From christian <@t> eisbo.dk Wed Jun 1 02:48:19 2005 From: christian <@t> eisbo.dk (christian@eisbo.dk) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Ki67. Message-ID: <758a26d4ec47df2ab23acf9597f0b69d@eisbo.dk> From christian <@t> eisbo.dk Wed Jun 1 02:50:45 2005 From: christian <@t> eisbo.dk (christian@eisbo.dk) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Ki67. Message-ID: <634ceeb7bf3c9de7df2bf51acb622dfe@eisbo.dk> Hey users of histonet. I am trying to stain kidneys for ki67! The sections have been in alcohol for 1 month and are now in parafin. The antibody I am using should be good enough. Does anyone have experience with this??? best regards. Christian EIsbo. From bruyntjes <@t> voeding.tno.nl Wed Jun 1 03:29:19 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Ki67. Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079DC6@ntexch1.voeding.tno.nl> Hi Christian Yes I do. May be it depends on the antibody you use. We use a rabbit monoclonal ki67 from Neomarkers (Labvision). Tissues fixed in alcohol don't react with the antibody using HIER in citrate buffer (pH 6,0). At least it didn't in my lab. It does when you use HIER in EDTA (pH 8.0) and in TRIS (pH 10). Hopes that will help you Joost Bruijntjes TNO Quality of Life Zeist Holland -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of christian@eisbo.dk Sent: woensdag 1 juni 2005 9:51 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Ki67. Hey users of histonet. I am trying to stain kidneys for ki67! The sections have been in alcohol for 1 month and are now in parafin. The antibody I am using should be good enough. Does anyone have experience with this??? best regards. Christian EIsbo. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From vincent.martin5 <@t> laposte.net Wed Jun 1 03:43:57 2005 From: vincent.martin5 <@t> laposte.net (Vincent Martin) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] visualization of muscle damage with light microscopy ? Message-ID: <002701c56686$0d65e6c0$0a4a7c8b@physiologie8> Dear list members, I would like to observe the effect of lengthening contractions on ultrastructural properties of rat tibialis anterior muscle. Is is possible to observe ultrastructural abnormalities with light microscopy with a x100 maginification ? If not, which are the minimal requirements to observe such abnormalities ? Thanks for your replies Best regards, Vincent Martin UPRES-EA 3285 D?terminants Physiologiques de l'Activit? Physique Facult? des Sciences du Sport de Marseille Universit? de la M?diterran?e BP910 - 163, avenue de Luminy 13288 Marseille cedex 09 France Phone : +33 (0)4-91-82-83-78 Fax : +33 (0)4-91-82-83-75 vincent.martin@staps.univ-mrs.fr www.physiologie.staps.univ-mrs.fr From shawnster73 <@t> aol.com Wed Jun 1 06:05:40 2005 From: shawnster73 <@t> aol.com (shawnster73@aol.com) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Histology of Frozen Tissue Message-ID: <8C734B0FB191BFF-620-23897@FWM-D40.sysops.aol.com> What would be the histological effect of allowing frozen OCT embedded tissue to come up to a temperature of 0 or -5oC?? From bboyce <@t> NEMOURS.ORG Wed Jun 1 07:08:46 2005 From: bboyce <@t> NEMOURS.ORG (Bobbie Boyce) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] BK Virus Message-ID: <6E41111281623B4B8A9AB8F9A7EA343737BFB5@wlmmsx02.nemours.org> I'm having problems locating a FFPE control for BKV. Can anyone tell me where I can get them? Bobbie Boyce duPont Hospital for Children Wilmington, DE From jackdodo <@t> msn.com Wed Jun 1 07:10:12 2005 From: jackdodo <@t> msn.com (WAYNE HOLLAND) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] HPV I and II Control Blocks Message-ID: Hello All, Help, can anyone give assistance to getting some blocks for HPV I and HPV II for controls. We are going to start In-Situ, we are in the testing phase. One block from and HPV I (LOW) and one block from HPV II (HIGH) would be extremely helpful. We would be willing to trade, Thanks in advance. From bucana <@t> audumla.mdacc.tmc.edu Wed Jun 1 08:44:09 2005 From: bucana <@t> audumla.mdacc.tmc.edu (Corazon D. Bucana) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] mouse endothelial cells in brain In-Reply-To: <16A0583FB1644E4DB8C0A0265028B6FD02B4E725@nihexchange13.nih .gov> Message-ID: <5.1.1.6.0.20050601084009.042c9c48@audumla.mdacc.tmc.edu> The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From doninn <@t> mail.nih.gov Wed Jun 1 09:08:33 2005 From: doninn <@t> mail.nih.gov (Donin, Nick (NIH/NCI)) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] staining with Solvent Blue 38 Message-ID: <16A0583FB1644E4DB8C0A0265028B6FD02B4E739@nihexchange13.nih.gov> Histonetters, I'm trying to stain the corpus callosum of formalin fixed paraffin embedded mouse brains using Solvent Blue 38. Could someone suggest a successful protocol? Thanks. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 From Luis.Chiriboga <@t> med.nyu.edu Wed Jun 1 10:23:20 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] mouse endothelial cells in brain In-Reply-To: <5.1.1.6.0.20050601084009.042c9c48@audumla.mdacc.tmc.edu> Message-ID: I have also used the pharmingen cd31 by frozen in mouse brain, muscle, and skin. It works extremely well in fresh system. FFPE on the other hand is much trickier. It works but very inconsistently, fixation and tissue processing are the key variables. Once you have obtained staining in FFPE, you need to follow the same sacrificing (perfusion etc..) grossing, processing procedure with "religious fanaticism" (no offense to anyone.....) L ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Corazon D. Bucana Sent: Wednesday, June 01, 2005 9:44 AM To: Donin, Nick (NIH/NCI) Cc: 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] mouse endothelial cells in brain The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From christiegowan <@t> msn.com Wed Jun 1 10:24:32 2005 From: christiegowan <@t> msn.com (CHRISTIE GOWAN) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Storage of Frozen Section slides Message-ID: I know this subject has been tossed around before, but I wonder if Patsy Ruegg or Chris Van der Loos or anyone else for that matter would share with me their protocol for storing frozen slides. The company I work for sells frozen TMA slides. We cut sections as requested by clients and mail them out on dry ice. We have found that after going into the block 10-12 times we start to see artifact. Pre-cutting slides would be much better for the life of the block. Has anyone tried storing the slides in a vacuum sealed (food saver) bag? Any help would be appreciated. Thanks, Christie Gowan HT (ASCP) From dellav <@t> musc.edu Wed Jun 1 10:18:17 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Will brain tissue shrink even more if storedin70%alcoholprior to processing? Message-ID: I hadn't considered the possibility of buffer salt crystals forming the vacuoles observed by the author and I don't know if she is on this list and would care to comment but this is an interesting premise worth investigating. I would think that a thorough water wash prior to going into alcohol would help to clear this up. thanks for the feedback. Vinnie Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> John Kiernan 05/27/05 04:27PM >>> In the HistoLogic May 2000 paper the duration of the initial formaldehyde fixation is not stated; only that the brains were "previously well fixed in neutral buffered formalin." Frequently specimens are not fixed for long enough to make them resistant to bad effects of solvents etc. It's also seems from the paper that the test pieces of brain were passed directly from the buffered fixative into 70% alcohol. This causes precipitation of sodium phosphate in the tissue (see Freida Carson 1996 "Histotechnology" 2nd edn, p.27). The control specimens in the HistoLogic paper were passed from buffered formalin to 60% ethanol (which safely extracts the phosphate buffer salts). Could the holes in the white matter be made by buffer salt crystals rather than forming slowly during storage in 70% ethanol? Just a thought! -- ------------------------------- 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 _______________________________ Vinnie Della Speranza wrote: > > You may wish to look at an article authored by Jane Chladny, published in HistoLogic, May 2000, entitled "Artifact in Tissues Held in 70% Ethanol" > > the author reported the appearance of 'vacuole' artifact in nervous tissues observed in a variety of mammalian tissues after storage in 70% ethanol. > > you can access the article by selecting the link for HistoLogic at www.sakuraus.com > > CM Bush wrote: > > > > Dear Histonet, > > > > Hello, here is my first post to the list, thank you in advance for your help. > > > > Summary: > > Having problems with mouse and human brain tissue shrinkage durning processing, but would like to better preserve antigens, concerned storage of tissue 70% ethanol will shrink tissue even further: > > > > We have lots of brain specimens, stored in formalin for a long time (months up to 10 years), both human and mouse. We perform immunohistochemistry on paraffin embedded brain. Of course, there is the massive over fixation. Also the tissue shrinks a lot during processing. > > > > In my last lab, after brain tissue had been fixed in formalin for 7 days we would store the tissue in 70% ethanol. I'd like to start storing the brain tissue I work with now in 70%, but I'm worried about the tissue possibly shinking too much, as the tissue already seems to shrink by greater than 60% after processing. > > > > (Previously, we would gradually increase the alcohols, 30% for 1 hour, then 50% for an hour in a bucket, room temperature on a rocker table, then put the cassettes into a bucket of 70% and store at room temp or 4C- does this process help any with controling the shrinkage factor?) > > > > Maybe this is a little bit long...thank you very much for your time. > > > > CM Bush > --------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Vinnie Della Speranza > Manager for Anatomic Pathology Services > Medical University of South Carolina > 165 Ashley Avenue Suite 309 > Charleston, SC 29425 > Ph: 843-792-6353 > fax: 843-792-8974 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Wed Jun 1 10:35:22 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] mouse endothelial cells in brain Message-ID: Nick, Fix the brains in Pharmingen Zinc Tris, process to paraffin and use the Pharmingen antibody. Works like a charm. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Corazon D. Bucana Sent: Wednesday, June 01, 2005 9:44 AM To: Donin, Nick (NIH/NCI) Cc: 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] mouse endothelial cells in brain The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From twheelock <@t> mclean.harvard.edu Wed Jun 1 11:19:17 2005 From: twheelock <@t> mclean.harvard.edu (Tim Wheelock) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] SOLVENT BLUE 38 ON PARAFFIN EMBEDDED MOUSE BRAIN Message-ID: <429DE005.1050905@mclean.harvard.edu> Hi Nick: I use Solvent Blue 38 (also known as Luxol Fast Blue) on formalin-fixed, paraffin embedded human brain, but have also seen it work well on mouse brain. The protocol is as follows. This particular recipe combines the myelin stain with an H+E, but it can be combined with a Nissl stain instead or simply by itself. Hope this helps. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 * LUXOL FAST BLUE**-HEMATOXYLIN-EOSIN (LHE) STAIN* * * 1. De-paraffinize 5 micron formalin-fixed sections and take them down to 95% ethanol. 2. Stain in Luxol Fast Blue solution for 2 hours at 60C. 3. Remove and allow to cool for 15 minutes. 4. Remove excess stain in 95% ethanol for 1 minute. 5. Wash in several changes of tap water. 6. Differentiate sections with 2 dips in the reducer solution, then *immediately* through 4 changes of tap water (10 dips each), then one more change of tap water. 7. Immerse sections in Gill 3 Hematoxylin for 10 minutes. 8. Rinse in several changes of tap water. 9. Differentiate sections in acid alcohol (7 dips). 10. Rinse in 4 changes of tap water (10 dips each) then one more change of tap water. 11. Blue sections in Scott?s Tap Water Substitute for 30 seconds. 12. Wash in 4 changes of tap water for 2 minutes each. 13. Immerse sections in 95% ethanol for 1 minute. 14. Immerse slides in alcoholic Eosin Y for 3 minute. 15. Differentiate in 95% ethanol (4 dips). 16 Dehydrate in first absolute ethanol for 20 dips. 17. Dehydrate in second absolute ethanol for 1 minute, and then clear, and mount. *SOLUTIONS*: Luxol Fast Blue: Luxol Fast Blue...............................................0.1 gram. 10% glacial acetic acid .......................................1 mls. 95% ethanol........................................................100mls. Reducer: Hydroquinone....................................................1 gram. Sodium Sulfite...................................................5 gram. Distilled water..................................................100 mls. Hematoxylin Differentiator: 1% Hydrochloric acid in 70% ethanol *RESULTS*: 1. Myelin: The myelin should be stained blue or greenish blue 2. Nuclei: The nuclear membrane should be sharply delineated in blue-black with the nucleoplasm clear except for the chromatin, also being blue-black. 3. Cytoplasm and background should be varying shades of red. NOTE 1. The thickness of the section, the type of hematoxylin, and all the times in hematoxylin, eosin, and dehydrating depend upon your requirements. 2. If the section thickness is more than 10 microns, you may want to use 3 or 4 dips in the Reducer, so as to avoid background myelin staining . 3. The so called "Reducer" (because it is used as a reducer in the Bodian silver protein stain) actually functions as a differentiator for the myelin stain. From dmccaig <@t> ckha.on.ca Wed Jun 1 11:18:44 2005 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] filtering hematoxylin Message-ID: <3E5A3F039F0BD8118B4700C00D0020240435BF@CKHA9> How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? From jqb7 <@t> cdc.gov Wed Jun 1 11:33:58 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] filtering hematoxylin Message-ID: We don't filter the hematoxylin we get from Richard-Allan. We replace it twice a month, but we have a light volume of H&E's. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 (404) 639-3590 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: Wednesday, June 01, 2005 12:19 PM To: Histonet (E-mail) Subject: [Histonet] filtering hematoxylin How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NMargaryan <@t> childrensmemorial.org Wed Jun 1 12:01:27 2005 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] filtering hematoxylin Message-ID: <63B8B599DE283148B92E83C78B32C15DB91C88@cmhexbe2.childrensmemorial.org> I am doing manual H&E and filtering the hematoxylin and Eosin Y (both from Richard-Allan) before use every other day. Naira -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine Sent: Wednesday, June 01, 2005 11:34 AM To: Diana McCaig; Histonet (E-mail) Subject: RE: [Histonet] filtering hematoxylin We don't filter the hematoxylin we get from Richard-Allan. We replace it twice a month, but we have a light volume of H&E's. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 (404) 639-3590 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: Wednesday, June 01, 2005 12:19 PM To: Histonet (E-mail) Subject: [Histonet] filtering hematoxylin How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? _______________________________________________ Histonet mailing list Histonet@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 contents of this e-mail message and any attachments are private and confidential communications intended solely for the addressee(s) named in this message. If you are not the intended recipient of this message, please 1) immediately notify the sender by reply e-mail and then delete this message and its attachments and 2) do not read, use, distribute disclose or copy this message and/or any attachments. From billingconsultants <@t> yahoo.com Wed Jun 1 12:07:21 2005 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Re: Histonet Digest, Vol 19, Issue 1 Message-ID: <20050601170721.74179.qmail@web54204.mail.yahoo.com> Hi everyone, I have a client interested in purchasing an IHC/ISH stainer. Any suggestions/recommendations and vendor replies would be welcome. Thank you in advance for your assistance. Louri Roberts --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour From jlf <@t> jax.org Wed Jun 1 12:10:54 2005 From: jlf <@t> jax.org (Judi Ford) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] GMA block question Message-ID: <6.0.3.0.0.20050601130056.01aee2b0@aretha.jax.org> Hi everyone, I have a question about gma blocks coming off the holder. We use Technovit 7100 to embed our tissue (mouse eyes). Once the resin is set, overnight @ room temperature, the blocks are popped out of the mold and dried in a room temperature oven overnight (once in awhile the temperature may exceed room temperature). The next day we cut them. The problem I see, is that every so often the tissue block is loose, even after being in the oven and in a dessicator. We end up having to cut the block off the backing and remount it using epoxy. There are others in the lab doing the same exact method as we are and they have had no problems. The tissue blocks are rock solid. We've tried to examine all the angles........ Any ideas about what could be causing this? Thanks, Judi Ford Histotechnologist The Jackson Laboratory Bar Harbor, ME From DEllenburg2 <@t> stfrancishealth.org Wed Jun 1 12:16:47 2005 From: DEllenburg2 <@t> stfrancishealth.org (DEllenburg2@stfrancishealth.org) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] RE: Histonet Digest, Vol 19, Issue 1 Message-ID: <0A502E8156DAA4468CB8979B27177555206135@bssmsx5501.intranet.stfrancishealth.com> In our lab we filter the hematoxylin and eosin daily. Old habits are hard to break. -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: None To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 19, Issue 1 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. Speaking of Gram's... (Breeden, Sara) 2. Freezing muscle biopsies (Osborn, Sharon) 3. Enzyme Staining and Billing for Controls (Mitchell, Guy) 4. RE: Number of Blocks Submitted by PA (Charles.Embrey) 5. Aquaporin immunohistochemistry (Anthony Reilly) 6. Ki67. (christian@eisbo.dk) 7. Ki67. (christian@eisbo.dk) 8. RE: Ki67. (Bruijntjes, J.P.) 9. visualization of muscle damage with light microscopy ? (Vincent Martin) 10. Histology of Frozen Tissue (shawnster73@aol.com) 11. BK Virus (Bobbie Boyce) 12. HPV I and II Control Blocks (WAYNE HOLLAND) 13. Re: mouse endothelial cells in brain (Corazon D. Bucana) 14. staining with Solvent Blue 38 (Donin, Nick (NIH/NCI)) 15. RE: mouse endothelial cells in brain (Luis Chiriboga) 16. Storage of Frozen Section slides (CHRISTIE GOWAN) 17. Re: Will brain tissue shrink even more if storedin70%alcoholprior to processing? (Vinnie Della Speranza) 18. RE: mouse endothelial cells in brain (Connolly, Brett M) 19. SOLVENT BLUE 38 ON PARAFFIN EMBEDDED MOUSE BRAIN (Tim Wheelock) 20. filtering hematoxylin (Diana McCaig) 21. RE: filtering hematoxylin (Bartlett, Jeanine) ---------------------------------------------------------------------- Message: 1 Date: Tue, 31 May 2005 12:33:01 -0600 From: "Breeden, Sara" Subject: [Histonet] Speaking of Gram's... To: Message-ID: Content-Type: text/plain; charset="us-ascii" I'm having trouble with my Gram's in that my positive bacteria are losing color. So, I'm wondering what the exact differentiation time should be for the step after the Gram's Iodine (I'm using the Sigma kit)? The instructions are not precise, so I'm thinkin' maybe I'm over differentiating or under-Safranin-ing or aliens (this is the State that's home to ROSWELL...) have taken over my kit. Anyone have anything a bit more specific? I'd appreciate any help... Sara A. Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX ------------------------------ Message: 2 Date: Tue, 31 May 2005 15:41:02 -0400 From: "Osborn, Sharon" Subject: [Histonet] Freezing muscle biopsies To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <29B25753F6B1D51196110002A589D44402397FB8@PALMSG30.us.schp.com> Content-Type: text/plain Liz, Our researchers often freeze tissue for later study. They use the plastic disposable embedding molds. These can be used with the dry ice as John Kiernan suggested. There is a thin layer of OCT placed in bottom of mold then the tissue is oriented in this with additional OCT to fill the mold. The mold can be labeled with a permanent felt tip pen. Each specimen is then placed into the labeled sterile sampling bags, packed in dry ice and shipped to respective location. We have experienced very little distortion of the tissue due to ice crystals, etc. The testing has worked out okay. The main tissues were liver, kidney, lung. You may wish to experiment with some tissues before actually tackling the research samples to get the right combination for you. Sharon Osborn DNAX ScheringPlough Biopharma Palo Alto, CA Hello Histonetters > > I have a unique question. We are currently starting to set up > procedure for collecting samples from a clinical trial. The clinical > trial involves taking multiple synovial biopsies at a surgery center. > Since portions of the samples need to be processed for frozen sections > we wanted to be able to freeze the specimens at the surgery center via > isopentane cooled liquid nitrogen. We really do not want to have to > transport the multiple specimens back to the main lab prior to > freezing due to the time involved it would probably be 1-2 hours post > biopsy before we could freeze the samples. The surgery center is > questioning the flammability of the isopentane. Has anyone > encountered anything like this? Any suggestions would be helpful. > Thanks in advance. > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > Manager > Premier Laboratory, LLC > P.O. Box 18592 > Boulder, Colorado 80308 > Office: (303) 735-5001 > Fax: (303) 735-3540 > liz@premierlab.com > www.premierlab.com > > Ship to Address: > Premier Laboratory > University of Colorado > MCDB, Room A3B40 > Boulder, Colorado 80309 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 3 Date: Tue, 31 May 2005 15:42:59 -0400 From: "Mitchell, Guy" Subject: [Histonet] Enzyme Staining and Billing for Controls To: "HistoNet Server" Message-ID: <4DCF5CA56B4AA04ABC383DE358BAD3820289100D@dcr-xchg-04.carolinas.org> Content-Type: text/plain; charset="US-ASCII" A question has come up concerning charges for muscle enzyme stains. We stain for Adenylate Deaminase and Phosphofructokinase as well as negatives for each. I assume they should be treated as controls without a charge but others disagree. Guy W. Mitchell (704) 379-5984 (704) 379-6144 Fax ----------------------------------------- This electronic message may contain information that is confidential and/or legally privileged. It is intended only for the use of the individual(s) and entity named as recipients in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from any computer. Do not deliver, distribute or copy this message, and do not disclose its contents or take any action in reliance on the information it contains. Thank you. ------------------------------ Message: 4 Date: Tue, 31 May 2005 15:42:58 -0500 From: "Charles.Embrey" Subject: RE: [Histonet] Number of Blocks Submitted by PA To: "Smith Wanda" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" I have been gone for a week and just got a chance to check my e-mails and found this question interesting. First, have you asked the PA? I know it seems like a simple question but you would be surprised at all the misunderstanding that can be cleared up with a simple question asked? Please know that I have NEVER met a pathologist that will gladly read more slides than he thinks are necessary for a good diagnosis on a routine basis. I gross for seven pathologists and it is tricky to satisfy all at the same time. Luckily, my pathologists and I here have great communication and we manage to keep the block count, as well a gross recut number down. As far as "rules of thumb" we do have certain guidelines that are basically thought of as "standards of care". Without knowing other details the best advice I could give is to ask the PA. Charles Embrey PA(ASCP) Urbana IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Smith Wanda Sent: Tuesday, May 24, 2005 7:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Number of Blocks Submitted by PA Good Morning Histonetters, I have a question for you this morning...Are there written standards or "Rules of Thumb" regarding number of blocks that should be submitted on certain tissues? The reason I ask is, "it seems to me" that sometimes the PA submits an extreme number of blocks on some specimens that could be accomplished with less. I'm not advocating cutting corners or not performing good medical practice, I just need to know. An example would be a uterus w/ fibroids, tubes and ovaries which we have gotten up to 24 blocks on. I'm not a PA, but it's interesting to me that when the Pathologist cut, we get alot less blocks. Please advise. Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Wed, 01 Jun 2005 14:13:15 +1000 From: "Anthony Reilly" Subject: [Histonet] Aquaporin immunohistochemistry To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hello All I have been requested to be involved in a research study demonstrating Aquaporin 1 (AQ1, AQP1) using IHC. To date I have found 2 antibodies available from Chemicon. My questions are: 1. Are there other commercial suppliers of this Ab. 2. If not, which of the 2 from Chemicon is the preferred option. 3. Do you realyy need to fix with paraformaldehyde as stated in most methods, and why? i would appreciate any advice forwarded. regards Tony Reilly Chief Scientist Anatomical Pathology Northside Pathology Prince Charles Hospital Rode rd Chermside Q 4032 Australia Ph: 07 3350 8543 Fax: 07 33508546 tony_reilly@health.qld.gov.au **************************************************************************** ******* This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is prohibited. It may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone or by return email. You should also delete this email and destroy any hard copies produced. **************************************************************************** ******* ------------------------------ Message: 6 Date: Wed, 01 Jun 2005 09:48:19 +0200 From: "christian@eisbo.dk" Subject: [Histonet] Ki67. To: histonet@lists.utsouthwestern.edu Message-ID: <758a26d4ec47df2ab23acf9597f0b69d@eisbo.dk> Content-Type: text/plain; charset=windows-1252 ------------------------------ Message: 7 Date: Wed, 01 Jun 2005 09:50:45 +0200 From: "christian@eisbo.dk" Subject: [Histonet] Ki67. To: histonet@lists.utsouthwestern.edu Message-ID: <634ceeb7bf3c9de7df2bf51acb622dfe@eisbo.dk> Content-Type: text/plain; charset=windows-1252 Hey users of histonet. I am trying to stain kidneys for ki67! The sections have been in alcohol for 1 month and are now in parafin. The antibody I am using should be good enough. Does anyone have experience with this??? best regards. Christian EIsbo. ------------------------------ Message: 8 Date: Wed, 1 Jun 2005 10:29:19 +0200 From: "Bruijntjes, J.P." Subject: RE: [Histonet] Ki67. To: Cc: Histonet@lists.utsouthwestern.edu Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079DC6@ntexch1.voeding.tno.nl> Content-Type: text/plain; charset="us-ascii" Hi Christian Yes I do. May be it depends on the antibody you use. We use a rabbit monoclonal ki67 from Neomarkers (Labvision). Tissues fixed in alcohol don't react with the antibody using HIER in citrate buffer (pH 6,0). At least it didn't in my lab. It does when you use HIER in EDTA (pH 8.0) and in TRIS (pH 10). Hopes that will help you Joost Bruijntjes TNO Quality of Life Zeist Holland -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of christian@eisbo.dk Sent: woensdag 1 juni 2005 9:51 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Ki67. Hey users of histonet. I am trying to stain kidneys for ki67! The sections have been in alcohol for 1 month and are now in parafin. The antibody I am using should be good enough. Does anyone have experience with this??? best regards. Christian EIsbo. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html ------------------------------ Message: 9 Date: Wed, 1 Jun 2005 10:43:57 +0200 From: "Vincent Martin" Subject: [Histonet] visualization of muscle damage with light microscopy ? To: Message-ID: <002701c56686$0d65e6c0$0a4a7c8b@physiologie8> Content-Type: text/plain; charset="iso-8859-1" Dear list members, I would like to observe the effect of lengthening contractions on ultrastructural properties of rat tibialis anterior muscle. Is is possible to observe ultrastructural abnormalities with light microscopy with a x100 maginification ? If not, which are the minimal requirements to observe such abnormalities ? Thanks for your replies Best regards, Vincent Martin UPRES-EA 3285 D?terminants Physiologiques de l'Activit? Physique Facult? des Sciences du Sport de Marseille Universit? de la M?diterran?e BP910 - 163, avenue de Luminy 13288 Marseille cedex 09 France Phone : +33 (0)4-91-82-83-78 Fax : +33 (0)4-91-82-83-75 vincent.martin@staps.univ-mrs.fr www.physiologie.staps.univ-mrs.fr ------------------------------ Message: 10 Date: Wed, 01 Jun 2005 07:05:40 -0400 From: shawnster73@aol.com Subject: [Histonet] Histology of Frozen Tissue To: histonet@lists.utsouthwestern.edu Message-ID: <8C734B0FB191BFF-620-23897@FWM-D40.sysops.aol.com> Content-Type: text/plain; charset="us-ascii" What would be the histological effect of allowing frozen OCT embedded tissue to come up to a temperature of 0 or -5oC?? ------------------------------ Message: 11 Date: Wed, 1 Jun 2005 08:08:46 -0400 From: "Bobbie Boyce" Subject: [Histonet] BK Virus To: "HistoNet Server" Message-ID: <6E41111281623B4B8A9AB8F9A7EA343737BFB5@wlmmsx02.nemours.org> Content-Type: text/plain; charset=iso-8859-1 I'm having problems locating a FFPE control for BKV. Can anyone tell me where I can get them? Bobbie Boyce duPont Hospital for Children Wilmington, DE ------------------------------ Message: 12 Date: Wed, 01 Jun 2005 08:10:12 -0400 From: "WAYNE HOLLAND" Subject: [Histonet] HPV I and II Control Blocks To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hello All, Help, can anyone give assistance to getting some blocks for HPV I and HPV II for controls. We are going to start In-Situ, we are in the testing phase. One block from and HPV I (LOW) and one block from HPV II (HIGH) would be extremely helpful. We would be willing to trade, Thanks in advance. ------------------------------ Message: 13 Date: Wed, 01 Jun 2005 08:44:09 -0500 From: "Corazon D. Bucana" Subject: Re: [Histonet] mouse endothelial cells in brain To: "Donin, Nick (NIH/NCI)" Cc: "'histonet@lists.utsouthwestern.edu'" Message-ID: <5.1.1.6.0.20050601084009.042c9c48@audumla.mdacc.tmc.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Wed, 1 Jun 2005 10:08:33 -0400 From: "Donin, Nick (NIH/NCI)" Subject: [Histonet] staining with Solvent Blue 38 To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <16A0583FB1644E4DB8C0A0265028B6FD02B4E739@nihexchange13.nih.gov> Content-Type: text/plain Histonetters, I'm trying to stain the corpus callosum of formalin fixed paraffin embedded mouse brains using Solvent Blue 38. Could someone suggest a successful protocol? Thanks. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 ------------------------------ Message: 15 Date: Wed, 01 Jun 2005 11:23:20 -0400 From: Luis Chiriboga Subject: RE: [Histonet] mouse endothelial cells in brain To: "Corazon D. Bucana" , "Donin, Nick (NIH/NCI)" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii I have also used the pharmingen cd31 by frozen in mouse brain, muscle, and skin. It works extremely well in fresh system. FFPE on the other hand is much trickier. It works but very inconsistently, fixation and tissue processing are the key variables. Once you have obtained staining in FFPE, you need to follow the same sacrificing (perfusion etc..) grossing, processing procedure with "religious fanaticism" (no offense to anyone.....) L ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Corazon D. Bucana Sent: Wednesday, June 01, 2005 9:44 AM To: Donin, Nick (NIH/NCI) Cc: 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] mouse endothelial cells in brain The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Wed, 01 Jun 2005 15:24:32 +0000 From: "CHRISTIE GOWAN" Subject: [Histonet] Storage of Frozen Section slides To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed I know this subject has been tossed around before, but I wonder if Patsy Ruegg or Chris Van der Loos or anyone else for that matter would share with me their protocol for storing frozen slides. The company I work for sells frozen TMA slides. We cut sections as requested by clients and mail them out on dry ice. We have found that after going into the block 10-12 times we start to see artifact. Pre-cutting slides would be much better for the life of the block. Has anyone tried storing the slides in a vacuum sealed (food saver) bag? Any help would be appreciated. Thanks, Christie Gowan HT (ASCP) ------------------------------ Message: 17 Date: Wed, 01 Jun 2005 11:18:17 -0400 From: "Vinnie Della Speranza" Subject: Re: [Histonet] Will brain tissue shrink even more if storedin70%alcoholprior to processing? To: Cc: jchladny@cvm.uiuc.edu, histonet@lists.utsouthwestern.edu, clarissabush@sbcglobal.net Message-ID: Content-Type: text/plain; charset=US-ASCII I hadn't considered the possibility of buffer salt crystals forming the vacuoles observed by the author and I don't know if she is on this list and would care to comment but this is an interesting premise worth investigating. I would think that a thorough water wash prior to going into alcohol would help to clear this up. thanks for the feedback. Vinnie Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> John Kiernan 05/27/05 04:27PM >>> In the HistoLogic May 2000 paper the duration of the initial formaldehyde fixation is not stated; only that the brains were "previously well fixed in neutral buffered formalin." Frequently specimens are not fixed for long enough to make them resistant to bad effects of solvents etc. It's also seems from the paper that the test pieces of brain were passed directly from the buffered fixative into 70% alcohol. This causes precipitation of sodium phosphate in the tissue (see Freida Carson 1996 "Histotechnology" 2nd edn, p.27). The control specimens in the HistoLogic paper were passed from buffered formalin to 60% ethanol (which safely extracts the phosphate buffer salts). Could the holes in the white matter be made by buffer salt crystals rather than forming slowly during storage in 70% ethanol? Just a thought! -- ------------------------------- 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 _______________________________ Vinnie Della Speranza wrote: > > You may wish to look at an article authored by Jane Chladny, published in HistoLogic, May 2000, entitled "Artifact in Tissues Held in 70% Ethanol" > > the author reported the appearance of 'vacuole' artifact in nervous tissues observed in a variety of mammalian tissues after storage in 70% ethanol. > > you can access the article by selecting the link for HistoLogic at www.sakuraus.com > > CM Bush wrote: > > > > Dear Histonet, > > > > Hello, here is my first post to the list, thank you in advance for your help. > > > > Summary: > > Having problems with mouse and human brain tissue shrinkage durning processing, but would like to better preserve antigens, concerned storage of tissue 70% ethanol will shrink tissue even further: > > > > We have lots of brain specimens, stored in formalin for a long time (months up to 10 years), both human and mouse. We perform immunohistochemistry on paraffin embedded brain. Of course, there is the massive over fixation. Also the tissue shrinks a lot during processing. > > > > In my last lab, after brain tissue had been fixed in formalin for 7 days we would store the tissue in 70% ethanol. I'd like to start storing the brain tissue I work with now in 70%, but I'm worried about the tissue possibly shinking too much, as the tissue already seems to shrink by greater than 60% after processing. > > > > (Previously, we would gradually increase the alcohols, 30% for 1 hour, then 50% for an hour in a bucket, room temperature on a rocker table, then put the cassettes into a bucket of 70% and store at room temp or 4C- does this process help any with controling the shrinkage factor?) > > > > Maybe this is a little bit long...thank you very much for your time. > > > > CM Bush > --------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Vinnie Della Speranza > Manager for Anatomic Pathology Services > Medical University of South Carolina > 165 Ashley Avenue Suite 309 > Charleston, SC 29425 > Ph: 843-792-6353 > fax: 843-792-8974 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Wed, 1 Jun 2005 11:35:22 -0400 From: "Connolly, Brett M" Subject: RE: [Histonet] mouse endothelial cells in brain To: "'Corazon D. Bucana'" , "Donin, Nick (NIH/NCI)" Cc: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Nick, Fix the brains in Pharmingen Zinc Tris, process to paraffin and use the Pharmingen antibody. Works like a charm. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Corazon D. Bucana Sent: Wednesday, June 01, 2005 9:44 AM To: Donin, Nick (NIH/NCI) Cc: 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] mouse endothelial cells in brain The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ---------------------------------------------------------------------------- -- ------------------------------ Message: 19 Date: Wed, 01 Jun 2005 12:19:17 -0400 From: Tim Wheelock Subject: [Histonet] SOLVENT BLUE 38 ON PARAFFIN EMBEDDED MOUSE BRAIN To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <429DE005.1050905@mclean.harvard.edu> Content-Type: text/plain; charset=windows-1252; format=flowed Hi Nick: I use Solvent Blue 38 (also known as Luxol Fast Blue) on formalin-fixed, paraffin embedded human brain, but have also seen it work well on mouse brain. The protocol is as follows. This particular recipe combines the myelin stain with an H+E, but it can be combined with a Nissl stain instead or simply by itself. Hope this helps. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 * LUXOL FAST BLUE**-HEMATOXYLIN-EOSIN (LHE) STAIN* * * 1. De-paraffinize 5 micron formalin-fixed sections and take them down to 95% ethanol. 2. Stain in Luxol Fast Blue solution for 2 hours at 60C. 3. Remove and allow to cool for 15 minutes. 4. Remove excess stain in 95% ethanol for 1 minute. 5. Wash in several changes of tap water. 6. Differentiate sections with 2 dips in the reducer solution, then *immediately* through 4 changes of tap water (10 dips each), then one more change of tap water. 7. Immerse sections in Gill 3 Hematoxylin for 10 minutes. 8. Rinse in several changes of tap water. 9. Differentiate sections in acid alcohol (7 dips). 10. Rinse in 4 changes of tap water (10 dips each) then one more change of tap water. 11. Blue sections in Scott's Tap Water Substitute for 30 seconds. 12. Wash in 4 changes of tap water for 2 minutes each. 13. Immerse sections in 95% ethanol for 1 minute. 14. Immerse slides in alcoholic Eosin Y for 3 minute. 15. Differentiate in 95% ethanol (4 dips). 