look them up and see what comparison was mentioned? I would also be interested in knowing about the tract tracing studies. Did they favor the Vibratome or the cryostat, and why? Certainly, if a freeze thaw cycle is to be used anyway, initial cutting with a Vibratome makes little sense as opposed to a cryostat, unless quite thick sections are needed. But Vibratomes are widely used for immunoreactions, and I have heard via the grapevine that they get better results. Why is that? Does everybody feel the need to permeablize? How big are the antibodies? Bigger or smaller than the target proteins? Antibodies get better access maybe, but the target protein gets a chance to leak out, and be found on a nearby cell that did not produce it. Vibratomes keep cell membranes more intact. I agree with your comments on HRP, clearly, no need to permabilze the cells. Keep the protein in, the little molecules will find their way. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Monday, April 19, 2004 5:17 PM To: Charles Scouten Cc: Mike King; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] vibratome or frozen sections for immuno Re: the Vibratome v. frozen sections for immunostaining discussion. I looked up the papers Charles mentions below. The first two deal with tract tracing and thus do not pertain to the original question about penetration of antibodies/immunostaining reagents into 50 micron sections. The second two papers use both Vibratome and frozen sections. Hartig et al., J. Neurosci Meth. 67:89-95, 1996 finds no difference between their results with the two types of sections. Patel et al., Neurosci. Lett. 63:185-190, 1986 uses both types of sections, but their Vibratome sections are 50-100 microns while the cryostat sections are 2-6 microns. This does not seem like a valid comparison to me. However, they did not mention any difference in results between one type of section and the other. I think confusions arises in the intended purpose of the sections. Certainly for immuno some sort of permeabilization is usually used to insure that the very large antibody molecules penetrate into the cell cytoplasm. This is usually freeze-thaw (usually after appropriate cryoprotection), Triton, Tween, buffered ethanol treatment or some combination of these. However, for neuronal tract tracing large molecules need not penetrate the cell membrane, the HRP (or whatever tracer) is already in the cytoplasm and only DAB and peroxide, relatively small molecules, need to penetrate the cell. At the same time, one does not want the tracer to leak out of the cytoplasm. Perhaps Vibratome sections are more appropriate for that task but I suspect other variables would need to be considered as well Geoff Charles Scouten wrote: > Numerous studies have made the comparison. See a selected few below. > >It has been popular wisdom for 20 years that Vibratome sections, by not facturing the cell membrane, preserve the cytosol better and give more intense staining of cytosol proteins. I have personally seen differences with HRP staining, but didn't try to prove that some other variable was not responsible. > >I have located the studies below, and many others, that have made the comparison. I do not have access to the complete text, and do not know if all of these say the vibratome results were better. I would not expect vibratome to do better with membrane bound proteins. > >The science is there and done, I would compile a complete list of recent references like those below (maybe about 20-30) if somebody will volunteer to look them all up in Science Direct or the original Journals and check whether Vibratome sectioning was better, worse or the same compared to cryostat sectioning. > > >Current concepts in neuroanatomical tracing C. K?bbert, R. Apps, I. >Bechmann, J.L. Lanciego, J. Mey & S. Thanos Progress in Neurobiology, >2000, 62:4:327-351 solon@uni-muenster.de . The success of labelling can >be estimated roughly, and the tissue can be sectioned with a vibratome. Cryosectioning of the tissue is not recommended, since some dye leakage occurs during the procedure. > > >Neural tract tracing using Di-I: a review and a new method to make fast >Di-I faster in human brain D. Larry Sparks, Lih-Fen Lue, Timothy A. >Martin & Joseph Rogers Journal of Neuroscience Methods, 2000, 103:1:3 - >10 lsparks@mail.sunhealth.org if a fluorescent Di-I product is desired, infiltrating and embedding brain tissues in polyacrylamide prior to sectioning by vibratome eliminates the necessity of dehydrating and clearing the tissue sections prior to coverslipping ( Hayaran and Bijlani, 1992). >(too thick) >Although the thickness of the vibratome sections (50 ?m) was less than >optimal for demonstrating individual axons, individual fiber tracts >could occasionally be traced for as much as 5 mm or more, as shown >here. Cryostat and other techniques were used to achieve thinner >sections, but requiring freezing of the tissue, resulted in diffuse >smearing of the fluorescence label > >Somebody let me know if this is favorable to my case. The comparison was made, but I can't get the full text to see how it came out. > > > Triple immunofluorescence labelling of parvalbumin, calbindin-D28k and >calretinin in rat and monkey brain Hartig, W. / Bruckner, G. / Brauer, >K. / Seeger, G. / Bigl, V., Journal of Neuroscience Methods, Aug 1996 ...Thereafter, two brains were cut on a Vibratome (TSE, Kronberg) at 50 1lm thickness...monkey brains the tissue was cryo-protected by equili- bration...about 4 days and subsequent cryo-protection. 2.3. Tnple immu7...results were obtained with Vibratome and frozen sections and with... > > > 53. Adenosine deaminase and histidine decarboxylase coexist in >certain neurons of the rat brain Patel, B.T. / Tudball, N. / Wada, H. / >Watanabe, T., Neuroscience Letters, Jan 1986 ...further 24 h at 4?C. Thick vibratome sections (50-100/~m) were obtained...required, the fixed blocks were cryo- protected with 301~:~ sucrose...and dried at 20~C for 24 h. Vibratome and cryostat sections were...immunoreactivity in rat brain vibratome sections was consistent with... > >Cordially, >Charles W. Scouten, Ph.D. >myNeuroLab.com >5918 Evergreen Blvd. >St. Louis, MO 63134 >Ph: 314 522 0300 >FAX 314 522 0377 >cwscouten@myneurolab.com >http://www.myneurolab.com > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mike >King >Sent: Wednesday, April 14, 2004 12:56 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] vibratome/frozen immuno, soap residue, >autofluorescence, and bad list posting habits > >re 9. RE: Vibratome Considerations (Charles Scouten) ...as for immunolabeling sensitivity differences between vibratome and frozen sections from cryoprotected tissue, is there any science to support this claim (references)? i've never seen a difference and doubt there is really any reason to expect one. especially if you need to permeabilize anyway... > >re Danielle Zalinski Vibratome Considerations post: >...50 micron sections of brain (vibratome or other) will label through the entire thickness depending on the fixation, permeabilization, and location of the antigen. if you're going to use the tissue for anything that demands full-thickness labeling, you should use a confocal microscope to check that the labeling is even through the entire thickness of the sections. papers in the histo literature will detail the procedures you need for your material, or post again for specifics. > > > > < other stuff deleted in the interest of brevity. -- -- ********************************************** 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 marytedo <@t> hotmail.com Sat Apr 24 20:20:55 2004 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Oven temp and time.... Message-ID: I usually leave my slices in the oven for O.N. between 42ºC and 47ºC. Mostly 42ºC. If I'm in a hurry I leave them for 30 min, or an hour at 58ºC. I'm afraid of burn the tissues... HT. MARIA T. DOMINGUEZ PATHOLOGY'S SERV. RIO GRANDE'S REG. HOSPITAL TIERRA DEL FUEGO, ARGENTINA. >From: Caroline Stott >To: Histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Oven temp and time.... >Date: Thu, 22 Apr 2004 11:00:52 +1200 > >37 degrees overnight. Or if in a real hurry, 60 degrees for >20minutes. >Caroline > >Caroline Stott > >Histology Service Unit >Medical School >University of Otago >Dunedin >(03) 479 7152 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Tired of spam? Get [1]advanced junk mail protection with MSN 8. References 1. http://g.msn.com/8HMAEN/2734??PS= From marjoh3 <@t> telus.net Sat Apr 24 21:14:54 2004 From: marjoh3 <@t> telus.net (Marilyn Johnson) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Immunohistochemistry on Poultry tissues Message-ID: <000e01c42a6b$19390110$6401a8c0@VALUED20606295> Hi Histonetters, I am searching for any Immunohistochemistry staining methods on poultry tissues. Please provide the diseases/tissues being tested and the method of staining (PAP, ABC) Any replies would be greatly appreciated Thanks in advance. Marilyn Johnson Alberta Agriculture Edmonton, Alberta, Canada From lpwenk <@t> covad.net Sun Apr 25 06:28:02 2004 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] PTAH Procedure References: <229A3566B9F0D311826E00D0B7441D79061A44E1@swedish_nt1.schosp.org> Message-ID: <00d101c42ab8$5f6dd8c0$c822d445@domainnotset.invalid> Instead of post-mordanting in Zenker or saturated mercuric chloride, try Bouin for 1 hour at 60 degree C., wash in running water 5-10 minutes to remove the yellow, and then stain as usual with PTAH. Usually works for us. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Cazares, Ruth" To: Sent: Friday, April 23, 2004 3:30 PM Subject: [Histonet] PTAH Procedure > Hello again Histonetters, > > > > I was wondering if anyone out there knows if there is a procedure for PTAH > that does not require the tissue to be treated in Zenker's? Any and all > information will be greatly appreciated. Thanks. > > > > Ruth Cazares > > > > > > > *** Confidentiality Statement *** > This e-mail is intended only for the use of the individual or entity to > which it is addressed and may contain information that is privileged and > confidential. If the reader of this message is not the intended recipient, > please notify the sender immediately by replying to this message and then > delete it from your system. Any review, dissemination, distribution, or > reproduction of this message by unintended recipients is strictly prohibited > and may be subject to legal restriction. > Thank you for your cooperation. > ************************* > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From lpwenk <@t> covad.net Sun Apr 25 06:51:01 2004 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Awards / Scholarships References: <314725EB.033E2430.3C887B41@aol.com> Message-ID: <00e501c42abb$94fee440$c822d445@domainnotset.invalid> To put in my 2 cents about the NSH Awards/Scholarships - APPLY, APPLY, APPLY! There are multiple awards/scholarships for taking classes (including attending conventions), obtaining information on IHC, mol. path., hard tissue, management, teaching, etc., ranging from $500 to $3000. The link to the NSH website for the scholarship/award applications (or to nominate someone else), criteria and application form is: http://www.nsh.org/membership/awardscholar.html Many scholarships/awards require that the recipient has been a member of NSH for at least 1 year. If you are not a member of NSH, join NOW, and apply NEXT year. For NSH membership form, go to: http://www.nsh.org/membership/application.html Summary of the awards below, if interested (skip if not interested): Student Scholarship Awards - three ($500.00) awards will be presented to deserving students in approved Schools of Histotechnology. Awards and sponsors are: - Sigma Diagnostics Student Scholarship - Sponsored by Sigma Diagnostics, Inc., St. Louis, Missouri - Sakura Finetek Student Scholarship - Sponsored by Sakura Finetek U.S.A. Inc., Torrance, California - Irwin S. Lerner Student Scholarship - Sponsored by Thermo Electron Corp., Pittsburgh, Pennsylvania Educational Scholarships - Five ($1000.00) Educational Scholarships will be presented. Awards and sponsors are: - Dezna C. Sheehan Memorial Educational Scholarship - Sponsored by National Society for Histotechnology - Robert A. Clark Memorial Educational Scholarship - Sponsored by Sakura Finetek, U.S.A. Inc., Torrance, California - Leonard Noble Educational Scholarship - Sponsored by Cardinal Health, Scientific Products Distribution, McGaw Park, Illinois - Richard-Allan Educational Scholarship - Sponsored by Richard-Allan Scientific, Kalamazoo, Michigan - Fisher Diagnostics/Fisher Healthcare Educational Scholarship - Fisher Diagnostics/Fisher Healthcare, Pittsburgh, PA Ventana Medical Systems Immunohistochemistry Award - Ventana Medical Systems, Tucson, AZ, sponsors this award with a $1000.00 award to a qualified applicant who is pursuing advanced education within the Histotechnology profession, with a specific interest in immunohistochemistry. Ventana Medical Systems In SITU Hybridization Award - Ventana Medical Systems Inc., Tucson, Arizona, sponsors this $1000 award to a qualified applicant who is pursuing advanced education within the histotechnology profession, with a specific interest in in situ hybridization. Ventana Medical Systems IHC/In Situ Laboratory Award - Ventana Medical Systems Inc., Tucson, Arizona, sponsors this $3000 award to a qualified Laboratory that is pursuing advanced education within the histotechnology profession, with a specific focus in immunohistochemistry and/or in situ hybridization. BioGenex Award for Excellence in In Situ Hybridization - BioGenex, San Ramon, California, sponsors this award with a $1000 grant. Criteria: NSH member certified in histology, who is pursuing advanced work in In-Situ Hybridization. BioGenex Award for Standardization in Immunohistochemistry - BioGenex, San Ramon, California, sponsors this award with a $1000 grant. Criteria: NSH member certified in histology, who has contributed to the advancement of histotechnology standardization in any facet of histopathology DAKO Award for Excellence in Standardization of the IHC Techniques -DAKO Corporation, Carpinteria, California, sponsors this award with a $1500 grant, OR the option of a seven day trip to DAKO Corporation in Santa Barbara, California for a five day training session on IHC procedures and techniques. Candidate must be familiar with and perform IHC techniques on a routine basis, as outlined in their institution's departmental manual on standardization (Quality Assurance). Ann Preece Award - PolySciences, Inc., Warrington, PA, sponsors this $750 award on to a qualified applicant who has excelled in working with calcified/undecalcified bone, other materials such as stents in vascular research or bioengineering development. Leica Leadership in Management Award - Leica Microsystems, Bannockburn, IL, sponsors this $3000 award to an experienced or recently promoted manager, assistant manager, or supervisor who is expected to provide management leadership within their current role. The individual exemplifies the qualities of a dedicated leader in communication and/or interpersonal skills. Leica Leadership in Teaching Award - Leica Mircosystems, Bannockburn, IL, sponsors this $1000 award to an individual dedicated to teaching Histotechnology, exemplifying the qualities of a dedicated teacher sharing their knowledge with others and advancing the growth of the profession of Histotechnology. The nominee can be an instructor, technician/technologist, or supervisor. J. B. McCormick, M.D. Award - Dr. James B. McCormick most generously sponsors this award for outstanding and exceptional service to the National Society for Histotechnology. Award consists of a functioning exact replica of an 1800 Liberkuhn Compass Microscope and an eight book boxed set of The Science Heritage Library, concerning the history of the microscope and histotechnology. Histotechnologist Of The Year Award - Thermo Electron Corp., Pittsburgh, Pennsylvania, sponsors this award with a $1000.00 grant to the Histologic Technician/Histotechnologist, who, in the opinion of the Awards committee, best exemplifies the qualities of dedication and service to the profession of Histotechnology. William J. Hacker Memorial Award - Hacker Instruments, Fairfield, New Jersey, sponsors this award with a $500.00 grant.to the best scientific paper or presentation utilizing microwave technology Lee G. Luna Foreign Travel Scholarship Award - This award is sponsored by Surgipath Medical Industries, Inc., Richmond, Illinois. $3000.00 to be used to support costs to travel, attend, or study abroad (outside the U.S.A.). 2. A copy of Mr. Lee Luna's book entitled, Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Rosemary and Donald Ostermeier Memorial Award - This award is sponsored by Mr. Mark E. Ostermeier of Naperville, Illinois with a $500.00 grant to the recipient that, in the opinion of the Awards Committee, embodies the qualities of dedication and devotion to their patients, profession, co-workers, staff and place of employment. I know this is rather long, but I think it is IMPRESSIVE that NSH has so many scholarships to help people in so many fields of histotechnology, and that so many companies are willing to sponsor these awards. Thank you from me, to NSH and to the companies. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Friday, April 23, 2004 5:35 PM Subject: [Histonet] Awards / Scholarships > Dear NSH Memebers, > > It is not too late!!! > Please take advantage of the generous benefits the NSH Awards / Scholarship program has to offer you as members, just remember you do not have to be nominated, "you" can just simply apply. Send your applications this next week, just submit the nomination and follow up with paperwork during the first 3 weeks in May. > > Thank you, > Susan McCoy > Awards Chairman > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From weimer_r_v <@t> hotmail.com Sun Apr 25 08:22:19 2004 From: weimer_r_v <@t> hotmail.com (r v weimer) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Antibodies - MSRS Message-ID: Maybe a little off topic, but the following may assist you in your work! Complimentary access to the MSRS PRIMARY ANTIBODY DATABASE ONLINE 200,000 PRIMARY ANTIBODIES and Where to Get Them The MSRS web site, www.antibodies-probes.com, offers an interactive searchable database containing a listing of 200,000+ PRIMARY ANTIBODIES!! We would like to give you a complimentary short term access in order that you can evaluate the database or due a search as regards your present request. USER NAME: tempor USER ID: msrs044abx The access codes are case sensitive - use only lower case. COMPLETE - Each primary antibody record contains 11 fields including antibody, host, antigen species, label, form, clone number, isotype, unit size, product number, specifications and supplier name. Each listing represents a mini-specifications sheet about the antibody and contains listings from 625+ manufacturers, laboratories and suppliers worldwide. The company index lists 5600+ immunology, molecular biology and biotechnology companies worldwide. Primary Antibody submissions are requested from companies as well as individuals/laboratories. Please inquire. MSRS/Aerie Corporation PO Box 1584 Birmingham, AL 35201 Tele: (800) 633-4931 Fax: (205) 995-1588 Subscriptions: msrs.antibodies@ebsco.com Technical: weimer@antibodies-probes.com Site: www.antibodies-probes.com TEMP >From: "Stylli, Stanley" <Stanley.Stylli@mh.org.au> >To: <histonet@lists.utsouthwestern.edu> >Subject: [Histonet] antibodies for FFPE human tissue >Date: Wed, 21 Apr 2004 08:24:18 +1000 > >Can someone please recommend a source for the following antibodies that they have used successfully (with or without antigen retrieval) on human tissue (preferably brain) > >p53 >bax >bid >bcl-2 >XIAP >bcl-XL >bcl-XS >caspase 3 > >Thanks > >Stan > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From badesuyi <@t> hotmail.com Sun Apr 25 10:56:59 2004 From: badesuyi <@t> hotmail.com (Banjo Adesuyi) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] PAN -KERATIN STAIN Message-ID: Hi, Please I want the best special stain technic for the demonstration of PAN-KERATIN. I will appreciate it, If I can be able to get this info. Thanking you all for the anticipated co-operation. Banjo Adesuyi. _________________________________________________________________ The new MSN 8: smart spam protection and 2 months FREE* http://join.msn.com/?page=features/junkmail From pam <@t> ategra.com Sun Apr 25 12:16:31 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Job Opportunities in Histology Latest Update 04/25/04 Message-ID: Hi Histonet, Here is the latest update on opportunities with some of my best clients throughout the US who are seeking Histology Supervisors, Histo Technologists and Histo Technicians. These are positions as direct employees of our client. There are fulltime 40 hour per week positions.. As a direct employee of one of our clients you will be provided with full benefits including Health Insurance, Vacation, Sick Pay, Relocation money and a lucrative sign-on bonus. I have part time and temporary positions as well. I have supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are some of my HOTTEST Histology Supevisory positions: 1. Colorado - Histology Lab Manager 2. Connecticut - Histology Manager 3. Connecticut - Histology Supervisor 4. Maryland - AP Supervisor (Histology/Cytology) 5. California - Histology Technical Coordinator Here are some of my HOTTEST Histo Tech bench positions: 1. Florida - Histo Tech (full time and part time positions) 2. Maine - Histo Tech 3. Georgia - Histo Tech 4. Pennsylvania - Histo Tech 5. Oregon - Histo Tech 6. Rhode Island - MOHS Tech 7. Wisconsin - MOHS Tech 8. California - Histology Trainer 9. California - Histo Tech 10. California - Lead Histo Tech 11. Nevada - Histo Tech 12. Maryland - Histo Tech 13. Massachusetts - Histo Tech (part time) 14. Colorado - Histo Tech 15. Massachusetts - MOHS Tech If you are interested in these jobs, please CALL ME ASAP at 800 466 9919 x234. To speed things up, please also send me a copy of your resume, (if you haven't already done so). If you are interested in jobs outside the above-mentioned areas, please send me your resume as well. I have clients throughout the US. I will keep your resume confidential and will not release it to anyone without your permission (This is Ategra policy as well as my own). My services are at no charge to you. Of course, you may be happy in your present job, but it never hurts to to keep an eye open. Also, if you have friends/peers who might be interested as well, if you could pass my query & name on to them I'd be very grateful. However, if you are interested in any of the jobs above, please call me. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Pam Barker Senior Lab Recruiter Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd. Winter Park, FL 32792 VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com To Learn More About Ategra: http://www.ategra.com -------------------------------------------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. -------------------------------------------------------------------------------------------------- From badesuyi <@t> hotmail.com Sun Apr 25 17:03:47 2004 From: badesuyi <@t> hotmail.com (Banjo Adesuyi) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] TISSUE SPECIMENS WANTED Message-ID: Hi, Please I will appreciate it, If you can assist me in getting the two tissue specimens listed below for my practical test. The tissue specimens are; 1) ARTERY - complete cross section. 2) OVARY - 1.0 x 1.0cm, to include cortical surface along one entire edge. Wishing you all God's guideance and protection, amen. BANJO ADESUYI, B.Sc, HT(ASCP) PATHOLOGY DEPARTMENT, VAL VERDE REGIONAL MEDICAL CENTER, 801 BEDELL AVENUE, DEL RIO, TEXAS 78840. _________________________________________________________________ Tired of spam? Get advanced junk mail protection with MSN 8. http://join.msn.com/?page=features/junkmail From jhnspam <@t> aol.com Sun Apr 25 20:10:26 2004 From: jhnspam <@t> aol.com (jhnspam@aol.com) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Biopsy processing schedule Message-ID: <1dc.1fde35e6.2dbdbb82@aol.com> What kind of tissue processor are you using? Pam Johnson From brucea <@t> unimelb.edu.au Sun Apr 25 21:09:51 2004 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] detect primordial germ cells in PFA-fixed marsupial embryos...? Message-ID: Hi I have been using alkaline phosphatase histochemistry to detect primordial germ cells in PFA-fixed marsupial embryos. I use 1mg/ml Fast Blue salt and 1mg/ml Naphthol AS phosphate sodium salt in 0.2M Tris buffer, pH 9.4. The staining is good but I have been told that it is not permanent. Ideally, I would like to dehydrate these stained specimens, embed them in paraffin and then section and stain them immunohistochemically for other germ cell markers and several growth factors. Does anyone have a protocol utilising alkaline phosphatase histochemistry that would allow me to do this? Thanks for any advice in advance, Dr Danielle Hickford Research Fellow Department of Zoology, University of Melbourne, Australia 3010. hickford@unimelb.edu.au -- 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 ******************************************************************************** This electronic message and all contents contain information 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 us immediately and destroy the original message and all copies. From Hitesh.Asarpota <@t> med.ge.com Mon Apr 26 04:32:47 2004 From: Hitesh.Asarpota <@t> med.ge.com (Asarpota, Hitesh (MED, Intern)) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Basic Info Message-ID: <6EDD9DC298BF9C48AC3EF66EA92596BC11819C2D@frbucmsx03medge> Hi.. I am a management student currently on an internship at GE Healthcare. Part of my work is researching on anti-bodies and reagents. Could someone please guide me to an appropriate URL wherein I could look for detailed (although comprehensible to a layman!) info on the same? Thank you! Regards, Hitesh Asarpota ---------------------------------------------------------- GE HealthCare Business Development Tel: +33 01 30 70 9224 Cell: +33 06 1171 4370 ---------------------------------------------------------- From c.m.vanderloos <@t> amc.uva.nl Mon Apr 26 04:37:33 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] RE: beta galactosidase nuclear counterstain Message-ID: <2d95662d5286.2d52862d9566@amc.uva.nl> Hi, Using X-gal as substrate and ferri/ferro-cyanide as chromogen for b-galactosidase activity you obtain a turquoise/blue reaction product that survives everything: alcohol, xylene, cooking in citrate, in situ procedure..... Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ---- Original Message ----- >From Shaumik Adhya Date Fri, 23 Apr 2004 10:46:59 +0100 To histonet@lists.utsouthwestern.edu Subject [Histonet] beta galactosidase nuclear counterstain Hi Histonetters, I'm trying to stain myocardium for beta galactosidase (resulting in a blue precipitate), and have been trying to use Nuclear Fast Red as a nuclear counterstain - whilst this gives me reasonable nuclear staining, the process of dehydrating and mounting makes me lose all the beta gal. If I use an aqueous mount such as gycergel, the Nuclear Fast Red dissolves out over the course of a week, which is less than ideal. My protocol for dehydrating and mounting is: 10 quick dips in 70% ethanol 2 min in 100% ethanol 5 min in 100% ethanol 5 min in Histoclear 2 min in Histoclear DPX to coverslip with. I'm wondering what people use as a nuclear counterstain for beta-gal, and if you had any tips or advice. Thanks From GDawson <@t> Milw.Dynacare.com Mon Apr 26 07:28:34 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] No Posting Ability Message-ID: Sorry, Just testing to see if anything I post makes it to the hist-net. G. Dawson From philip.bergin <@t> microbio.gu.se Mon Apr 26 08:28:13 2004 From: philip.bergin <@t> microbio.gu.se (Phil Bergin) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] OCT definition Message-ID: Hi everyone, Does anyone know what OCT stands for, and do people ever define it in papers or just call it OCT compound? Seems a silly question- but what do people normally do? We normally just call it OCT compound, but I have been asked to define it, by the copy editor for a paper. Thanks Phil From Terry.Marshall <@t> rothgen.nhs.uk Mon Apr 26 08:40:08 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] OCT definition Message-ID: Optimum cutting temperature 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: Phil Bergin [mailto:philip.bergin@microbio.gu.se] Sent: 26 April 2004 14:28 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] OCT definition Hi everyone, Does anyone know what OCT stands for, and do people ever define it in papers or just call it OCT compound? Seems a silly question- but what do people normally do? We normally just call it OCT compound, but I have been asked to define it, by the copy editor for a paper. Thanks Phil _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pstefanova <@t> sten.sunnybrook.utoronto.ca Mon Apr 26 08:41:45 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] OCT definition References: <200404261332.i3QDWU1e022591@sten.sunnybrook.utoronto.ca> Message-ID: <005d01c42b94$3aad5a10$9d194c8e@WS21203> HI Phil, O.C.T. means optimal cutting temperature. Good luck! Petia ----- Original Message ----- From: "Phil Bergin" To: Sent: Monday, April 26, 2004 9:28 AM Subject: [Histonet] OCT definition > Hi everyone, > > > > Does anyone know what OCT stands for, and do people ever define it in papers > or just call it OCT compound? Seems a silly question- but what do people > normally do? We normally just call it OCT compound, but I have been asked > to define it, by the copy editor for a paper. > > > > Thanks > > > > Phil > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From bhewlett <@t> cogeco.ca Mon Apr 26 08:48:09 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] OCT definition References: <20040426133139.87F6F284E@fep2.cogeco.net> Message-ID: <000701c42b95$30865d10$6500a8c0@mainbox> Hi Phil, OCT stands for Optimal Cuttting Temperature compound. Check out the following; http://www.tedpella.com/msds_html/27050msd.htm Regards, Bryan ----- Original Message ----- From: "Phil Bergin" To: Sent: Monday, April 26, 2004 9:28 AM Subject: [Histonet] OCT definition > Hi everyone, > > > > Does anyone know what OCT stands for, and do people ever define it in papers > or just call it OCT compound? Seems a silly question- but what do people > normally do? We normally just call it OCT compound, but I have been asked > to define it, by the copy editor for a paper. > > > > Thanks > > > > Phil > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From kspencer <@t> utmem.edu Mon Apr 26 09:38:17 2004 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] OCT definition In-Reply-To: <20040426133152.3F0652C051@mail1.utmem.edu> Message-ID: <5B7FAC88-978F-11D8-A6E7-000393967904@utmem.edu> Optimal Cutting Temperature. Never did make sense to me. -Kathleen On Monday, April 26, 2004, at 08:28 AM, Phil Bergin wrote: > Hi everyone, > > > > Does anyone know what OCT stands for, and do people ever define it in > papers > or just call it OCT compound? Seems a silly question- but what do > people > normally do? We normally just call it OCT compound, but I have been > asked > to define it, by the copy editor for a paper. > > > > Thanks > > > > Phil > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SUSANLMCCOY <@t> aol.com Mon Apr 26 10:25:07 2004 From: SUSANLMCCOY <@t> aol.com (SUSANLMCCOY@aol.com) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Awards / Scholarships Message-ID: <01F6E1D9.6450FC86.3C887B41@aol.com> HI PEGGY, I AGREE, APPLY! APPLY! APPLY! THANK YOU FOR YOUR MESSAHE TO THE MEMBERS. SUSAN From gcallis <@t> montana.edu Mon Apr 26 10:37:44 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] beta galactosidase nuclear counterstain In-Reply-To: <1082713619.4088e613a2bb6@www.webmail.ucl.ac.uk> Message-ID: <3.0.6.32.20040426093744.00be7e08@gemini.msu.montana.edu> Just air dry sections after final water rinse (after nuclear fast red counterstain). You can do this in front of a fan and up to 2 hours. Mount a permanent coverslip and avoid dehydration through alcohols, etc. At 10:46 AM 4/23/2004 +0100, you wrote: > > >Hi Histonetters, > >I'm trying to stain myocardium for beta galactosidase (resulting in a blue >precipitate), and have been trying to use Nuclear Fast Red as a nuclear >counterstain - whilst this gives me reasonable nuclear staining, the process >of dehydrating and mounting makes me lose all the beta gal. If I use an >aqueous mount such as gycergel, the Nuclear Fast Red dissolves out over the >course of a week, which is less than ideal. > >My protocol for dehydrating and mounting is: > >10 quick dips in 70% ethanol >2 min in 100% ethanol >5 min in 100% ethanol >5 min in Histoclear >2 min in Histoclear >DPX to coverslip with. > >I'm wondering what people use as a nuclear counterstain for beta-gal, and if >you had any tips or advice. Thanks > >Shaumik > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Apr 26 11:08:58 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] OCT definition In-Reply-To: <200404261333.i3QDXdpV022277@mail1.msu.montana.edu> Message-ID: <3.0.6.32.20040426100858.00be7e08@gemini.msu.montana.edu> Optimal Cutting Temperature, clearly stated on the dispensing bottle. Dr. McCormick was in on the development of this cryoembedding media with Miles aka Tissue Tek, now Sakura Finetek. The bottle also says it contains polyvinyl alcohol, polyethylene glycol and inert ingredients. The PVA and PEG concentration and molecular weights are proprietary. Via private discussion with him, he said one could adjust the PVA and/or PEG to give different cutting qualities at different temperatures and different hardness to the media. The mixture we buy today is what they came up with. It has a good temperature range (we use anywhere from -16C to -35C). It is designed to surround the tissue for support and not infiltrate (as one does with paraffin). Some do this, but it must be pointed out the if the MW of the major ingredients is large, infiltration would probably be minimal. Interesting, I have never been asked to define OCT in great detail for any publication! Usually it is stated tissue placed in O.C.T. cryoembedding media (be sure to put in trademark, Sakura Finetek, Torrance CA) and snap frozen at certain temperature. Methinks your editor asketh for too much when it is not really necessary to explain OCT is such detail. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Don.Birgerson <@t> leica-microsystems.com Mon Apr 26 11:16:59 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] OCT definition Message-ID: Kathleen, The original compound came in three temperature ranges. O.C.T. Roman numeral( l) for 0 to -15 degrees, , O.C.T.( II ) for -15 to -30, and O.C.T.( III ) for -30 to -60. Today you have only one choice. I think "O.C.T." Compound was a copyright name at one time and competitors now refer to "freezing compound" for their products. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 Kathleen Spencer To: Phil Bergin Sent by: cc: histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth Subject: Re: [Histonet] OCT definition western.edu 04/26/2004 09:38 AM Optimal Cutting Temperature. Never did make sense to me. -Kathleen On Monday, April 26, 2004, at 08:28 AM, Phil Bergin wrote: > Hi everyone, > > > > Does anyone know what OCT stands for, and do people ever define it in > papers > or just call it OCT compound? Seems a silly question- but what do > people > normally do? We normally just call it OCT compound, but I have been > asked > to define it, by the copy editor for a paper. > > > > Thanks > > > > Phil > > > > _______________________________________________ > Histonet mailing list > Histonet@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 weneng <@t> hotmail.com Mon Apr 26 12:09:19 2004 From: weneng <@t> hotmail.com (Wendy England) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Zinc formalin and 10% NBF Message-ID: Hello, One of my coworkers fixed some samples in Zinc formalin and some in 10% NBF but forgot to lable them. So now we don't know which is Zinc formalin and which is 10% NBF. The tissue are going to be stained by H&E. His question is if it's ok to put all of them in 10% NBF now. I think it's fine. But I want to hear you expert's opinion please. Wendy _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar – get it now! http://toolbar.msn.com/go/onm00200415ave/direct/01/ From Sue.Kapoor <@t> uhsi.org Mon Apr 26 12:16:30 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Tissue Processor - Vendors Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F55B1@khmcexch.uhsi.org> I am interested in getting quotes to purchase a new/used tissue processor. Please contact me off line: Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 sue.kapoor@uhsi.org From mcauliff <@t> umdnj.edu Mon Apr 26 15:21:18 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] vibratome or frozen sections for immuno In-Reply-To: References: Message-ID: <408D6F3E.6010607@umdnj.edu> Hi Charles: Charles Scouten wrote: >Geoff, if I sent 30 or 40 refereces that used both frozen cryostat sections and Vibratome sections, would you have time look them up and see what comparison was mentioned? I would also be interested in knowing about the tract tracing studies. Did they favor the Vibratome or the cryostat, and why? > No, keeping up with my own library work is difficult enough. >Certainly, if a freeze thaw cycle is to be used anyway, initial cutting with a Vibratome makes little sense as opposed to a cryostat, unless quite thick sections are needed. > >But Vibratomes are widely used for immunoreactions, and I have heard via the grapevine that they get better results. Why is that? > I don't know, ask the people who claim better results! >Does everybody feel the need to permeablize? How big are the antibodies? > The MW of an antibody varies from 150,000 to about 850,000 depending on which antibody class it belongs to. > Bigger or smaller than the target proteins? > Often larger than the target, but it does depend on the target. > Antibodies get better access maybe, but the target protein gets a chance to leak out, and be found on a nearby cell that did not produce it. > One of the goals of fixation is to stabilize proteins so they do not 'migrate'. > Vibratomes keep cell membranes more intact. > >I agree with your comments on HRP, clearly, no need to permabilze the cells. Keep the protein in, the little molecules will find their way. > >Cordially, >Charles W. Scouten, Ph.D. >myNeuroLab.com >5918 Evergreen Blvd. >St. Louis, MO 63134 >Ph: 314 522 0300 >FAX 314 522 0377 >cwscouten@myneurolab.com >http://www.myneurolab.com > Best, Geoff >-----Original Message----- >From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] >Sent: Monday, April 19, 2004 5:17 PM >To: Charles Scouten >Cc: Mike King; histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] vibratome or frozen sections for immuno > >Re: the Vibratome v. frozen sections for immunostaining discussion. > > I looked up the papers Charles mentions below. The first two deal with tract tracing and thus do not pertain to the original question about penetration of antibodies/immunostaining reagents into 50 micron sections. The second two papers use both Vibratome and frozen sections. >Hartig et al., J. Neurosci Meth. 67:89-95, 1996 finds no difference between their results with the two types of sections. Patel et al., Neurosci. Lett. 63:185-190, 1986 uses both types of sections, but their Vibratome sections are 50-100 microns while the cryostat sections are >2-6 microns. This does not seem like a valid comparison to me. However, they did not mention any difference in results between one type of section and the other. > I think confusions arises in the intended purpose of the sections. >Certainly for immuno some sort of permeabilization is usually used to insure that the very large antibody molecules penetrate into the cell cytoplasm. This is usually freeze-thaw (usually after appropriate cryoprotection), Triton, Tween, buffered ethanol treatment or some combination of these. However, for neuronal tract tracing large molecules need not penetrate the cell membrane, the HRP (or whatever >tracer) is already in the cytoplasm and only DAB and peroxide, relatively small molecules, need to penetrate the cell. At the same time, one does not want the tracer to leak out of the cytoplasm. Perhaps Vibratome sections are more appropriate for that task but I suspect other variables would need to be considered as well > >Geoff > >Charles Scouten wrote: > > > >>Numerous studies have made the comparison. See a selected few below. >> >>It has been popular wisdom for 20 years that Vibratome sections, by not facturing the cell membrane, preserve the cytosol better and give more intense staining of cytosol proteins. I have personally seen differences with HRP staining, but didn't try to prove that some other variable was not responsible. >> >>I have located the studies below, and many others, that have made the comparison. I do not have access to the complete text, and do not know if all of these say the vibratome results were better. I would not expect vibratome to do better with membrane bound proteins. >> >>The science is there and done, I would compile a complete list of recent references like those below (maybe about 20-30) if somebody will volunteer to look them all up in Science Direct or the original Journals and check whether Vibratome sectioning was better, worse or the same compared to cryostat sectioning. >> >> >>Current concepts in neuroanatomical tracing C. K?bbert, R. Apps, I. >>Bechmann, J.L. Lanciego, J. Mey & S. Thanos Progress in Neurobiology, >>2000, 62:4:327-351 solon@uni-muenster.de . The success of labelling can >>be estimated roughly, and the tissue can be sectioned with a vibratome. Cryosectioning of the tissue is not recommended, since some dye leakage occurs during the procedure. >> >> >>Neural tract tracing using Di-I: a review and a new method to make fast >>Di-I faster in human brain D. Larry Sparks, Lih-Fen Lue, Timothy A. >>Martin & Joseph Rogers Journal of Neuroscience Methods, 2000, 103:1:3 - >>10 lsparks@mail.sunhealth.org if a fluorescent Di-I product is desired, infiltrating and embedding brain tissues in polyacrylamide prior to sectioning by vibratome eliminates the necessity of dehydrating and clearing the tissue sections prior to coverslipping ( Hayaran and Bijlani, 1992). >>(too thick) >>Although the thickness of the vibratome sections (50 ?m) was less than >>optimal for demonstrating individual axons, individual fiber tracts >>could occasionally be traced for as much as 5 mm or more, as shown >>here. Cryostat and other techniques were used to achieve thinner >>sections, but requiring freezing of the tissue, resulted in diffuse >>smearing of the fluorescence label >> >>Somebody let me know if this is favorable to my case. The comparison was made, but I can't get the full text to see how it came out. >> >> >>Triple immunofluorescence labelling of parvalbumin, calbindin-D28k and >>calretinin in rat and monkey brain Hartig, W. / Bruckner, G. / Brauer, >>K. / Seeger, G. / Bigl, V., Journal of Neuroscience Methods, Aug 1996 ...Thereafter, two brains were cut on a Vibratome (TSE, Kronberg) at 50 1lm thickness...monkey brains the tissue was cryo-protected by equili- bration...about 4 days and subsequent cryo-protection. 2.3. Tnple immu7...results were obtained with Vibratome and frozen sections and with... >> >> >> 53. Adenosine deaminase and histidine decarboxylase coexist in >>certain neurons of the rat brain Patel, B.T. / Tudball, N. / Wada, H. / >>Watanabe, T., Neuroscience Letters, Jan 1986 ...further 24 h at 4?C. Thick vibratome sections (50-100/~m) were obtained...required, the fixed blocks were cryo- protected with 301~:~ sucrose...and dried at 20~C for 24 h. Vibratome and cryostat sections were...immunoreactivity in rat brain vibratome sections was consistent with... >> >>Cordially, >>Charles W. Scouten, Ph.D. >>myNeuroLab.com >>5918 Evergreen Blvd. >>St. Louis, MO 63134 >>Ph: 314 522 0300 >>FAX 314 522 0377 >>cwscouten@myneurolab.com >>http://www.myneurolab.com >> >> >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mike >>King >>Sent: Wednesday, April 14, 2004 12:56 PM >>To: histonet@lists.utsouthwestern.edu >>Subject: [Histonet] vibratome/frozen immuno, soap residue, >>autofluorescence, and bad list posting habits >> >>re 9. RE: Vibratome Considerations (Charles Scouten) ...as for immunolabeling sensitivity differences between vibratome and frozen sections from cryoprotected tissue, is there any science to support this claim (references)? i've never seen a difference and doubt there is really any reason to expect one. especially if you need to permeabilize anyway... >> >>re Danielle Zalinski Vibratome Considerations post: >>...50 micron sections of brain (vibratome or other) will label through the entire thickness depending on the fixation, permeabilization, and location of the antigen. if you're going to use the tissue for anything that demands full-thickness labeling, you should use a confocal microscope to check that the labeling is even through the entire thickness of the sections. papers in the histo literature will detail the procedures you need for your material, or post again for specifics. >> >> >> >> >> >> >< other stuff deleted in the interest of brevity. > > >-- >-- >********************************************** >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 >********************************************** > > > -- -- ********************************************** 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 secretGurl <@t> swmed.edu Mon Apr 26 13:42:09 2004 From: secretGurl <@t> swmed.edu (secretGurl@swmed.edu) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Hello! Message-ID: Dear Histonet, It's me ;-) [cid:photo.jpeg] i am honest, responsible, romantic person. iwould like to find my only love,to find my destiny. Attached file will tell you everything. Cheers, SecretGurl From weneng <@t> hotmail.com Mon Apr 26 13:51:20 2004 From: weneng <@t> hotmail.com (Wendy England) Date: Fri Sep 16 15:22:52 2005 Subject: Thanks! RE: [Histonet] Zinc formalin and 10% NBF Message-ID: Thanks to all who helped me on the question about the fixative! I forwarded all your email to him and he is so happy now. And I learned a lot too. Thanks again! Wendy >From: "Wendy England" >To: histonet@pathology.swmed.edu >Subject: [Histonet] Zinc formalin and 10% NBF >Date: Mon, 26 Apr 2004 10:09:19 -0700 > >Hello, >One of my coworkers fixed some samples in Zinc formalin and some in 10% NBF >but forgot to lable them. So now we don't know which is Zinc formalin and >which is 10% NBF. The tissue are going to be stained by H&E. His question >is if it's ok to put all of them in 10% NBF now. I think it's fine. But I >want to hear you expert's opinion please. > >Wendy > >_________________________________________________________________ >FREE pop-up blocking with the new MSN Toolbar – get it now! >http://toolbar.msn.com/go/onm00200415ave/direct/01/ > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ MSN Toolbar provides one-click access to Hotmail from any Web page – FREE download! http://toolbar.msn.com/go/onm00200413ave/direct/01/ From DDittus787 <@t> aol.com Mon Apr 26 13:51:47 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Zinc formalin and 10% NBF Message-ID: <7B6381B1.5A4D1B7E.0A1F969F@aol.com> I think putting both in 10%nbf is fine, but if you want to do a little detective work to see which was in which fixative ,then before putting both in you could a: cut fresh tissue stain and see if any have formalin deposits b. cut fresh and stain both for say cd20-bet you the zinc formalin is cleaner, stronger and better stained. good luck-just a little csi in me coming out. dana From pruegg <@t> colobio.com Mon Apr 26 13:36:19 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Zinc formalin and 10% NBF In-Reply-To: Message-ID: Wendy it would be ok to put the samples in 10%NBF after zinc formalin fixation (for processing on the processor I assume?) but the best way to store samples after they have been aldehyde fixed is in 70% alcohol, this is more important for avoiding over fixation for IHC so if you are just doing H&E on these samples it should not be of concern to you. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Wendy England Sent: Monday, April 26, 2004 11:09 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Zinc formalin and 10% NBF Hello, One of my coworkers fixed some samples in Zinc formalin and some in 10% NBF but forgot to lable them. So now we don't know which is Zinc formalin and which is 10% NBF. The tissue are going to be stained by H&E. His question is if it's ok to put all of them in 10% NBF now. I think it's fine. But I want to hear you expert's opinion please. Wendy _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar ? get it now! http://toolbar.msn.com/go/onm00200415ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From marytedo <@t> hotmail.com Mon Apr 26 14:10:47 2004 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] About Respiratory cytologies Message-ID: Hi all! Today I received a Bronchial washing specimen (3 ml) in the lab. We don't usually performe these kind of specimens. The sample was collected tree days ago in the Ushuaia's Hospital ,300km away,and sent it to this Hospital (RíoGrande's Hospital), this morning without any fixative. Dam! I said... What should I do first? I put the sample in the cytocentrifuge chambers with Saccommanno's fixative, and let them there for 10 min at 1500rpm. Now my quiestion is "What happen if I would fill the chambers with the bronchial washing and Salt Solution equal parts? How come avoid the cellular distortion? The result was not so bad. My boss saw the cells, but just a few of them. I don't know if it was the right amount ,or if it was just the very resistents cells. A comment it will be apreciated. Ht. Maria Teresa Dominguez Pathology Serv. Río Grande Reg. Hospital, Tierra del Fuego, Argentina. _________________________________________________________________ Tired of spam? Get [1]advanced junk mail protection with MSN 8. References 1. http://g.msn.com/8HMAEN/2734??PS= From SYates <@t> agr.state.il.us Mon Apr 26 14:14:05 2004 From: SYates <@t> agr.state.il.us (SYates@agr.state.il.us) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Leica CV5030 Message-ID: Hi Histonetters My lab recently acquired the Leica CV 5030 coverslipper and have continuosly had problems with it dropping slides. Usually it drops the slides when the arm picks it up from the xylene bath or, as soon as the coverslip is placed on it, the arm releases it before the slide is put in the output rack. Does anyone else have these problems with this machine? Any suggestions? Sarah Yates Illinois Department of Agriculture Animal Disease Laboratory 9732 Shattuc Rd. Centralia, Il 62881 618-532-6701 From Rcartun <@t> harthosp.org Mon Apr 26 14:33:05 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] OCT definition Message-ID: I believe it is "Optimal Cutting Temperature". Richard Cartun >>> "Phil Bergin" 04/26/04 09:28AM >>> Hi everyone, Does anyone know what OCT stands for, and do people ever define it in papers or just call it OCT compound? Seems a silly question- but what do people normally do? We normally just call it OCT compound, but I have been asked to define it, by the copy editor for a paper. Thanks Phil _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From burch007 <@t> mc.duke.edu Mon Apr 26 14:44:23 2004 From: burch007 <@t> mc.duke.edu (James L Burchette) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Anti-MAGE B57 antibody Message-ID: Does anyone know of a source for anti-MAGE B57 antibody? Kindest regards, Jim Burchette From MDiCarlo <@t> KaleidaHealth.Org Mon Apr 26 14:56:19 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] need drawers for filing Message-ID: Hello Histonetters, I need to purchase drawers that 2 x 3 inch slides can fit in. I have the cabinet so I only need the drawers. The ones that I have are 19" long x 2 1/4" wide x 1 3/4" deep. Does anyone know where I can purchase these? Thanks for your help. Peggy DiCarlo HT (ASCP) Ortho Bone Lab Buffalo General Hospital 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 Ian.Bernard <@t> LACKLAND.AF.MIL Mon Apr 26 15:33:29 2004 From: Ian.Bernard <@t> LACKLAND.AF.MIL (Bernard Ian R SSgt 59 CRES/MSROP) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Hello! Message-ID: Not here sister -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of secretGurl@swmed.edu Sent: Monday, April 26, 2004 1:42 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hello! Dear Histonet, It's me ;-) [cid:photo.jpeg] i am honest, responsible, romantic person. iwould like to find my only love,to find my destiny. Attached file will tell you everything. Cheers, SecretGurl From Jackie.O'Connor <@t> abbott.com Mon Apr 26 15:55:48 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Hello! Message-ID: Could be a brother . . . . .secretly. Bernard Ian R SSgt 59 CRES/MSROP Sent by: histonet-bounces@lists.utsouthwestern.edu 04/26/2004 03:33 PM To: "'secretGurl@swmed.edu'" , histonet@pathology.swmed.edu cc: Subject: RE: [Histonet] Hello! Not here sister -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of secretGurl@swmed.edu Sent: Monday, April 26, 2004 1:42 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hello! Dear Histonet, It's me ;-) [cid:photo.jpeg] i am honest, responsible, romantic person. iwould like to find my only love,to find my destiny. Attached file will tell you everything. Cheers, SecretGurl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From degaboh <@t> rice.edu Mon Apr 26 16:20:01 2004 From: degaboh <@t> rice.edu (ZP) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] General: formalin & OCT? Message-ID: <20040426212002.4A3731DB31@handler11.mail.rice.edu> I am trying to do IHC for gelatin microparticles that are loaded with a growth factor. Can I use formalin to fix them, and then embed them in OCT for cryosectioning for later IHC? Better yet...can I use acetone instead of formalin? Which will give the best results for the IHC? thanks! From Ian.Bernard <@t> LACKLAND.AF.MIL Mon Apr 26 16:35:08 2004 From: Ian.Bernard <@t> LACKLAND.AF.MIL (Bernard Ian R SSgt 59 CRES/MSROP) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Hello! Message-ID: Just kidding you'all -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Monday, April 26, 2004 3:56 PM To: Bernard Ian R SSgt 59 CRES/MSROP Cc: histonet@pathology.swmed.edu Subject: RE: [Histonet] Hello! Could be a brother . . . . .secretly. Bernard Ian R SSgt 59 CRES/MSROP Sent by: histonet-bounces@lists.utsouthwestern.edu 04/26/2004 03:33 PM To: "'secretGurl@swmed.edu'" , histonet@pathology.swmed.edu cc: Subject: RE: [Histonet] Hello! Not here sister -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of secretGurl@swmed.edu Sent: Monday, April 26, 2004 1:42 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hello! Dear Histonet, It's me ;-) [cid:photo.jpeg] i am honest, responsible, romantic person. iwould like to find my only love,to find my destiny. Attached file will tell you everything. Cheers, SecretGurl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Robert.Fauck <@t> ccdhb.org.nz Mon Apr 26 18:17:51 2004 From: Robert.Fauck <@t> ccdhb.org.nz (Robert Fauck) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] RE: Histonet Digest, Vol 5, Issue 40 Message-ID: <9952FA35C61B4F4F90DC0B2D58FE68A958444F@WN0NTEML01.wn0.hiq.net.nz> Hi Pam, Do you know by chance also some job vacancies in the Middle East? Or a good website for searching for jobs in the Middle East? Thanking you in advance for some info. Cheers, Robert Fauck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, 27 April 2004 5:03 a.m. To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 5, Issue 40 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Job Opportunities in Histology Latest Update 04/25/04 (Pam Barker (extension 234)) 2. TISSUE SPECIMENS WANTED (Banjo Adesuyi) 3. Re: Biopsy processing schedule (jhnspam@aol.com) 4. detect primordial germ cells in PFA-fixed marsupial embryos...? (Bruce Abaloz) 5. Basic Info (Asarpota, Hitesh (MED, Intern)) 6. RE: beta galactosidase nuclear counterstain (C.M. van der Loos) 7. No Posting Ability (Dawson, Glen) 8. OCT definition (Phil Bergin) 9. RE: OCT definition (Marshall Terry Dr, Consultant Histopathologist) 10. Re: OCT definition (Petia P Stefanova) 11. Re: OCT definition (Bryan Hewlett) 12. Re: OCT definition (Kathleen Spencer) 13. Re: Awards / Scholarships (SUSANLMCCOY@aol.com) 14. Re: beta galactosidase nuclear counterstain (Gayle Callis) 15. OCT definition (Gayle Callis) 16. Re: OCT definition (Don.Birgerson@leica-microsystems.com) ---------------------------------------------------------------------- Message: 1 Date: Sun, 25 Apr 2004 13:16:31 -0400 From: Pam Barker (extension 234) Subject: [Histonet] Job Opportunities in Histology Latest Update 04/25/04 To: Histonetters Message-ID: Content-Type: Text/Plain Hi Histonet, Here is the latest update on opportunities with some of my best clients throughout the US who are seeking Histology Supervisors, Histo Technologists and Histo Technicians. These are positions as direct employees of our client. There are fulltime 40 hour per week positions.. As a direct employee of one of our clients you will be provided with full benefits including Health Insurance, Vacation, Sick Pay, Relocation money and a lucrative sign-on bonus. I have part time and temporary positions as well. I have supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are some of my HOTTEST Histology Supevisory positions: 1. Colorado - Histology Lab Manager 2. Connecticut - Histology Manager 3. Connecticut - Histology Supervisor 4. Maryland - AP Supervisor (Histology/Cytology) 5. California - Histology Technical Coordinator Here are some of my HOTTEST Histo Tech bench positions: 1. Florida - Histo Tech (full time and part time positions) 2. Maine - Histo Tech 3. Georgia - Histo Tech 4. Pennsylvania - Histo Tech 5. Oregon - Histo Tech 6. Rhode Island - MOHS Tech 7. Wisconsin - MOHS Tech 8. California - Histology Trainer 9. California - Histo Tech 10. California - Lead Histo Tech 11. Nevada - Histo Tech 12. Maryland - Histo Tech 13. Massachusetts - Histo Tech (part time) 14. Colorado - Histo Tech 15. Massachusetts - MOHS Tech If you are interested in these jobs, please CALL ME ASAP at 800 466 9919 x234. To speed things up, please also send me a copy of your resume, (if you haven't already done so). If you are interested in jobs outside the above-mentioned areas, please send me your resume as well. I have clients throughout the US. I will keep your resume confidential and will not release it to anyone without your permission (This is Ategra policy as well as my own). My services are at no charge to you. Of course, you may be happy in your present job, but it never hurts to to keep an eye open. Also, if you have friends/peers who might be interested as well, if you could pass my query & name on to them I'd be very grateful. However, if you are interested in any of the jobs above, please call me. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Pam Barker Senior Lab Recruiter Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd. Winter Park, FL 32792 VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com To Learn More About Ategra: http://www.ategra.com ------------------------------------------------------------------------ -------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ------------------------------------------------------------------------ -------------------------- ------------------------------ Message: 2 Date: Mon, 26 Apr 2004 01:03:47 +0300 From: "Banjo Adesuyi" Subject: [Histonet] TISSUE SPECIMENS WANTED To: histonet@lists.utsouthwestern.edu Cc: badesuyi@hotmail.com Message-ID: Content-Type: text/plain; format=flowed Hi, Please I will appreciate it, If you can assist me in getting the two tissue specimens listed below for my practical test. The tissue specimens are; 1) ARTERY - complete cross section. 2) OVARY - 1.0 x 1.0cm, to include cortical surface along one entire edge. Wishing you all God's guideance and protection, amen. BANJO ADESUYI, B.Sc, HT(ASCP) PATHOLOGY DEPARTMENT, VAL VERDE REGIONAL MEDICAL CENTER, 801 BEDELL AVENUE, DEL RIO, TEXAS 78840. _________________________________________________________________ Tired of spam? Get advanced junk mail protection with MSN 8. http://join.msn.com/?page=features/junkmail ------------------------------ Message: 3 Date: Sun, 25 Apr 2004 21:10:26 EDT From: jhnspam@aol.com Subject: Re: [Histonet] Biopsy processing schedule To: histonet@lists.utsouthwestern.edu Message-ID: <1dc.1fde35e6.2dbdbb82@aol.com> Content-Type: text/plain; charset="US-ASCII" What kind of tissue processor are you using? Pam Johnson ------------------------------ Message: 4 Date: Mon, 26 Apr 2004 12:09:51 +1000 From: Bruce Abaloz Subject: [Histonet] detect primordial germ cells in PFA-fixed marsupial embryos...? To: histonet-bounces@lists.utsouthwestern.edu, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Hi I have been using alkaline phosphatase histochemistry to detect primordial germ cells in PFA-fixed marsupial embryos. I use 1mg/ml Fast Blue salt and 1mg/ml Naphthol AS phosphate sodium salt in 0.2M Tris buffer, pH 9.4. The staining is good but I have been told that it is not permanent. Ideally, I would like to dehydrate these stained specimens, embed them in paraffin and then section and stain them immunohistochemically for other germ cell markers and several growth factors. Does anyone have a protocol utilising alkaline phosphatase histochemistry that would allow me to do this? Thanks for any advice in advance, Dr Danielle Hickford Research Fellow Department of Zoology, University of Melbourne, Australia 3010. hickford@unimelb.edu.au -- 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 ************************************************************************ ******** This electronic message and all contents contain information 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 us immediately and destroy the original message and all copies. ------------------------------ Message: 5 Date: Mon, 26 Apr 2004 11:32:47 +0200 From: "Asarpota, Hitesh (MED, Intern)" Subject: [Histonet] Basic Info To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <6EDD9DC298BF9C48AC3EF66EA92596BC11819C2D@frbucmsx03medge> Content-Type: text/plain; charset="iso-8859-1" Hi.. I am a management student currently on an internship at GE Healthcare. Part of my work is researching on anti-bodies and reagents. Could someone please guide me to an appropriate URL wherein I could look for detailed (although comprehensible to a layman!) info on the same? Thank you! Regards, Hitesh Asarpota ---------------------------------------------------------- GE HealthCare Business Development Tel: +33 01 30 70 9224 Cell: +33 06 1171 4370 ---------------------------------------------------------- ------------------------------ Message: 6 Date: Mon, 26 Apr 2004 11:37:33 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: beta galactosidase nuclear counterstain To: histonet@lists.utsouthwestern.edu Message-ID: <2d95662d5286.2d52862d9566@amc.uva.nl> Content-Type: text/plain; charset=us-ascii Hi, Using X-gal as substrate and ferri/ferro-cyanide as chromogen for b-galactosidase activity you obtain a turquoise/blue reaction product that survives everything: alcohol, xylene, cooking in citrate, in situ procedure..... Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ---- Original Message ----- >From Shaumik Adhya Date Fri, 23 Apr 2004 10:46:59 +0100 To histonet@lists.utsouthwestern.edu Subject [Histonet] beta galactosidase nuclear counterstain Hi Histonetters, I'm trying to stain myocardium for beta galactosidase (resulting in a blue precipitate), and have been trying to use Nuclear Fast Red as a nuclear counterstain - whilst this gives me reasonable nuclear staining, the process of dehydrating and mounting makes me lose all the beta gal. If I use an aqueous mount such as gycergel, the Nuclear Fast Red dissolves out over the course of a week, which is less than ideal. My protocol for dehydrating and mounting is: 10 quick dips in 70% ethanol 2 min in 100% ethanol 5 min in 100% ethanol 5 min in Histoclear 2 min in Histoclear DPX to coverslip with. I'm wondering what people use as a nuclear counterstain for beta-gal, and if you had any tips or advice. Thanks ------------------------------ Message: 7 Date: Mon, 26 Apr 2004 07:28:34 -0500 From: "Dawson, Glen" Subject: [Histonet] No Posting Ability To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Sorry, Just testing to see if anything I post makes it to the hist-net. G. Dawson ------------------------------ Message: 8 Date: Mon, 26 Apr 2004 15:28:13 +0200 From: "Phil Bergin" Subject: [Histonet] OCT definition To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi everyone, Does anyone know what OCT stands for, and do people ever define it in papers or just call it OCT compound? Seems a silly question- but what do people normally do? We normally just call it OCT compound, but I have been asked to define it, by the copy editor for a paper. Thanks Phil ------------------------------ Message: 9 Date: Mon, 26 Apr 2004 14:40:08 +0100 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] OCT definition To: "Phil Bergin" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Optimum cutting temperature 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: Phil Bergin [mailto:philip.bergin@microbio.gu.se] Sent: 26 April 2004 14:28 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] OCT definition Hi everyone, Does anyone know what OCT stands for, and do people ever define it in papers or just call it OCT compound? Seems a silly question- but what do people normally do? We normally just call it OCT compound, but I have been asked to define it, by the copy editor for a paper. Thanks Phil _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Mon, 26 Apr 2004 09:41:45 -0400 From: "Petia P Stefanova" Subject: Re: [Histonet] OCT definition To: "Phil Bergin" , Message-ID: <005d01c42b94$3aad5a10$9d194c8e@WS21203> Content-Type: text/plain; charset="iso-8859-1" HI Phil, O.C.T. means optimal cutting temperature. Good luck! Petia ----- Original Message ----- From: "Phil Bergin" To: Sent: Monday, April 26, 2004 9:28 AM Subject: [Histonet] OCT definition > Hi everyone, > > > > Does anyone know what OCT stands for, and do people ever define it in papers > or just call it OCT compound? Seems a silly question- but what do people > normally do? We normally just call it OCT compound, but I have been asked > to define it, by the copy editor for a paper. > > > > Thanks > > > > Phil > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 11 Date: Mon, 26 Apr 2004 09:48:09 -0400 From: "Bryan Hewlett" Subject: Re: [Histonet] OCT definition To: "Phil Bergin" , Message-ID: <000701c42b95$30865d10$6500a8c0@mainbox> Content-Type: text/plain; charset="iso-8859-1" Hi Phil, OCT stands for Optimal Cuttting Temperature compound. Check out the following; http://www.tedpella.com/msds_html/27050msd.htm Regards, Bryan ----- Original Message ----- From: "Phil Bergin" To: Sent: Monday, April 26, 2004 9:28 AM Subject: [Histonet] OCT definition > Hi everyone, > > > > Does anyone know what OCT stands for, and do people ever define it in papers > or just call it OCT compound? Seems a silly question- but what do people > normally do? We normally just call it OCT compound, but I have been asked > to define it, by the copy editor for a paper. > > > > Thanks > > > > Phil > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 12 Date: Mon, 26 Apr 2004 09:38:17 -0500 From: Kathleen Spencer Subject: Re: [Histonet] OCT definition To: Phil Bergin Cc: histonet@lists.utsouthwestern.edu Message-ID: <5B7FAC88-978F-11D8-A6E7-000393967904@utmem.edu> Content-Type: text/plain; format=flowed; charset=US-ASCII Optimal Cutting Temperature. Never did make sense to me. -Kathleen On Monday, April 26, 2004, at 08:28 AM, Phil Bergin wrote: > Hi everyone, > > > > Does anyone know what OCT stands for, and do people ever define it in > papers > or just call it OCT compound? Seems a silly question- but what do > people > normally do? We normally just call it OCT compound, but I have been > asked > to define it, by the copy editor for a paper. > > > > Thanks > > > > Phil > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Mon, 26 Apr 2004 11:25:07 -0400 From: SUSANLMCCOY@aol.com Subject: Re: [Histonet] Awards / Scholarships To: lpwenk@covad.net, histonet@lists.utsouthwestern.edu Message-ID: <01F6E1D9.6450FC86.3C887B41@aol.com> Content-Type: text/plain; charset=iso-8859-1 HI PEGGY, I AGREE, APPLY! APPLY! APPLY! THANK YOU FOR YOUR MESSAHE TO THE MEMBERS. SUSAN ------------------------------ Message: 14 Date: Mon, 26 Apr 2004 09:37:44 -0600 From: Gayle Callis Subject: Re: [Histonet] beta galactosidase nuclear counterstain To: Shaumik Adhya , Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040426093744.00be7e08@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" Just air dry sections after final water rinse (after nuclear fast red counterstain). You can do this in front of a fan and up to 2 hours. Mount a permanent coverslip and avoid dehydration through alcohols, etc. At 10:46 AM 4/23/2004 +0100, you wrote: > > >Hi Histonetters, > >I'm trying to stain myocardium for beta galactosidase (resulting in a blue >precipitate), and have been trying to use Nuclear Fast Red as a nuclear >counterstain - whilst this gives me reasonable nuclear staining, the process >of dehydrating and mounting makes me lose all the beta gal. If I use an >aqueous mount such as gycergel, the Nuclear Fast Red dissolves out over the >course of a week, which is less than ideal. > >My protocol for dehydrating and mounting is: > >10 quick dips in 70% ethanol >2 min in 100% ethanol >5 min in 100% ethanol >5 min in Histoclear >2 min in Histoclear >DPX to coverslip with. > >I'm wondering what people use as a nuclear counterstain for beta-gal, and if >you had any tips or advice. Thanks > >Shaumik > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 15 Date: Mon, 26 Apr 2004 10:08:58 -0600 From: Gayle Callis Subject: [Histonet] OCT definition To: "Phil Bergin" , Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040426100858.00be7e08@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" Optimal Cutting Temperature, clearly stated on the dispensing bottle. Dr. McCormick was in on the development of this cryoembedding media with Miles aka Tissue Tek, now Sakura Finetek. The bottle also says it contains polyvinyl alcohol, polyethylene glycol and inert ingredients. The PVA and PEG concentration and molecular weights are proprietary. Via private discussion with him, he said one could adjust the PVA and/or PEG to give different cutting qualities at different temperatures and different hardness to the media. The mixture we buy today is what they came up with. It has a good temperature range (we use anywhere from -16C to -35C). It is designed to surround the tissue for support and not infiltrate (as one does with paraffin). Some do this, but it must be pointed out the if the MW of the major ingredients is large, infiltration would probably be minimal. Interesting, I have never been asked to define OCT in great detail for any publication! Usually it is stated tissue placed in O.C.T. cryoembedding media (be sure to put in trademark, Sakura Finetek, Torrance CA) and snap frozen at certain temperature. Methinks your editor asketh for too much when it is not really necessary to explain OCT is such detail. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 16 Date: Mon, 26 Apr 2004 11:16:59 -0500 From: Don.Birgerson@leica-microsystems.com Subject: Re: [Histonet] OCT definition To: Kathleen Spencer Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu, Phil Bergin Message-ID: Content-Type: text/plain; charset=us-ascii Kathleen, The original compound came in three temperature ranges. O.C.T. Roman numeral( l) for 0 to -15 degrees, , O.C.T.( II ) for -15 to -30, and O.C.T.( III ) for -30 to -60. Today you have only one choice. I think "O.C.T." Compound was a copyright name at one time and competitors now refer to "freezing compound" for their products. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 Kathleen Spencer To: Phil Bergin Sent by: cc: histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth Subject: Re: [Histonet] OCT definition western.edu 04/26/2004 09:38 AM Optimal Cutting Temperature. Never did make sense to me. -Kathleen On Monday, April 26, 2004, at 08:28 AM, Phil Bergin wrote: > Hi everyone, > > > > Does anyone know what OCT stands for, and do people ever define it in > papers > or just call it OCT compound? Seems a silly question- but what do > people > normally do? We normally just call it OCT compound, but I have been > asked > to define it, by the copy editor for a paper. > > > > Thanks > > > > Phil > > > > _______________________________________________ > Histonet mailing list > Histonet@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 End of Histonet Digest, Vol 5, Issue 40 *************************************** C&C DHB Secure Mail Server. ************************************************************************ ******** No Viruses were detected in this message. ************************************************************************ ******** C&C DHB Secure Mail Server. ******************************************************************************** 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. (AC_S001) No Viruses were detected in this message. ******************************************************************************** From rowani.mohdrawi <@t> student.adelaide.edu.au Mon Apr 26 18:38:21 2004 From: rowani.mohdrawi <@t> student.adelaide.edu.au (Rowani) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Best Way to obtain specific area of mouse foetal and neonatal Brain for EM and light microscopy References: <3.0.6.32.20040420090357.00bc2778@gemini.msu.montana.edu> Message-ID: <003301c42be7$8fd95040$87997f81@LaptopRowani> Yes, its electron microscopy but what I meant is to cut specific area of the brain (ie. hippocampus, motor cortex etc) which could then be used for EM and various other purposes. By the way, could anyone share with me the best method to obtain specific area of foetal/neonatal mouse brain for further high resolution analysis (ie. dentate gyrus, hippocampus, thalamus, motor cortex etc). Thank You Rowani From AnthonyH <@t> chw.edu.au Mon Apr 26 19:24:26 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] PTAH Procedure Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1A3@simba.kids> Ruth, we use the Cherukian Method: Regards Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html Document Procedure: Phosphotungstic Acid Haematoxylin Principle: Cherukian's modification employs an eosin solution that stains the erythrocytes red and differentiates them from the blue fibrin. Fixation: 10% buffered formalin. Microtomy: paraffin sections at 5um. Controls: Use brain sections and section of muscle. A good stain will demonstrate the dendrites as blue where as in a bad stain they appear light grey to salmon in colour. Nuclei, fibrin, platelets and muscle will be blue, red cells and collagen appear red. Muscle striations should be well defined. Reagents: 1. Eosin: - Warning: Flammable - see MSDS Eosin Y, water soluble (CI 145380) 0.5g Distilled Water 10ml 80% Ethanol 190ml Working: 10ml stock and Before use add 50ul glacial acetic acid. 2. 1% Periodic Acid 3. PTAH solution Haematoxylin (CI 75290) 0.5g Phosphotungstic Acid 10g Distilled water 500ml Dissolve solid ingredients in separate portions of the water. Use gentle heat for Haematoxylin. Combine solutions when cool. Add 0.088g potassium permanganate to ripen. The stain is ready to use. Procedure: 1. Dewax and hydrate sections to 80% alcohol. 2. Place slides in eosin for 30 seconds. 3. Wash slides in distilled water for a few seconds. 4. Place slides in 1% periodic acid for 20 minutes. 5. Wash slides in water for 3 minutes. 6. Place slides in PTAH for 30-90 minutes in 60oC oven. Check from 30 minutes on. 7. Dehydrate, clear and mount. Results: Dendrites, nuclei, fibrin, platelets and muscle - blue Red blood cells and collagen - red. Notes: Reference: 1. Cherukian, C.J., Histologic. 8(4); 105, (1977). 2. Luna, L., Histologic. 5(2); 66, (1975). -----Original Message----- From: Cazares, Ruth [mailto:RCazares@schosp.org] Sent: Saturday, 24 April 2004 5:31 AM To: histonet@pathology.swmed.edu Subject: [Histonet] PTAH Procedure Hello again Histonetters, I was wondering if anyone out there knows if there is a procedure for PTAH that does not require the tissue to be treated in Zenker's? Any and all information will be greatly appreciated. Thanks. Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From WilsonL <@t> jwci.org Mon Apr 26 20:17:54 2004 From: WilsonL <@t> jwci.org (Lori Wilson) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Re: H&E staining of frozen section Message-ID: <1CD765A8D6B6DB4F887110A8EEE09CE502BB65A5@smexsvr1.jwci.org> Does anyone have a protocol for H&E staining of frozen sections? I am particularly interested in breast, but any tissue type would be useful. The specimens were prepared in a cryomold using OCT. From WilsonL <@t> jwci.org Mon Apr 26 20:19:59 2004 From: WilsonL <@t> jwci.org (Lori Wilson) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] RE: Cryojane cutting and LCM Message-ID: <1CD765A8D6B6DB4F887110A8EEE09CE502BB65A6@smexsvr1.jwci.org> Has anyone used the cryojane with OCT frozen sections to perform LCM? We are using it for protein extraction and mass spectrometry analysis. I am having incomplete microdissection and there is an odd background staining. Does anyone have experience with this? From brucea <@t> unimelb.edu.au Mon Apr 26 20:29:49 2004 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee & Peggy/Tony, SOMEONE....) Message-ID: Hi, my name is Danielle & I need some advice PLEASE - I have been using alkaline phosphatase histochemistry to detect primordial germ cells in PFA-fixed marsupial embryos. I use 1mg/ml Fast Blue salt and 1mg/ml Naphthol AS phosphate sodium salt in 0.2M Tris buffer, pH 9.4. The staining is good but I have been told that it is not permanent. Ideally, I would like to dehydrate these stained specimens, embed them in paraffin and then section and stain them immunohistochemically for other germ cell markers and several growth factors. Does anyone have a protocol utilising alkaline phosphatase histochemistry that would allow me to do this? Thanks for any advice in advance, Dr Danielle Hickford Research Fellow Department of Zoology, University of Melbourne, Australia 3010.>hickford@unimelb.edu.au -- 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 ******************************************************************************** This electronic message and all contents contain information 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 us immediately and destroy the original message and all copies. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- 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 ******************************************************************************** This electronic message and all contents contain information 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 us immediately and destroy the original message and all copies. From jkiernan <@t> uwo.ca Tue Apr 27 00:08:19 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Zinc formalin and 10% NBF References: Message-ID: <408DEAC3.DF3A01C7@uwo.ca> After fixing the specimens in either NBF or a "zinc-formalin" mixture you should put them in water to dissolve out the sodium phosphate or zinc salts (chloride or sulphate?) before moving the specimens into the dehydrating alcohols. John Kiernan London, Canada ________________________________________________________ Wendy England wrote: > > Hello, > One of my coworkers fixed some samples in Zinc formalin and some in 10% NBF > but forgot to lable them. So now we don't know which is Zinc formalin and > which is 10% NBF. The tissue are going to be stained by H&E. His question is > if it's ok to put all of them in 10% NBF now. I think it's fine. But I want > to hear you expert's opinion please. > > Wendy > > _________________________________________________________________ > FREE pop-up blocking with the new MSN Toolbar ? get it now! > http://toolbar.msn.com/go/onm00200415ave/direct/01/ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo <@t> tiscali.co.uk Tue Apr 27 01:52:29 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Re: H&E staining of frozen section In-Reply-To: <1CD765A8D6B6DB4F887110A8EEE09CE502BB65A5@smexsvr1.jwci.org> Message-ID: <408BBDAE00005F74@mk-cpfrontend-2.mail.uk.tiscali.com> We used to stain frozen sections, unmounted on slides, in little coplin jars. I concede these were sections of brain etc that were rather thick, but it seemed to work. I'd try cutting the sections at 10 mu and then staining in fresh (not over-oxidised) haematoxylin for 30s, blue in tap water, counterstain in eosin for 30s, then dehydrate. I can't remember when we attached to slide, but it was either before or after dehydration but before clearing, as the sections went hard and wouldn't flatten. Breast may be a problem due to the fat, and you may want to put the sections through alcohol and then a clearing agent to remove the fat, then back to water. I'd be interested to hear how you get on. TTFN __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From mab70 <@t> medschl.cam.ac.uk Tue Apr 27 01:54:50 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] need drawers for filing Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16A4@mius.medlan.cam.ac.uk> Hi, Margaret, why not try Raymond A. Lamb? They manufacture a wide range of slide filing drawers and have a website: http://www.ralamb.net/ They usually come up with the goods for histology. Mr Lamb the founder was a histologist before setting up the company. Good luck margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of DiCarlo, Margaret Sent: Monday, April 26, 2004 8:56 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] need drawers for filing Hello Histonetters, I need to purchase drawers that 2 x 3 inch slides can fit in. I have the cabinet so I only need the drawers. The ones that I have are 19" long x 2 1/4" wide x 1 3/4" deep. Does anyone know where I can purchase these? Thanks for your help. Peggy DiCarlo HT (ASCP) Ortho Bone Lab Buffalo General Hospital 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 kemlo <@t> tiscali.co.uk Tue Apr 27 01:58:45 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] About Respiratory cytologies In-Reply-To: Message-ID: <408BBDAE00005FBC@mk-cpfrontend-2.mail.uk.tiscali.com> 'Stable(g) door and Horse'? I assume any damage done en route is done; 'We are, where we are' (fashionable saying in management speak), so I don't expect intensive care with Saccy's will do owt. What you have left is what you have left, so interprete with caution. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From kemlo <@t> tiscali.co.uk Tue Apr 27 02:02:12 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Hello! In-Reply-To: Message-ID: <408BBDAE00005FE1@mk-cpfrontend-2.mail.uk.tiscali.com> Geez this Listserv is looking up. Spam is very nice with chips, especially if battered. Anyone tried battered, deep fried Mars bar? __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From kemlo <@t> tiscali.co.uk Tue Apr 27 01:43:29 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Zinc formalin and 10% NBF In-Reply-To: <408DEAC3.DF3A01C7@uwo.ca> Message-ID: <408BBDAE00005F17@mk-cpfrontend-2.mail.uk.tiscali.com> Can't you test the residual fixative for the presence or absence of zinc? My Chemistry is not what it was but can't you do something with an element and the double decomposition of a salt? Something to do with an element higher up or lower down the Periodic Table that displaces an element from a salt that is the other way round, can't remember. But NBF doesn't contain the heavy metal does it? Why is it important to know? All I can remember is that zinc formalin made the tissue brittle, or was that lead? The nuclear stain was rather deep too. Hope that helps. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From mariatere <@t> infovia.com.ar Tue Apr 27 04:37:56 2004 From: mariatere <@t> infovia.com.ar (Teresa Dominguez) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] About Respiratory cytologies References: <408BBDAE00005FBC@mk-cpfrontend-2.mail.uk.tiscali.com> Message-ID: <007d01c42c3b$5410b450$417f46c8@tere> Thanks a lot! I will try to be careful the next time. ----- Original Message ----- From: "Kemlo" To: "Mar?a Teresa Dom?nguez" ; Sent: Tuesday, April 27, 2004 3:58 AM Subject: RE: [Histonet] About Respiratory cytologies 'Stable(g) door and Horse'? I assume any damage done en route is done; 'We are, where we are' (fashionable saying in management speak), so I don't expect intensive care with Saccy's will do owt. What you have left is what you have left, so interprete with caution. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mab70 <@t> medschl.cam.ac.uk Tue Apr 27 04:46:25 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Le e & Peggy/Tony, SOMEONE....) Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16A6@mius.medlan.cam.ac.uk> Hi, Everyone, With regard to this problem, I was wondering if the alk phos enzyme would survive processing, I rather think it would otherwise we wouldn't need to add levamisole when immunostaining paraffin sections using alk phos as a label. So why not process into wax then do the detection of alkaline phosphatase and the other markers you wish to identify. What do other histonetters think? Is it worth a try? Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bruce Abaloz Sent: Tuesday, April 27, 2004 2:30 AM To: histonet-bounces@lists.utsouthwestern.edu; histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee & Peggy/Tony, SOMEONE....) Hi, my name is Danielle & I need some advice PLEASE - I have been using alkaline phosphatase histochemistry to detect primordial germ cells in PFA-fixed marsupial embryos. I use 1mg/ml Fast Blue salt and 1mg/ml Naphthol AS phosphate sodium salt in 0.2M Tris buffer, pH 9.4. The staining is good but I have been told that it is not permanent. Ideally, I would like to dehydrate these stained specimens, embed them in paraffin and then section and stain them immunohistochemically for other germ cell markers and several growth factors. Does anyone have a protocol utilising alkaline phosphatase histochemistry that would allow me to do this? Thanks for any advice in advance, Dr Danielle Hickford Research Fellow Department of Zoology, University of Melbourne, Australia 3010.>hickford@unimelb.edu.au -- 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 **************************************************************************** **** This electronic message and all contents contain information 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 us immediately and destroy the original message and all copies. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- 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 **************************************************************************** **** This electronic message and all contents contain information 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 us immediately and destroy the original message and all copies. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Diane.Gladney <@t> se.amedd.army.mil Tue Apr 27 06:12:03 2004 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Re: H&E staining of frozen section Message-ID: <9D41AB7C56F8304F98537ABD87B249F66261DA@DASMTHGBZ001> Lori, I do H&E staining on frozen sections using sections that are 6 - 8 microns thick. We pick the sections up with a room temp. slide which is then placed in a coplin jar containing 95% Ethanol (ethyl alcohol). This technique requires some practice so I suggest practicing with a block of OCT compound without any tissue to perfect your technique before you attempt this with any tissue. The following staining protocol is used to stain the slides and they turn out beautiful (I have used this protocol for more years than I care to think about): 1. 95% Ethanol (brought from Cryostat) - Dip until clear 2. Running tap water (gentle stream of water running into a bowl or appropriate container) - dip until clear 3. Distilled water - dip until clear 4. Hematoxylin 2 (Richard Allen Scientific) - 30 seconds 5. Running tap water - until clear 6. Bluing Reagent (Richard Allen Scientific) - dip 3 - 4 times 7. Running tap water - 30 seconds 8. 95% Ethanol - 3 - 4 dips 9. Eosin - Y (Richard Allen Scientific) - 1 - 2 dips 10. 95% Ethanol - 2 changes - 5 - 10 dips 11. 100% Ethanol - 3 changes - 5 - 10 dips 12. Xylene - 2 changes - 5 - 10 dips 13. Mount with resinous medium (coverslip) I have a staining series set-up for doing frozen sections. The tissues are already mounted on the slide before you do the staining series, therefore you just need to coverslip when you finish staining the slides. I hope that this has helped you and meets your needs. Happy Cutting, Diane Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Lori Wilson Sent: Monday, April 26, 2004 9:18 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Re: H&E staining of frozen section Does anyone have a protocol for H&E staining of frozen sections? I am particularly interested in breast, but any tissue type would be useful. The specimens were prepared in a cryomold using OCT. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From froyer <@t> bitstream.net Tue Apr 27 08:04:51 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Hello! In-Reply-To: References: Message-ID: <408E5A73.6050104@bitstream.net> Wow... I've always said that the HistoNet was a great service! Now my subscription can even help me find a cyber girl friend! (It's okay! I'm single) And some people think that histology is monotonous ...... ~ Ford ;-) Ford M. Royer, MT(ASCP) Analytical Instruments, llc Minneapolis, MN secretGurl@swmed.edu wrote: > Dear Histonet, > It's me ;-) > [cid:photo.jpeg] > i am honest, responsible, romantic person. iwould like to find my only > love,to find my destiny. > Attached file will tell you everything. > Cheers, SecretGurl > > >------------------------------------------------------------------------ > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From HETZER <@t> surgery.wisc.edu Tue Apr 27 08:13:00 2004 From: HETZER <@t> surgery.wisc.edu (Michael Hetzer) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] staining frozen sections Message-ID: Lori, Staining frozen sections is a lot like staining paraffin sections except that there is no need to dewax. (OCT will rinse off during the staining process.) Dehyrdration is the critical factor with fresh frozen tissue. For fatty sections, be sure that the slides are dry before fixing them in 95% ETOH for three minutes. (You can fix the slides in 10% NBF but then you must really be sure to thoroughly rinse the slides before proceeding with the staining process.) We use Harris Hematoxylin and alcoholic Eosin and stain regressively but you can use your own H&E protocol. Mike Hetzer Mohs Surgery Madison, WI From scoop <@t> mail.nih.gov Tue Apr 27 08:01:03 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] mouse erythroid lineage subtypes Message-ID: Hi Histonetters, Does anyone know of any antibodies to the various stages of erythroid development that work in mice? I just use ter-119 now, but would like to get more specific. Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From barbara.wright2 <@t> dnax.org Tue Apr 27 08:34:42 2004 From: barbara.wright2 <@t> dnax.org (Wright, Barbara (SPRI 2)) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] RE: Cryojane cutting and LCM Message-ID: <29B25753F6B1D51196110002A589D4440105C897@PALMSG30.us.schp.com> Lori, I believe you need to use 1/2x slides for cryosjane sectioning (instead of the 1x) if you are going to use them for LCM. Barb -----Original Message----- From: Lori Wilson [mailto:WilsonL@jwci.org] Sent: Monday, April 26, 2004 6:20 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Cryojane cutting and LCM Has anyone used the cryojane with OCT frozen sections to perform LCM? We are using it for protein extraction and mass spectrometry analysis. I am having incomplete microdissection and there is an odd background staining. Does anyone have experience with this? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* 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 juan.gutierrez <@t> christushealth.org Tue Apr 27 08:44:38 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Leica CV5030 Message-ID: Yes, and I wish they(Leica) would take the piece of .... back! Stuck in the same boat, Juan -----Original Message----- From: SYates@agr.state.il.us [mailto:SYates@agr.state.il.us] Sent: Monday, April 26, 2004 2:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Leica CV5030 Hi Histonetters My lab recently acquired the Leica CV 5030 coverslipper and have continuosly had problems with it dropping slides. Usually it drops the slides when the arm picks it up from the xylene bath or, as soon as the coverslip is placed on it, the arm releases it before the slide is put in the output rack. Does anyone else have these problems with this machine? Any suggestions? Sarah Yates Illinois Department of Agriculture Animal Disease Laboratory 9732 Shattuc Rd. Centralia, Il 62881 618-532-6701 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Lizbeth_Kelly <@t> hgsi.com Tue Apr 27 08:23:00 2004 From: Lizbeth_Kelly <@t> hgsi.com (Lizbeth_Kelly@hgsi.com) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] NSH Region II Symposium (June 3,4, & 5th) Message-ID: The Maryland Society of Histotechnologists (MSH) is sponsoring the Region II Symposium June 3, 4, and 1/2 day on the 5th at the Holiday Inn Select, North-Baltimore, Maryland. For information, you may contact Terri DeCarli at 410-787-4546 or Renate Jaacks at 410-879-9012. Lizbeth Kelly, HT (ASCP), QIHC Board Member, MSH From juan.gutierrez <@t> christushealth.org Tue Apr 27 08:48:43 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Hello! Message-ID: I don't think viruses have genders. -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Monday, April 26, 2004 3:56 PM To: Bernard Ian R SSgt 59 CRES/MSROP Cc: histonet@pathology.swmed.edu Subject: RE: [Histonet] Hello! Could be a brother . . . . .secretly. Bernard Ian R SSgt 59 CRES/MSROP Sent by: histonet-bounces@lists.utsouthwestern.edu 04/26/2004 03:33 PM To: "'secretGurl@swmed.edu'" , histonet@pathology.swmed.edu cc: Subject: RE: [Histonet] Hello! Not here sister -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of secretGurl@swmed.edu Sent: Monday, April 26, 2004 1:42 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hello! Dear Histonet, It's me ;-) [cid:photo.jpeg] i am honest, responsible, romantic person. iwould like to find my only love,to find my destiny. Attached file will tell you everything. Cheers, SecretGurl _______________________________________________ Histonet mailing list Histonet@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 RCazares <@t> schosp.org Tue Apr 27 08:47:10 2004 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Re: PTAH Message-ID: <229A3566B9F0D311826E00D0B7441D79061A4559@swedish_nt1.schosp.org> Thank you for all the suggestions on the PTAH procedure! Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ************************* From philip.bergin <@t> microbio.gu.se Tue Apr 27 09:07:39 2004 From: philip.bergin <@t> microbio.gu.se (Phil Bergin) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] OCT definition- THANKS In-Reply-To: Message-ID: Thanks to everyone for the definition- one of those things that you remember when you read! Phil From Terry.Marshall <@t> rothgen.nhs.uk Tue Apr 27 09:00:01 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Zinc formalin and 10% NBF Message-ID: Crisp to cut and crisp to look at - yes. The phosphate in formalin buffer precipitates zinc, as, naturally, zinc phosphate, so washing between one and the other is desirable. If not, one sometimes sees haematoxyphil smudges in the tissue, and of course, cloudiness in the solution before processing. (A little acetic acid takes care of that). Of course, the easiest course, if you want to go from zinc to formalin, is to go to unbuffered formalin. (Why is there such a fixation for buffered fixatives - no pun intended?) 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: Kemlo [mailto:kemlo@tiscali.co.uk] Sent: 27 April 2004 07:43 To: jkiernan@uwo.ca; Wendy England Cc: histonet@pathology.swmed.edu Subject: Re: [Histonet] Zinc formalin and 10% NBF Can't you test the residual fixative for the presence or absence of zinc? My Chemistry is not what it was but can't you do something with an element and the double decomposition of a salt? Something to do with an element higher up or lower down the Periodic Table that displaces an element from a salt that is the other way round, can't remember. But NBF doesn't contain the heavy metal does it? Why is it important to know? All I can remember is that zinc formalin made the tissue brittle, or was that lead? The nuclear stain was rather deep too. Hope that helps. From mprice26 <@t> juno.com Tue Apr 27 09:39:34 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Leitz 1512/Grease Message-ID: <20040427.074033.26044.657@webmail08.nyc.untd.com> Histonetters, Does anyone know where I can order the grease and oil for the Leitz 1512 microtome? Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From gcallis <@t> montana.edu Tue Apr 27 09:46:22 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Re: H&E staining of frozen section In-Reply-To: <1CD765A8D6B6DB4F887110A8EEE09CE502BB65A5@smexsvr1.jwci.org > Message-ID: <3.0.6.32.20040427084622.00bcefe8@gemini.msu.montana.edu> This is the only time we do not air dry frozen section. Cut section and go immediately to neutral buffered formalin, a minute or so fixation, rinse with tap water. Richard Allan Hematoxylin 1 for 30 dips, rinse in tap water (count to 30 or so) Bluing solution until section visibly turns blue rinse in tap water (count to 30 or so) Dip in 70% alcohol 20 times Eosin 10 to 20 dips (use amount of time you need for your favorite eosin quality of staining) 95% X 2 100% X 2 Clearant X 2 Coverslip It all takes less than 10 minutes for staining. We have used 95% ethanol for fixation, flood slide or dip for 30 seconds, rinse and do same staining, but our morphology is better with NBF. At 06:17 PM 4/26/2004 -0700, you wrote: >Does anyone have a protocol for H&E staining of frozen sections? I am >particularly interested in breast, but any tissue type would be useful. The >specimens were prepared in a cryomold using OCT. > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue Apr 27 10:21:13 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Zinc formalin and 10% NBF In-Reply-To: Message-ID: <3.0.6.32.20040427092113.00bfba50@gemini.msu.montana.edu> Have used zinc formalin mixture, but not in house preparation, we purchased it from Anatech and never experienced crisp nor difficult to section paraffin embedded tissues. We never went back into NBF though (on processor) and did routine tissue processing starting in 70% ethanol. Pre-rinsing was very short, 5 minutes to remove fixative on outside of tissues, doubtful it was extensive enough to remove zinc formalin further into tissues. Our human kidney biopsies were fixed in Zinc formalin, hand processed through a very short biopsy schedule - the sectioning was wonderful, 1 to 2 um sections were possible, without tears, biopsy was never crispy! Staining was crisp, sectioning was normal. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Don.Birgerson <@t> leica-microsystems.com Tue Apr 27 10:27:41 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Leitz 1512/Grease Message-ID: Hi Marsha, The oil and grease for the 1512 is still available from Leica-Microsystems. 14033621783 Oil #601 14033625075 #411 White grease 14033624601 #410 Brown grease Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 mprice26@juno.com Sent by: To: histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] Leitz 1512/Grease 04/27/2004 09:39 AM Histonetters, Does anyone know where I can order the grease and oil for the Leitz 1512 microtome? Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sue.Kapoor <@t> uhsi.org Tue Apr 27 09:59:20 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] slide stainers - vendors Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F55BC@khmcexch.uhsi.org> Hi, I'm putting through my budget and I'm interested in slide stainers with the capacity to stain more than one rack at a time. I currently have a (working)Tissue-Tek stainer, possible trade-in? Vendors please contact me off-line. Thank you, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 sue.kapoor@uhsi.org From Terry.Marshall <@t> rothgen.nhs.uk Tue Apr 27 10:36:48 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Zinc formalin and 10% NBF Message-ID: Having only once used zinc as a primary fixative (in NZ), and having used it for 30 years as a secondary fixative, I can say with some confidence that it just isn't as good as a secondary fixative, but does improve the sections - i.e., better than not having it. However, the only time I've not had complaints about the tissue being crisp, brittle and lifting off is when used as a primary fixative. Conclusion: Zinc and pathologists who use it as a secondary fixative are tiresome. 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: 27 April 2004 16:21 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Zinc formalin and 10% NBF Have used zinc formalin mixture, but not in house preparation, we purchased it from Anatech and never experienced crisp nor difficult to section paraffin embedded tissues. We never went back into NBF though (on processor) and did routine tissue processing starting in 70% ethanol. Pre-rinsing was very short, 5 minutes to remove fixative on outside of tissues, doubtful it was extensive enough to remove zinc formalin further into tissues. Our human kidney biopsies were fixed in Zinc formalin, hand processed through a very short biopsy schedule - the sectioning was wonderful, 1 to 2 um sections were possible, without tears, biopsy was never crispy! Staining was crisp, sectioning was normal. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From susan.wells <@t> bms.com Tue Apr 27 10:46:52 2004 From: susan.wells <@t> bms.com (Susan Q Wells) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Human on human staining Message-ID: <408E806B.3257C408@bms.com> Good morning- Anyone out there doing IHC with a fully human antibody on human tissue ? I'd like to purchase a kit if there is one? Otherwise I may have to biotinylate my antibody? I'd appreciate any tips, suggestions. Thanks, Sue Wells From jfish <@t> gladstone.ucsf.edu Tue Apr 27 11:22:03 2004 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Best Way to obtain specific area of mouse foetal and neonatal Brain for EM and light microscopy In-Reply-To: <003301c42be7$8fd95040$87997f81@LaptopRowani> References: <3.0.6.32.20040420090357.00bc2778@gemini.msu.montana.edu> <003301c42be7$8fd95040$87997f81@LaptopRowani> Message-ID: Dear Rowani, Here is what I do to select specific areas of the brain for EM. First, cut 100um vibratome sections after the initial glutaraldehyde fixation. Then process these sections through dehydration and infiltration with Eponate 12. Embed them between Aclar sheets (I buy mine from Ted Pella) with just a small drop of resin for each section. I have embedded at least 50 sections between two 8x6" sheets. After they have polymerized you can look at the sections with an ordinary upright microscope. You can then select specific areas and mount them onto empty or "blank" resin blocks (that you have made ahead of time). Just remove the area you have chosen with a razor blade or scalpel. You might practice first before going for the important and rare specimen! I use regular gel type Super Glue to glue the areas to the blocks. After they have dried you can trim and section just like any other EM block. The nice benefit of this method is that the embedded sheets can be stored in an ordinary binder until you need them again. This works great for me, hope it works great for you. Take care and good luck, Jo Dee At 9:08 AM +0930 4/27/04, Rowani wrote: >Yes, its electron microscopy but what I meant is to cut specific area of the >brain (ie. hippocampus, motor cortex etc) which could then be used for EM >and various other purposes. > >Thank You >Rowani > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** ********** Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 695-3720 Fax: (415) 285-5632 E-mail: jfish@gladstone.ucsf.edu Mailing address: Gladstone Institutes P.O. Box 419100 San Francisco, CA 94141-9100 From Kristopher.Kalleberg <@t> unilever.com Tue Apr 27 12:07:09 2004 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher.Kalleberg@unilever.com) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] (no subject) Message-ID: When performing IHC, and the stain is combined with water, are better results achieved when rinsed with water or should PBS be used From mwhitesi <@t> adventisthealthcare.com Tue Apr 27 12:18:37 2004 From: mwhitesi <@t> adventisthealthcare.com (Marnie Whiteside) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Job Opening Message-ID: Hello Currently we have a histo tech position opening here at Washington Adventist Hospital. We are located inside the beltway in Takoma Park Maryland. If you would like more information please call me at 301-891-6306. Marnie Whiteside HT From TJJ <@t> Stowers-Institute.org Tue Apr 27 13:15:02 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Re: Zinc formalin and 10% NBF Message-ID: In working through our processing schedule dilemma, we fixed some pancreas in NBF (overnight), and fixed some in Zinc Formalin for 3 hours, followed by water wash and storage in 70% ethanol. We found that using a variety of processing schedules, the NBF fixed pancreas looked good with routine 30 min/station (with 2 hours total time in paraffin), all the way down to 10 min/station (with 80 min total time in paraffin). The Zinc Formalin fixed pancreas looks dry and brittle still, even with the short processing schedule. It was amazing to see how firm and grayish the tissue was as compared to the NBF fixed tissue, within even an hour of fixation time. We will try a hand-processing schedule for these to see if we can get the processing perfect for the ZnF pancreas. Our answer to this problem could be that we need to limit the amount of time it is in fixative. Gayle, how long were the kidney biopsies fixed before processing? Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From Jan.Minshew <@t> leica-microsystems.com Tue Apr 27 13:21:52 2004 From: Jan.Minshew <@t> leica-microsystems.com (Jan.Minshew@leica-microsystems.com) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] RE: Final version for Histonet Message-ID: Dear Histonetters, This is in response to the posting sent by Sara Yates regarding her problem with slides being dropped by the Leica CV5030 Coverslipper. After speaking with Sara, I learned that the problem only occurs on slides that are pre-labeled before staining. We have found that if the label is exposed to xylene during the staining or coverslipping process, the adhesive material will cause the coverslipper gripper arm to stick to the sides of the slide, fail in its attempt to place the slide in the output rack and eventually allow the slide to fall. To significantly reduce the problem, the label must be perfectly centered so that no adhesive areas extend over the edges of the slide, and the xylene levels must be lowered in the staining and holding containers to prevent exposure of the labels to the adhesive solvent. If Sara had experienced problems with unlabeled slides being dropped, it would probably indicate that the gripper arm needed to be optimized using our new "teach-in" to insure that it recognized the size of the slide being used. Since manufacturers offer various sizes of slides, it's possible that the factory settings might not be appropriate. This update can be accomplished by any Leica Sales Representative or Field Service Engineer. If any other individual is experiencing problems with a Leica product, please call us at 1-800-248-0123 and ask for assistance from our Technical Application Center (TAC). Thank you for keeping us aware of issues as our main concern is customer satisfaction. Carolyn Earley Marketing Manager Leica Microsystems 847-405-7014 Carolyn.Earley@Leica-Microsystems.com From ploykasek <@t> phenopath.com Tue Apr 27 13:35:56 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] NBT/BCIP Message-ID: I have a question to post from a research colleague. For those of you using NBT/BCIP as a substrate, which company/formulation do you prefer for stability? It will be used as a substrate in a tyramide amplification procedure. Thanks for your help. Patti Loykasek PhenoPath Laboratories Seattle, WA From sladd <@t> hsc.usf.edu Tue Apr 27 13:49:47 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] searching the archives for a specific posting Message-ID: I have a paper copy of a posting from Jan. 23, 2003. The subject line says "Cell Blocks". The posting contains a protocol for embedding cells using histogel specimen processing gel and I wanted to send the posting to someone else. No matter what combination of keywords I use, or how I look, I cannot find this posting in the archives. Any suggestions? Sharron From gcallis <@t> montana.edu Tue Apr 27 14:33:20 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Zinc formalin on human renal biopsies In-Reply-To: Message-ID: <3.0.6.32.20040427133320.00bc1090@gemini.msu.montana.edu> Teri, The renal biopsies (thin, needle bx) were fixed 12 hours, were sent overnight to our lab and on occasion were fixed longer. Well fixed and then we hand processed these to avoid any overexposure to dehydrants. Pathologist did not like VIP short schedule, but we did changes using vacuum for each change. You did not say IF you made up your zinc formalin in house or bought it from Anatech? We never made this in house as some people complain about Zn formalin precipitating out during processing. We never had this problem, and attributed this to a proper buffering, making up of commercial solution by Anatech - whatever they do/did was superb for our needs. Remember we were working with human tissue, although Diane Sterchi and I gave a workshop and compared Zinc formalin to NBF on animal tissues without problems. Hmmm full moon causes problems!!??? Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 lab At 01:15 PM 4/27/2004 -0500, you wrote: >In working through our processing schedule dilemma, we fixed some >pancreas in NBF (overnight), and fixed some in Zinc Formalin for 3 >hours, followed by water wash and storage in 70% ethanol. We found that >using a variety of processing schedules, the NBF fixed pancreas looked >good with routine 30 min/station (with 2 hours total time in paraffin), >all the way down to 10 min/station (with 80 min total time in paraffin). >The Zinc Formalin fixed pancreas looks dry and brittle still, even with >the short processing schedule. > >It was amazing to see how firm and grayish the tissue was as compared to >the NBF fixed tissue, within even an hour of fixation time. > >We will try a hand-processing schedule for these to see if we can get >the processing perfect for the ZnF pancreas. Our answer to this problem >could be that we need to limit the amount of time it is in fixative. >Gayle, how long were the kidney biopsies fixed before processing? > > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, Missouri 64110 >tjj@stowers-institute.org > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From cfavara <@t> niaid.nih.gov Tue Apr 27 14:43:53 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] searching the archives for a specific posting Message-ID: I can tell you how I do this. Cells usually arrive in a centrifuge tube. Spin down add some warm histo gel and viola you have a nice pellet that will go through the processor with ease. If you have a very particular client and they want a specific dilution that can be done as well! Good luck! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: S Ladd [mailto:sladd@hsc.usf.edu] Sent: Tuesday, April 27, 2004 12:50 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] searching the archives for a specific posting I have a paper copy of a posting from Jan. 23, 2003. The subject line says "Cell Blocks". The posting contains a protocol for embedding cells using histogel specimen processing gel and I wanted to send the posting to someone else. No matter what combination of keywords I use, or how I look, I cannot find this posting in the archives. Any suggestions? Sharron _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sandra.Etheridge <@t> gems8.gov.bc.ca Tue Apr 27 14:48:54 2004 From: Sandra.Etheridge <@t> gems8.gov.bc.ca (Etheridge, Sandra AGF:EX) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Fibrin and Myoglobin Stains Message-ID: <424FAC08A8CDFE4BADD6497AE75B5E2F0E0A866D@atlas.gov.bc.ca> Hello fellow Histonetters, I work in an provincial animal health monitoring lab (mostly agricultural animals) and one of the pathologists was asking if there were specific special stains for fibrin and myoglobin. We currently use PTAH for fibrin and muscle fibers, and Okajima for hemoglobin and myoglobin. Does anyone else know of more specific stains for these two constituents? I can't seem to find much in the limited reference library here. Thanks for the help. Sandra Etheridge Animal Health Center BC Ministry of Agriculture Abbotsford, BC Canada From siksik03 <@t> comcast.net Tue Apr 27 14:52:42 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Re: H&E staining of frozen section In-Reply-To: <3.0.6.32.20040427084622.00bcefe8@gemini.msu.montana.edu> References: <3.0.6.32.20040427084622.00bcefe8@gemini.msu.montana.edu> Message-ID: Hi HistoNetters I have suggested this many times in the past, but I'll say it once again... Nuclear details will be much better preserved if the cryostat (frozen) sections are first fixed in a microwave (see Chapter 14 of Kok & Boon, "Combining microwave and freeze techniques"). The easiest way to do this is quickly put the slide in a horizontal position on a polystyrene platform in the microwave and cover the section with a few drops of Kryofix or equivalent PEG/ethyl alcohol fixative. Microwave at 450W for 20 seconds. Alternatively, you can place the slide vertically in a Coplin jar filled with fixative and microwave at 55?C for 20 seconds. The slide should still be wet, in any case, after microwaving. Other methods have used one part 40% formaldehyde and 3 parts 96% alcohol, or Wolman's fluid (5% acetic acid in industrial alcohol), but I prefer the PEG/ethyl alcohol methods. After fixing, stain as suggested by Gayle Callis. best regards, Steven Slap Microwave Consultant From pstefanova <@t> sten.sunnybrook.utoronto.ca Tue Apr 27 14:41:38 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] 4% PFA References: Message-ID: <008301c42c8f$a83704e0$9d194c8e@WS21203> Hi to all, Does anybody know the duration of fixation with PFA? My samples are mice ovaries and testis so I guess it won't take long. I appreciate your hepl! Petia From lagehan <@t> hotmail.com Tue Apr 27 15:05:15 2004 From: lagehan <@t> hotmail.com (LORALEE GEHAN) Date: Fri Sep 16 15:22:52 2005 Subject: [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee & Peggy/Tony, SOMEONE....) Message-ID: With my experience it won't survive the processing. Try it plastic. The processing is a bit longer but the stain works nicely. Hope that helps. Loralee Gehan, HTL >From: Margaret Blount >To: 'Bruce Abaloz' , histonet-bounces@lists.utsouthwestern.edu, histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee & Peggy/Tony, SOMEONE....) >Date: Tue, 27 Apr 2004 10:46:25 +0100 > >Hi, Everyone, > >With regard to this problem, I was wondering if the alk phos enzyme would >survive processing, I rather think it would otherwise we wouldn't need to >add levamisole when immunostaining paraffin sections using alk phos as a >label. So why not process into wax then do the detection of alkaline >phosphatase and the other markers you wish to identify. > >What do other histonetters think? Is it worth a try? > >Margaret > >Margaret Blount >Chief Technician >Clinical Biochemistry >University of Cambridge >Addenbrooke's Hospital >Hills Road >Cambridge >CB2 2QR > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bruce >Abaloz >Sent: Tuesday, April 27, 2004 2:30 AM >To: histonet-bounces@lists.utsouthwestern.edu; >histonet@lists.utsouthwestern.edu >Subject: [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee >& Peggy/Tony, SOMEONE....) > > >Hi, my name is Danielle & I need some advice PLEASE - >I have been using alkaline phosphatase histochemistry to detect primordial >germ cells in PFA-fixed marsupial embryos. I use 1mg/ml Fast Blue salt and >1mg/ml Naphthol AS phosphate sodium salt in 0.2M Tris buffer, pH 9.4. The >staining is good but I have been told that it is not permanent. Ideally, I >would like to dehydrate these stained specimens, embed them in paraffin and >then section and stain them immunohistochemically for other germ cell >markers and several growth factors. Does anyone have a protocol utilising >alkaline phosphatase histochemistry that would allow me to do this? Thanks >for any advice in advance, > >Dr Danielle Hickford >Research Fellow >Department of Zoology, University of Melbourne, >Australia 3010.