16 Dehydrate in first absolute ethanol for 20 dips. 17. Dehydrate in second absolute ethanol for 1 minute, and then clear, and mount. *SOLUTIONS*: Luxol Fast Blue: Luxol Fast Blue...............................................0.1 gram. 10% glacial acetic acid .......................................1 mls. 95% ethanol........................................................100mls. Reducer: Hydroquinone....................................................1 gram. Sodium Sulfite...................................................5 gram. Distilled water..................................................100 mls. Hematoxylin Differentiator: 1% Hydrochloric acid in 70% ethanol *RESULTS*: 1. Myelin: The myelin should be stained blue or greenish blue 2. Nuclei: The nuclear membrane should be sharply delineated in blue-black with the nucleoplasm clear except for the chromatin, also being blue-black. 3. Cytoplasm and background should be varying shades of red. NOTE 1. The thickness of the section, the type of hematoxylin, and all the times in hematoxylin, eosin, and dehydrating depend upon your requirements. 2. If the section thickness is more than 10 microns, you may want to use 3 or 4 dips in the Reducer, so as to avoid background myelin staining . 3. The so called "Reducer" (because it is used as a reducer in the Bodian silver protein stain) actually functions as a differentiator for the myelin stain. ------------------------------ Message: 20 Date: Wed, 1 Jun 2005 12:18:44 -0400 From: Diana McCaig Subject: [Histonet] filtering hematoxylin To: "Histonet (E-mail)" Message-ID: <3E5A3F039F0BD8118B4700C00D0020240435BF@CKHA9> Content-Type: text/plain How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? ------------------------------ Message: 21 Date: Wed, 1 Jun 2005 12:33:58 -0400 From: "Bartlett, Jeanine" Subject: RE: [Histonet] filtering hematoxylin To: "Diana McCaig" , "Histonet \(E-mail\)" Message-ID: Content-Type: text/plain; charset="us-ascii" We don't filter the hematoxylin we get from Richard-Allan. We replace it twice a month, but we have a light volume of H&E's. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 (404) 639-3590 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: Wednesday, June 01, 2005 12:19 PM To: Histonet (E-mail) Subject: [Histonet] filtering hematoxylin How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? _______________________________________________ Histonet mailing list Histonet@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 19, Issue 1 *************************************** The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at (864) 255-1000, and permanently delete the original e-mail, attachment(s), and any copies. From funderwood <@t> mcohio.org Wed Jun 1 12:20:36 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] filtering hematoxylin Message-ID: Weekly. A film forms on the hematoxylin over the weekend. Fred >>> Diana McCaig 06/01/05 12:18PM >>> How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Wed Jun 1 12:28:27 2005 From: pmarcum <@t> vet.upenn.edu (pmarcum@vet.upenn.edu) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] filtering hematoxylin In-Reply-To: References: Message-ID: <1117646907.429df03b5af0d@imp.vet.upenn.edu> It is dependent on the formula of the Harris Modified Hematoxylin used. Some commercial formulas will form less precipitate and film than others. If it is made in the laboratory generally I filter each day. Commercial formulas are checked each day for a film on the top or if I hear solid material when I shake or stir the solution. Pam Marcum Quoting Fred Underwood : > Weekly. A film forms on the hematoxylin over the weekend. > > Fred > > >>> Diana McCaig 06/01/05 12:18PM >>> > How often do you filter commercially prepared Harris Hematoxylin (used > on > autostainer)? > > _______________________________________________ > Histonet mailing list > Histonet@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 Thomas.Crowell <@t> biogenidec.com Wed Jun 1 12:34:54 2005 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Frozen TMA's In-Reply-To: Message-ID: Hello all, I'm hoping that there are some experts among the histonetters who have mastered the technique of creating frozen tissue microarray blocks, and would be willing to share some of the details with our laboratory. We are currently using a 2.5mm Harris uni-core to make a 4X5 template, and using the same core size to sample tissues - the cores fit nicely into the templates, however the interface between the tissue core and the OCT block is creating much havoc in trying to obtain reproducible sequential cryosections (sections curl up, drag along the blade, compress). Smearing a layer of OCT onto the block surface does help to fill in the interface irregularities, but only alleviates the problem for 3- 5 sections. Any suggestions for improvement would be greatly appreciated! Tom Crowell BiogenIdec Cambridge, MA From sharon.osborn <@t> dnax.org Wed Jun 1 12:44:51 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Filtering Harris hematoxylin Message-ID: <29B25753F6B1D51196110002A589D44402397FC0@PALMSG30.us.schp.com> We filter Harris Hematoxylin everytime or daily that it is being used. It forms a film sheen across the top of container that coats the slides creating blue spots on the slides if not filtered often. Other types of hematoxyling preparations do not have such requirements. sharon osborn DNAX ScheringPlough Biopharma Palo Alto, CA On Behalf Of Diana McCaig Sent: Wednesday, June 01, 2005 12:19 PM To: Histonet (E-mail) Subject: [Histonet] filtering hematoxylin How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? ********************************************************************* 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 christiegowan <@t> msn.com Wed Jun 1 12:52:31 2005 From: christiegowan <@t> msn.com (CHRISTIE GOWAN) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Storing frozen TMA sections Message-ID: I know this is an old problem but I was wondering if Patsy Ruegg or Chris Van der Loos or anyone for that matter could send me their protocol for storing frozen TMA sections. Has anyone ever tried to store them in vacuum sealed (Food Saver) bags? I work for a company that provides frozen TMA slides for customers. We section them as needed and have discovered that the tissue starts showing freezer artifact after going into the block about 10-12 times. Any help would be greatly appreciated. Thanks, Christie Gowan From JWEEMS <@t> sjha.org Wed Jun 1 12:54:17 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Muscle and Nerve Biopsies Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45E58@sjhaexc02.sjha.org> Hello, Can anyone recommmend a good reference laboratory for muscle and nerve biopsies, cheap and with quick TAT?!!! Thanks, Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From christiegowan <@t> msn.com Wed Jun 1 13:02:48 2005 From: christiegowan <@t> msn.com (CHRISTIE GOWAN) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Frozen TMA's In-Reply-To: Message-ID: Thomas, Do you build your TMA's in a cryostat or on dry ice. Sometimes this has an effect on the tissue interface. Also, we use the cryojane tape system by Intrumedics. This allows us to get consistent sections each time. If you build your TMA's on dry ice let me know and I will be happy to send you our protocol. Christie Gowan Thomas Crowell wrote... >Hello all, > >I'm hoping that there are some experts among the histonetters who have >mastered the technique of creating frozen tissue microarray blocks, and >would be willing to share some of the details with our laboratory. We are >currently using a 2.5mm Harris uni-core to make a 4X5 template, and using >the same core size to sample tissues - the cores fit nicely into the >templates, however the interface between the tissue core and the OCT block >is creating much havoc in trying to obtain reproducible sequential >cryosections (sections curl up, drag along the blade, compress). Smearing >a layer of OCT onto the block surface does help to fill in the interface >irregularities, but only alleviates the problem for 3- 5 sections. > >Any suggestions for improvement would be greatly appreciated! > >Tom Crowell >BiogenIdec >Cambridge, MA > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Jun 1 13:38:24 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Storing frozen TMA sections In-Reply-To: Message-ID: <200506011838.j51IcO1C013736@chip.viawest.net> Christie, I store frozen tissue blocks which have been carefully resealed with OCT after sectioning at -80 in freezer quality zip locks. I put the tissue in one zip lock seal it and then store that bag inside another zip lock with some dessicant in it. I can imagine that taking the tissue from -80 to cyrostat temps (usually around -20) repeatedly may cause some degradation. Although, I have stored tissues this way for years for tonsil controls for instance and gone back and forth serveral times with no apparent ill effects. As for storing the cut slides, I usally airdry overnight and store the section without fixation wrapped in foil and double bagged as for the frozen blocks. I fix just before staining. I take the bag out of the freezer which also has dessicant in it and let it come to rt (about 15-30 min.) before opening the sealed bag to prevent water forming on the section. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of CHRISTIE GOWAN Sent: Wednesday, June 01, 2005 10:53 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Storing frozen TMA sections I know this is an old problem but I was wondering if Patsy Ruegg or Chris Van der Loos or anyone for that matter could send me their protocol for storing frozen TMA sections. Has anyone ever tried to store them in vacuum sealed (Food Saver) bags? I work for a company that provides frozen TMA slides for customers. We section them as needed and have discovered that the tissue starts showing freezer artifact after going into the block about 10-12 times. Any help would be greatly appreciated. Thanks, Christie Gowan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Jun 1 13:40:53 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] GMA block question In-Reply-To: <6.0.3.0.0.20050601130056.01aee2b0@aretha.jax.org> Message-ID: <200506011840.j51Ieq1C014564@chip.viawest.net> Judi are you using metal or plastic tissue block holders? Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Judi Ford Sent: Wednesday, June 01, 2005 10:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] GMA block question Hi everyone, I have a question about gma blocks coming off the holder. We use Technovit 7100 to embed our tissue (mouse eyes). Once the resin is set, overnight @ room temperature, the blocks are popped out of the mold and dried in a room temperature oven overnight (once in awhile the temperature may exceed room temperature). The next day we cut them. The problem I see, is that every so often the tissue block is loose, even after being in the oven and in a dessicator. We end up having to cut the block off the backing and remount it using epoxy. There are others in the lab doing the same exact method as we are and they have had no problems. The tissue blocks are rock solid. We've tried to examine all the angles........ Any ideas about what could be causing this? Thanks, Judi Ford Histotechnologist The Jackson Laboratory Bar Harbor, ME _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mclarke <@t> allsaintshealthcare.org Wed Jun 1 13:53:59 2005 From: mclarke <@t> allsaintshealthcare.org (Clarke, Mary) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] cryostats Message-ID: <6FE2A453DA437F4D96FAAF363F20ABB20FC84F@WFEXBE04.wfsi.priv> We are in the process of purchasing a new cryostat. Has anyone used the Richard-Allan 525 or the Leica CM 1850? We are looking at both and wonder how people like or dislike them? Thanks in advance for your help. Terri Clarke Histology Supervisor All Saints Laboratory 3801 Spring Street Racine, Wi 53405 mclarke@allsaintshealthcare.org 262-687-6609 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, June 01, 2005 12:01 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 19, Issue 1 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. Speaking of Gram's... (Breeden, Sara) 2. Freezing muscle biopsies (Osborn, Sharon) 3. Enzyme Staining and Billing for Controls (Mitchell, Guy) 4. RE: Number of Blocks Submitted by PA (Charles.Embrey) 5. Aquaporin immunohistochemistry (Anthony Reilly) 6. Ki67. (christian@eisbo.dk) 7. Ki67. (christian@eisbo.dk) 8. RE: Ki67. (Bruijntjes, J.P.) 9. visualization of muscle damage with light microscopy ? (Vincent Martin) 10. Histology of Frozen Tissue (shawnster73@aol.com) 11. BK Virus (Bobbie Boyce) 12. HPV I and II Control Blocks (WAYNE HOLLAND) 13. Re: mouse endothelial cells in brain (Corazon D. Bucana) 14. staining with Solvent Blue 38 (Donin, Nick (NIH/NCI)) 15. RE: mouse endothelial cells in brain (Luis Chiriboga) 16. Storage of Frozen Section slides (CHRISTIE GOWAN) 17. Re: Will brain tissue shrink even more if storedin70%alcoholprior to processing? (Vinnie Della Speranza) 18. RE: mouse endothelial cells in brain (Connolly, Brett M) 19. SOLVENT BLUE 38 ON PARAFFIN EMBEDDED MOUSE BRAIN (Tim Wheelock) 20. filtering hematoxylin (Diana McCaig) 21. RE: filtering hematoxylin (Bartlett, Jeanine) ---------------------------------------------------------------------- Message: 1 Date: Tue, 31 May 2005 12:33:01 -0600 From: "Breeden, Sara" Subject: [Histonet] Speaking of Gram's... To: Message-ID: Content-Type: text/plain; charset="us-ascii" I'm having trouble with my Gram's in that my positive bacteria are losing color. So, I'm wondering what the exact differentiation time should be for the step after the Gram's Iodine (I'm using the Sigma kit)? The instructions are not precise, so I'm thinkin' maybe I'm over differentiating or under-Safranin-ing or aliens (this is the State that's home to ROSWELL...) have taken over my kit. Anyone have anything a bit more specific? I'd appreciate any help... Sara A. Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX ------------------------------ Message: 2 Date: Tue, 31 May 2005 15:41:02 -0400 From: "Osborn, Sharon" Subject: [Histonet] Freezing muscle biopsies To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <29B25753F6B1D51196110002A589D44402397FB8@PALMSG30.us.schp.com> Content-Type: text/plain Liz, Our researchers often freeze tissue for later study. They use the plastic disposable embedding molds. These can be used with the dry ice as John Kiernan suggested. There is a thin layer of OCT placed in bottom of mold then the tissue is oriented in this with additional OCT to fill the mold. The mold can be labeled with a permanent felt tip pen. Each specimen is then placed into the labeled sterile sampling bags, packed in dry ice and shipped to respective location. We have experienced very little distortion of the tissue due to ice crystals, etc. The testing has worked out okay. The main tissues were liver, kidney, lung. You may wish to experiment with some tissues before actually tackling the research samples to get the right combination for you. Sharon Osborn DNAX ScheringPlough Biopharma Palo Alto, CA Hello Histonetters > > I have a unique question. We are currently starting to set up > procedure for collecting samples from a clinical trial. The clinical > trial involves taking multiple synovial biopsies at a surgery center. > Since portions of the samples need to be processed for frozen sections > we wanted to be able to freeze the specimens at the surgery center via > isopentane cooled liquid nitrogen. We really do not want to have to > transport the multiple specimens back to the main lab prior to > freezing due to the time involved it would probably be 1-2 hours post > biopsy before we could freeze the samples. The surgery center is > questioning the flammability of the isopentane. Has anyone > encountered anything like this? Any suggestions would be helpful. > Thanks in advance. > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > Manager > Premier Laboratory, LLC > P.O. Box 18592 > Boulder, Colorado 80308 > Office: (303) 735-5001 > Fax: (303) 735-3540 > liz@premierlab.com > www.premierlab.com > > Ship to Address: > Premier Laboratory > University of Colorado > MCDB, Room A3B40 > Boulder, Colorado 80309 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 3 Date: Tue, 31 May 2005 15:42:59 -0400 From: "Mitchell, Guy" Subject: [Histonet] Enzyme Staining and Billing for Controls To: "HistoNet Server" Message-ID: <4DCF5CA56B4AA04ABC383DE358BAD3820289100D@dcr-xchg-04.carolinas.org> Content-Type: text/plain; charset="US-ASCII" A question has come up concerning charges for muscle enzyme stains. We stain for Adenylate Deaminase and Phosphofructokinase as well as negatives for each. I assume they should be treated as controls without a charge but others disagree. Guy W. Mitchell (704) 379-5984 (704) 379-6144 Fax ----------------------------------------- This electronic message may contain information that is confidential and/or legally privileged. It is intended only for the use of the individual(s) and entity named as recipients in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from any computer. Do not deliver, distribute or copy this message, and do not disclose its contents or take any action in reliance on the information it contains. Thank you. ------------------------------ Message: 4 Date: Tue, 31 May 2005 15:42:58 -0500 From: "Charles.Embrey" Subject: RE: [Histonet] Number of Blocks Submitted by PA To: "Smith Wanda" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" I have been gone for a week and just got a chance to check my e-mails and found this question interesting. First, have you asked the PA? I know it seems like a simple question but you would be surprised at all the misunderstanding that can be cleared up with a simple question asked? Please know that I have NEVER met a pathologist that will gladly read more slides than he thinks are necessary for a good diagnosis on a routine basis. I gross for seven pathologists and it is tricky to satisfy all at the same time. Luckily, my pathologists and I here have great communication and we manage to keep the block count, as well a gross recut number down. As far as "rules of thumb" we do have certain guidelines that are basically thought of as "standards of care". Without knowing other details the best advice I could give is to ask the PA. Charles Embrey PA(ASCP) Urbana IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Smith Wanda Sent: Tuesday, May 24, 2005 7:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Number of Blocks Submitted by PA Good Morning Histonetters, I have a question for you this morning...Are there written standards or "Rules of Thumb" regarding number of blocks that should be submitted on certain tissues? The reason I ask is, "it seems to me" that sometimes the PA submits an extreme number of blocks on some specimens that could be accomplished with less. I'm not advocating cutting corners or not performing good medical practice, I just need to know. An example would be a uterus w/ fibroids, tubes and ovaries which we have gotten up to 24 blocks on. I'm not a PA, but it's interesting to me that when the Pathologist cut, we get alot less blocks. Please advise. Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Wed, 01 Jun 2005 14:13:15 +1000 From: "Anthony Reilly" Subject: [Histonet] Aquaporin immunohistochemistry To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hello All I have been requested to be involved in a research study demonstrating Aquaporin 1 (AQ1, AQP1) using IHC. To date I have found 2 antibodies available from Chemicon. My questions are: 1. Are there other commercial suppliers of this Ab. 2. If not, which of the 2 from Chemicon is the preferred option. 3. Do you realyy need to fix with paraformaldehyde as stated in most methods, and why? i would appreciate any advice forwarded. regards Tony Reilly Chief Scientist Anatomical Pathology Northside Pathology Prince Charles Hospital Rode rd Chermside Q 4032 Australia Ph: 07 3350 8543 Fax: 07 33508546 tony_reilly@health.qld.gov.au *********************************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is prohibited. It may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone or by return email. You should also delete this email and destroy any hard copies produced. *********************************************************************************** ------------------------------ Message: 6 Date: Wed, 01 Jun 2005 09:48:19 +0200 From: "christian@eisbo.dk" Subject: [Histonet] Ki67. To: histonet@lists.utsouthwestern.edu Message-ID: <758a26d4ec47df2ab23acf9597f0b69d@eisbo.dk> Content-Type: text/plain; charset=windows-1252 ------------------------------ Message: 7 Date: Wed, 01 Jun 2005 09:50:45 +0200 From: "christian@eisbo.dk" Subject: [Histonet] Ki67. To: histonet@lists.utsouthwestern.edu Message-ID: <634ceeb7bf3c9de7df2bf51acb622dfe@eisbo.dk> Content-Type: text/plain; charset=windows-1252 Hey users of histonet. I am trying to stain kidneys for ki67! The sections have been in alcohol for 1 month and are now in parafin. The antibody I am using should be good enough. Does anyone have experience with this??? best regards. Christian EIsbo. ------------------------------ Message: 8 Date: Wed, 1 Jun 2005 10:29:19 +0200 From: "Bruijntjes, J.P." Subject: RE: [Histonet] Ki67. To: Cc: Histonet@lists.utsouthwestern.edu Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079DC6@ntexch1.voeding.tno.nl> Content-Type: text/plain; charset="us-ascii" Hi Christian Yes I do. May be it depends on the antibody you use. We use a rabbit monoclonal ki67 from Neomarkers (Labvision). Tissues fixed in alcohol don't react with the antibody using HIER in citrate buffer (pH 6,0). At least it didn't in my lab. It does when you use HIER in EDTA (pH 8.0) and in TRIS (pH 10). Hopes that will help you Joost Bruijntjes TNO Quality of Life Zeist Holland -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of christian@eisbo.dk Sent: woensdag 1 juni 2005 9:51 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Ki67. Hey users of histonet. I am trying to stain kidneys for ki67! The sections have been in alcohol for 1 month and are now in parafin. The antibody I am using should be good enough. Does anyone have experience with this??? best regards. Christian EIsbo. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html ------------------------------ Message: 9 Date: Wed, 1 Jun 2005 10:43:57 +0200 From: "Vincent Martin" Subject: [Histonet] visualization of muscle damage with light microscopy ? To: Message-ID: <002701c56686$0d65e6c0$0a4a7c8b@physiologie8> Content-Type: text/plain; charset="iso-8859-1" Dear list members, I would like to observe the effect of lengthening contractions on ultrastructural properties of rat tibialis anterior muscle. Is is possible to observe ultrastructural abnormalities with light microscopy with a x100 maginification ? If not, which are the minimal requirements to observe such abnormalities ? Thanks for your replies Best regards, Vincent Martin UPRES-EA 3285 D?terminants Physiologiques de l'Activit? Physique Facult? des Sciences du Sport de Marseille Universit? de la M?diterran?e BP910 - 163, avenue de Luminy 13288 Marseille cedex 09 France Phone : +33 (0)4-91-82-83-78 Fax : +33 (0)4-91-82-83-75 vincent.martin@staps.univ-mrs.fr www.physiologie.staps.univ-mrs.fr ------------------------------ Message: 10 Date: Wed, 01 Jun 2005 07:05:40 -0400 From: shawnster73@aol.com Subject: [Histonet] Histology of Frozen Tissue To: histonet@lists.utsouthwestern.edu Message-ID: <8C734B0FB191BFF-620-23897@FWM-D40.sysops.aol.com> Content-Type: text/plain; charset="us-ascii" What would be the histological effect of allowing frozen OCT embedded tissue to come up to a temperature of 0 or -5oC?? ------------------------------ Message: 11 Date: Wed, 1 Jun 2005 08:08:46 -0400 From: "Bobbie Boyce" Subject: [Histonet] BK Virus To: "HistoNet Server" Message-ID: <6E41111281623B4B8A9AB8F9A7EA343737BFB5@wlmmsx02.nemours.org> Content-Type: text/plain; charset=iso-8859-1 I'm having problems locating a FFPE control for BKV. Can anyone tell me where I can get them? Bobbie Boyce duPont Hospital for Children Wilmington, DE ------------------------------ Message: 12 Date: Wed, 01 Jun 2005 08:10:12 -0400 From: "WAYNE HOLLAND" Subject: [Histonet] HPV I and II Control Blocks To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hello All, Help, can anyone give assistance to getting some blocks for HPV I and HPV II for controls. We are going to start In-Situ, we are in the testing phase. One block from and HPV I (LOW) and one block from HPV II (HIGH) would be extremely helpful. We would be willing to trade, Thanks in advance. ------------------------------ Message: 13 Date: Wed, 01 Jun 2005 08:44:09 -0500 From: "Corazon D. Bucana" Subject: Re: [Histonet] mouse endothelial cells in brain To: "Donin, Nick (NIH/NCI)" Cc: "'histonet@lists.utsouthwestern.edu'" Message-ID: <5.1.1.6.0.20050601084009.042c9c48@audumla.mdacc.tmc.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Wed, 1 Jun 2005 10:08:33 -0400 From: "Donin, Nick (NIH/NCI)" Subject: [Histonet] staining with Solvent Blue 38 To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <16A0583FB1644E4DB8C0A0265028B6FD02B4E739@nihexchange13.nih.gov> Content-Type: text/plain Histonetters, I'm trying to stain the corpus callosum of formalin fixed paraffin embedded mouse brains using Solvent Blue 38. Could someone suggest a successful protocol? Thanks. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 ------------------------------ Message: 15 Date: Wed, 01 Jun 2005 11:23:20 -0400 From: Luis Chiriboga Subject: RE: [Histonet] mouse endothelial cells in brain To: "Corazon D. Bucana" , "Donin, Nick (NIH/NCI)" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii I have also used the pharmingen cd31 by frozen in mouse brain, muscle, and skin. It works extremely well in fresh system. FFPE on the other hand is much trickier. It works but very inconsistently, fixation and tissue processing are the key variables. Once you have obtained staining in FFPE, you need to follow the same sacrificing (perfusion etc..) grossing, processing procedure with "religious fanaticism" (no offense to anyone.....) L ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Corazon D. Bucana Sent: Wednesday, June 01, 2005 9:44 AM To: Donin, Nick (NIH/NCI) Cc: 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] mouse endothelial cells in brain The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Wed, 01 Jun 2005 15:24:32 +0000 From: "CHRISTIE GOWAN" Subject: [Histonet] Storage of Frozen Section slides To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed I know this subject has been tossed around before, but I wonder if Patsy Ruegg or Chris Van der Loos or anyone else for that matter would share with me their protocol for storing frozen slides. The company I work for sells frozen TMA slides. We cut sections as requested by clients and mail them out on dry ice. We have found that after going into the block 10-12 times we start to see artifact. Pre-cutting slides would be much better for the life of the block. Has anyone tried storing the slides in a vacuum sealed (food saver) bag? Any help would be appreciated. Thanks, Christie Gowan HT (ASCP) ------------------------------ Message: 17 Date: Wed, 01 Jun 2005 11:18:17 -0400 From: "Vinnie Della Speranza" Subject: Re: [Histonet] Will brain tissue shrink even more if storedin70%alcoholprior to processing? To: Cc: jchladny@cvm.uiuc.edu, histonet@lists.utsouthwestern.edu, clarissabush@sbcglobal.net Message-ID: Content-Type: text/plain; charset=US-ASCII I hadn't considered the possibility of buffer salt crystals forming the vacuoles observed by the author and I don't know if she is on this list and would care to comment but this is an interesting premise worth investigating. I would think that a thorough water wash prior to going into alcohol would help to clear this up. thanks for the feedback. Vinnie Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> John Kiernan 05/27/05 04:27PM >>> In the HistoLogic May 2000 paper the duration of the initial formaldehyde fixation is not stated; only that the brains were "previously well fixed in neutral buffered formalin." Frequently specimens are not fixed for long enough to make them resistant to bad effects of solvents etc. It's also seems from the paper that the test pieces of brain were passed directly from the buffered fixative into 70% alcohol. This causes precipitation of sodium phosphate in the tissue (see Freida Carson 1996 "Histotechnology" 2nd edn, p.27). The control specimens in the HistoLogic paper were passed from buffered formalin to 60% ethanol (which safely extracts the phosphate buffer salts). Could the holes in the white matter be made by buffer salt crystals rather than forming slowly during storage in 70% ethanol? Just a thought! -- ------------------------------- 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 _______________________________ Vinnie Della Speranza wrote: > > You may wish to look at an article authored by Jane Chladny, published in HistoLogic, May 2000, entitled "Artifact in Tissues Held in 70% Ethanol" > > the author reported the appearance of 'vacuole' artifact in nervous tissues observed in a variety of mammalian tissues after storage in 70% ethanol. > > you can access the article by selecting the link for HistoLogic at www.sakuraus.com > > CM Bush wrote: > > > > Dear Histonet, > > > > Hello, here is my first post to the list, thank you in advance for your help. > > > > Summary: > > Having problems with mouse and human brain tissue shrinkage durning processing, but would like to better preserve antigens, concerned storage of tissue 70% ethanol will shrink tissue even further: > > > > We have lots of brain specimens, stored in formalin for a long time (months up to 10 years), both human and mouse. We perform immunohistochemistry on paraffin embedded brain. Of course, there is the massive over fixation. Also the tissue shrinks a lot during processing. > > > > In my last lab, after brain tissue had been fixed in formalin for 7 days we would store the tissue in 70% ethanol. I'd like to start storing the brain tissue I work with now in 70%, but I'm worried about the tissue possibly shinking too much, as the tissue already seems to shrink by greater than 60% after processing. > > > > (Previously, we would gradually increase the alcohols, 30% for 1 hour, then 50% for an hour in a bucket, room temperature on a rocker table, then put the cassettes into a bucket of 70% and store at room temp or 4C- does this process help any with controling the shrinkage factor?) > > > > Maybe this is a little bit long...thank you very much for your time. > > > > CM Bush > --------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Vinnie Della Speranza > Manager for Anatomic Pathology Services > Medical University of South Carolina > 165 Ashley Avenue Suite 309 > Charleston, SC 29425 > Ph: 843-792-6353 > fax: 843-792-8974 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Wed, 1 Jun 2005 11:35:22 -0400 From: "Connolly, Brett M" Subject: RE: [Histonet] mouse endothelial cells in brain To: "'Corazon D. Bucana'" , "Donin, Nick (NIH/NCI)" Cc: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Nick, Fix the brains in Pharmingen Zinc Tris, process to paraffin and use the Pharmingen antibody. Works like a charm. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Corazon D. Bucana Sent: Wednesday, June 01, 2005 9:44 AM To: Donin, Nick (NIH/NCI) Cc: 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] mouse endothelial cells in brain The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. 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If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ ------------------------------ Message: 19 Date: Wed, 01 Jun 2005 12:19:17 -0400 From: Tim Wheelock Subject: [Histonet] SOLVENT BLUE 38 ON PARAFFIN EMBEDDED MOUSE BRAIN To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <429DE005.1050905@mclean.harvard.edu> Content-Type: text/plain; charset=windows-1252; format=flowed Hi Nick: I use Solvent Blue 38 (also known as Luxol Fast Blue) on formalin-fixed, paraffin embedded human brain, but have also seen it work well on mouse brain. The protocol is as follows. This particular recipe combines the myelin stain with an H+E, but it can be combined with a Nissl stain instead or simply by itself. Hope this helps. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 * LUXOL FAST BLUE**-HEMATOXYLIN-EOSIN (LHE) STAIN* * * 1. De-paraffinize 5 micron formalin-fixed sections and take them down to 95% ethanol. 2. Stain in Luxol Fast Blue solution for 2 hours at 60C. 3. Remove and allow to cool for 15 minutes. 4. Remove excess stain in 95% ethanol for 1 minute. 5. Wash in several changes of tap water. 6. Differentiate sections with 2 dips in the reducer solution, then *immediately* through 4 changes of tap water (10 dips each), then one more change of tap water. 7. Immerse sections in Gill 3 Hematoxylin for 10 minutes. 8. Rinse in several changes of tap water. 9. Differentiate sections in acid alcohol (7 dips). 10. Rinse in 4 changes of tap water (10 dips each) then one more change of tap water. 11. Blue sections in Scott's Tap Water Substitute for 30 seconds. 12. Wash in 4 changes of tap water for 2 minutes each. 13. Immerse sections in 95% ethanol for 1 minute. 14. Immerse slides in alcoholic Eosin Y for 3 minute. 15. Differentiate in 95% ethanol (4 dips). 16 Dehydrate in first absolute ethanol for 20 dips. 17. Dehydrate in second absolute ethanol for 1 minute, and then clear, and mount. *SOLUTIONS*: Luxol Fast Blue: Luxol Fast Blue...............................................0.1 gram. 10% glacial acetic acid .......................................1 mls. 95% ethanol........................................................100mls. Reducer: Hydroquinone....................................................1 gram. Sodium Sulfite...................................................5 gram. Distilled water..................................................100 mls. Hematoxylin Differentiator: 1% Hydrochloric acid in 70% ethanol *RESULTS*: 1. Myelin: The myelin should be stained blue or greenish blue 2. Nuclei: The nuclear membrane should be sharply delineated in blue-black with the nucleoplasm clear except for the chromatin, also being blue-black. 3. Cytoplasm and background should be varying shades of red. NOTE 1. The thickness of the section, the type of hematoxylin, and all the times in hematoxylin, eosin, and dehydrating depend upon your requirements. 2. If the section thickness is more than 10 microns, you may want to use 3 or 4 dips in the Reducer, so as to avoid background myelin staining . 3. The so called "Reducer" (because it is used as a reducer in the Bodian silver protein stain) actually functions as a differentiator for the myelin stain. ------------------------------ Message: 20 Date: Wed, 1 Jun 2005 12:18:44 -0400 From: Diana McCaig Subject: [Histonet] filtering hematoxylin To: "Histonet (E-mail)" Message-ID: <3E5A3F039F0BD8118B4700C00D0020240435BF@CKHA9> Content-Type: text/plain How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? ------------------------------ Message: 21 Date: Wed, 1 Jun 2005 12:33:58 -0400 From: "Bartlett, Jeanine" Subject: RE: [Histonet] filtering hematoxylin To: "Diana McCaig" , "Histonet \(E-mail\)" Message-ID: Content-Type: text/plain; charset="us-ascii" We don't filter the hematoxylin we get from Richard-Allan. We replace it twice a month, but we have a light volume of H&E's. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 (404) 639-3590 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: Wednesday, June 01, 2005 12:19 PM To: Histonet (E-mail) Subject: [Histonet] filtering hematoxylin How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? _______________________________________________ Histonet mailing list Histonet@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 19, Issue 1 *************************************** Privileged/Confidential information may be contained in this message. 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Opinions, conclusions and other information in this message that do not relate to the official business of the firm shall be understood as neither given nor endorsed by it. From info <@t> instrumedics.com Wed Jun 1 14:45:33 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Frozen TMA's References: Message-ID: <01d701c566e2$7fc29db0$6401a8c0@INSTRUMEDICS22> Thomas, We have direct experience with a lab that prepared frozen tissue arrays. However, the quality of the section of cores was very poor until they used the CryoJane Tape-Transfer system. The morphology was preserved. Sectioning the block did not raise the problems you describe. See publication, M .Schoenberg- Fejzo and Dennis J. Slamen "Frozen Tumor Tissue Microarray Technology for Analysis of Tumor RNA, DNA, and Proteins, . American Journal of Pathology November, 2001, pg 1645. The paper discusses the CryoJane and says "The tape transfer system (Instrumedics, Inc) was critical to maintaining the integrity of the sample" Please visit our web site and click on "products" and go to "CryoJane" to see how the system works! Bernice Instrumedics ----- Original Message ----- From: "Thomas Crowell" To: Sent: Wednesday, June 01, 2005 1:34 PM Subject: [Histonet] Frozen TMA's > Hello all, > > I'm hoping that there are some experts among the histonetters who have > mastered the technique of creating frozen tissue microarray blocks, and > would be willing to share some of the details with our laboratory. We are > currently using a 2.5mm Harris uni-core to make a 4X5 template, and using > the same core size to sample tissues - the cores fit nicely into the > templates, however the interface between the tissue core and the OCT block > is creating much havoc in trying to obtain reproducible sequential > cryosections (sections curl up, drag along the blade, compress). Smearing > a layer of OCT onto the block surface does help to fill in the interface > irregularities, but only alleviates the problem for 3- 5 sections. > > Any suggestions for improvement would be greatly appreciated! > > Tom Crowell > BiogenIdec > Cambridge, MA > > > From info <@t> instrumedics.com Wed Jun 1 15:02:24 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Storing frozen TMA sections References: Message-ID: <01e701c566e4$da12f7e0$6401a8c0@INSTRUMEDICS22> Christie, We coat our blocks with Instrumedcis Protective Oil that has a freezing point of -10C. It will not melt the tissue cores in the block and will prevent dehydration when storing in a freezer for even long periods. We also coat the blockface when we leave the block in the microtome for next day sectioning The frozen oil can be sectioned, just like embedding media, so it can removed and the tissue cores are ready to be sectioned. Bernice Instrumedics ----- Original Message ----- From: "CHRISTIE GOWAN" To: Sent: Wednesday, June 01, 2005 1:52 PM Subject: [Histonet] Storing frozen TMA sections >I know this is an old problem but I was wondering if Patsy Ruegg or Chris >Van der Loos or anyone for that matter could send me their protocol for >storing frozen TMA sections. Has anyone ever tried to store them in vacuum >sealed (Food Saver) bags? I work for a company that provides frozen TMA >slides for customers. We section them as needed and have discovered that >the tissue starts showing freezer artifact after going into the block about >10-12 times. Any help would be greatly appreciated. > Thanks, > Christie Gowan > > > > > From petepath <@t> yahoo.com Wed Jun 1 15:56:43 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Frozen TMA's Message-ID: <20050601205643.26577.qmail@web30405.mail.mud.yahoo.com> Tom, I have a few customers making frozen microarrays simply by adhering the tissue pieces to the floor of my embedding well bars. You can use frozen cores or pieces by putting a touch of OCT on them to adhere the tissue to the well floor. You can fill it with cold OCT to avoid rewarming. Works great with fresh tissue if snap freezing is not a requirement. If you are not familiar with my well bars have a visit to my web site. Depending on the tissue it should cut like any other frozen. Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From ynwang <@t> u.washington.edu Wed Jun 1 16:28:33 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] resin embedded tissue and sirius red staining Message-ID: Dear histonetters, I have a question regarding the feasibility of staining resin embedded skin (in technovit) with Picosirius red (for collagen). We have some old samples embedded in resin that we would like to stain for collagen if at all possible. We normally do our sirius red staining on frozen and paraffin embedded tissue with good results. I tried running the same protocol (1hr sirius red followed by quick wash in acidified water) as well as at longer times (sirius red for 3 hours) with the resin sections but I get very dull staining (under polarized light I do not get the nice red/green/orange/yellow I normally see, instead it's rather murky). I haven't found anything online in terms of staining resin embedded samples with sirius red so I was wondering if this was one of those things that can't be done. Any comments would be much appreciated. Thank you Yak-Nam Wang Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 From pruegg <@t> ihctech.net Wed Jun 1 16:38:52 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Will brain tissue shrink even more ifstoredin70%alcoholprior to processing? In-Reply-To: Message-ID: <200506012138.j51Lcq1C017708@chip.viawest.net> For IHC it is usually recommended that tissues be optimally fixed in NBF then transferred to 70% alcohol for storage before processing to limit the ill effects of cross linking of proteins by over fixation with aldehyde fixatives. If buffer salt crystals can form using these methods we we reconsider this? I have always been a believer in washing after fixation but most of us do not have the luxury of having the time to do so. As I researched a presentation I was doing on IHC I ran across this: "A little known fact is that if tissues were washed in running tap h202 (10-20 min.) after aldehyde fixation most epitope retrieval procedures would be unnecessary." Now there may be another reason to wash after aldehyde fixation???? Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vinnie Della Speranza Sent: Wednesday, June 01, 2005 8:18 AM To: jkiernan@uwo.ca Cc: jchladny@cvm.uiuc.edu; histonet@lists.utsouthwestern.edu; clarissabush@sbcglobal.net Subject: Re: [Histonet] Will brain tissue shrink even more ifstoredin70%alcoholprior to processing? I hadn't considered the possibility of buffer salt crystals forming the vacuoles observed by the author and I don't know if she is on this list and would care to comment but this is an interesting premise worth investigating. I would think that a thorough water wash prior to going into alcohol would help to clear this up. thanks for the feedback. Vinnie Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> John Kiernan 05/27/05 04:27PM >>> In the HistoLogic May 2000 paper the duration of the initial formaldehyde fixation is not stated; only that the brains were "previously well fixed in neutral buffered formalin." Frequently specimens are not fixed for long enough to make them resistant to bad effects of solvents etc. It's also seems from the paper that the test pieces of brain were passed directly from the buffered fixative into 70% alcohol. This causes precipitation of sodium phosphate in the tissue (see Freida Carson 1996 "Histotechnology" 2nd edn, p.27). The control specimens in the HistoLogic paper were passed from buffered formalin to 60% ethanol (which safely extracts the phosphate buffer salts). Could the holes in the white matter be made by buffer salt crystals rather than forming slowly during storage in 70% ethanol? Just a thought! -- ------------------------------- 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 _______________________________ Vinnie Della Speranza wrote: > > You may wish to look at an article authored by Jane Chladny, published in HistoLogic, May 2000, entitled "Artifact in Tissues Held in 70% Ethanol" > > the author reported the appearance of 'vacuole' artifact in nervous tissues observed in a variety of mammalian tissues after storage in 70% ethanol. > > you can access the article by selecting the link for HistoLogic at > www.sakuraus.com > > CM Bush wrote: > > > > Dear Histonet, > > > > Hello, here is my first post to the list, thank you in advance for your help. > > > > Summary: > > Having problems with mouse and human brain tissue shrinkage durning processing, but would like to better preserve antigens, concerned storage of tissue 70% ethanol will shrink tissue even further: > > > > We have lots of brain specimens, stored in formalin for a long time (months up to 10 years), both human and mouse. We perform immunohistochemistry on paraffin embedded brain. Of course, there is the massive over fixation. Also the tissue shrinks a lot during processing. > > > > In my last lab, after brain tissue had been fixed in formalin for 7 days we would store the tissue in 70% ethanol. I'd like to start storing the brain tissue I work with now in 70%, but I'm worried about the tissue possibly shinking too much, as the tissue already seems to shrink by greater than 60% after processing. > > > > (Previously, we would gradually increase the alcohols, 30% for 1 > > hour, then 50% for an hour in a bucket, room temperature on a rocker > > table, then put the cassettes into a bucket of 70% and store at room > > temp or 4C- does this process help any with controling the shrinkage > > factor?) > > > > Maybe this is a little bit long...thank you very much for your time. > > > > CM Bush > --------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Vinnie Della Speranza > Manager for Anatomic Pathology Services Medical University of South > Carolina > 165 Ashley Avenue Suite 309 > Charleston, SC 29425 > Ph: 843-792-6353 > fax: 843-792-8974 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Jun 1 16:40:56 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] resin embedded tissue and sirius red staining In-Reply-To: Message-ID: <200506012140.j51Leu1C018299@chip.viawest.net> I have done sirrus red staining on GMA sections with good results. How thick are your sections. I used 5 micron thick sections and stained with SR for 30 min. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Y. Wang Sent: Wednesday, June 01, 2005 2:29 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] resin embedded tissue and sirius red staining Dear histonetters, I have a question regarding the feasibility of staining resin embedded skin (in technovit) with Picosirius red (for collagen). We have some old samples embedded in resin that we would like to stain for collagen if at all possible. We normally do our sirius red staining on frozen and paraffin embedded tissue with good results. I tried running the same protocol (1hr sirius red followed by quick wash in acidified water) as well as at longer times (sirius red for 3 hours) with the resin sections but I get very dull staining (under polarized light I do not get the nice red/green/orange/yellow I normally see, instead it's rather murky). I haven't found anything online in terms of staining resin embedded samples with sirius red so I was wondering if this was one of those things that can't be done. Any comments would be much appreciated. Thank you Yak-Nam Wang Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From petepath <@t> yahoo.com Wed Jun 1 16:42:08 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Frozen TMA's Message-ID: <20050601214208.81166.qmail@web30411.mail.mud.yahoo.com> Tom, Had to run for a frozen. If you make frozen microarrays in my well bars by adhering the tissue directly to the well floor and then filling it with OCT, you will not have the problem of "curling away" of the small pieces because of crevices left between the cores and the OCT. As I said it will cut like any other frozen. I am happy to let users demo this technique and return the apparatus without obligation if not satisfied. Please visit my web site http://pathologyinnovations.com/index.html to learn more about my " Precision Cryoembedding system". Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From ynwang <@t> u.washington.edu Wed Jun 1 17:14:56 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] resin embedded tissue and sirius red staining In-Reply-To: <200506012140.j51Leu1C018299@chip.viawest.net> References: <200506012140.j51Leu1C018299@chip.viawest.net> Message-ID: Patsy, We cut our sections at 6 microns although I think our microtome cuts thicker than it says. Someone from the list also recommended incubating at 60 deg C before staining or using microwave heat. I'll try thinner sections and heat and let you know of the results (hopefully good). Thank you both for your suggestions. Yak-Nam > I have done sirrus red staining on GMA sections with good results. How > thick are your sections. I used 5 micron thick sections and stained with SR > for 30 min. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Y. Wang > Sent: Wednesday, June 01, 2005 2:29 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] resin embedded tissue and sirius red staining > > Dear histonetters, > > I have a question regarding the feasibility of staining resin embedded skin > (in technovit) with Picosirius red (for collagen). We have some old samples > embedded in resin that we would like to stain for collagen if at all > possible. We normally do our sirius red staining on frozen and paraffin > embedded tissue with good results. I tried running the same protocol (1hr > sirius red followed by quick wash in acidified water) as well as at longer > times (sirius red for 3 hours) with the resin sections but I get very dull > staining (under polarized light I do not get the nice > red/green/orange/yellow I normally see, instead it's rather murky). I > haven't found anything online in terms of staining resin embedded samples > with sirius red so I was wondering if this was one of those things that > can't be done. Any comments would be much appreciated. > > Thank you > Yak-Nam Wang > > Senior Fellow > Department of Bioengineering > University of Washington > Box 357962 > Seattle, WA 98195 > > Tel.: (206)-221-5873 > Fax.: (206)-221-5874 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From lyarrow <@t> shaw.ca Wed Jun 1 20:54:08 2005 From: lyarrow <@t> shaw.ca (lyarrow) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Fw: ethanol versus denatured alcohol Message-ID: <001401c56715$f71a5780$e8f79244@cg.shawcable.net> ----- Original Message ----- From: lyarrow To: histonet@list.utsouthwesternedu Sent: Wednesday, June 01, 2005 7:25 PM Subject: ethanol versus denatured alcohol Hello, At our Laboratory we are considering using denatured alcohol versus the present of ethanol. Has anyone experienced this reversal and what impact did it have on the quality and the follow-up work of Immuno and other tests? I would appreciate the comments because the price difference is a real saving. Louise Yarrow TechII Histology Foothills Medical Centre Calgary Lab Services louise.yarrow2@cls.ab.ca From Kemlo.Rogerson <@t> elht.nhs.uk Thu Jun 2 02:23:53 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] filtering hematoxylin Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD417@elht-exch1.xelht.nhs.uk> Interesting question. Do you filter to remove haematoxylin/ mordant precipitant or cells/ tissue? As a recovering Cytologist I would suggest you filter at least daily, if not 2x daily, to remove cells/ tissue that can cause 'carry over'. Cells from 'fatty organs' such as breast or small celled undiff., from my experience can do this damage. It's easier to filter than for the 'sword of Damocles' to hang over the Patient. -----Original Message----- From: Diana McCaig [mailto:dmccaig@ckha.on.ca] Sent: 01 June 2005 17:19 To: Histonet (E-mail) Subject: [Histonet] filtering hematoxylin How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Thu Jun 2 02:25:25 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Fw: ethanol versus denatured alcohol Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD21E@elht-exch1.xelht.nhs.uk> Sometimes I have noticed a haziness with 'denatured' alcohol but then I tend to look at nuclei. -----Original Message----- From: lyarrow [mailto:lyarrow@shaw.ca] Sent: 02 June 2005 02:54 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fw: ethanol versus denatured alcohol ----- Original Message ----- From: lyarrow To: histonet@list.utsouthwesternedu Sent: Wednesday, June 01, 2005 7:25 PM Subject: ethanol versus denatured alcohol Hello, At our Laboratory we are considering using denatured alcohol versus the present of ethanol. Has anyone experienced this reversal and what impact did it have on the quality and the follow-up work of Immuno and other tests? I would appreciate the comments because the price difference is a real saving. Louise Yarrow TechII Histology Foothills Medical Centre Calgary Lab Services louise.yarrow2@cls.ab.ca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Thu Jun 2 06:00:29 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] cryostats Message-ID: I have the Leica and it has worked very well for me. Betsy Molinari HT(ASCP) Texas heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Clarke, Mary Sent: Wednesday, June 01, 2005 1:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cryostats We are in the process of purchasing a new cryostat. Has anyone used the Richard-Allan 525 or the Leica CM 1850? We are looking at both and wonder how people like or dislike them? Thanks in advance for your help. Terri Clarke Histology Supervisor All Saints Laboratory 3801 Spring Street Racine, Wi 53405 mclarke@allsaintshealthcare.org 262-687-6609 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, June 01, 2005 12:01 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 19, Issue 1 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. Speaking of Gram's... (Breeden, Sara) 2. Freezing muscle biopsies (Osborn, Sharon) 3. Enzyme Staining and Billing for Controls (Mitchell, Guy) 4. RE: Number of Blocks Submitted by PA (Charles.Embrey) 5. Aquaporin immunohistochemistry (Anthony Reilly) 6. Ki67. (christian@eisbo.dk) 7. Ki67. (christian@eisbo.dk) 8. RE: Ki67. (Bruijntjes, J.P.) 9. visualization of muscle damage with light microscopy ? (Vincent Martin) 10. Histology of Frozen Tissue (shawnster73@aol.com) 11. BK Virus (Bobbie Boyce) 12. HPV I and II Control Blocks (WAYNE HOLLAND) 13. Re: mouse endothelial cells in brain (Corazon D. Bucana) 14. staining with Solvent Blue 38 (Donin, Nick (NIH/NCI)) 15. RE: mouse endothelial cells in brain (Luis Chiriboga) 16. Storage of Frozen Section slides (CHRISTIE GOWAN) 17. Re: Will brain tissue shrink even more if storedin70%alcoholprior to processing? (Vinnie Della Speranza) 18. RE: mouse endothelial cells in brain (Connolly, Brett M) 19. SOLVENT BLUE 38 ON PARAFFIN EMBEDDED MOUSE BRAIN (Tim Wheelock) 20. filtering hematoxylin (Diana McCaig) 21. RE: filtering hematoxylin (Bartlett, Jeanine) ---------------------------------------------------------------------- Message: 1 Date: Tue, 31 May 2005 12:33:01 -0600 From: "Breeden, Sara" Subject: [Histonet] Speaking of Gram's... To: Message-ID: Content-Type: text/plain; charset="us-ascii" I'm having trouble with my Gram's in that my positive bacteria are losing color. So, I'm wondering what the exact differentiation time should be for the step after the Gram's Iodine (I'm using the Sigma kit)? The instructions are not precise, so I'm thinkin' maybe I'm over differentiating or under-Safranin-ing or aliens (this is the State that's home to ROSWELL...) have taken over my kit. Anyone have anything a bit more specific? I'd appreciate any help... Sara A. Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX ------------------------------ Message: 2 Date: Tue, 31 May 2005 15:41:02 -0400 From: "Osborn, Sharon" Subject: [Histonet] Freezing muscle biopsies To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <29B25753F6B1D51196110002A589D44402397FB8@PALMSG30.us.schp.com> Content-Type: text/plain Liz, Our researchers often freeze tissue for later study. They use the plastic disposable embedding molds. These can be used with the dry ice as John Kiernan suggested. There is a thin layer of OCT placed in bottom of mold then the tissue is oriented in this with additional OCT to fill the mold. The mold can be labeled with a permanent felt tip pen. Each specimen is then placed into the labeled sterile sampling bags, packed in dry ice and shipped to respective location. We have experienced very little distortion of the tissue due to ice crystals, etc. The testing has worked out okay. The main tissues were liver, kidney, lung. You may wish to experiment with some tissues before actually tackling the research samples to get the right combination for you. Sharon Osborn DNAX ScheringPlough Biopharma Palo Alto, CA Hello Histonetters > > I have a unique question. We are currently starting to set up > procedure for collecting samples from a clinical trial. The clinical > trial involves taking multiple synovial biopsies at a surgery center. > Since portions of the samples need to be processed for frozen sections > we wanted to be able to freeze the specimens at the surgery center via > isopentane cooled liquid nitrogen. We really do not want to have to > transport the multiple specimens back to the main lab prior to > freezing due to the time involved it would probably be 1-2 hours post > biopsy before we could freeze the samples. The surgery center is > questioning the flammability of the isopentane. Has anyone > encountered anything like this? Any suggestions would be helpful. > Thanks in advance. > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > Manager > Premier Laboratory, LLC > P.O. Box 18592 > Boulder, Colorado 80308 > Office: (303) 735-5001 > Fax: (303) 735-3540 > liz@premierlab.com > www.premierlab.com > > Ship to Address: > Premier Laboratory > University of Colorado > MCDB, Room A3B40 > Boulder, Colorado 80309 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 3 Date: Tue, 31 May 2005 15:42:59 -0400 From: "Mitchell, Guy" Subject: [Histonet] Enzyme Staining and Billing for Controls To: "HistoNet Server" Message-ID: <4DCF5CA56B4AA04ABC383DE358BAD3820289100D@dcr-xchg-04.carolinas.org> Content-Type: text/plain; charset="US-ASCII" A question has come up concerning charges for muscle enzyme stains. We stain for Adenylate Deaminase and Phosphofructokinase as well as negatives for each. I assume they should be treated as controls without a charge but others disagree. Guy W. Mitchell (704) 379-5984 (704) 379-6144 Fax ----------------------------------------- This electronic message may contain information that is confidential and/or legally privileged. It is intended only for the use of the individual(s) and entity named as recipients in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from any computer. Do not deliver, distribute or copy this message, and do not disclose its contents or take any action in reliance on the information it contains. Thank you. ------------------------------ Message: 4 Date: Tue, 31 May 2005 15:42:58 -0500 From: "Charles.Embrey" Subject: RE: [Histonet] Number of Blocks Submitted by PA To: "Smith Wanda" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" I have been gone for a week and just got a chance to check my e-mails and found this question interesting. First, have you asked the PA? I know it seems like a simple question but you would be surprised at all the misunderstanding that can be cleared up with a simple question asked? Please know that I have NEVER met a pathologist that will gladly read more slides than he thinks are necessary for a good diagnosis on a routine basis. I gross for seven pathologists and it is tricky to satisfy all at the same time. Luckily, my pathologists and I here have great communication and we manage to keep the block count, as well a gross recut number down. As far as "rules of thumb" we do have certain guidelines that are basically thought of as "standards of care". Without knowing other details the best advice I could give is to ask the PA. Charles Embrey PA(ASCP) Urbana IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Smith Wanda Sent: Tuesday, May 24, 2005 7:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Number of Blocks Submitted by PA Good Morning Histonetters, I have a question for you this morning...Are there written standards or "Rules of Thumb" regarding number of blocks that should be submitted on certain tissues? The reason I ask is, "it seems to me" that sometimes the PA submits an extreme number of blocks on some specimens that could be accomplished with less. I'm not advocating cutting corners or not performing good medical practice, I just need to know. An example would be a uterus w/ fibroids, tubes and ovaries which we have gotten up to 24 blocks on. I'm not a PA, but it's interesting to me that when the Pathologist cut, we get alot less blocks. Please advise. Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Wed, 01 Jun 2005 14:13:15 +1000 From: "Anthony Reilly" Subject: [Histonet] Aquaporin immunohistochemistry To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hello All I have been requested to be involved in a research study demonstrating Aquaporin 1 (AQ1, AQP1) using IHC. To date I have found 2 antibodies available from Chemicon. My questions are: 1. Are there other commercial suppliers of this Ab. 2. If not, which of the 2 from Chemicon is the preferred option. 3. Do you realyy need to fix with paraformaldehyde as stated in most methods, and why? i would appreciate any advice forwarded. regards Tony Reilly Chief Scientist Anatomical Pathology Northside Pathology Prince Charles Hospital Rode rd Chermside Q 4032 Australia Ph: 07 3350 8543 Fax: 07 33508546 tony_reilly@health.qld.gov.au *********************************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is prohibited. It may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone or by return email. You should also delete this email and destroy any hard copies produced. *********************************************************************************** ------------------------------ Message: 6 Date: Wed, 01 Jun 2005 09:48:19 +0200 From: "christian@eisbo.dk" Subject: [Histonet] Ki67. To: histonet@lists.utsouthwestern.edu Message-ID: <758a26d4ec47df2ab23acf9597f0b69d@eisbo.dk> Content-Type: text/plain; charset=windows-1252 ------------------------------ Message: 7 Date: Wed, 01 Jun 2005 09:50:45 +0200 From: "christian@eisbo.dk" Subject: [Histonet] Ki67. To: histonet@lists.utsouthwestern.edu Message-ID: <634ceeb7bf3c9de7df2bf51acb622dfe@eisbo.dk> Content-Type: text/plain; charset=windows-1252 Hey users of histonet. I am trying to stain kidneys for ki67! The sections have been in alcohol for 1 month and are now in parafin. The antibody I am using should be good enough. Does anyone have experience with this??? best regards. Christian EIsbo. ------------------------------ Message: 8 Date: Wed, 1 Jun 2005 10:29:19 +0200 From: "Bruijntjes, J.P." Subject: RE: [Histonet] Ki67. To: Cc: Histonet@lists.utsouthwestern.edu Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079DC6@ntexch1.voeding.tno.nl> Content-Type: text/plain; charset="us-ascii" Hi Christian Yes I do. May be it depends on the antibody you use. We use a rabbit monoclonal ki67 from Neomarkers (Labvision). Tissues fixed in alcohol don't react with the antibody using HIER in citrate buffer (pH 6,0). At least it didn't in my lab. It does when you use HIER in EDTA (pH 8.0) and in TRIS (pH 10). Hopes that will help you Joost Bruijntjes TNO Quality of Life Zeist Holland -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of christian@eisbo.dk Sent: woensdag 1 juni 2005 9:51 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Ki67. Hey users of histonet. I am trying to stain kidneys for ki67! The sections have been in alcohol for 1 month and are now in parafin. The antibody I am using should be good enough. Does anyone have experience with this??? best regards. Christian EIsbo. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html ------------------------------ Message: 9 Date: Wed, 1 Jun 2005 10:43:57 +0200 From: "Vincent Martin" Subject: [Histonet] visualization of muscle damage with light microscopy ? To: Message-ID: <002701c56686$0d65e6c0$0a4a7c8b@physiologie8> Content-Type: text/plain; charset="iso-8859-1" Dear list members, I would like to observe the effect of lengthening contractions on ultrastructural properties of rat tibialis anterior muscle. Is is possible to observe ultrastructural abnormalities with light microscopy with a x100 maginification ? If not, which are the minimal requirements to observe such abnormalities ? Thanks for your replies Best regards, Vincent Martin UPRES-EA 3285 D?terminants Physiologiques de l'Activit? Physique Facult? des Sciences du Sport de Marseille Universit? de la M?diterran?e BP910 - 163, avenue de Luminy 13288 Marseille cedex 09 France Phone : +33 (0)4-91-82-83-78 Fax : +33 (0)4-91-82-83-75 vincent.martin@staps.univ-mrs.fr www.physiologie.staps.univ-mrs.fr ------------------------------ Message: 10 Date: Wed, 01 Jun 2005 07:05:40 -0400 From: shawnster73@aol.com Subject: [Histonet] Histology of Frozen Tissue To: histonet@lists.utsouthwestern.edu Message-ID: <8C734B0FB191BFF-620-23897@FWM-D40.sysops.aol.com> Content-Type: text/plain; charset="us-ascii" What would be the histological effect of allowing frozen OCT embedded tissue to come up to a temperature of 0 or -5oC?? ------------------------------ Message: 11 Date: Wed, 1 Jun 2005 08:08:46 -0400 From: "Bobbie Boyce" Subject: [Histonet] BK Virus To: "HistoNet Server" Message-ID: <6E41111281623B4B8A9AB8F9A7EA343737BFB5@wlmmsx02.nemours.org> Content-Type: text/plain; charset=iso-8859-1 I'm having problems locating a FFPE control for BKV. Can anyone tell me where I can get them? Bobbie Boyce duPont Hospital for Children Wilmington, DE ------------------------------ Message: 12 Date: Wed, 01 Jun 2005 08:10:12 -0400 From: "WAYNE HOLLAND" Subject: [Histonet] HPV I and II Control Blocks To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hello All, Help, can anyone give assistance to getting some blocks for HPV I and HPV II for controls. We are going to start In-Situ, we are in the testing phase. One block from and HPV I (LOW) and one block from HPV II (HIGH) would be extremely helpful. We would be willing to trade, Thanks in advance. ------------------------------ Message: 13 Date: Wed, 01 Jun 2005 08:44:09 -0500 From: "Corazon D. Bucana" Subject: Re: [Histonet] mouse endothelial cells in brain To: "Donin, Nick (NIH/NCI)" Cc: "'histonet@lists.utsouthwestern.edu'" Message-ID: <5.1.1.6.0.20050601084009.042c9c48@audumla.mdacc.tmc.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Wed, 1 Jun 2005 10:08:33 -0400 From: "Donin, Nick (NIH/NCI)" Subject: [Histonet] staining with Solvent Blue 38 To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <16A0583FB1644E4DB8C0A0265028B6FD02B4E739@nihexchange13.nih.gov> Content-Type: text/plain Histonetters, I'm trying to stain the corpus callosum of formalin fixed paraffin embedded mouse brains using Solvent Blue 38. Could someone suggest a successful protocol? Thanks. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 ------------------------------ Message: 15 Date: Wed, 01 Jun 2005 11:23:20 -0400 From: Luis Chiriboga Subject: RE: [Histonet] mouse endothelial cells in brain To: "Corazon D. Bucana" , "Donin, Nick (NIH/NCI)" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii I have also used the pharmingen cd31 by frozen in mouse brain, muscle, and skin. It works extremely well in fresh system. FFPE on the other hand is much trickier. It works but very inconsistently, fixation and tissue processing are the key variables. Once you have obtained staining in FFPE, you need to follow the same sacrificing (perfusion etc..) grossing, processing procedure with "religious fanaticism" (no offense to anyone.....) L ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Corazon D. Bucana Sent: Wednesday, June 01, 2005 9:44 AM To: Donin, Nick (NIH/NCI) Cc: 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] mouse endothelial cells in brain The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Wed, 01 Jun 2005 15:24:32 +0000 From: "CHRISTIE GOWAN" Subject: [Histonet] Storage of Frozen Section slides To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed I know this subject has been tossed around before, but I wonder if Patsy Ruegg or Chris Van der Loos or anyone else for that matter would share with me their protocol for storing frozen slides. The company I work for sells frozen TMA slides. We cut sections as requested by clients and mail them out on dry ice. We have found that after going into the block 10-12 times we start to see artifact. Pre-cutting slides would be much better for the life of the block. Has anyone tried storing the slides in a vacuum sealed (food saver) bag? Any help would be appreciated. Thanks, Christie Gowan HT (ASCP) ------------------------------ Message: 17 Date: Wed, 01 Jun 2005 11:18:17 -0400 From: "Vinnie Della Speranza" Subject: Re: [Histonet] Will brain tissue shrink even more if storedin70%alcoholprior to processing? To: Cc: jchladny@cvm.uiuc.edu, histonet@lists.utsouthwestern.edu, clarissabush@sbcglobal.net Message-ID: Content-Type: text/plain; charset=US-ASCII I hadn't considered the possibility of buffer salt crystals forming the vacuoles observed by the author and I don't know if she is on this list and would care to comment but this is an interesting premise worth investigating. I would think that a thorough water wash prior to going into alcohol would help to clear this up. thanks for the feedback. Vinnie Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> John Kiernan 05/27/05 04:27PM >>> In the HistoLogic May 2000 paper the duration of the initial formaldehyde fixation is not stated; only that the brains were "previously well fixed in neutral buffered formalin." Frequently specimens are not fixed for long enough to make them resistant to bad effects of solvents etc. It's also seems from the paper that the test pieces of brain were passed directly from the buffered fixative into 70% alcohol. This causes precipitation of sodium phosphate in the tissue (see Freida Carson 1996 "Histotechnology" 2nd edn, p.27). The control specimens in the HistoLogic paper were passed from buffered formalin to 60% ethanol (which safely extracts the phosphate buffer salts). Could the holes in the white matter be made by buffer salt crystals rather than forming slowly during storage in 70% ethanol? Just a thought! -- ------------------------------- 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 _______________________________ Vinnie Della Speranza wrote: > > You may wish to look at an article authored by Jane Chladny, published in HistoLogic, May 2000, entitled "Artifact in Tissues Held in 70% Ethanol" > > the author reported the appearance of 'vacuole' artifact in nervous tissues observed in a variety of mammalian tissues after storage in 70% ethanol. > > you can access the article by selecting the link for HistoLogic at www.sakuraus.com > > CM Bush wrote: > > > > Dear Histonet, > > > > Hello, here is my first post to the list, thank you in advance for your help. > > > > Summary: > > Having problems with mouse and human brain tissue shrinkage durning processing, but would like to better preserve antigens, concerned storage of tissue 70% ethanol will shrink tissue even further: > > > > We have lots of brain specimens, stored in formalin for a long time (months up to 10 years), both human and mouse. We perform immunohistochemistry on paraffin embedded brain. Of course, there is the massive over fixation. Also the tissue shrinks a lot during processing. > > > > In my last lab, after brain tissue had been fixed in formalin for 7 days we would store the tissue in 70% ethanol. I'd like to start storing the brain tissue I work with now in 70%, but I'm worried about the tissue possibly shinking too much, as the tissue already seems to shrink by greater than 60% after processing. > > > > (Previously, we would gradually increase the alcohols, 30% for 1 hour, then 50% for an hour in a bucket, room temperature on a rocker table, then put the cassettes into a bucket of 70% and store at room temp or 4C- does this process help any with controling the shrinkage factor?) > > > > Maybe this is a little bit long...thank you very much for your time. > > > > CM Bush > --------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Vinnie Della Speranza > Manager for Anatomic Pathology Services > Medical University of South Carolina > 165 Ashley Avenue Suite 309 > Charleston, SC 29425 > Ph: 843-792-6353 > fax: 843-792-8974 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Wed, 1 Jun 2005 11:35:22 -0400 From: "Connolly, Brett M" Subject: RE: [Histonet] mouse endothelial cells in brain To: "'Corazon D. Bucana'" , "Donin, Nick (NIH/NCI)" Cc: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Nick, Fix the brains in Pharmingen Zinc Tris, process to paraffin and use the Pharmingen antibody. Works like a charm. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Corazon D. Bucana Sent: Wednesday, June 01, 2005 9:44 AM To: Donin, Nick (NIH/NCI) Cc: 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] mouse endothelial cells in brain The only time we got good staining is on frozen sections using anti-mouse CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB or AEC staining. At 11:54 AM 5/31/2005 -0400, you wrote: >Histonetters, > >This is my first posting, so I'm excited that someone out there might be >able to help me. > > >I am trying to stain endothelial cells in brain tumors in formalin fixed >paraffin embedded mouse tissues. I have tried several antibodies, and none >of them seem to work. I am consistently getting high background and no true >staining. I have used the following antibodies: > > > >BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 > >Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this >works, but my protocol didn't produce good results) > >Cedarlane mouse CD31 cat#CL8930 AP > >Cymbus Biotechnology anti-ms CD31 cat# CBL1337 > > > >The protocol I use is shown below. > > > > > >I'm confident that out of these antibodies, at least one will be able to >stain the endothelial cells in these tumors, but I think that my protocol >may be the problem. Could someone possibly suggest and antibody or protocol >that would work for me? Thanks so much for your help, much appreciated. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ ------------------------------ Message: 19 Date: Wed, 01 Jun 2005 12:19:17 -0400 From: Tim Wheelock Subject: [Histonet] SOLVENT BLUE 38 ON PARAFFIN EMBEDDED MOUSE BRAIN To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <429DE005.1050905@mclean.harvard.edu> Content-Type: text/plain; charset=windows-1252; format=flowed Hi Nick: I use Solvent Blue 38 (also known as Luxol Fast Blue) on formalin-fixed, paraffin embedded human brain, but have also seen it work well on mouse brain. The protocol is as follows. This particular recipe combines the myelin stain with an H+E, but it can be combined with a Nissl stain instead or simply by itself. Hope this helps. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 * LUXOL FAST BLUE**-HEMATOXYLIN-EOSIN (LHE) STAIN* * * 1. De-paraffinize 5 micron formalin-fixed sections and take them down to 95% ethanol. 2. Stain in Luxol Fast Blue solution for 2 hours at 60C. 3. Remove and allow to cool for 15 minutes. 4. Remove excess stain in 95% ethanol for 1 minute. 5. Wash in several changes of tap water. 6. Differentiate sections with 2 dips in the reducer solution, then *immediately* through 4 changes of tap water (10 dips each), then one more change of tap water. 7. Immerse sections in Gill 3 Hematoxylin for 10 minutes. 8. Rinse in several changes of tap water. 9. Differentiate sections in acid alcohol (7 dips). 10. Rinse in 4 changes of tap water (10 dips each) then one more change of tap water. 11. Blue sections in Scott's Tap Water Substitute for 30 seconds. 12. Wash in 4 changes of tap water for 2 minutes each. 13. Immerse sections in 95% ethanol for 1 minute. 14. Immerse slides in alcoholic Eosin Y for 3 minute. 15. Differentiate in 95% ethanol (4 dips). 16 Dehydrate in first absolute ethanol for 20 dips. 17. Dehydrate in second absolute ethanol for 1 minute, and then clear, and mount. *SOLUTIONS*: Luxol Fast Blue: Luxol Fast Blue...............................................0.1 gram. 10% glacial acetic acid .......................................1 mls. 95% ethanol........................................................100mls. Reducer: Hydroquinone....................................................1 gram. Sodium Sulfite...................................................5 gram. Distilled water..................................................100 mls. Hematoxylin Differentiator: 1% Hydrochloric acid in 70% ethanol *RESULTS*: 1. Myelin: The myelin should be stained blue or greenish blue 2. Nuclei: The nuclear membrane should be sharply delineated in blue-black with the nucleoplasm clear except for the chromatin, also being blue-black. 3. Cytoplasm and background should be varying shades of red. NOTE 1. The thickness of the section, the type of hematoxylin, and all the times in hematoxylin, eosin, and dehydrating depend upon your requirements. 2. If the section thickness is more than 10 microns, you may want to use 3 or 4 dips in the Reducer, so as to avoid background myelin staining . 3. The so called "Reducer" (because it is used as a reducer in the Bodian silver protein stain) actually functions as a differentiator for the myelin stain. ------------------------------ Message: 20 Date: Wed, 1 Jun 2005 12:18:44 -0400 From: Diana McCaig Subject: [Histonet] filtering hematoxylin To: "Histonet (E-mail)" Message-ID: <3E5A3F039F0BD8118B4700C00D0020240435BF@CKHA9> Content-Type: text/plain How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? ------------------------------ Message: 21 Date: Wed, 1 Jun 2005 12:33:58 -0400 From: "Bartlett, Jeanine" Subject: RE: [Histonet] filtering hematoxylin To: "Diana McCaig" , "Histonet \(E-mail\)" Message-ID: Content-Type: text/plain; charset="us-ascii" We don't filter the hematoxylin we get from Richard-Allan. We replace it twice a month, but we have a light volume of H&E's. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 (404) 639-3590 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: Wednesday, June 01, 2005 12:19 PM To: Histonet (E-mail) Subject: [Histonet] filtering hematoxylin How often do you filter commercially prepared Harris Hematoxylin (used on autostainer)? _______________________________________________ Histonet mailing list Histonet@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 19, Issue 1 *************************************** Privileged/Confidential information may be contained in this message. 