>hickford@unimelb.edu.au > >-- >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 >********************************************************************* ******* >**** This electronic message and all contents contain information 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 us immediately and destroy the original message and all >copies. > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >-- >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 >********************************************************************* ******* >**** This >electronic message and all contents contain information 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 us immediately and destroy the original message and all >copies. > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ [1]MSN Toolbar provides one-click access to Hotmail from any Web page FREE download! References 1. http://g.msn.com/8HMBENUS/2731??PS= From pruegg <@t> colobio.com Tue Apr 27 16:04:26 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee &Peggy/Tony, SOMEONE....) In-Reply-To: Message-ID: In my experience with trying to do alk. phos. on ffpe decalcified bone for osteoblasts the enzyme reactivity did not survive. I had to do it on bone embedded in GMA without decal. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of LORALEE GEHAN Sent: Tuesday, April 27, 2004 2:05 PM To: mab70@medschl.cam.ac.uk; brucea@unimelb.edu.au; histonet-bounces@lists.utsouthwestern.edu; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee &Peggy/Tony, SOMEONE....) With my experience it won't survive the processing. Try it plastic. The processing is a bit longer but the stain works nicely. Hope that helps. Loralee Gehan, HTL >From: Margaret Blount >To: 'Bruce Abaloz' , histonet-bounces@lists.utsouthwestern.edu, histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee & Peggy/Tony, SOMEONE....) >Date: Tue, 27 Apr 2004 10:46:25 +0100 > >Hi, Everyone, > >With regard to this problem, I was wondering if the alk phos enzyme would >survive processing, I rather think it would otherwise we wouldn't need to >add levamisole when immunostaining paraffin sections using alk phos as a >label. So why not process into wax then do the detection of alkaline >phosphatase and the other markers you wish to identify. > >What do other histonetters think? Is it worth a try? > >Margaret > >Margaret Blount >Chief Technician >Clinical Biochemistry >University of Cambridge >Addenbrooke's Hospital >Hills Road >Cambridge >CB2 2QR > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bruce >Abaloz >Sent: Tuesday, April 27, 2004 2:30 AM >To: histonet-bounces@lists.utsouthwestern.edu; >histonet@lists.utsouthwestern.edu >Subject: [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee >& Peggy/Tony, SOMEONE....) > > >Hi, my name is Danielle & I need some advice PLEASE - >I have been using alkaline phosphatase histochemistry to detect primordial >germ cells in PFA-fixed marsupial embryos. I use 1mg/ml Fast Blue salt and >1mg/ml Naphthol AS phosphate sodium salt in 0.2M Tris buffer, pH 9.4. The >staining is good but I have been told that it is not permanent. Ideally, I >would like to dehydrate these stained specimens, embed them in paraffin and >then section and stain them immunohistochemically for other germ cell >markers and several growth factors. Does anyone have a protocol utilising >alkaline phosphatase histochemistry that would allow me to do this? Thanks >for any advice in advance, > >Dr Danielle Hickford >Research Fellow >Department of Zoology, University of Melbourne, >Australia 3010.>hickford@unimelb.edu.au > >-- >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 >********************************************************************* ******* >**** This electronic message and all contents contain information 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 us immediately and destroy the original message and all >copies. > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >-- >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 >********************************************************************* ******* >**** This >electronic message and all contents contain information 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 us immediately and destroy the original message and all >copies. > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ [1]MSN Toolbar provides one-click access to Hotmail from any Web page FREE download! References 1. http://g.msn.com/8HMBENUS/2731??PS= _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From caroline.stott <@t> anatomy.otago.ac.nz Tue Apr 27 16:26:49 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Re: H&E staining of frozen section Message-ID: <5.2.1.1.0.20040428092250.00a14760@anatomy.otago.ac.nz> Hi there, When I am staining H&E from frozen..... I take the section onto a slide let it dry for a few minutes and immerse in formal calcium for a few minutes. Then straight into water for a quick wash, then into Harris (or whichever Haematoxylin) for about 30 seconds. Back to water, then blue, water, then up through 70%, 95% and 100% into Alcoholic eosin for 10 or so seconds and through 2 changes of 100% alc. Finally into 3 changes of xylene and mount in DPX. Hope that helps Caroline Caroline Stott Histology Service Unit Medical School University of Otago Dunedin (03) 479 7152 From bhewlett <@t> cogeco.ca Tue Apr 27 19:22:05 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] 4% PFA References: <008301c42c8f$a83704e0$9d194c8e@WS21203> Message-ID: <005301c42cb6$d7510e20$6500a8c0@mainbox> Petia, The duration of fixation with PFA is the same as NBF, i.e. 24 hours minimum, full fixation takes 5-7 days. Bryan ----- Original Message ----- From: "Petia P Stefanova" To: Sent: Tuesday, April 27, 2004 3:41 PM Subject: [Histonet] 4% PFA > Hi to all, > > Does anybody know the duration of fixation with PFA? > My samples are mice ovaries and testis so I guess it won't take long. > > I appreciate your hepl! > Petia > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From cmalc <@t> unimelb.edu.au Tue Apr 27 19:42:05 2004 From: cmalc <@t> unimelb.edu.au (Cathy Malcontenti-Wilson) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Microtomes Message-ID: <6.0.0.22.2.20040428103813.01f25db8@mail.staff.unimelb.edu.au> Dear All, I would like to hear from people who have had some experience with more than one brand of rotary microtome. We would like to purchase one that is automatic, or semi-automatic. We are basically only cutting paraffin sections, at 3-4micron, and want one that is reliable. If you have an opinion on which ones you have found to be really good or really bad, I would appreciate your point of view to help us with our purchase. Thank you. Cat From immuno33 <@t> yahoo.com Tue Apr 27 21:43:27 2004 From: immuno33 <@t> yahoo.com (immuno33) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Two questions Message-ID: <20040428024327.29462.qmail@web61208.mail.yahoo.com> Hello Everyone: I have two questions. 1. Is anyone running p27 on a Techmate stainer and getting consistent clean results? If so, do you do anything special for them to come out clean? 2. Anyone preparing Tissue MicroArray slides that contain adhesive? If so, are you able to rid the slide of debris that becomes attached to the adhesive backing? Thanks for your help. Ken Chapman __________________________________ Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs http://hotjobs.sweepstakes.yahoo.com/careermakeover From c.m.vanderloos <@t> amc.uva.nl Wed Apr 28 05:31:47 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Re: HELP PLEASE/alk. phosph. Message-ID: <55c4d155a0b4.55a0b455c4d1@amc.uva.nl> Hi, I am missing something here? Why would you perform Fast Blue staining first and then embed in paraffin and have them next stained immunohistochemically with several markers??? Using New Fuchsin kit (DakoCyto) you will obtain a more permanent AP reaction product. It just dissolves a little bit in alcohol. Furthermore, there is Vector Red AP chromogen kit from Vector Labs. Its reaction product is also dissolving a little bit in alcohol. If you have strong staining from your immuno there is no problem processing the slides through alcohol-xylene and mount up organically. If you change your marker enzyme to b-galactosidase and using X-gal as substrate/ferri-ferrocyanide as chromogen one yields a reaction product that can stand everything without being affected or dissolved. Hope this helps Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Bruce Abaloz Date Tue, 27 Apr 2004 11:29:49 +1000 To histonet-bounces@lists.utsouthwestern.edu, histonet@lists.utsouthwestern.edu Subject [Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee & Peggy/Tony, SOMEONE....) Hi, my name is Danielle & I need some advice PLEASE - I have been using alkaline phosphatase histochemistry to detect primordial germ cells in PFA-fixed marsupial embryos. I use 1mg/ml Fast Blue salt and 1mg/ml Naphthol AS phosphate sodium salt in 0.2M Tris buffer, pH 9.4. The staining is good but I have been told that it is not permanent. Ideally, I would like to dehydrate these stained specimens, embed them in paraffin and then section and stain them immunohistochemically for other germ cell markers and several growth factors. Does anyone have a protocol utilising alkaline phosphatase histochemistry that would allow me to do this? Thanks for any advice in advance, Dr Danielle Hickford Research Fellow Department of Zoology, University of Melbourne, Australia 3010.>hickford@unimelb.edu.au From pstefanova <@t> sten.sunnybrook.utoronto.ca Wed Apr 28 07:49:37 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] 4% PFA References: <008301c42c8f$a83704e0$9d194c8e@WS21203> <005301c42cb6$d7510e20$6500a8c0@mainbox> Message-ID: <000d01c42d1f$471b63f0$9d194c8e@WS21203> Thanks to all for the answers!! have a good day Petia From anicoll <@t> cellpath.co.uk Wed Apr 28 07:51:38 2004 From: anicoll <@t> cellpath.co.uk (Alastair Nicoll) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] need drawers for filing - vendor posting details Message-ID: <0F42D9ABA325D811BBE300902712436D0A4612@cellmail.internal.cellpath.co.uk> CellPath yesterday displayed the drawer you require for the very first time at the European Histopathology Forum in Northampton. It has been designed for use within the Omnistor 15 metal cabinet. Cardboard storage solutions for mega blocks and slides have also just been launched. Please contact CellPath customer services on should you require further details. Export Customer Services Manager: Name: Stacey Mitchell Tel: +44 1686 611 333 Fax: +44 1686 622 946 Email: sales@cellpath.co.uk Ali CellPath Plc. Please visit our improved website for safety data sheets, news and information. www.cellpath.co.uk Buy CellPath products online: www.cellpath-store.co.uk Stuck for archiving space? Click for space www.cellnass.com Medical Equipment & consumables www.practice-direct.co.uk -----Original Message----- From: DiCarlo, Margaret [mailto:MDiCarlo@KaleidaHealth.Org] Sent: 26 April 2004 20:56 To: 'histonet@pathology.swmed.edu' Subject: [Histonet] need drawers for filing Hello Histonetters, I need to purchase drawers that 2 x 3 inch slides can fit in. I have the cabinet so I only need the drawers. The ones that I have are 19" long x 2 1/4" wide x 1 3/4" deep. Does anyone know where I can purchase these? Thanks for your help. Peggy DiCarlo HT (ASCP) Ortho Bone Lab Buffalo General Hospital 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 shive003 <@t> umn.edu Wed Apr 28 08:20:33 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Fibrin and Myoglobin Stains References: <424FAC08A8CDFE4BADD6497AE75B5E2F0E0A866D@atlas.gov.bc.ca> Message-ID: <003501c42d23$96424fd0$78065486@vdl220FAC> I stain for myoglobin using immunohistochemistry. I use the Dako polyclonal antibody and it works very well. No tissue pretreatment is necessary. Jan Shivers Univ. of Minnesota Veterinary Diagnostic Lab St. Paul, MN 55108 USA ----- Original Message ----- From: "Etheridge, Sandra AGF:EX" To: Sent: Tuesday, April 27, 2004 2:48 PM Subject: [Histonet] Fibrin and Myoglobin Stains > Hello fellow Histonetters, > > I work in an provincial animal health monitoring lab (mostly agricultural > animals) and one of the pathologists was asking if there were specific > special stains for fibrin and myoglobin. We currently use PTAH for fibrin > and muscle fibers, and Okajima for hemoglobin and myoglobin. Does anyone > else know of more specific stains for these two constituents? I can't seem > to find much in the limited reference library here. > > Thanks for the help. > > Sandra Etheridge > Animal Health Center > BC Ministry of Agriculture > Abbotsford, BC > Canada > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ASelf <@t> gmhsc.com Wed Apr 28 08:51:27 2004 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] CJD Message-ID: <39836CD6DB61654E8F95A35898C921860A86C8@exchange.gmhpost.com> Dear Netters, Does anyone have any info. or point me in the direction where I can find some info. on CJD and how it should be handled in the histopathology lab or during autopsies. I need to update my policy/procedure... Thanks in advance, amy Amy Self Georgetown Hospital Systems 843-527-7179 (phone) 843-520-7882 (fax) NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. From mari.ann.mailhiot <@t> leica-microsystems.com Wed Apr 28 09:27:13 2004 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] CJD Message-ID: Amy Contact the CDC. They can give you some important information on handling CJD. 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 "Amy Self" Sent by: To: histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] CJD 04/28/2004 08:51 AM Dear Netters, Does anyone have any info. or point me in the direction where I can find some info. on CJD and how it should be handled in the histopathology lab or during autopsies. I need to update my policy/procedure... Thanks in advance, amy Amy Self Georgetown Hospital Systems 843-527-7179 (phone) 843-520-7882 (fax) NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Apr 28 09:43:10 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] CJD and other prion disease website/links In-Reply-To: <39836CD6DB61654E8F95A35898C921860A86C8@exchange.gmhpost.co m> Message-ID: <3.0.6.32.20040428084310.00bc9580@gemini.msu.montana.edu> Amy, Go to this website, other links are there http://www.cjdsurveillance.com/ Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From pxs76 <@t> po.cwru.edu Wed Apr 28 10:03:33 2004 From: pxs76 <@t> po.cwru.edu (phyllis) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] CJD Message-ID: <5.1.0.14.0.20040428105750.00b858f0@pop.cwru.edu> The National Prion Disease Pathology Surveillance Center (www.cjdsurvelliance.org) provides a full service to handle suspected CJD cases. You can call 216--368-0587 or visit our website Phyllis Scalzo, HT(ASCP) Case Western Reserve University 2085 Adelbert Rd. Cleveland, Ohio 44106 Ph: 216-368-0822 Fax: 216-368-2546 From Terry.Marshall <@t> rothgen.nhs.uk Wed Apr 28 10:09:23 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Re: Zinc formalin and 10% NBF Message-ID: "It was amazing to see how firm and grayish the tissue was as compared to the NBF fixed tissue, within even an hour of fixation time." I too am eager to learn what formulation you are using. The first time I tried zinc, (in 1971), I used about 8% (because I had no idea what strength to use - just trying a poor man's mercury), and the tissue went the colour and consistency of concrete within half an hour. In fact, it only takes homeopathic doses of zinc to do the trick. 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: Johnson, Teri [mailto:TJJ@Stowers-Institute.org] Sent: 27 April 2004 19:15 To: histonet@pathology.swmed.edu Subject: [Histonet] Re: Zinc formalin and 10% NBF In working through our processing schedule dilemma, we fixed some pancreas in NBF (overnight), and fixed some in Zinc Formalin for 3 hours, followed by water wash and storage in 70% ethanol. We found that using a variety of processing schedules, the NBF fixed pancreas looked good with routine 30 min/station (with 2 hours total time in paraffin), all the way down to 10 min/station (with 80 min total time in paraffin). The Zinc Formalin fixed pancreas looks dry and brittle still, even with the short processing schedule. We will try a hand-processing schedule for these to see if we can get the processing perfect for the ZnF pancreas. Our answer to this problem could be that we need to limit the amount of time it is in fixative. Gayle, how long were the kidney biopsies fixed before processing? Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Wed Apr 28 10:13:31 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Re: Zinc formalin and 10% NBF Message-ID: I am using Richard Allan's commercial formulation. They'd tell you what concentration of Zinc they use, but then they'd have to kill you. This could be fodder for the next Mission: Impossible movie. Think Tom Cruise would go for it? -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Wednesday, April 28, 2004 10:09 AM To: Johnson, Teri; histonet@pathology.swmed.edu Subject: RE: [Histonet] Re: Zinc formalin and 10% NBF "It was amazing to see how firm and grayish the tissue was as compared to the NBF fixed tissue, within even an hour of fixation time." I too am eager to learn what formulation you are using. The first time I tried zinc, (in 1971), I used about 8% (because I had no idea what strength to use - just trying a poor man's mercury), and the tissue went the colour and consistency of concrete within half an hour. In fact, it only takes homeopathic doses of zinc to do the trick. 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: Johnson, Teri [mailto:TJJ@Stowers-Institute.org] Sent: 27 April 2004 19:15 To: histonet@pathology.swmed.edu Subject: [Histonet] Re: Zinc formalin and 10% NBF In working through our processing schedule dilemma, we fixed some pancreas in NBF (overnight), and fixed some in Zinc Formalin for 3 hours, followed by water wash and storage in 70% ethanol. We found that using a variety of processing schedules, the NBF fixed pancreas looked good with routine 30 min/station (with 2 hours total time in paraffin), all the way down to 10 min/station (with 80 min total time in paraffin). The Zinc Formalin fixed pancreas looks dry and brittle still, even with the short processing schedule. We will try a hand-processing schedule for these to see if we can get the processing perfect for the ZnF pancreas. Our answer to this problem could be that we need to limit the amount of time it is in fixative. Gayle, how long were the kidney biopsies fixed before processing? Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From siksik03 <@t> comcast.net Wed Apr 28 10:15:40 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:53 2005 Subject: Subject: [Histonet] Zinc formalin on human renal biopsies Message-ID: Hi Teri & HistoNetters Given that you only fixed the pancreas for three hours in the Zinc Formalin, I wonder whether your problem isn't just underfixation, as opposed to the overnight fixation in formalin. The dryness and brittleness could be due to exposure to alcohols before the tissue was properly fixed. Just a thought... Steven Slap >At 01:15 PM 4/27/2004 -0500, you wrote: > >In working through our processing schedule dilemma, we fixed some >>pancreas in NBF (overnight), and fixed some in Zinc Formalin for 3 >>hours, followed by water wash and storage in 70% ethanol. We found that >>using a variety of processing schedules, the NBF fixed pancreas looked >>good with routine 30 min/station (with 2 hours total time in paraffin), >>all the way down to 10 min/station (with 80 min total time in paraffin). >>The Zinc Formalin fixed pancreas looks dry and brittle still, even with >>the short processing schedule. >> >>It was amazing to see how firm and grayish the tissue was as compared to >>the NBF fixed tissue, within even an hour of fixation time. >> >>We will try a hand-processing schedule for these to see if we can get >>the processing perfect for the ZnF pancreas. Our answer to this problem >>could be that we need to limit the amount of time it is in fixative. >>Gayle, how long were the kidney biopsies fixed before processing? >> > From jkiernan <@t> uwo.ca Wed Apr 28 10:15:30 2004 From: jkiernan <@t> uwo.ca (J. A. Kiernan) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] 4% PFA References: <008301c42c8f$a83704e0$9d194c8e@WS21203> Message-ID: <408FCA92.2EC11EEF@uwo.ca> You do not say what "PFA" is. If the concentration is 4% a fair gues is that it is formaldehyde, perhaps generated by depolymerizing paraformaldehyde. If so, the fixation time is exactly the same for a 2 mm mouse's ovary as for a 10 mm cube of elephant's liver: 24 hours for reasonable structural preservation, but a week or two will stabilize both intra- and extracellular details more thoroughly. Formaldehyde (whether generated from the low polymers in formalin or the high polymers of paraformaldehyde) penetrates tissues nearly as rapidly as water, but its chemical reactions with proteins are quite slow. Minimal fixation (minutes to a few hours) by formaldehyde is necessary for some purposes, and is followed by cryoprotection, quick freezing and cutting of frozen sections. The structure, especially in cytoplasm viewed at high magnification, is rather poorly preserved. This doesn't matter for many investigations, especially with the central nervous system. Probably your Histonet enquiry will collect many answers like this one, and they won't all give you the same advice. Fixation has to be matched to the needs of the work being done. There are plenty of books that explain about fixatives and how to choose the right one for a particular job. Every histo lab should have a shelf with at least a dozen. In a research lab (you did say mouse ovaries!) you cannot be far from a library, full of textbooks and also the journals that contain the papers cited in textbooks. Go to the original source if you are repeating someone else's technique and hope to get similar results. John Kiernan. London, Canada. ____________________________ Petia P Stefanova wrote: > > Hi to all, > > Does anybody know the duration of fixation with PFA? > My samples are mice ovaries and testis so I guess it won't take long. > > I appreciate your hepl! > Petia > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Wed Apr 28 10:25:00 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:53 2005 Subject: Subject: [Histonet] Zinc formalin on human renal biopsies Message-ID: Steven, I was wondering that too. Although the tissues had been stored in 70% alcohol, I had post-fixed one sample another 4 hours in Zinc Formalin. I ran the post-fixed tissues in the processor at the same time as the originally Zinc Formalin fixed material, and the morphology was worse with the post-fixed sample. Again, this was a sub-optimal test in that it would be more helpful to use fresh pancreas for the fixation test, but it's the material that I had available. Having used Zinc Formalin extensively on human lymph node specimens in the past, I've seen what prolonged fixation (overnight) in Zinc Formalin can do to harden them. Maybe the formulation is the key here, as Gayle has used Anatech's formulation for 15 hours with tiny kidney biopsies with no ill effects. Thanks for your input! Teri -----Original Message----- From: Steven E. Slap [mailto:siksik03@comcast.net] Sent: Wednesday, April 28, 2004 10:16 AM To: Johnson, Teri Cc: HistoNet Server Subject: Subject: [Histonet] Zinc formalin on human renal biopsies Hi Teri & HistoNetters Given that you only fixed the pancreas for three hours in the Zinc Formalin, I wonder whether your problem isn't just underfixation, as opposed to the overnight fixation in formalin. The dryness and brittleness could be due to exposure to alcohols before the tissue was properly fixed. Just a thought... Steven Slap >At 01:15 PM 4/27/2004 -0500, you wrote: > >In working through our processing schedule dilemma, we fixed some >>pancreas in NBF (overnight), and fixed some in Zinc Formalin for 3 >>hours, followed by water wash and storage in 70% ethanol. We found >>that using a variety of processing schedules, the NBF fixed pancreas >>looked good with routine 30 min/station (with 2 hours total time in >>paraffin), all the way down to 10 min/station (with 80 min total time >>in paraffin). The Zinc Formalin fixed pancreas looks dry and brittle >>still, even with the short processing schedule. >> >>It was amazing to see how firm and grayish the tissue was as compared >>to the NBF fixed tissue, within even an hour of fixation time. >> >>We will try a hand-processing schedule for these to see if we can get >>the processing perfect for the ZnF pancreas. Our answer to this >>problem could be that we need to limit the amount of time it is in >>fixative. Gayle, how long were the kidney biopsies fixed before >>processing? >> > From tpmorken <@t> labvision.com Wed Apr 28 10:36:56 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] CJD Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2177560@usca0082k08.labvision.apogent.com> Amy, here are some places to get CJD info. Journal article: Handling Creutzveldt-Jakcob disease Tissues in the Histology Laboratory, Titford, M. and Bastian, F.O., J Histotech, V12, No.3 Sept 1989 If you have access to the internet you can go to these sites: CAP: Detailed information on handling CJD in the autopsy suite and the lab, with references. Search for "CJD" http://www.cap.org/ National Prion Disease Pathology Surveillance Center in Cleveland, OH. Their telephone number is (216) 368-0587; their website is: www.cjdsurveillance.com. For all CDC information on CJD, go to to the main CDC web page and search for "CJD" : http://www.cdc.gov Updated info available at: http://www.cdc.gov/od/ohs/biosfty/bmbl/sect7e.htm#Trans AGENT: Transmissible Spongiform Encephalopathies (Creutzfeldt-Jakob, kuru and related agents) Laboratory-associated infections with the transmissible spongiform encephalopathies (prion diseases) have not been documented. However, there is evidence that Creutzfeldt-Jakob disease (CJD) has been transmitted iatrogenically to patients by corneal transplants, dura mater grafts and growth hormone extracted from human pituitary glands, and by exposure to contaminated electroencephalographic electrodes (99). Infection is always fatal. There is no known nonhuman reservoir for CJD or kuru. Nonhuman primates and other laboratory animals have been infected by inoculation, but there is no evidence of secondary transmission. Scrapie of sheep and goats, bovine spongiform encephalopathy and mink encephalopathy are transmissible spongiform encephalopathies of animals that are similar to the human transmissible diseases. However, there is no evidence that the animal diseases can be transmitted to man. LABORATORY HAZARDS: High titers of a transmissible agent have been demonstrated in the brain and spinal cord of persons with kuru. In persons with Creutzfeldt-Jakob disease and its Gerstmann-Str?ussler-Schenker Syndrome variants, a similar transmissible agent has been demonstrated in the brain, spleen, liver, lymph nodes, lungs, spinal cord, kidneys, cornea and lens, and in spinal fluid and blood. Accidental parenteral inoculation, especially of nerve tissues, including formalin-fixed specimens, is extremely hazardous. Although non-nerve tissues are less often infectious, all tissues of humans and animals infected with these agents should be considered potentially hazardous. The risk of infection from aerosols, droplets, and exposure to intact skin, gastric and mucous membranes is not known; however, there is no evidence of contact or aerosol transmission. These agents are characterized by extreme resistance to conventional inactivation procedures including irradiation, boiling, dry heat and chemicals (formalin, betapropiolactone, alcohols); however, they are inactivated by 1 N NaOH, sodium hypochlorite (>2% free chlorine concentration) and