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Privileged/Confidential information may be contained in this message. The information contained in this message is intended only for the use of the recipient(s) named above and their co-workers who are working on the same matter. The recipient of this information is prohibited from disclosing the information to any other party unless this disclosure has been authorized in advance. If you are not intended recipient of this message or any agent responsible for delivery of the message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or action taken in reliance on the contents of this message is strictly prohibited. You should immediately destroy this message and kindly notify the sender by reply E-Mail. Please advise immediately if you or your employer does not consent to Internet E-Mail for messages of this kind. Opinions, conclusions and other information in this message that do not relate to the official business of the firm shall be understood as neither given nor endorsed by it. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Susan.Walzer <@t> HCAHealthcare.com Thu Jun 2 07:26:52 2005 From: Susan.Walzer <@t> HCAHealthcare.com (Walzer Susan) Date: Fri Sep 16 15:25:10 2005 Subject: **SPAM** [Histonet] gram stain control Message-ID: <471953BC63077941B82C26A4338272B42F0475@ORLEV03.hca.corpad.net> We use fresh tissue (umbilical cord works well)and give 2 pieces to micro to incubate, one in gram +, one in gram-. After a couple of days we retrieve it and put in formalin, process and embed a piece of each in a block. Works great. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Steven Coakley Sent: Friday, May 27, 2005 3:32 PM To: Histonet@lists.utsouthwestern.edu Subject: **SPAM** [Histonet] gram stain control I'm looking for a good gram stain control with both gram +/-. Any good suppliers out there. Steve --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Thu Jun 2 08:02:04 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] gram control Message-ID: <20050602130204.64758.qmail@web90204.mail.scd.yahoo.com> Any chance of picking up a used blk? Steve --------------------------------- Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. From susan.wells <@t> bms.com Thu Jun 2 08:02:16 2005 From: susan.wells <@t> bms.com (Susan Q Wells) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Cryostat Frozen Sectioning Aid Message-ID: <429F0358.6000103@bms.com> Anyone know what the shelf life of the adhesive coated slides for the Instrumedics system is? Thanks in advance, Susan Wells From cwscouten <@t> myneurolab.com Thu Jun 2 09:26:28 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] frozen sections Message-ID: <5784D843593D874C93E9BADCB87342AB44F9F8@tpiserver03.Coretech-holdings.com> See the following link: http://www.myneurolab.com/global/Manuals/Tips%20and%20Techniques%20Freezing%20Artifact.pdf 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 Emerson, Rachael Sent: Friday, May 27, 2005 2:51 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] frozen sections Hello. I am in need of some help with trying to freeze mouse embryos for frozen sectioning. I am dissecting mouse embryos out in PBS (Embryonic day 8.5-14.5) and freezing them in VWR's Neg 50 Embedding Media. Initially I put a small amount of Neg 50 in the bottom of a Polyscience Plastic Peel-Away Mold, oriented the embryo, and then added more Neg 50. I then put the molds in the -80 Freezer and let them solidify. They seem to cut OK, but the morphology was terrible. Not so much in the E14.5, but other embryos looked very degraded. Next, I tried the same procedure but froze them by floating the mold in ethanol with dry ice. Still the morphology was terrible. I tried to freeze the embryos directly and then embed them, but they just turned to mush. On my final attempt I repeated the same procedure, but froze the mold by holding the bottom in liquid nitrogen. The block froze in seconds, but when I took them it out of the mold it was cracked and very hard to cut. The few sections I did manage were terrible and you could see the ice crystals in the embryos. I would really appreciate any advice or suggestions you have to offer. Thank you Rachael Emerson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From shive003 <@t> umn.edu Thu Jun 2 10:44:18 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Pig C-Reactive Protein IHC Message-ID: <003f01c56789$f0976000$41065486@auxs.umn.edu> Does anyone do IHC for Pig C-Reactive Protein? If so, could you please let me know who your supplier is? Thanks. Jan Shivers U of MN Vet Diag Lab From funderwood <@t> mcohio.org Thu Jun 2 10:58:08 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Stain Racks Message-ID: Hi Cindy, Cardinal Health carries them. The catalog number is s7640 15. Their phone number is 1.888.444.5440. Price is $216 for a package of 10. Fred >>> Cindy DuBois 05/31/05 10:19AM >>> Does anyone know where I can purchase slide racks for the TissueTek DRS ? We currently run about 250 slides a day and only have 6 racks (one which is broken). Sakura is not currently manufacturing them due to a production problem. Any suggestions is greatly appreciated, Cindy Dubois Delta Pathology Assoc. __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> instrumedics.com Thu Jun 2 11:23:53 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:25:10 2005 Subject: Fw: [Histonet] Cryostat Frozen Sectioning Aid Message-ID: <01c501c5678f$7caddb00$6401a8c0@INSTRUMEDICS22> ----- Original Message ----- From: "Instrumedics" To: "Susan Q Wells" Sent: Thursday, June 02, 2005 12:15 PM Subject: Re: [Histonet] Cryostat Frozen Sectioning Aid > Susan > The shelf life of the adhesive coated slides is years as long as the > slides > are not exposed to UV light. > > Bernice > Instrumedics > > > ----- Original Message ----- > From: "Susan Q Wells" > To: > Sent: Thursday, June 02, 2005 9:02 AM > Subject: [Histonet] Cryostat Frozen Sectioning Aid > > >> Anyone know what the shelf life of the adhesive coated slides for the >> Instrumedics system is? >> Thanks in advance, >> Susan Wells >> >> >> > From Nancy.Lowen <@t> med.va.gov Thu Jun 2 11:33:54 2005 From: Nancy.Lowen <@t> med.va.gov (Lowen, Nancy) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Info on Cryojane Message-ID: If anyone is using the Cryojane tape transfer system for cutting undecalcified bone for frozen sections, we would appreciate your imput. What temperature do you cut bone at and does the tape transfer the complete section of bone to the slide? We are getting shattering of the bone on most sections and are really having some problems with the bubbles not rolling out. We are still in the testing process, but we would love to have any tips from anyone who has more experience on how to obtain the best sections. Should we expect to see totally intact sections when cutting bone? Also, what freezing method have you found to be the best..Thanks in advance Nancy From pex0220 <@t> yahoo.com.cn Thu Jun 2 11:46:45 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] different antigens need different methods for antigen retrieval? Message-ID: <20050602164645.15307.qmail@web15502.mail.cnb.yahoo.com> Hello, all, I feel confused for immunofluorescence. I am doing immunofluorescence in vertebra sections. I use the same protocol for immunostaining, but the results are very different, the one is very nice, but the other is weak, I do not know the reasons. Do different antigens need different methods for antigen retrieval? Generally, which methods for antigen retrieval in bone sections are better? (Enzyme digestion? or HIER? or both?) Any suggestions will be helpful for me! Thank you! --------------------------------- DO YOU YAHOO!? ÑÅ»¢Ãâ·ÑGÓÊÏ䣭ÖйúµÚÒ»¾øÎÞÀ¬»øÓʼþɧÈų¬´óÓÊÏä From MadaryJ <@t> MedImmune.com Thu Jun 2 11:47:00 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Congo Red/Toludine Blue for Eosinaphils and Mast Message-ID: <746FDB897740814EA52BDDCB5ED1DDBCBB1583@medimmune4.medimmune.com> Anyone hear of a combo CR/TB for Eo's and Mast Cells? From petepath <@t> yahoo.com Thu Jun 2 12:17:59 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Cutting undecalcified bone Message-ID: <20050602171759.81401.qmail@web30415.mail.mud.yahoo.com> Nancy, I have experience cutting frozens on undecacified bone specimens for routine surg path cases using conventional technique but have no experience with the tape transfer system. I routinely use disposible low profile blades which in not optimum for bone. I imagine a stronger high profile blade or knife will work better. My success has varied with the hardness of the bone. Trebecular bone coming from older patients can cut fairly easily porvided the blade is sharp, and the section is taken with a fluid continuous motion. The result is a surprisingly intact section. The harder the bone ( typically the younger the patient ) there will be more damage to the blade in each pass. The sections will be streaked and splitting requiring one or more blade changes by time I get a resonable section. It is quite a juggling act. Cutting hard cortical bone is often very frustrating and very difficult to get any resonable section. From what you describe as bubbles, I am guessing you are getting sections of bone thicker than you are hoping for and as a result when you roll it out, the thicker bone pieces are protecting the rest of the section from rolling. Often the first section of any tissue will be considerably thicker than those that follow after a few fluid turns of the wheel. If you are not letting a few pass your first section may be considerably thicker and as a result will shatter more and create thicker trabeculae which may be leading to your problem. The equivalent using conventional technique will give me a section that my coverslip will not lie flat against the slide and my mounting medium will not spread. I look foreward to other peoples experience on this subject. Stephen From liz <@t> premierlab.com Thu Jun 2 12:22:40 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Info on Cryojane In-Reply-To: Message-ID: <000101c56797$b0d3a470$a7d48a80@AMY> Nancy John Tarpley wrote a nice article in the Journal of Histotechnology about undecalcifed bone sections and the tape transfer system. I have minimal experience but I found that the coated slides with the least amount of adhesive worked the best (the 1X). I would only expose the slides once to the UV light and gently remove the tape starting at one corner. Patsy helped me out when I was first doing this. I remember her setting the angle of the knife holder different and I can't recall what that was or what the temp was. She will probably respond. When I was cutting human cortical bone I did get a few fracture marks, but I think that this artifact is difficult to remove. A nice sharp tungsten carbide knife works also and when sectioning an even smooth motion is helpful. Good Luck Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lowen, Nancy Sent: Thursday, June 02, 2005 9:34 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Info on Cryojane If anyone is using the Cryojane tape transfer system for cutting undecalcified bone for frozen sections, we would appreciate your imput. What temperature do you cut bone at and does the tape transfer the complete section of bone to the slide? We are getting shattering of the bone on most sections and are really having some problems with the bubbles not rolling out. We are still in the testing process, but we would love to have any tips from anyone who has more experience on how to obtain the best sections. Should we expect to see totally intact sections when cutting bone? Also, what freezing method have you found to be the best..Thanks in advance Nancy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lfidgen <@t> vt.edu Thu Jun 2 12:42:52 2005 From: lfidgen <@t> vt.edu (Laura Fidgen) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Ventana Benchmark Message-ID: <6.0.0.22.0.20050602133527.0250bbe8@pop.vt.edu> We are a veterinary teaching hospital currently evaluating a Ventana Benchmark LT. I have a few questions: 1. Do protocols for antibodies vary among different animal species? 2. For cytology specimens (FNA or wet preps), where did you get your protocols? 3. For cytology specimens (fna or wet preps), what fixation, if any, do these slides need? Thank you in advance to those that respond. Have a great day! Laura L. Fidgen, MScF, BSc, HT(ASCP) Laboratory and Research Practioner VMRCVM, Histopatholgy/Clinical Pathology Dept. Blacksburg, VA 24060-0443 From JEllin <@t> yumaregional.org Thu Jun 2 13:32:48 2005 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Refence Labs Message-ID: I was wondering how many Hospitals have lost revenue or specimens to refence labs because of turn around time and reports? To be specific an outreach program. Also what are you doing within your facility to stop this issue from occuring? Jesus Ellin Yuma Regional Medical Center From sbreeden <@t> nmda.nmsu.edu Thu Jun 2 14:18:33 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Automatic Coverslipper Comparison Message-ID: If you were to purchase an automatic coverslipper today, which one would you buy? If you already have one, did you buy a glass coverslipper - and, if you did, do you like it? If you aren't happy, why? Did you buy a tape coverslipper - and, if you did, do you like it? If you aren't happy, why? If you'd care to cite a specific brand but have some not-so-positive remarks, you may email me directly. I'm appreciating your comments and my coverslip-ragged fingertips thank you! Sara A. Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From MadaryJ <@t> MedImmune.com Thu Jun 2 14:23:02 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Summer Job at Medimmune Message-ID: <746FDB897740814EA52BDDCB5ED1DDBCBB1585@medimmune4.medimmune.com> We are looking for a person to work this summer who is willing to forego benefits, just straight pay for histology work. We prefer a person who can cut good frozens and do IHC. For the right person we could train, but of course at less pay! Even someone really good at paraffin histology who needs no training. A retired person might be perfect for this job, as it is temporary. Email me or call Nick Madary, HT/HTL(ASCP)QIHC Histology Manager, Medimmune 3013986113/4745 From Janet.Bonner <@t> FLHOSP.ORG Thu Jun 2 15:11:55 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Automatic Coverslipper Comparison Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42AA@fh2k093.fhmis.net> We are very happy with our Leica cv5030. Under no circumstances would we buy a "tape" coverslipper - after five years the tape is lifting on all of our slides and the tissue section comes off with the tape! We do at least 300,000 slides per year so you can imagine the mess that we have. Good luck in your search! -Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Breeden, Sara Sent: Thursday, June 02, 2005 3:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automatic Coverslipper Comparison If you were to purchase an automatic coverslipper today, which one would you buy? If you already have one, did you buy a glass coverslipper - and, if you did, do you like it? If you aren't happy, why? Did you buy a tape coverslipper - and, if you did, do you like it? If you aren't happy, why? If you'd care to cite a specific brand but have some not-so-positive remarks, you may email me directly. I'm appreciating your comments and my coverslip-ragged fingertips thank you! Sara A. Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Thu Jun 2 15:37:03 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Reference Labs Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42AB@fh2k093.fhmis.net> We have a routine 24 hour turn-around-time, no more than a week for special studies. We run a pending list on Monday, Wednesday, and Friday to catch any cases that the Pathologist overlooked or got skipped just because someone was talking and put the case in the "done" cupboard. We've lost no business, in fact we've had to hire four more Pathologists in the last three years and go 24-7 with five more techs! ----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jesus Ellin Sent: Thursday, June 02, 2005 2:33 PM To: histonet@lists.utsouthwestern.edu; ppma-request@mailman.u.washington.edu Subject: [Histonet] Refence Labs I was wondering how many Hospitals have lost revenue or specimens to refence labs because of turn around time and reports? To be specific an outreach program. Also what are you doing within your facility to stop this issue from occuring? Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emerald_lake77 <@t> yahoo.com Thu Jun 2 15:45:54 2005 From: emerald_lake77 <@t> yahoo.com (- -) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] My Negative is now Positive ... IHC assistance needed? Message-ID: <20050602204554.63616.qmail@web31709.mail.mud.yahoo.com> Hello, I recently worked up a new antibody for my lab. It is a rabbit anti-mouse (antigen). The antigen is found throughout the kidney and has been pinpointed by ISH to be located in the proximal tubules. For the IHC, I have determined the best concentration for my antibody was 7 ug/ml. I used Jackson's ChromPure Rabbit IgG, whole molecule as a negative control at the same concentration, diluted in the same buffer as my primary. I am also using the Envision system by DAKO. Lately I've noticed a lot of background coming up with my negative. I re-ran a titre for my primary and had negatives at the corresponding concentrations. To my surprise, I got a lot of background on the negative, and moreover, it appears to be decreasing as the dilutions get higher. OK -- what's going on? Have I contaminated my negative somehow?? I keep the glass jar it originally came in at 4 degree C. When needed, I aliquot what I need under the Biosafety Cabinet (sterile conditions). If it has been contaminated, what has it been contaminated with and why is it giving me background? How can I avoid this in the future (can it be aliquoted and frozen)? Other suggestions will be very helpful. My positive looks good. Very clear staining and background only seen when Ab concentration is very high. I need a clear negative though to support my primary Ab results. Thank you all again for your help. --------------------------------- Discover Yahoo! Use Yahoo! to plan a weekend, have fun online & more. Check it out! From emerald_lake77 <@t> yahoo.com Thu Jun 2 16:14:06 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] My Negative is now Positive ... IHC assistance needed? Message-ID: <20050602211406.20825.qmail@web31705.mail.mud.yahoo.com> Hello, I recently worked up a new antibody for my lab. It is a rabbit anti-mouse (antigen). The antigen is found throughout the kidney and has been pinpointed by ISH to be located in the proximal tubules. For the IHC, I have determined the best concentration for my antibody was 7 ug/ml. I used Jackson's ChromPure Rabbit IgG, whole molecule as a negative control at the same concentration, diluted in the same buffer as my primary. I am also using the Envision system by DAKO. Lately I've noticed a lot of background coming up with my negative. I re-ran a titre for my primary and had negatives at the corresponding concentrations. To my surprise, I got a lot of background on the negative, and moreover, it appears to be decreasing as the dilutions get higher. OK -- what's going on? Have I contaminated my negative somehow?? I keep the glass jar it originally came in at 4 degree C. When needed, I aliquot what I need under the Biosafety Cabinet (sterile conditions). If it has been contaminated, what has it