steam autoclaving at 134 degrees C for 1 hour. RECOMMENDED PRECAUTIONS: Biosafety Level 2 practices and facilities are recommended for all activities utilizing known or potentially infectious tissues and fluids from naturally-infected humans and from experimentally infected animals. Extreme care must be taken to avoid accidental autoinoculation, or other traumatic parenteral inoculations of infectious tissues and fluids (76). Although there is no evidence to suggest that aerosol transmission occurs in the natural disease, it is prudent to avoid the generation of aerosols or droplets during the manipulation of tissues or fluids, and during the necropsy of experimental animals. It is further strongly recommended that gloves be worn for activities that provide the opportunity for skin contact with infectious tissues and fluids. Formaldehyde-fixed and paraffin-embedded tissues, especially of the brain, remain infectious. It is recommended that formalin-fixed tissues from suspected cases of transmissible encephalopathy be immersed in 96% formic acid for 30 minutes before histopathologic processing (15). Vaccines are not available for use in humans (51). _________________________ From: lpwenk@netquest.com (Wenk, Lee & Peggy) Save Address Block Sender To: "Carson, Karla" CC: "'Histonet'" Subject: Re: CJD Date: Wed, 21 Jul 1999 21:52:34 -0400 NSH just had a teleconference on this TODAY - Wed. July 21, 1999. Contact the NSH office at 301-262-6221. Ask them for Jennifer's phone number, the person who gave the teleconference today. I forget her last name, and I'm at home, and that's at work. You really need a good procedure. THIS IS NOTHING TO GOOF UP ON. Basically, you will need to fix the tissue in 10% NBF for several days, place it in 90% formic acid for an hour, go back into NBF, hand process in plastic dishes, collect all the fixation and processing solutions and dishes for special sterilization and incineration, wear all the personal protective equipment you can, cover your microtomes with plastic wrap or aluminum foil to collect all the shavings (to be sterilized and incinerated), hand stain in plastic disposable containers (and again collect all staining solutions to be sterilized and incinerated). I hope someone will be able to contact Jennifer early tomorrow, or that someone will be able to email you their procedure. (Sorry, I got a new computer at work, and literally cannot do a thing with it. But that's another story.) Also, maybe someone can email you the address of the Histonet archives. I can't find my bookmark on it. I know this has been discussed before, so you might be able to find a procedure from previous postings. Good luck. Peggy A. Wenk, HTL(ASCP) William Beaumont Hospital Royal Oak, MI 48073 CJD References for NaOH use There are numerous references for using sodium hydroxide as a decontamination tool. Two that first come to mind are "Tissue Handling in Suspected Creutzfeldt - Jakob Disease and Other Human Spongiform Encephalopathies (Prion Diseases)" published in Brain Pathology 1995 by Herbert Budka et al . Another good reference is Dr. Barbara Crain's article in CAP Today, January 1996: "Safety Tips for anatomic studies of possible CJD. There are different opinions on what concentration of NaOH to use, but many agree on its effectiveness. I agree, it is definitely better to deactivate prion prior to processing with formic acid treatment of tissues. My reason for encouraging sodium hydroxide use is as an alternative to bleach. Sodium Hydroxide is more stable than bleach which must be used FRESH> Hope this helped, Jennifer Jennifer Hofecker, HT (ASCP) University of Rochester Pathology (716)273-4129 CJD Decon -- Original Message -- I am wondering if someone is willing to share a procedure for decontaminating a tissue processor after processing a Creutzfeldt-Jacob case (unknowingly). Do I use formic acid or should I use fresh 10% bleach? I am updating manual for JCAHO inspection and any information would be appreciated. Thanks in advance!! > Cathy Goeden HT(ASCP) Histology Technician VAMC Sioux Falls, SD REPLY From: Rose Richardson [mailto:rrichar3@iupui.edu] Sent: Tuesday, August 14, 2001 10:23 AM For this situation I would recomend to use an autoclave if you have metal buckets, or if the parts are not stable to the heat use 2M NaOH or 10% bleach, wipe down and let stand for an hour then rinse. Included is the CAP protocol for known CJD. I have worked with CJD for 15 years and if you have further questions please feel free to contact me. Rose Richardson Creutzfeldt-Jakob Disease Safety tips for anatomic studies of possible CJD * Precautions for tissue handling in the autopsy room Decontaminating the autopsy room Decontaminating the tissue * Precautions for tissue handling in the histology lab Handling slides and blocks * References Barbara J. Crain, MD, PhD Identifying cases at risk for Creutzfeldt-Jakob Disease (CJD) Surgical pathology All brain biopsies for dementia should be handled as possible CJD cases. No frozen sections should be done. Autopsy pathology 1. In cases of neurodegenerative disease, examine the chart carefully for specific mention of CJD, spongiform encephalopathy, or other prion diseases. Irrespective of the clinical diagnosis, examine the chart for any of the following: rapidly progressive dementia; dementia less than three years total duration; signs or symptoms of cerebellar disease; dementia accompanied by lower motor neuron findings; demential accompanied by any focal neurologic deficit not explainable on the basis of documented structural disease (e.g., infarct, tumor); dementia with seizures, especially myoclonic seizures early in the disease course; previous dural implants; or human growth hormone treatment. If any of these warning signs is present, discuss the case with a neuropathologist or the attending neurologist. 2. If there is any suspicion of CJD, the autopsy should be limited to the brain only and the tissue treated as outlined below. Exceptions to this rule should be very few. Precautions for tissue handling in the autopsy room 1. Follow universal precautions against conventional bloodborne agents; cut-resistant gloves are preferable. 2. Wear a mask and eye shield, although there is no evidence that CJD is transmitted by aerosols or by nonpenetrating mucosal contamination. 3. Confine all tissues and fluids (including running water) to the table. 4. This may be facilitated by placing a plastic sheet over the table. Remove the calvarium with a hand saw if possible. If a Stryker saw is used, use some form of shielding (such as a plastic bag) to contain small drops of blood and tissue. 5. Do not contaminate the outer surface of the specimen container. 6. Clearly, label the container as infectious and place in a similarly labeled secondary container. Notify the funeral home of the infectious nature of the case. Decontaminating the autopsy room 1. Wash the area of the incision and any other contaminated skin surfaces with freshly opened undiluted commercial bleach (sodium hypochlorite). After 10 minutes, wash off the bleach with water. 2. Place all gowns, gloves, plastic sheets, and other disposable supplies in "biohazard" bags and incinerate them. Alternatively, autoclave (132 degrees Celsius steam) the disposables and discard them. Disinfect any liquids on the autopsy table with an equal volume of bleach or 2 normal sodium hydroxide (2N NaOH) before disposing. 3. Decontaminate hard surfaces and surgical instruments with bleach or 2N NaOH. These two treatments are equally efficacious; the choice between them depends on convenience and the material being decontaminated. NaOH is preferred for steel instruments because it is less corrosive than bleach. Leave the disinfectant in contact with the surface for at least 15 and preferably 60 minutes. Decontaminating the tissue. 1. The strongly preferred approach is formalin fixation followed by formic acid treatment of tissue blocks. i. Fix the intact brain in formalin for at least 10 days prior to cutting. Agitate the tissue blocks (including at least one section from each cortical lobe, basal ganglia plus cerebellum) in at least 50-100 mL of 95 percent-100 percent formic acid for one hour and then return them to formalin for two days prior to embedding. ii. Alternatively, take the necessary diagnostic sections from the fresh brain; fix them in formalin for two to seven days (as one would a surgical biopsy for dementia), treat with formic acid for one hour, and fix again in formalin for two days. iii. The formic acid treatment significantly reduces infectivity, although it does interfere with some silver stains for the plaques and tangles of Alzheimer's disease. 2. Retain the remaining brain tissue until a diagnosis has been made. 3. If the initial sections show CJD, proceed with a workup for Alzheimer's disease and other dementias. Precautions for tissue handling in the histology lab Tissue processing and sectioning. 1. Tissue treated with formic acid is essentially decontaminated and may be processed routinely, although many histology laboratories still prefer hand processing. Treat hand-processed material as potentially infectious. 2. Wear double gloves at all times. 3. Treat all solutions with equal volumes of fresh undiluted bleach for 60 minutes before disposal. 4. Handle disposables, glassware, tools, etc. according to the procedures used in the autopsy room (see preceding comments). 5. Collect all scraps of paraffin and unused sections on a disposable sheet. 6. Use disposable microtome blades. 7. The microtome itself may be wiped with bleach or sodium hydroxide solution, but it obviously cannot be thoroughly decontaminated. Laboratories that frequently handle possible CJD cases may wish to dedicate an old microtome to this purpose. Handling slides and blocks 1. No special precautions are needed in handling intact glass slides once they have been coverslipped. Decontaminate and discard broken slides. 2. Paraffin blocks should be stored in a properly labeled bag or box. References 1. Brown P. Guidelines for high-risk autopsy cases: special precautions for Creutzfeldt-Jakob disease. In: Hutchins G, ed. Autopsy Performance and Reporting, Northfield, Ill.: College of American Pathologists; 1990:68-74. 2. Brown P, Wolff A, Gajdusek DC. A simple and effective method for inactivating virus infectivity in formalin-fixed tissue samples from patients with Creutzfeldt-Jakob disease. Neurology. 1990;40:887-890. Dr. Crain, of the Department of Pathology, Johns Hopkins Hospital, Baltimore, is a member of the CAP Neuropathology Committee. Her article is one of a periodic series of articles written by the members of committees composing the CAP Commission on Anatomic Pathology. Tim Morken Lab Vision - Neomarkers www.labvision.com -----Original Message----- From: Amy Self [mailto:ASelf@gmhsc.com] Sent: Wednesday, April 28, 2004 6:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD Dear Netters, Does anyone have any info. or point me in the direction where I can find some info. on CJD and how it should be handled in the histopathology lab or during autopsies. I need to update my policy/procedure... Thanks in advance, amy Amy Self Georgetown Hospital Systems 843-527-7179 (phone) 843-520-7882 (fax) NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Solverboy <@t> aol.com Wed Apr 28 10:51:34 2004 From: Solverboy <@t> aol.com (Solverboy@aol.com) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Help With Cutting Heart Message-ID: <4F0BD035.19BAAE7B.0293A525@aol.com> Hello Histonetters, I have been cutting transverse sections of rat heart using the following protocol: 1) Dissect heart, cut in 3mm disks 2) Fix overnight 4% PFA @ 4 deg. 3) Cryoprotect in 30% sucrose around 5 days. 4) Embed in OCT and freeze blocks in isopentane that has been cooled in liqid N2 for 1min. 5) Store 6 hours at -80 deg. 6) Store o.n. at -20 deg and cut 16uM on cryostat also at this temp. The quality of the resulting histology is o.k but not great, specifically the endocardium is a bit torn. I cannot parraffin embed because of downstream in-situ hybridization signal intensity problems so i must use a frozen or perhaps fresh tissue histology protocol. Does anyone have any suggestions or experience with good frozen heart histology, any changes i could make to my protocol? maybe a new protocol? Please help! Thanks in advance, Fred Grau Dept. Neurbiology & Behavior SUNY Stony Brook From cwscouten <@t> myneurolab.com Wed Apr 28 11:21:18 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Help With Cutting Heart Message-ID: Sacrifice perfusion of the heart in advance would help. Formaldehyde pentrates slowly, and fixes slowly. Meanwhile, the tissue quality can be deteriorating. Fixing by immersion is inferior to getting fixative to every cell through the open capillaries almost instantantly. Try this. Heavily anesthetize the animal, and open the chest cavity. Enter the returning vena cava with a needle. Cut the ascending aorta to allow outflow. Flow 5% sucrose for a minute to wash out blood and extracellular sodium. Flow PFA for 5 mintues or more to thoroughly penetrate tissue. Let sit for 4 hours, with very slow flow, or no flow, of fixative. Flow 30% sucrose for 1 minute to thoroughly penetrate throughout the muscle. For immunology where overfixation can be a problem, this is enough. If for Nissl or other simple stain, add some PFA to the 30% sucrose. Remove heart, and let sit in 30% sucrose (with or without fixative, depending on intended use) for a day or so. It will already be cryoprotected and throughly penetrated with sucrose, so 5 days is unnecessary. An instrument to readily handle the fluids can be found at: http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Solverboy@aol.com Sent: Wednesday, April 28, 2004 10:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Help With Cutting Heart Hello Histonetters, I have been cutting transverse sections of rat heart using the following protocol: 1) Dissect heart, cut in 3mm disks 2) Fix overnight 4% PFA @ 4 deg. 3) Cryoprotect in 30% sucrose around 5 days. 4) Embed in OCT and freeze blocks in isopentane that has been cooled in liqid N2 for 1min. 5) Store 6 hours at -80 deg. 6) Store o.n. at -20 deg and cut 16uM on cryostat also at this temp. The quality of the resulting histology is o.k but not great, specifically the endocardium is a bit torn. I cannot parraffin embed because of downstream in-situ hybridization signal intensity problems so i must use a frozen or perhaps fresh tissue histology protocol. Does anyone have any suggestions or experience with good frozen heart histology, any changes i could make to my protocol? maybe a new protocol? Please help! Thanks in advance, Fred Grau Dept. Neurbiology & Behavior SUNY Stony Brook _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From psanquin <@t> lugo.usc.es Wed Apr 28 11:50:13 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Help with Smooth Muscle In-Reply-To: <4F0BD035.19BAAE7B.0293A525@aol.com> Message-ID: <3.0.6.32.20040428185013.00816630@pop.lugo.usc.es> Hello Histonetters, Could you advice me regarding a good marker for smooth muscle in veins and arteries? Thanks in advance, Pablo Sanchez Lugo (Spain) From mcauliff <@t> umdnj.edu Wed Apr 28 16:13:37 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Help With Cutting Heart In-Reply-To: References: Message-ID: <40901E81.5000702@umdnj.edu> Hi Fred: While it might be possible to perfuse the heart with the method Charles describes below, the perfusate will take a rather lengthy course to get to the heart muscle. If one perfuses via the vena cava (either inferior or superior) the perfusate will enter the r. atrium then the r. ventricle, the pulmonary trunk, the pulmonary arteries, pass through the vascular beds of the lungs, enter the l. artium, the l. ventricle, out the aorta and finally to the coronary arteries. True, there will be some backfilling of the vasculature via the coronary sinus and Thebesian veins but only a small amount of fixative will reach the heart muscle via those routes. A more direct route would be to enter the left ventricle with a needle and then cut the right atrium or inf. vena cava. Ten or fifteen seconds of perfusion will be sufficient to wash enough blood out of the coronary vessles to get a good perfusion, you can see the heart change color very quickly during this process. However, I don't think perfusion mechanics are your problem. Try fixing longer and at room temp and make sure your knife is very sharp. Small pieces of heart should not need 5 days in sucrose to be cryoprotected, 1-2 days should be plenty. Perhaps some gelatin in the ventricular cavity would help support the endocardium? Geoff Charles Scouten wrote: > Sacrifice perfusion of the heart in advance would help. Formaldehyde pentrates slowly, and fixes slowly. Meanwhile, the tissue quality can be deteriorating. Fixing by immersion is inferior to getting fixative to every cell through the open capillaries almost instantantly. Try this. > >Heavily anesthetize the animal, and open the chest cavity. >Enter the returning vena cava with a needle. Cut the ascending aorta to allow outflow. Flow 5% sucrose for a minute to wash out blood and extracellular sodium. Flow PFA for 5 mintues or more to thoroughly penetrate tissue. Let sit for 4 hours, with very slow flow, or no flow, of fixative. Flow 30% sucrose for 1 minute to thoroughly penetrate throughout the muscle. For immunology where overfixation can be a problem, this is enough. If for Nissl or other simple stain, add some PFA to the 30% sucrose. Remove heart, and let sit in 30% sucrose (with or without fixative, depending on intended use) for a day or so. It will already be cryoprotected and throughly penetrated with sucrose, so 5 days is unnecessary. > >An instrument to readily handle the fluids can be found at: > >http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 > > > >Cordially, >Charles W. Scouten, Ph.D. >myNeuroLab.com >5918 Evergreen Blvd. >St. Louis, MO 63134 >Ph: 314 522 0300 >FAX 314 522 0377 >cwscouten@myneurolab.com >http://www.myneurolab.com > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Solverboy@aol.com >Sent: Wednesday, April 28, 2004 10:52 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Help With Cutting Heart > >Hello Histonetters, > I have been cutting transverse sections of rat heart using the following protocol: >1) Dissect heart, cut in 3mm disks >2) Fix overnight 4% PFA @ 4 deg. >3) Cryoprotect in 30% sucrose around 5 days. >4) Embed in OCT and freeze blocks in isopentane that has been > cooled in liqid N2 for 1min. >5) Store 6 hours at -80 deg. >6) Store o.n. at -20 deg and cut 16uM on cryostat also at this > temp. > > The quality of the resulting histology is o.k but not great, specifically the endocardium is a bit torn. I cannot parraffin embed because of downstream in-situ hybridization signal intensity problems so i must use a frozen or perhaps fresh tissue histology protocol. Does anyone have any suggestions or experience with good frozen heart histology, any changes i could make to my protocol? maybe a new protocol? Please help! >Thanks in advance, >Fred Grau >Dept. Neurbiology & Behavior >SUNY Stony Brook > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >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 cwscouten <@t> myneurolab.com Wed Apr 28 13:35:21 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Help With Cutting Heart Message-ID: Thanks for the anatomy correction, blood returning to the right ventricle is not what feeds the heart muscle. Traditional perfusion route gets the heart just fine. I was trying to be creative, but needlessly so. However, I stand by my comments about the value of perfusion over immersion for better preservation of the tissue. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Wednesday, April 28, 2004 4:14 PM To: Charles Scouten Cc: Solverboy@aol.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Help With Cutting Heart Hi Fred: While it might be possible to perfuse the heart with the method Charles describes below, the perfusate will take a rather lengthy course to get to the heart muscle. If one perfuses via the vena cava (either inferior or superior) the perfusate will enter the r. atrium then the r. ventricle, the pulmonary trunk, the pulmonary arteries, pass through the vascular beds of the lungs, enter the l. artium, the l. ventricle, out the aorta and finally to the coronary arteries. True, there will be some backfilling of the vasculature via the coronary sinus and Thebesian veins but only a small amount of fixative will reach the heart muscle via those routes. A more direct route would be to enter the left ventricle with a needle and then cut the right atrium or inf. vena cava. Ten or fifteen seconds of perfusion will be sufficient to wash enough blood out of the coronary vessles to get a good perfusion, you can see the heart change color very quickly during this process. However, I don't think perfusion mechanics are your problem. Try fixing longer and at room temp and make sure your knife is very sharp. Small pieces of heart should not need 5 days in sucrose to be cryoprotected, 1-2 days should be plenty. Perhaps some gelatin in the ventricular cavity would help support the endocardium? Geoff Charles Scouten wrote: > Sacrifice perfusion of the heart in advance would help. Formaldehyde pentrates slowly, and fixes slowly. Meanwhile, the tissue quality can be deteriorating. Fixing by immersion is inferior to getting fixative to every cell through the open capillaries almost instantantly. Try this. > >Heavily anesthetize the animal, and open the chest cavity. >Enter the returning vena cava with a needle. Cut the ascending aorta to allow outflow. Flow 5% sucrose for a minute to wash out blood and extracellular sodium. Flow PFA for 5 mintues or more to thoroughly penetrate tissue. Let sit for 4 hours, with very slow flow, or no flow, of fixative. Flow 30% sucrose for 1 minute to thoroughly penetrate throughout the muscle. For immunology where overfixation can be a problem, this is enough. If for Nissl or other simple stain, add some PFA to the 30% sucrose. Remove heart, and let sit in 30% sucrose (with or without fixative, depending on intended use) for a day or so. It will already be cryoprotected and throughly penetrated with sucrose, so 5 days is unnecessary. > >An instrument to readily handle the fluids can be found at: > >http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct= >471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc >=Sacrifice+Equipment&idsubcategory=21 > > > >Cordially, >Charles W. Scouten, Ph.D. >myNeuroLab.com >5918 Evergreen Blvd. >St. Louis, MO 63134 >Ph: 314 522 0300 >FAX 314 522 0377 >cwscouten@myneurolab.com >http://www.myneurolab.com > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >Solverboy@aol.com >Sent: Wednesday, April 28, 2004 10:52 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Help With Cutting Heart > >Hello Histonetters, > I have been cutting transverse sections of rat heart using the following protocol: >1) Dissect heart, cut in 3mm disks >2) Fix overnight 4% PFA @ 4 deg. >3) Cryoprotect in 30% sucrose around 5 days. >4) Embed in OCT and freeze blocks in isopentane that has been > cooled in liqid N2 for 1min. >5) Store 6 hours at -80 deg. >6) Store o.n. at -20 deg and cut 16uM on cryostat also at this > temp. > > The quality of the resulting histology is o.k but not great, specifically the endocardium is a bit torn. I cannot parraffin embed because of downstream in-situ hybridization signal intensity problems so i must use a frozen or perhaps fresh tissue histology protocol. Does anyone have any suggestions or experience with good frozen heart histology, any changes i could make to my protocol? maybe a new protocol? Please help! >Thanks in advance, >Fred Grau >Dept. Neurbiology & Behavior >SUNY Stony Brook > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >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 KarBieber <@t> aol.com Wed Apr 28 14:16:16 2004 From: KarBieber <@t> aol.com (KarBieber@aol.com) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] NC Job Opening Message-ID: We have a position open for a full-time Histotechnician in Greensboro, NC. You will be responsible for sectioning skin and surgical tissue using a rotary microtome as well as the staining of tissue sections for microscopic evaluation. All histotechs actively participate in the daily quality control and equipment troubleshooting in this fast paced histology laboratory. Additional duties include tissue processor upkeep and general laboratory maintainance. Qualifications: Current HT(ASCP) certification a plus - a desire to become certified required. Associate's degree in related field required; one year working experience desired. Please forward resume/CV to: Greensboro Pathology Attn: HR Manager PO Box 13508 Greensboro, NC 27415 Fax: 336-510-0192 epancoast@gsopath.com www.greensboropathology.com From tytell <@t> fas.harvard.edu Wed Apr 28 14:40:39 2004 From: tytell <@t> fas.harvard.edu (Eric Tytell) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] thick sections in zebrafish Message-ID: <001c01c42d58$c544f820$5561f78c@TURBODOG> Hi all -- I'm trying to get thick (between 50 and 100 micron) transverse sections of adult zebrafish (15 to 20mm long). I'm concerned about the 3D structure of the muscle tissue, so I'd like to have the specimens embedded in some supporting medium. I've had good luck using plastic (JB-4) sectioned with a saw, but the trouble with that method is that I lose 300microns to the saw each time I cut. I'd like to try embedding in a different medium, but I haven't been able to find a protocol. Has anyone tried to do something like this? I'm currently experimenting with paraffin embedding. Specifically, does anyone have suggestions for how long and in what to decalcify? I currently have both EDTA and Poly-NoCal available. Also, how long should I infiltrate in paraffin? Alternatively, I've tried using a cryostat microtome, but the muscle tissue seems to be damaged by the freezing or possibly by desiccation. Is it necessary/possible to decalcify for frozen sections? Any suggestions for avoiding freezing damage? Also, does anyone know of a good polyclonal antibody for zebrafish skeletal muscle? I'm not looking for anything too specific -- just enough to distinguish skeletal muscle from bone and collagen under low magnification confocal imaging. I'd appreciate any suggestions. Thanks in advance, Eric Tytell ------------------------------------------ Eric Tytell Museum of Comparative Zoology Laboratories Harvard University From jlinda <@t> ces.clemson.edu Wed Apr 28 15:49:34 2004 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Keratin structure of the fingernail Message-ID: <5.2.1.1.0.20040428164129.03328468@mailhost.ces.clemson.edu> Dear colleagues, Could someone direct me to an atlas containing a microscopic picture of a fingernail? I have just completed doing the Ayoub-Sklar and Fraser-Lendrum stains on numerous sections of Nair softened nails. Interesting stains...BUT...I have no idea what the nail should like and am interested in the keratin structure of the nail. Internet searches led me to an overabundance of cosmetology sites! Thanks! Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo/histo.htm From amosbrooks <@t> earthlink.net Wed Apr 28 17:06:24 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Buffer vs Water In-Reply-To: <200404280814.1biQLo45z3NZFji0@eagle> References: <200404280814.1biQLo45z3NZFji0@eagle> Message-ID: <6.0.0.22.0.20040428170042.01ba51f8@pop.earthlink.net> Hi, You would be better off using buffer (PBS or TBS). The reason as it was described to me was that the antibodies are more likely to locate their binding sites if they don't have a lot of tonicity and VanDerwalls force induced agitation of the solutions. The pH of the washes should be as close to the pH of the antibody solution. If the antibody has a strong affinity to the antigen then you may not notice any difference. It becomes important when there isn't a strong attraction between the two. Hope this helps, Amos Brooks At 10:14 AM 4/28/2004, you wrote: >Message: 1 >Date: Tue, 27 Apr 2004 13:07:09 -0400 >From: Kristopher.Kalleberg@unilever.com >Subject: [Histonet] (no subject) >To: histonet@lists.utsouthwestern.edu >Message-ID: >Content-Type: text/plain; charset=iso-8859-1 > >When performing IHC, and the stain is combined with water, are better results >achieved when rinsed with water or should PBS be used From caroline.stott <@t> anatomy.otago.ac.nz Wed Apr 28 16:22:50 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] 4% PFA Message-ID: <5.2.1.1.0.20040429092047.00a02ec0@anatomy.otago.ac.nz> We use Paraformaldehyde and other fixatives on mouse ovaries regularly. We generally tell people to leave them in Para for approximately 24hrs. If they were interested in structure rather than immuno work, we'd tell them to leave them a little longer. Caroline Caroline Stott Histology Service Unit Medical School University of Otago Dunedin (03) 479 7152 From pruegg <@t> colobio.com Wed Apr 28 16:41:48 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Help with Smooth Muscle In-Reply-To: <3.0.6.32.20040428185013.00816630@pop.lugo.usc.es> Message-ID: I have always liked Desmin or Muscle Specific Actin (MSA) Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Pablo Sanchez Quinteiro Sent: Wednesday, April 28, 2004 10:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Help with Smooth Muscle Hello Histonetters, Could you advice me regarding a good marker for smooth muscle in veins and arteries? Thanks in advance, Pablo Sanchez Lugo (Spain) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From longoae <@t> biopur.com.ar Wed Apr 28 17:27:09 2004 From: longoae <@t> biopur.com.ar (Antonio Enzo Longo) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] unsuscribe Message-ID: <000e01c42d6f$f2ea07e0$bf2d2fc8@INEYJAVIS> From marytedo <@t> hotmail.com Wed Apr 28 18:53:26 2004 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Muscle and Nerve biop. Methods Message-ID: Yes, do it! I would like to learn more about that, because since I left the Histotechnology School, I have never seen that kind of biopsies again... This would be a great oportunity to update my knowledges. >From: "George Cole" >To: 'María Teresa Domínguez' >Subject: RE: [Histonet] Oven temp and time.... >Date: Sat, 24 Apr 2004 20:04:03 -0700 > >Maria; >If muscle and nerve biopsies are done at your hospital there in Tierra >Del Fuego, I would like to send you one of the packets that I have been >offering on the Histonet since last July. I've sent 118 of them out so >far--AT NO CHARGE to anybody---to 34 states and many countries over >seas. Its 85 minutes of video on a disc---meant to be used on computers >fitted with the ability to read Video CD's---and 21 pages of procedures >for muscle and nerve biopsy methods that make it possible to find much, >much more of the information hidden in muscle and nerve biopsy tissues >than the standard procedures. All I need is your complete mailing >address---our Post Office here is very particular---and I will get it >right in the mail to you. >georgecole@ev1.net > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of María >Teresa Domínguez >Sent: Saturday, April 24, 2004 6:21 PM >To: caroline.stott@anatomy.otago.ac.nz; >Histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Oven temp and time.... > > > I usually leave my slices in the oven for O.N. between 42ºC and >47ºC. > Mostly 42ºC. > > If I'm in a hurry I leave them for 30 min, or an hour at 58ºC. >I'm > afraid of burn the tissues... > > HT. MARIA T. DOMINGUEZ > > PATHOLOGY'S SERV. > > RIO GRANDE'S REG. HOSPITAL > > TIERRA DEL FUEGO, ARGENTINA. > >From: Caroline Stott > >To: > Histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Oven >temp > and time.... >Date: Thu, 22 Apr 2004 11:00:52 +1200 > >37 >degrees > overnight. Or if in a real hurry, 60 degrees for >20minutes. > >Caroline > > >Caroline Stott > >Histology Service Unit >Medical >School > >University of Otago >Dunedin >(03) 479 7152 > > > > >_______________________________________________ >Histonet >mailing > list >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _________________________________________________________________ > > Tired of spam? Get [1]advanced junk mail protection with MSN 8. > >References > > 1. http://g.msn.com/8HMAEN/2734??PS= >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _________________________________________________________________ The new [1]MSN 8: smart spam protection and 2 months FREE* References 1. http://g.msn.com/8HMBEN/2737??PS= From nlouisea <@t> gis.net Tue Apr 27 19:28:09 2004 From: nlouisea <@t> gis.net (Nancy Adams) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] water bath policy Message-ID: <000e01c42cb7$afbc3140$ac81fea9@eds-desk> I'm preparing for CAP inspection. Question ANP.23350 asks about a "policy" for preventing cross-contamination of paraffin sections . I understand the need to clean it off between each block but can't find a reference. Can anyone direct me to a source? Thanks Nancy Adams Falmouth Hospital Falmouth, MA From kildee6093 <@t> msn.com Wed Apr 28 20:24:25 2004 From: kildee6093 <@t> msn.com (MARILYN MCDONALD) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Don Hammer Message-ID: Don if you are reading this will you please call Dr. Pat Dean @ 1-888-2GIPATH. Thanks, Marilyn From Megan.Kear <@t> hunter.health.nsw.gov.au Wed Apr 28 23:31:04 2004 From: Megan.Kear <@t> hunter.health.nsw.gov.au (Megan Kear) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] laminin Message-ID: Hi Histonetters Anybody using a good laminin polyclonal antibody. To be used on formalin fixed paraffin fixed embedded sections. thank you for your help. Megan Clarke HAPS IHC dept Newcastle Australia From kemlo <@t> tiscali.co.uk Thu Apr 29 02:49:29 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Re: Zinc formalin and 10% NBF In-Reply-To: Message-ID: <408BBDAE0000FE27@mk-cpfrontend-2.mail.uk.tiscali.com> 1971? First time YOU used zinc? I have that time period etched on my poor sorry soul. Whenever I see galvanised metal or a lead pipe I twitch. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From johannes.korn <@t> anat.uni-freiburg.de Thu Apr 29 03:37:53 2004 From: johannes.korn <@t> anat.uni-freiburg.de (Johannes Korn) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Re: Help with Smooth Muscle In-Reply-To: References: Message-ID: <4090BEE1.8080105@anat.uni-freiburg.de> Hi Pablo, I don't know what kind of tissue you have, but in mouse, chicken and quail tissues I have got excellent results with a monoclonal antibody from Sigma against smooth muscle actin. Strong signal, almost no background staining. Yours, Johannes > > Message: 7 > Date: Wed, 28 Apr 2004 18:50:13 +0200 > From: Pablo S?nchez Quinteiro > Subject: [Histonet] Help with Smooth Muscle > To: histonet@lists.utsouthwestern.edu > Message-ID: <3.0.6.32.20040428185013.00816630@pop.lugo.usc.es> > Content-Type: text/plain; charset="us-ascii" > > Hello Histonetters, > > Could you advice me regarding a good marker for smooth muscle in veins and > arteries? > > Thanks in advance, > > Pablo Sanchez > > Lugo (Spain) > > -- Dr. Johannes Korn Institute for Anatomy and Cell Biology II Albert-Ludwigs-Universit?t Freiburg johannes.korn@anat.uni-freiburg.de tel: +49-761-203-5092 fax: +49-761-203-5091 From cormier <@t> MIT.EDU Thu Apr 29 05:05:51 2004 From: cormier <@t> MIT.EDU (Kathleen Cormier) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Re: [IHCRG] B and T-cell markers on mouse In-Reply-To: <25040710.1083192833663.JavaMail.root@rowlf.psp.pas.earthli nk.net> Message-ID: <5.2.1.1.2.20040429055757.00adab38@hesiod> Rosalia, We have used Dako's CD 3, (A 0452) Sigma CD 3 (C 7930) for T cell and eBioscience CD45R/b220 (14-0452) Dako CD79 (M 7051) and Novacastra's Ncl-bla36 for B cell. They all work well with mouse. Kathy Cormier Histology/Necropsy Supervisor Div Comparative Medicine MIT At 03:53 PM 4/28/2004 -0700, you wrote: >Hello, > >Has anyone out there ever worked with B-cell and T-cell markers on mouse >and guinea pigs? If so, who are the manufacturers that you have used? > >Thanks a million, > >Rosalia Soriano, HT, HTL (ASCP) >Biopathology Sciences >South San Francisco, CA 94080 > > > > >Yahoo! Groups Links > ><*> To visit your group on the web, go to: > http://groups.yahoo.com/group/IHCRG/ > ><*> To unsubscribe from this group, send an email to: > IHCRG-unsubscribe@yahoogroups.com > ><*> Your use of Yahoo! Groups is subject to: > http://docs.yahoo.com/info/terms/ > From k.brand <@t> erasmusmc.nl Thu Apr 29 07:40:25 2004 From: k.brand <@t> erasmusmc.nl (Karl Brand) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] quickest freezing method AND maintain morphology of mouse brains for LCM then microarray??? Message-ID: Histonetters, Like to hear your methods for the most rapid method of freezing tissue for subsequent cryosectioning, whilst maintaining best possible morphology. We using the PALM LCM scope to capture a small region of mouse brain, from fresh, rapid heamatoxylin stained ~8um cryosections. After this microarray analysis will be performed, so- **minimising RNA degradation whilst maintaining morphology are the critical goals here** Until now i have dropped the brain into TissueTek OCT medium and freezing on a bed of dry ice which typically takes ~4 mins or so. The morphology is satisfactory using this method, but working with such small amounts of tissue makes every second critical i feel. I just tried dropping the specimen (in OCT in mould) into isopentane, but a big crack formed in the OCT which i fear may also cause a fissure in the specimen! Suggestions? Experience? Greatly appreciated! yours, Karl Karl Brand k.brand@erasmusmc.nl Department of Genetics Erasmus MC Dr Molewaterplein 50 3015 GE Rotterdam lab +31 (0)10 408 7409 fax +31 (0)10 408 9468 mob +31 (0)621 446 504 From la.sebree <@t> hosp.wisc.edu Thu Apr 29 08:22:35 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] laminin Message-ID: Megan, We're currently using polyclonal laminin from NeoMarkers (Lab Vision) at 1:50 with protease pretreatment and stained on Ventana instruments. We were using it clinically for basement membrane staining but it seems to be inconsistent in its staining so now we use it as a research tool. Our pathologists don't feel its a problem with the antibody itself but rather the nature of laminin. 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: Megan Kear [mailto:Megan.Kear@hunter.health.nsw.gov.au] Sent: Wednesday, April 28, 2004 11:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] laminin Hi Histonetters Anybody using a good laminin polyclonal antibody. To be used on formalin fixed paraffin fixed embedded sections. thank you for your help. Megan Clarke HAPS IHC dept Newcastle Australia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Apr 29 08:58:04 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] B and T-cell markers on mouse In-Reply-To: <5.2.1.1.2.20040429055757.00adab38@hesiod> References: <25040710.1083192833663.JavaMail.root@rowlf.psp.pas.earthlink.net> Message-ID: <3.0.6.32.20040429075804.00bc8cf8@gemini.msu.montana.edu> We work almost exclusively with monoclonal antibodies from BD Pharmingen Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From info <@t> instrumedics.com Thu Apr 29 09:08:58 2004 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Re: Histonet Digest, Vol 5, Issue 42 References: Message-ID: <010c01c42df3$84133290$6501a8c0@INSTRUMEDICS22> Histonetters, If anyone has the Instrumedics' video narrated by Dr. Richard Zarbo they no longer need, can they please return it to us. We have requests for the video and have none in stock at this time. Thank you for your help. Bernice Instrumedics,Inc 61 South State St. Hackensack, NJ 07601 ----- Original Message ----- From: To: Sent: Tuesday, April 27, 2004 12:26 PM Subject: Histonet Digest, Vol 5, Issue 42 > 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: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee & > Peggy/Tony, SOMEONE....) (Bruce Abaloz) > 2. Re: Zinc formalin and 10% NBF (John Kiernan) > 3. RE: Re: H&E staining of frozen section (Kemlo) > 4. RE: need drawers for filing (Margaret Blount) > 5. RE: About Respiratory cytologies (Kemlo) > 6. RE: Hello! (Kemlo) > 7. Re: Zinc formalin and 10% NBF (Kemlo) > 8. Re: About Respiratory cytologies (Teresa Dominguez) > 9. RE: Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Le e & > Peggy/Tony, SOMEONE....) (Margaret Blount) > 10. RE: Re: H&E staining of frozen section > (Gladney, Diane C Ms MACH) > 11. Re: Hello! (Ford Royer) > 12. staining frozen sections (Michael Hetzer) > 13. mouse erythroid lineage subtypes (Sharon Cooperman) > 14. RE: RE: Cryojane cutting and LCM (Wright, Barbara (SPRI 2)) > 15. RE: Leica CV5030 (GUTIERREZ, JUAN) > 16. NSH Region II Symposium (June 3,4, & 5th) (Lizbeth_Kelly@hgsi.com) > 17. RE: Hello! (GUTIERREZ, JUAN) > 18. Re: PTAH (Cazares, Ruth) > 19. RE: OCT definition- THANKS (Phil Bergin) > 20. RE: Zinc formalin and 10% NBF > (Marshall Terry Dr, Consultant Histopathologist) > 21. Leitz 1512/Grease (mprice26@juno.com) > 22. Re: Re: H&E staining of frozen section (Gayle Callis) > 23. RE: Zinc formalin and 10% NBF (Gayle Callis) > 24. Re: Leitz 1512/Grease (Don.Birgerson@leica-microsystems.com) > 25. slide stainers - vendors (Kapoor, Sue) > 26. RE: Zinc formalin and 10% NBF > (Marshall Terry Dr, Consultant Histopathologist) > 27. Human on human staining (Susan Q Wells) > 28. Re: Best Way to obtain specific area of mouse foetal and > neonatal Brain for EM and light microscopy (Jo Dee Fish) > ---------------------------------------------------------------------------- ---- > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From m.vankempen <@t> erasmusmc.nl Thu Apr 29 09:14:01 2004 From: m.vankempen <@t> erasmusmc.nl (m. van mkempen) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] anti-CD45 antibody for mouse parrafin sections Message-ID: <40910DA9.1F89DFD5@erasmusmc.nl> Hello, Are there any people who have experience with the following anti-CD45 antibody: mouse monoclonal, clone OX-1 from BD #550566. I am trying to get this antibody to work on MOUSE spleen parrafin sections (It works in RAT cryosections). The tissue has been fixed in 4% paraformaldehyde during 24h at 4 C. After making the sections (5-6 microns) and deparrafinizing I did the following steps: Endogenous peroxidase blocking with 3% H2O2 in methanol for 20 min Rinse with PBS HIER with a citrate buffer (pH 6,0 and microwave 450W, 9 min) Rinse with PBS Block with PBS 5%BSA (30 min, RT) Incubate with primairy antibody dilluted 1:50 in PBS 5%BSA at (O/N, 4 C) Rinse with PBS Incubate with rabbit anti-mouse immunoglbulins (DAKO #Z 0259) dilluted 1:50 in PBS 5%BSA (30min, RT) Rinse with PBS Incubate with monoclonal mouse PAP (DAKO #P 0850) dilluted 1:50 in PBS 5%BSA (30min, RT) Rinse with PBS Enzyme detection with DAB substrate (5-7 min) Afterwards I can not detect anything besides some background from the endogenous peroxidase. Should I try another antibody or can I improve my protocol? Best regards, Marion van Kempen (Erasmus MC, Rotterdam The Netherlands) -- ---------------------------- *- mkempen -* MAILTO:m.vankempen@erasmusmc.nl ----------------------------------------- From histosci <@t> shentel.net Thu Apr 29 09:15:54 2004 From: histosci <@t> shentel.net (HSRL) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Finding Plain Slides Message-ID: <001701c42df4$80f9c820$0200a8c0@HSRLMAIN> Dear Netters, Could you please tell where everyone else buys their "plain" white labeled microscope slides? Our normal vendor for these slides have QC issues and we need to go elsewhere to buy them. These slides need to have a white label, 90 degree corners and not have "Superfrost" or "snowcoat" or "bond-rite" or "adhesive" etc. written below the label! It is tougher than expected to find such slides. Thanks for your help. Sincerely, Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org From gcallis <@t> montana.edu Thu Apr 29 09:17:01 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] freezing method AND maintain morphology ofmouse brains for LCM In-Reply-To: Message-ID: <3.0.6.32.20040429081701.00bc8cf8@gemini.msu.montana.edu> A good friend who does lots of LCM uses dry ice cooled hexane, just sits a beaker in dry ice and cools the hexane in a beaker. Dry ice can be added to hexane or 2 methyl butane, but she liked cooled hexane without dry ice better. Dry ice, as you are doing it is actually too slow, and you can get freezing artifact. YOu should freeze faster than 4 minutes! You can either embed the tissue in OCT OR freeze the tissue without embedding and embed it later in OCT, but still immerse in the dry ice cooled hexane. These solvents are not good to breathe, work in a hood and NEVER store 2 methylbutane in a nonexplosive freezer, it is total disaster. To freeze in dry ice/2 methylbutane slurry, we cut outer edges of plastic cryomolds (only Tissue Tek molds as other molds plastic is too thick, block will twist out) away, leaving one end to grasp, this give a better heat sink, or less plastic to cool, plus easier storage in 50 ml conical tubes. Embed tissue in OCT, then hold mold so you have bottom enter dry ice slurry, it starts to freeze, immerse and remove. Takes only seconds and OCT will not crack. Do overfill mold with OCT. Freezing bottom first, then allowing to immerse and not overfilling mold will help prevent cracks. It is a bit of technic involved here. One can also float a petri dish on liquid nitrogen, place mold inside dish but do NOT allow liquid nitrogen to enter dish, and freeze, this is colder than dry ice. Support petri dish with some type of holder to keep dish from tipping over into Liq N2. This results in the perfectly flat faced block and you can freeze many blocks at one sitting. At 02:40 PM 4/29/2004 +0200, you wrote: >Histonetters, > >Like to hear your methods for the most rapid method of freezing tissue for >subsequent cryosectioning, whilst maintaining best possible morphology. > >We using the PALM LCM scope to capture a small region of mouse brain, from >fresh, rapid heamatoxylin stained ~8um cryosections. After this microarray >analysis will be performed, so- > >**minimising RNA degradation whilst maintaining morphology are the critical >goals here** > >Until now i have dropped the brain into TissueTek OCT medium and freezing on >a bed of dry ice which typically takes ~4 mins or so. The morphology is >satisfactory using this method, but working with such small amounts of >tissue makes every second critical i feel. I just tried dropping the >specimen (in OCT in mould) into isopentane, but a big crack formed in the >OCT which i fear may also cause a fissure in the specimen! > >Suggestions? Experience? Greatly appreciated! > >yours, > >Karl > >Karl Brand k.brand@erasmusmc.nl >Department of Genetics >Erasmus MC >Dr Molewaterplein 50 >3015 GE Rotterdam >lab +31 (0)10 408 7409 fax +31 (0)10 408 9468 mob +31 (0)621 446 504 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Jackie.O'Connor <@t> abbott.com Thu Apr 29 09:22:18 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Finding Plain Slides Message-ID: I purchase superfrost plus slides from VWR (cat 48311-703). They have no writing except for the + signs at the bottom of the slide, and of course they are positively charged - I don't know if that is an issue for you. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics 847.938.4919 Fax 847.938.3266 "HSRL" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/29/2004 09:15 AM Please respond to histosci To: cc: Subject: [Histonet] Finding Plain Slides Dear Netters, Could you please tell where everyone else buys their "plain" white labeled microscope slides? Our normal vendor for these slides have QC issues and we need to go elsewhere to buy them. These slides need to have a white label, 90 degree corners and not have "Superfrost" or "snowcoat" or "bond-rite" or "adhesive" etc. written below the label! It is tougher than expected to find such slides. Thanks for your help. Sincerely, Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Thu Apr 29 12:51:59 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Finding Plain Slides In-Reply-To: <001701c42df4$80f9c820$0200a8c0@HSRLMAIN> References: <001701c42df4$80f9c820$0200a8c0@HSRLMAIN> Message-ID: <409140BF.8000001@umdnj.edu> How about Fisher Scientific? Geoff HSRL wrote: >Dear Netters, > > > >Could you please tell where everyone else buys their "plain" white >labeled microscope slides? Our normal vendor for these slides have QC >issues and we need to go elsewhere to buy them. These slides need to >have a white label, 90 degree corners and not have "Superfrost" or >"snowcoat" or "bond-rite" or "adhesive" etc. written below the label! >It is tougher than expected to find such slides. Thanks for your help. > > > >Sincerely, > > > >Tom Galati > >Laboratory Director > >HSRL, Inc.- A GLP Compliant Laboratory > >137 South Main Street > >Woodstock, Virginia 22664 > >(540)459-8211 > >Fax: (540)459-8217 > >tomgalati@hsrl.org > >www.hsrl.org > > > >_______________________________________________ >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 richalarson <@t> yahoo.com Thu Apr 29 10:03:08 2004 From: richalarson <@t> yahoo.com (Richard Larson) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Job interview questions Message-ID: <20040429150308.27225.qmail@web50710.mail.yahoo.com> I have a job interview next week with a private lab as a histotech. I have my ht certification plus four years of experience doing embedding, staining and some microtomy. I have been out of the lab the past three years doing other work but would now like to return to the histology profession. If anyone has had a recent interview, I would be interested in learning what sort of questions interviewers will likely ask, so that I can be more prepared. Thanks for any replies. --------------------------------- Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs From Terry.Marshall <@t> rothgen.nhs.uk Thu Apr 29 09:57:07 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Re: Zinc formalin and 10% NBF Message-ID: Don't be ridiculous. You have always walked like that. 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: Kemlo [mailto:kemlo@tiscali.co.uk] Sent: 29 April 2004 08:49 To: Marshall Terry Dr, Consultant Histopathologist; Johnson, Teri; histonet@pathology.swmed.edu Subject: RE: [Histonet] Re: Zinc formalin and 10% NBF 1971? First time YOU used zinc? I have that time period etched on my poor sorry soul. Whenever I see galvanised metal or a lead pipe I twitch. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From MAUGER <@t> email.chop.edu Thu Apr 29 09:50:12 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Finding Plain Slides Message-ID: Tom, I heard about the slide problem yesterday. Fisher Scientific should be able to get you slides. Fisher just bought Erie Glass, the company that makes most of them. Jo >>> HSRL 04/29/04 10:15AM >>> Dear Netters, Could you please tell where everyone else buys their "plain" white labeled microscope slides? Our normal vendor for these slides have QC issues and we need to go elsewhere to buy them. These slides need to have a white label, 90 degree corners and not have "Superfrost" or "snowcoat" or "bond-rite" or "adhesive" etc. written below the label! It is tougher than expected to find such slides. Thanks for your help. Sincerely, Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kosmicdog <@t> hotmail.com Thu Apr 29 10:29:33 2004 From: kosmicdog <@t> hotmail.com (jason madore) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] IHC camera and scope set up? Message-ID: Can anyone recommend a reasonable camera/scope/computer setup for taking microphotographs of standard IHC slides and TMA slides? _________________________________________________________________ Tired of spam? Get advanced junk mail protection with MSN Premium http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines From pstefanova <@t> sten.sunnybrook.utoronto.ca Thu Apr 29 10:43:11 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] anti-CD45 antibody for mouse parrafin sections References: <40910DA9.1F89DFD5@erasmusmc.nl> Message-ID: <005d01c42e00$b4daeaa0$9d194c8e@WS21203> Hi Marion, I think you should do antigen retrieval after deparaffination and than apply your immunostaining protocol. Fixation and impregnation with paraffin mask the epitopes and it decreases the antigenicity. You can find some information for antigen retrieval. http://herpesvirus.tripod.com/research/protoc.htm#Antigen%20Retrieval%20for Good luck! Petia From ploykasek <@t> phenopath.com Thu Apr 29 11:45:18 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Mounting media Message-ID: Thanks for all the help with the BCIP/NBT question. Now I have another question from the same research coworker. She would like to use an aqueous based mounting media. The one we used to use is no longer available. What is a good one to purchase for this purpose? Thanks again. Patti Loykasek PhenoPath Laboratories Seattle, WA From rschoon <@t> email.unc.edu Thu Apr 29 12:08:13 2004 From: rschoon <@t> email.unc.edu (Robert Schoonhoven) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] IHC camera and scope set up? References: Message-ID: <4091367D.1040804@email.unc.edu> This is a question with MANY answers and all of them have merit. For my light microscope I have an Olympus BH-2 with a Qimaging Micropulisher 5.0camera. My inverted Flou. microscope is an Olympus IX 71 with the Olympus MicroFire 5 MP camera. I also have it setup so that I can move the MicroFire to a second (light) microscope. Both of these cameras are 5 megapixil cameras the MicroFire will take 16 bit images and the Qimaging takes 12 bit images. In addition to the digital cameras ( well actually I had it before the digitals') we have a dedicated Olympus setup for traditional film. Ok, that tells you what I have but not the really important stuff - like why??? I'll try to make this short but there is a lot of information to take into consideration (which means that I will skip over a few things). First one needs to take into account the available funds and where to allocate them within your proposed system, as well as whether to go digital or stay with the traditional film type of system. Funds - spend as much as possible on the optics of the microscope, the best camera in the world will not correct for bad glass (optics). It really is a matter of user prefferance as to what the make of the scope is but do get the best objectives that your budget will allow for. Film vs Digital - As good as digital is with today's technology (16 MP + cameras are available), film still gives the best resolution. That said.... it's pretty darn hard to tell the difference between the two when looking at a printed micrograph in a publication. We've been submitting digital micrographs for several years with no complaints. Archived film will last for centuries , digital storage media does not. Photomicroscopy using a film camera actually takes a fair amount of skill and knowledge without even going into the darkroom part of it, where as I can show a student how to take a digital image in less then an hour and have them get good micrographs each time (I said good, not great). It should also be noted that while the old saying is "that film is cheap take lots of pictures", electrons are cheaper take more". The preceding is true but................ the initial cost of a dedicated digital camera, software and computer is MUCH higher than that of a dedicated film camera. Another good point is that the computer is your darkroom for digital images and there are incredible things that you can do with programs like Adobe Photoshop. I could go on for many pages regarding the other important factors like camera bit depth (8, 12, & 16), image analysis printing, software, ad nausium but I'm running out of time. Bob, Robert Schoonhoven, UNC-CH jason madore wrote: > Can anyone recommend a reasonable camera/scope/computer setup for > taking microphotographs of standard IHC slides and TMA slides? > > _________________________________________________________________ > Tired of spam? Get advanced junk mail protection with MSN Premium > http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Apr 29 12:44:33 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] anti-CD45 antibody for mouse parrafin sections In-Reply-To: <40910DA9.1F89DFD5@erasmusmc.nl> Message-ID: <3.0.6.32.20040429114433.00bc8cf8@gemini.msu.montana.edu> I suggest you buy the rat antiMouse anti CD45 monoclonal, the one you are attempting to work with detects CD45 in rat tissues. BD Pharmingen carries the rat antiMouse. I am not sure if the OX-1 clone even cross reacts to mouse antigens, plus you will have mouse to mouse problems. Any clone with OX-1 is usually a rat clone. At 04:14 PM 4/29/2004 +0200, you wrote: >Hello, > >Are there any people who have experience with the following anti-CD45 >antibody: mouse monoclonal, clone OX-1 from BD #550566. > >I am trying to get this antibody to work on MOUSE spleen parrafin >sections (It works in RAT cryosections). The tissue has been fixed in 4% >paraformaldehyde during 24h at 4 C. After making the sections (5-6 >microns) and deparrafinizing I did the following steps: > >Endogenous peroxidase blocking with 3% H2O2 in methanol for 20 min >Rinse with PBS >HIER with a citrate buffer (pH 6,0 and microwave 450W, 9 min) >Rinse with PBS >Block with PBS 5%BSA (30 min, RT) >Incubate with primairy antibody dilluted 1:50 in PBS 5%BSA at (O/N, 4 C) >Rinse with PBS >Incubate with rabbit anti-mouse immunoglbulins (DAKO #Z 0259) dilluted >1:50 in PBS 5%BSA (30min, RT) >Rinse with PBS >Incubate with monoclonal mouse PAP (DAKO #P 0850) dilluted 1:50 in PBS >5%BSA (30min, RT) >Rinse with PBS > >Enzyme detection with DAB substrate (5-7 min) > >Afterwards I can not detect anything besides some background from the >endogenous peroxidase. Should I try another antibody or can I improve my >protocol? > >Best regards, Marion van Kempen (Erasmus MC, Rotterdam The Netherlands) > > > > > > > >-- >---------------------------- >*- mkempen -* >MAILTO:m.vankempen@erasmusmc.nl >----------------------------------------- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From ASelf <@t> gmhsc.com Thu Apr 29 12:51:35 2004 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] PA Job Descriptions Message-ID: <39836CD6DB61654E8F95A35898C921860A86CF@exchange.gmhpost.com> Dear Histonetters, I need your help again......... Does anyone have a "Pathologist Assistant Job Description" that they would be willing to share with me..... Thanks in advance....amy Amy Self Georgetown Hospital Systems 843-527-7179 (phone) 843-520-7882 (fax) NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. From garygill <@t> dcla.com Thu Apr 29 13:05:18 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] PA Job Descriptions Message-ID: Go to: http://www.pathologistsassistants.org/Public_content/General_info/what%20is. htm -----Original Message----- From: Amy Self [mailto:ASelf@gmhsc.com] Sent: Thursday, April 29, 2004 12:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PA Job Descriptions Dear Histonetters, I need your help again......... Does anyone have a "Pathologist Assistant Job Description" that they would be willing to share with me..... Thanks in advance....amy Amy Self Georgetown Hospital Systems 843-527-7179 (phone) 843-520-7882 (fax) NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Thu Apr 29 14:08:35 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] re anti laminin Ab reagent Message-ID: <002301c42e1d$5f66abf0$82412cd9@home> Sigma L9393 polyclonal. For me, works on human, mouse, chick, with proteolytic or M/W pretreatment ( I prefer M/W; less variable). --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.668 / Virus Database: 430 - Release Date: 24/04/2004 From yangpw <@t> umich.edu Thu Apr 29 14:12:30 2004 From: yangpw <@t> umich.edu (yangpw@umich.edu) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Finding Plain Slides Message-ID: <1083265950.4091539e2fb50@mail.umich.edu> Hi, Usually I use superfrost for slide-mounted sections. And for free floating IHC sections, I buy Gold Seal precleaned slide from Fishersci (cat # 3064) and gel coat them. Pengwei Yang University of Michigan From histo <@t> compbio.com Thu Apr 29 15:09:34 2004 From: histo <@t> compbio.com (Histology) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] grinding bone Message-ID: <000d01c42e25$e53f1200$fda55742@ca.sprintbbd.net> Does anybody have a procedure for grinding bone on a buehler ecomet-3. Thank you connie From bill501 <@t> mindspring.com Thu Apr 29 15:32:15 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Microscope parts In-Reply-To: References: Message-ID: Does anyone know where I can get a halogen lamp socket for A Reichert Microstar IV scope? TIA, Bill -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board From gcallis <@t> montana.edu Thu Apr 29 16:41:44 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] NSH Veterinary, Industry and Research Committee webpage update Message-ID: <3.0.6.32.20040429154144.00bc6a30@gemini.msu.montana.edu> Dear All, The National Society for Histotechnology, VIR committee has an updated their webpage. Go to www.NSH.org and click on VIR. Updates are: Corrections to the Animal Processing Manual's first edition are posted for those of you who purchased the manual from NSH. Message from the new chairman of VIR Committee. Contact information Watch for VIR related Literature and Links and comments from chairman of Hard Tissue Committee. A call to all VIR histotechnologists to join the committee (membership form available on the VIR page) and update member personal information. Gayle Callis Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From mari.ann.mailhiot <@t> leica-microsystems.com Thu Apr 29 16:43:30 2004 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Microscope parts Message-ID: Bill Please contact Mark Streebel at 716 686 3166. He should be able to help you. 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 Bill Blank To: histonet@pathology.swmed.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Microscope parts western.edu 04/29/2004 03:32 PM Does anyone know where I can get a halogen lamp socket for A Reichert Microstar IV scope? TIA, Bill -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mary.bliss <@t> northwestpathology.com Thu Apr 29 17:07:08 2004 From: mary.bliss <@t> northwestpathology.com (Bliss, Mary E.) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] nail sectioning Message-ID: <27066863AD02214B81FE49477F3E71AD0D6F2C3C@hinet1.hinet.org> Hi to All, Does anyone have a good procedure for fixing and cutting in good fingernail and toenail sections? We are trying to come up with something which will allow us to produce a good stainable slide with ALL of the tissue still intact on it! I have heard about using trichloroacetic acid but can't seem to find a procedure anywhere. Have also heard about /tried Nair and soapy water. Does anyone have any information to share? Humbly yours, Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX From histo <@t> compbio.com Thu Apr 29 18:04:40 2004 From: histo <@t> compbio.com (Histology) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] grinding bone Message-ID: <000d01c42e3e$5b558d80$fda55742@ca.sprintbbd.net> Sorry, I'am processing the bones and embedding in (MMA) Methy methacrylate. thanks connie From karen <@t> gateslinger.com Thu Apr 29 19:19:31 2004 From: karen <@t> gateslinger.com (Karen Lahti) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Arizona Society conference Message-ID: The Arizona Society for Histotechnology and Region VII conference is planned for June 3-5, 2004 in Phoenix, Arizona. The complete booklet along with registration and membership forms can be obtained from our website - www.arizonahisto.org. We have many workshop planned to include PCR, Forensic Anthropology, HT Readiness, QIHC, H&E's from A to Z, IHC, Chemical Hazards, ISH, Yoga, Infectious Diseases and TEM/SEM. Please Do Not submit funds online. The forms are available to be downloaded then printed off. Please contact me if you have any problems or questions. We are having a drawing for a free room for three nights for paid registrants by May 10, 2004. So far, we have 22 vendors coming to support our society. The Best Western Grace Inn in Ahwatukee has free airport shuttle and is providing great room rates - $59/night for a single up to $79/night for a quad. Hospitality suites and one-bedroom suites are available. www.graceinn.com for more information. Thank You, Karen Lahti ASH President From KAELPERS <@t> aol.com Thu Apr 29 20:04:12 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] water bath policy Message-ID: <78.55a0f4c3.2dc3000c@aol.com> I have 2 references that mention cleaning the water bath, however the word cross contamination is not mentioned. If I find something I will send it to you - Good Luck with your CAP inspection! Theory and Practice of Histotechnology Sheehan, Hrapchak, second edition - "Processing of Tissue", Chapter 3, p.83 important point number 9 in the Tissue-Handling Chart For Sectioning. Theory and Practice of Histological Techniques Bancroft, Gamblin, fifth edition - "Tissue Processing and Microtomy", Chapter 6, p.97 last sentence under the subheading floating out sections. lge From KAELPERS <@t> aol.com Thu Apr 29 20:14:05 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] slide stainers - vendors Message-ID: <6b.283d2342.2dc3025d@aol.com> Hacker Linear Stainers...you can load until your heart is content. The only difficulty may be matching your current stain schedule to one that will be able to work on the hacker system. There is a timer you set permanently on the instrument and each rack will stay in that solution for that set time then prompt the instrument to lift and move to the next solution container. It is a great system, the only other factor is that it is a relatively long instrument and requires some space. lge From gliuygao <@t> hotmail.com Thu Apr 29 21:11:00 2004 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Tissue array Message-ID: Hi, Histonet. I am interested to get a few tissue array for different human cancers and cell array from human tumor cell lines. Any recommendation? Yan Gao Ph.D Norvatis _________________________________________________________________ [1]Stop worrying about overloading your inbox - get MSN Hotmail Extra Storage! References 1. http://g.msn.com/8HMBENUS/2737??PS= From kemlo <@t> tiscali.co.uk Fri Apr 30 02:48:42 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:53 2005 Subject: =?iso-8859-1?Q?RE=3A=20=5BHistonet=5D=20quickest=20freezing=20method=20AND=20maintain=20morphology=20of=09mouse=20brains=20for=20LCM=20then=20microarray=3F=3F=3F?= In-Reply-To: Message-ID: <40800C1000030126@mk-cpfrontend-4.mail.uk.tiscali.com> London on a January day. Quickest way to freeze anyone's brain; luckily these Southeners are immune! __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From tissuearray <@t> hotmail.com Fri Apr 30 05:14:30 2004 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] nail sectioning Message-ID: Have you tried 15% ammonia water. We have had success at softing the nail before processing and after if needed for sectioning. I have used this technique for horse hoff as well. Thom >From: "Bliss, Mary E." >To: >Subject: [Histonet] nail sectioning >Date: Thu, 29 Apr 2004 15:07:08 -0700 > >Hi to All, > > > >Does anyone have a good procedure for fixing and cutting in good >fingernail and toenail sections? We are trying to come up with >something which will allow us to produce a good stainable slide with ALL >of the tissue still intact on it! I have heard about using >trichloroacetic acid but can't seem to find a procedure anywhere. Have >also heard about /tried Nair and soapy water. Does anyone have any >information to share? > > > >Humbly yours, > > > >Mary E. Bliss > >Lead Histologist > >Northwest Pathology, P.S. > >3614 Meridian St. Suite 100 > >Bellingham, WA 98225 > >(360)734-2800 x601 > >(360)734-3818 FAX > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ [1]Stop worrying about overloading your inbox - get MSN Hotmail Extra Storage! References 1. http://g.msn.com/8HMBENUS/2737??PS= From mark.lewis <@t> thermo.com Fri Apr 30 07:48:20 2004 From: mark.lewis <@t> thermo.com (mark.lewis@thermo.com) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] nail sectioning Message-ID: Mary, Have you ever tried cutting Frozen sections of the unfixed toenail and fingernail ? You may be surprised to find they cut quite easily as a frozen section compared to a paraffin processed block. Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com "Bliss, Mary E." .com> cc: Sent by: Subject: [Histonet] nail sectioning histonet-bounces@lists.utsouth western.edu 04/29/2004 06:07 PM Hi to All, Does anyone have a good procedure for fixing and cutting in good fingernail and toenail sections? We are trying to come up with something which will allow us to produce a good stainable slide with ALL of the tissue still intact on it! I have heard about using trichloroacetic acid but can't seem to find a procedure anywhere. Have also heard about /tried Nair and soapy water. Does anyone have any information to share? Humbly yours, Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri Apr 30 08:45:50 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] laminin Message-ID: Does it have to be polyclonal? We use DAKO's monoclonal with very good results. (1:20, no pretreatment). They also have a polyclonal, but I have not tried it. Good luck, Juan -----Original Message----- From: Megan Kear [mailto:Megan.Kear@hunter.health.nsw.gov.au] Sent: Wednesday, April 28, 2004 11:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] laminin Hi Histonetters Anybody using a good laminin polyclonal antibody. To be used on formalin fixed paraffin fixed embedded sections. thank you for your help. Megan Clarke HAPS IHC dept Newcastle Australia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JosefaNava <@t> texashealth.org Fri Apr 30 09:22:17 2004 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Looking for ZAP-70and CD38 antibodies Message-ID: <7A5E02BB0E0A2E409F971D889B200C298A53DB@phdex05.txhealth.org> Hello to everyone! Can someone tell me where I can get ZAP -70 and CD38 antibodies tnat will work well with paraffin sections and also with Ventana Benchmark/ XT stainer. I thank you very much. Josie The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From RBARNHART <@t> summithealth.org Fri Apr 30 09:28:35 2004 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Container Message-ID: Our OR has asked that we get different containers for the larger specimens. Our current container is the basic container with a snap lid, they would like a screw top lid. The concern is if the container is dropped it will leak with the containers we have now. The only problem I am having is trying to find a gallon screw top container. Any ideas? Again thank you in advance. Becky From Diane.Gladney <@t> se.amedd.army.mil Fri Apr 30 09:44:50 2004 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Container Message-ID: <9D41AB7C56F8304F98537ABD87B249F66261EF@DASMTHGBZ001> You may have a hard time finding gallon size specimen containers with screw tops. Our OR has a large deep tray with a handle that they use to transport specimens to the Histology Dept. The containers that they are now using have lids that snap on snuggly and I sometimes have difficulty getting these lids off. How do they transport the containers to your lab? I have found that screw top containers will leak like crazy if the lids are not put on straight. We have trouble from our clinics even with the small screw top containers leaking because they won't take the time to put the lids on straight. Diane Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rebecca Barnhart Sent: Friday, April 30, 2004 10:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Container Our OR has asked that we get different containers for the larger specimens. Our current container is the basic container with a snap lid, they would like a screw top lid. The concern is if the container is dropped it will leak with the containers we have now. The only problem I am having is trying to find a gallon screw top container. Any ideas? Again thank you in advance. Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kbradshaw <@t> lcpath.com Fri Apr 30 09:49:24 2004 From: kbradshaw <@t> lcpath.com (Kari Bradshaw) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Job interview questions In-Reply-To: <20040429150308.27225.qmail@web50710.mail.yahoo.com> Message-ID: Dear Histonetters, Here is a list that I have developed over the last few years. I hope it's helpful. Kari Bradshaw, HT(ASCP) Laboratory Manager Lower Columbia Pathologists 1217 14th Ave Longview, WA 98632 (360)425-5620 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Larson Sent: Thursday, April 29, 2004 8:03 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Job interview questions I have a job interview next week with a private lab as a histotech. I have my ht certification plus four years of experience doing embedding, staining and some microtomy. I have been out of the lab the past three years doing other work but would now like to return to the histology profession. If anyone has had a recent interview, I would be interested in learning what sort of questions interviewers will likely ask, so that I can be more prepared. Thanks for any replies. --------------------------------- Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lwhite <@t> lakeridgehealth.on.ca Fri Apr 30 10:11:03 2004 From: lwhite <@t> lakeridgehealth.on.ca (White, Lori) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Contamination of processor alcohols Message-ID: Hi, We use an alcohol/xylene recycler and routinely test the alcohol for xylene contamination before recycling. We have noticed a significant proportion of our alcohols are contaminated when tested. Through process of elimination, we have determined that our contamination is coming from the alcohols on the processor. We replaced all of our alcohols with clean, non-recycled alcohol and ran a cycle. The alcohols were tested the following morning and one of the alcohols was contaminated. Has anyone had experience with this? Thanks, Lori Lakeridge Health Corporation Ontario, Canada From pwg1 <@t> cdc.gov Fri Apr 30 10:26:09 2004 From: pwg1 <@t> cdc.gov (Greer, Patricia) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Container Message-ID: Becky, Nalgene makes a 1 gallon wide-mouth container with screw top lid - we often get specimens sent to us in these containers. I'm sure any of the supply catalogs would have them. Actually I just looked at the Fisher catalog and they have several sizes of wide mouth bottles listed. You can go to their web site (www.fishersci.com) and search for bottles in their general laboratory section. Pat Greer Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road Atlanta, GA 30333 Our OR has asked that we get different containers for the larger specimens. Our current container is the basic container with a snap lid, they would like a screw top lid. The concern is if the container is dropped it will leak with the containers we have now. The only problem I am having is trying to find a gallon screw top container. Any ideas? Again thank you in advance. Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Fri Apr 30 10:49:33 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Zinc formalin and 10% NBF Message-ID: <494304423C63E246A5CF87A3AEEB577011B5E7@bumail01.barrynet.barry.edu> It's not the periodic table, but the electromotive series. Ions of elements at the top of the series will replace free elements (only on the surface of any but the tiniest particles) lower in the series. The ion is reduced to the free element and the lower down free element is oxidized to the ion. Zinc is pretty low in the electromotive series, so many ions can be reduced by zinc metal, converting the zinc metal to zinc ions. Unfortunately, the zinc in tissues is there as ions already. Any metal low enough in the series to reduce zinc ions to zinc metal would be such a good reducing agent that it would reduce water to hydrogen. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kemlo Sent: Tuesday, April 27, 2004 2:43 AM To: jkiernan@uwo.ca; Wendy England Cc: histonet@pathology.swmed.edu Subject: Re: [Histonet] Zinc formalin and 10% NBF Can't you test the residual fixative for the presence or absence of zinc? My Chemistry is not what it was but can't you do something with an element and the double decomposition of a salt? Something to do with an element higher up or lower down the Periodic Table that displaces an element from a salt that is the other way round, can't remember. But NBF doesn't contain the heavy metal does it? Why is it important to know? All I can remember is that zinc formalin made the tissue brittle, or was that lead? The nuclear stain was rather deep too. Hope that helps. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From Don.Birgerson <@t> leica-microsystems.com Fri Apr 30 11:21:14 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] thick sections in zebrafish Message-ID: Hi Eric, I have read your question with some curiosity. If the JB4 supports your specimen without distortion, why not use a motorized rotary microtome to section? If you have any questions, please feel free to phone me. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 7:00 - 4:00 CDT "Eric Tytell" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] thick sections in zebrafish western.edu 04/28/2004 02:40 PM Hi all -- I'm trying to get thick (between 50 and 100 micron) transverse sections of adult zebrafish (15 to 20mm long). I'm concerned about the 3D structure of the muscle tissue, so I'd like to have the specimens embedded in some supporting medium. I've had good luck using plastic (JB-4) sectioned with a saw, but the trouble with that method is that I lose 300microns to the saw each time I cut. I'd like to try embedding in a different medium, but I haven't been able to find a protocol. Has anyone tried to do something like this? I'm currently experimenting with paraffin embedding. Specifically, does anyone have suggestions for how long and in what to decalcify? I currently have both EDTA and Poly-NoCal available. Also, how long should I infiltrate in paraffin? Alternatively, I've tried using a cryostat microtome, but the muscle tissue seems to be damaged by the freezing or possibly by desiccation. Is it necessary/possible to decalcify for frozen sections? Any suggestions for avoiding freezing damage? Also, does anyone know of a good polyclonal antibody for zebrafish skeletal muscle? I'm not looking for anything too specific -- just enough to distinguish skeletal muscle from bone and collagen under low magnification confocal imaging. I'd appreciate any suggestions. Thanks in advance, Eric Tytell ------------------------------------------ Eric Tytell Museum of Comparative Zoology Laboratories Harvard University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From i_stain <@t> yahoo.com Fri Apr 30 11:44:16 2004 From: i_stain <@t> yahoo.com (Scott Mordue) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Tissue array Message-ID: <20040430164416.15176.qmail@web42002.mail.yahoo.com> Hi Yan Gao, I think you should contact Zymed. They have very high quality Tissue Microarray, especially the human ones. 800-874-4494 Scott CSU -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of yan gao Sent: Thursday, April 29, 2004 7:11 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Tissue array Hi, Histonet. I am interested to get a few tissue array for different human cancers and cell array from human tumor cell lines. Any recommendation? Yan Gao Ph.D Norvatis _________________________________________________________________ [1]Stop worrying about __________________________________ Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs http://hotjobs.sweepstakes.yahoo.com/careermakeover From scott.turner <@t> dnax.org Fri Apr 30 11:47:12 2004 From: scott.turner <@t> dnax.org (Turner, Scott) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Job opening in Palo Alto, CA Message-ID: <29B25753F6B1D51196110002A589D44401B721CA@PALMSG30.us.schp.com> Hi all-- We're looking to fill a histotech position here at DNAX in Palo Alto, CA (see job description below). Anyone interested in this job should apply online at http://www.dnaxresearch.com . Scott Turner DNAX Research Inc. Palo Alto, CA ---------------- DNAX Research Inc. is a biotechnology company located within the San Francisco Bay Area Biotechnology sector, and adjacent to Stanford University. DNAX carries out translational research leading to drug development in areas of Immunology and Oncology in a highly collaborative environment. We are seeking Scientist I-Histotechnologist PRIMARY RESPONSIBILITIES: * Perform routine and advanced histological techniques in a research environment, including gross examinations, trimming of tissue, tissue processing and embedding, and sectioning of paraffin and frozen samples from experimental studies, both human and animal. * Maintain and run a dynamic histology lab within an industrial setting, perform routine maintenance of automated equipment, order supplies, ensure quality of work meeting standards set by pathologists, and archive specimens. * Coordinate multiple studies from a variety of investigators and work with pathologists to deliver results. QUALIFICATIONS: * Candidate must have AS/HT (ASCP) and 10 years experience or BS/HT or HTL (ASCP) and 7 years of experience in a histology lab performing gross examinations, processing and embedding of human, non-human primate, and rodent tissues, preferably in a research environment. * Excellent microtomy skills, both paraffin and frozen, and capable of preparing serial sections. * Knowledge of routine and special histological stains required. Knowledge of immunohistochemistry a plus. * Self-motivated, possess excellent organizational skills and strong computer skills, and ability to manage their own time and deliver results on time. * Candidate must be a team player in a small fast paced pathology department and possess the ability to work with a wide variety of people, including various researchers and multiple pathologists. Candidates applying for this position should have a strong commitment to working in a collaborative and dynamic environment. www.dnaxresearch.com DNAX is an equal opportunity employer. DNAX Research, Inc. is a biotechnology subsidiary of Schering-Plough Corp. ********************************************************************* 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 jcline <@t> wchsys.org Fri Apr 30 11:59:51 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Mary Bliss/ nail sectioning Message-ID: <002401c42ed4$8e1805e0$1d2a14ac@wchsys.org> I use Nair. Place the nail into a pile of Nair and cover over. The nail usually takes 2 days to get soft enough, I check it each day. Use coated slides to mount sections. ***** 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 degaboh <@t> rice.edu Fri Apr 30 12:07:51 2004 From: degaboh <@t> rice.edu (ZP) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Embedding in OCT and subsequent steps In-Reply-To: <20040430161325.15F6219958@fungible1.mail.rice.edu> Message-ID: <20040430170752.8B6191DB87@handler10.mail.rice.edu> If I embed my sample in OCT, do I need to use xylene in later steps (i.e. when staining or before mounting)? I have a protocol that does that but I thought xylene was really more for paraffin embedding. Thanks!! Zarana Patel degaboh@rice.edu From JPCOLEMA <@t> sentara.com Fri Apr 30 12:33:17 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Zap-70 and CD38 Message-ID: I use CD38 from Novo Castra (NCL-CD38-290) and have just ordered ZAP- 70 from CellMarque ( who makes antibodies for Ventana). Cat #CMC 825. I have yet to receive the Zap, but the CD38 works beautifully- Citrate 6.0 retrieval 25 min, 1:100-200 dilution, 30 minute primary inc, Biogenex automation, Supersensitive concentrate kit, DAB chromogen. Questions? E-me. JPJC(QIHC) From JPCOLEMA <@t> sentara.com Fri Apr 30 12:46:43 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Xylene contamination in alcohol Message-ID: Lori- the alcohol's job in the staining line is to remove xylene before going to water so the contamination here is expected. The alcohol's job in the processor however, is to remove water from tissue before xylene is introdced so the only contamination of alcohol with xylene out of the processor will be in the "cleaning" alcohol, which "chases " xylene in the retort after processing is done. If your alcohol is contaminated after recycling, your recycler's temperature controls should be adjusted, since recycling is based on the boiling point of different solvents as a means to separate them. If your processor uses a rotary valve for fluid handling, this valve may be worn if the above conditions are not true and the contaminated alcohol is in the PRE Xylene portion of the processing schedule. Questions? E-me JPJC (QIHC) From JPCOLEMA <@t> sentara.com Fri Apr 30 13:00:02 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Zebra fish Message-ID: Eric- as a Zebra fish fan and having worked in Muscle labs, vet labs and research labs, I would try on some practice fish first, and would go for throwing the whole fish into a tissue cassette and processing em whole on a routine histology program. Are the skeletons significantly calcific? If not decal might not be necessary. I'd stay away from HCl decal solutions in any event. I would also stain with a trichrome or PTAH stain instead of using an antibody, or even just H&E to visualise the muscle tissue. Freezing does by the way injure muscle tissue unless the fish were to be frozen in liquid N2 chilled isopentane. Maybe I'll try this on guppies and see how it goes-- I don't know your experience with routine histo procedures though, so excuse my assumptions. Questions? E-me. JPJC(QIHC) From cwscouten <@t> myneurolab.com Fri Apr 30 13:52:27 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Embedding in OCT and subsequent steps Message-ID: Xylene infiltrates tissue, and prepares it for coverslipping (ie, with Permount). You still need xylene or a substitute if you plan to coverslip Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of ZP Sent: Friday, April 30, 2004 12:08 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Embedding in OCT and subsequent steps If I embed my sample in OCT, do I need to use xylene in later steps (i.e. when staining or before mounting)? I have a protocol that does that but I thought xylene was really more for paraffin embedding. Thanks!! Zarana Patel degaboh@rice.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Gillian.Barlow <@t> cshs.org Fri Apr 30 21:10:41 2004 From: Gillian.Barlow <@t> cshs.org (Barlow, Gillian) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] Problems with Fluorojade staining Message-ID: <3CFAA0108952D111A5BF00805FA6FB0F07947569@PEDSNTAS.csmc.edu> Hi there Does anyone out there have experience with Fluorojade staining of paraformaldehyde-fixed, paraffin-embedded mouse brains? We are getting quite high background following the published protocol, can anyone recommend changes/modifications? Sections are only 4 microns. Many thanks! Gillian Gillian M. Barlow, PhD Postdoctoral Fellow Laboratory of Julie Korenberg, PhD, MD Cedars-Sinai Medical Center Davis Bldg, Lab 2007 110 George Burns Rd Los Angeles, CA 90048 Phone: (310) 423 7650 Fax: (310) 423 0302