been contaminated with and why is it giving me background? How can I avoid this in the future (can it be aliquoted and frozen)? Other suggestions will be very helpful. My positive looks good. Very clear staining and background only seen when Ab concentration is very high. I need a clear negative though to support my primary Ab results. Thank you all again for your help. __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From galinadeyneko <@t> yahoo.com Thu Jun 2 16:49:36 2005 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Re: stain rack Message-ID: <20050602214936.90940.qmail@web33103.mail.mud.yahoo.com> Cindy, Look VWR catalog, p.750, cat # 25608-868 called Slide holder, 24 place. Galina Deyneko Novartis Cambridge histonet-request@lists.utsouthwestern.edu wrote: 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: Stain Racks (Fred Underwood) 2. Fw: [Histonet] Cryostat Frozen Sectioning Aid (Instrumedics) 3. Info on Cryojane (Lowen, Nancy) 4. different antigens need different methods for antigen retrieval? (pex) 5. Congo Red/Toludine Blue for Eosinaphils and Mast (Madary, Joseph) ---------------------------------------------------------------------- Message: 1 Date: Thu, 02 Jun 2005 11:58:08 -0400 From: "Fred Underwood" Subject: Re: [Histonet] Stain Racks To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Cindy, Cardinal Health carries them. The catalog number is s7640 15. Their phone number is 1.888.444.5440. Price is $216 for a package of 10. Fred >>> Cindy DuBois 05/31/05 10:19AM >>> Does anyone know where I can purchase slide racks for the TissueTek DRS ? We currently run about 250 slides a day and only have 6 racks (one which is broken). Sakura is not currently manufacturing them due to a production problem. Any suggestions is greatly appreciated, Cindy Dubois Delta Pathology Assoc. __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Thu, 2 Jun 2005 12:23:53 -0400 From: "Instrumedics" Subject: Fw: [Histonet] Cryostat Frozen Sectioning Aid To: "HistoNet Server" Message-ID: <01c501c5678f$7caddb00$6401a8c0@INSTRUMEDICS22> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=response ----- Original Message ----- From: "Instrumedics" To: "Susan Q Wells" Sent: Thursday, June 02, 2005 12:15 PM Subject: Re: [Histonet] Cryostat Frozen Sectioning Aid > Susan > The shelf life of the adhesive coated slides is years as long as the > slides > are not exposed to UV light. > > Bernice > Instrumedics > > > ----- Original Message ----- > From: "Susan Q Wells" > To: > Sent: Thursday, June 02, 2005 9:02 AM > Subject: [Histonet] Cryostat Frozen Sectioning Aid > > >> Anyone know what the shelf life of the adhesive coated slides for the >> Instrumedics system is? >> Thanks in advance, >> Susan Wells >> >> >> > ------------------------------ Message: 3 Date: Thu, 2 Jun 2005 09:33:54 -0700 From: "Lowen, Nancy" Subject: [Histonet] Info on Cryojane To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain If anyone is using the Cryojane tape transfer system for cutting undecalcified bone for frozen sections, we would appreciate your imput. What temperature do you cut bone at and does the tape transfer the complete section of bone to the slide? We are getting shattering of the bone on most sections and are really having some problems with the bubbles not rolling out. We are still in the testing process, but we would love to have any tips from anyone who has more experience on how to obtain the best sections. Should we expect to see totally intact sections when cutting bone? Also, what freezing method have you found to be the best..Thanks in advance Nancy ------------------------------ Message: 4 Date: Fri, 3 Jun 2005 00:46:45 +0800 (CST) From: pex Subject: [Histonet] different antigens need different methods for antigen retrieval? To: Histonet@lists.utsouthwestern.edu Message-ID: <20050602164645.15307.qmail@web15502.mail.cnb.yahoo.com> Content-Type: text/plain; charset=gb2312 Hello, all, I feel confused for immunofluorescence. I am doing immunofluorescence in vertebra sections. I use the same protocol for immunostaining, but the results are very different, the one is very nice, but the other is weak, I do not know the reasons. Do different antigens need different methods for antigen retrieval? Generally, which methods for antigen retrieval in bone sections are better? (Enzyme digestion? or HIER? or both?) Any suggestions will be helpful for me! Thank you! --------------------------------- DO YOU YAHOO!? ????????G?????­???????????????????????????????? ------------------------------ Message: 5 Date: Thu, 2 Jun 2005 12:47:00 -0400 From: "Madary, Joseph" Subject: [Histonet] Congo Red/Toludine Blue for Eosinaphils and Mast To: Message-ID: <746FDB897740814EA52BDDCB5ED1DDBCBB1583@medimmune4.medimmune.com> Content-Type: text/plain; charset="iso-8859-1" Anyone hear of a combo CR/TB for Eo's and Mast Cells? ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 19, Issue 4 *************************************** __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From anh2006 <@t> med.cornell.edu Thu Jun 2 17:16:47 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] My Negative is now Positive ... IHC assistance needed? In-Reply-To: <20050602211406.20825.qmail@web31705.mail.mud.yahoo.com> References: <20050602211406.20825.qmail@web31705.mail.mud.yahoo.com> Message-ID: From what I understand from reading and through my own personal experience, over time this Chrompure Rabbit IgG reagent from Jackson gives background. I am not sure why. To avoid this I have heard people say they aliquot and freeze it and only keep a working aliquot at 4 deg C. I have this problem so much so with the Chrompure Reagents that I am considering looking for another source for my control IgGs. >Hello, > >I recently worked up a new antibody for my lab. It is a rabbit >anti-mouse (antigen). The antigen is found throughout the kidney and has been >pinpointed by ISH to be located in the proximal tubules. For the IHC, >I have determined the best concentration for my antibody was 7 ug/ml. >I used Jackson's ChromPure Rabbit IgG, whole molecule as a negative >control at the same concentration, diluted in the same buffer as my >primary. I am also using the Envision system by DAKO. > >Lately I've noticed a lot of background coming up with my negative. I >re-ran a titre for my primary and had negatives at the corresponding >concentrations. To my surprise, I got a lot of background on the >negative, and moreover, it appears to be decreasing as the dilutions get >higher. > >OK -- what's going on? Have I contaminated my negative somehow?? I >keep the glass jar it originally came in at 4 degree C. When needed, I >aliquot what I need under the Biosafety Cabinet (sterile conditions). >If it has been contaminated, what has it been contaminated with and why >is it giving me background? How can I avoid this in the future (can it >be aliquoted and frozen)? Other suggestions will be very helpful. > >My positive looks good. Very clear staining and background only seen >when Ab concentration is very high. I need a clear negative though to >support my primary Ab results. > >Thank you all again for your help. -- From katri <@t> cogeco.ca Thu Jun 2 19:10:04 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] different antigens need different methods for antigenretrieval? References: <20050602164645.15307.qmail@web15502.mail.cnb.yahoo.com> Message-ID: <001f01c567d0$987b19b0$6a9a9618@Katri> Generally speaking each antibody is optimized by the manufacturer (if not= ,=20 it should be) as far as, application, antigen retrieval and approximate=20 dilution. Their specification sheet gives you that information. That is y= our=20 starting point to set it up in your detection system. If your test material is outside the application recommended for that=20 antibody, it still may work, but you will have to do a lot more trials,=20 before you know that. For instance, if the application is said to be for=20 frozen section, but you want to use it with formalin fixed paraffin embed= ded=20 tissue, you'll have to do as complete work-up as you can. ie. try various= =20 enzymatic digestions, HIER in different buffers or no retrieval at all. I= t=20 can sometimes take days and even weeks of concentrated effort and even th= en=20 you may not be able to make an antibody to work for something it was not=20 meant for. So, when selecting an antibody, try to match it for the=20 application you will need it for. Hope this helps a bit... Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message -----=20 From: "pex" To: Sent: Thursday, June 02, 2005 12:46 PM Subject: [Histonet] different antigens need different methods for=20 antigenretrieval? > Hello, all, > > I feel confused for immunofluorescence. > I am doing immunofluorescence in vertebra sections. I use the same=20 > protocol for immunostaining, but the results are very different, the on= e=20 > is very nice, but the other is weak, I do not know the reasons. Do=20 > different antigens need different methods for antigen retrieval? > Generally, which methods for antigen retrieval in bone sections are=20 > better? (Enzyme digestion? or HIER? or both?) > Any suggestions will be helpful for me! > > Thank you! > > > > > > --------------------------------- > DO YOU YAHOO!? > =D1=C5=BB=A2=C3=E2=B7=D1G=D3=CA=CF=E4=A3=AD=D6=D0=B9=FA=B5=DA=D2=BB=BE= =F8=CE=DE=C0=AC=BB=F8=D3=CA=BC=FE=C9=A7=C8=C5=B3=AC=B4=F3=D3=CA=CF=E4 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20 From lyarrow <@t> shaw.ca Thu Jun 2 20:24:03 2005 From: lyarrow <@t> shaw.ca (lyarrow) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] reference books for plastic re kidneys Message-ID: <002601c567da$ef2f19a0$e8f79244@cg.shawcable.net> Hello, We are looking for good reference books on kidneys done by plastics. Our lab has outdated books. Any ideas would be appreciated. Thank you, Louise Yarrow Tech II Foothills Medical Centre Calgary Lab Services Histology louise.yarrow2@cls.ab.ca From haldana <@t> unimoron.edu.ar Thu Jun 2 20:55:10 2005 From: haldana <@t> unimoron.edu.ar (Hernan Aldana) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Mouse anti-human androgen receptor Message-ID: <002701c567df$46e3f680$9d25e818@b3w6zzmtb6juvbs> Dear Histonetters I need to work with Mouse anti-human androgen receptor from BD PharMingen applied in tissues of human testicles. Does anyone have any advices for me? Fixation, paraffin processing, antibody dilution, times, antigen recovering, etc. Thanks in advance Dr. Hern?n J. Aldana Marcos Secretario Acad?mico Facultad de Medicina. Universidad de Mor?n Machado 914. B1708JPD. Buenos Aires. Argentina e-mail alternativo hernanjavier@yahoo.com web:http://www.ht.org.ar/histologia/NUEVAS%20UNIDADES/home.htm From Kemlo.Rogerson <@t> elht.nhs.uk Fri Jun 3 02:15:34 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Ventana Benchmark Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD423@elht-exch1.xelht.nhs.uk> Logically, but having no knowledge of this at all, I would assume that you would have to use different protocols for differing animal species. Everything else various, rbc count, wbc count, different reactions to drugs, etc. Very interesting question but how you would find them out eludes me. Usually fix preps with methanol for air dried or spray fix for 'fixed'. -----Original Message----- From: Laura Fidgen [mailto:lfidgen@vt.edu] Sent: 02 June 2005 18:43 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Ventana Benchmark We are a veterinary teaching hospital currently evaluating a Ventana Benchmark LT. I have a few questions: 1. Do protocols for antibodies vary among different animal species? 2. For cytology specimens (FNA or wet preps), where did you get your protocols? 3. For cytology specimens (fna or wet preps), what fixation, if any, do these slides need? Thank you in advance to those that respond. Have a great day! Laura L. Fidgen, MScF, BSc, HT(ASCP) Laboratory and Research Practioner VMRCVM, Histopatholgy/Clinical Pathology Dept. Blacksburg, VA 24060-0443 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mikael.niku <@t> helsinki.fi Fri Jun 3 03:25:43 2005 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Olympus DP70 - opinions? Message-ID: <42A01407.7000609@helsinki.fi> Hello! We are planning to get a new digital camera for our Leica DM4000 microscope. The camera will be used for photographing both normal light microscopy and standard epifluorescence microscopy of various histological samples. One of the cameras we have tested is Olympus DP70, which seems to be pretty good. However, I'm a bit uncertain of the specific technology used in the camera (it has a 1.4 megapixel CCD which is slightly moved around to produce up to 12 megapixel images). So, if anyone has experience in this system, I'd welcome any comments. Of course, recommendations for other systems will also be appreciated. Briefly, we need a camera for colour work, sensitive enough for epifluorescence, and let's say at least 4-5 megapixels. -- //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// From BMolinari <@t> heart.thi.tmc.edu Fri Jun 3 06:13:18 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Automatic Coverslipper Comparison Message-ID: Sakura glass coverslipper..love it. Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Thursday, June 02, 2005 3:12 PM To: 'Breeden, Sara'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Automatic Coverslipper Comparison We are very happy with our Leica cv5030. Under no circumstances would we buy a "tape" coverslipper - after five years the tape is lifting on all of our slides and the tissue section comes off with the tape! We do at least 300,000 slides per year so you can imagine the mess that we have. Good luck in your search! -Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Breeden, Sara Sent: Thursday, June 02, 2005 3:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automatic Coverslipper Comparison If you were to purchase an automatic coverslipper today, which one would you buy? If you already have one, did you buy a glass coverslipper - and, if you did, do you like it? If you aren't happy, why? Did you buy a tape coverslipper - and, if you did, do you like it? If you aren't happy, why? If you'd care to cite a specific brand but have some not-so-positive remarks, you may email me directly. I'm appreciating your comments and my coverslip-ragged fingertips thank you! Sara A. Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 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 jestrong <@t> comcast.net Fri Jun 3 09:05:46 2005 From: jestrong <@t> comcast.net (Jes Strong) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Caveat Emptor Message-ID: Dear Histonetters, While we would never use this forum for commercial marketing purposes, we feel that this would be the best platform to set the record straight and to clear up any misunderstandings in the marketplace. As of November 20, 2004, Milestone s.r.l. (manufacturers of microwave tissue processors and digital imaging systems) ended its distribution arrangement with Hacker Instruments and set up a Direct US Sales and Support Office in Shelton, Connecticut known as Milestone Medical Inc. As of November 20, 2004, no company other than Milestone Medical is authorized to sell Milestone products in the United States; nor will Milestone provide warranties, parts, service or application support to any company that attempts to sell Milestone products without authorization. All Milestone products sold by authorized representatives (before and after November 20, 2004) will continue to be supported according to the manufacturer?s warranties and extended service agreements. This message is also intended to reach current Milestone users who may need any technical or applications support. You can get in touch with us using the information below and we will promptly return your inquiry. Thank you for your understanding. We look forward to serving the needs of the histology community. Milestone Medical Inc. 25 Controls Dr. Shelton, CT 06484 866-995-5300 (toll free) info@milestonemed.com www.milestonemed.com From emerald_lake77 <@t> yahoo.com Fri Jun 3 09:36:34 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] My Negative is now Positive ... IHC assistance needed? In-Reply-To: <000c01c5683e$81d5ca20$41065486@auxs.umn.edu> Message-ID: <20050603143635.64927.qmail@web31706.mail.mud.yahoo.com> Jan, How do you know the initial concentration of IgG in the Normal Rabbit Serum (is it given to you by DAKO?)? Sounds like a good switch if I actually can dilute the serum IgG to the same concentration as my primary. Let me know. Thanks. Jan Shivers wrote: Try DAKO's Normal Rabbit Serum (X0902). Works nicely for me as negative control to accompany my rabbit polyclonals, when diluted to the same Ig concentration as the primary is used at. Jan Shivers U of MN Vet Diag Lab ----- Original Message ----- From: "GT Hebert" To: Sent: Thursday, June 02, 2005 4:14 PM Subject: [Histonet] My Negative is now Positive ... IHC assistance needed? > Hello, > > I recently worked up a new antibody for my lab. It is a rabbit > anti-mouse (antigen). The antigen is found throughout the kidney and has been > pinpointed by ISH to be located in the proximal tubules. For the IHC, > I have determined the best concentration for my antibody was 7 ug/ml. > I used Jackson's ChromPure Rabbit IgG, whole molecule as a negative > control at the same concentration, diluted in the same buffer as my > primary. I am also using the Envision system by DAKO. > > Lately I've noticed a lot of background coming up with my negative. I > re-ran a titre for my primary and had negatives at the corresponding > concentrations. To my surprise, I got a lot of background on the > negative, and moreover, it appears to be decreasing as the dilutions get > higher. > > OK -- what's going on? Have I contaminated my negative somehow?? I > keep the glass jar it originally came in at 4 degree C. When needed, I > aliquot what I need under the Biosafety Cabinet (sterile conditions). > If it has been contaminated, what has it been contaminated with and why > is it giving me background? How can I avoid this in the future (can it > be aliquoted and frozen)? Other suggestions will be very helpful. > > My positive looks good. Very clear staining and background only seen > when Ab concentration is very high. I need a clear negative though to > support my primary Ab results. > > Thank you all again for your help. > > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From POWELL_SA <@t> Mercer.edu Fri Jun 3 09:39:44 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Olympus DP70 - opinions? In-Reply-To: <42A01407.7000609@helsinki.fi> Message-ID: <01LP0S8QMVGS8WWTRF@Macon2.Mercer.edu> Mikael, I use a Sony Cybershot, 5 megapixel camera mounted (the mount was developed by Bobby Martin of www.martinmicroscope.com) on our Olympus BX40 which has served us very well here at Mercer. You may contact him via the web site. They offer several other cameras as well. Shirley Powell -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mikael Niku Sent: Friday, June 03, 2005 3:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Olympus DP70 - opinions? Hello! We are planning to get a new digital camera for our Leica DM4000 microscope. The camera will be used for photographing both normal light microscopy and standard epifluorescence microscopy of various histological samples. One of the cameras we have tested is Olympus DP70, which seems to be pretty good. However, I'm a bit uncertain of the specific technology used in the camera (it has a 1.4 megapixel CCD which is slightly moved around to produce up to 12 megapixel images). So, if anyone has experience in this system, I'd welcome any comments. Of course, recommendations for other systems will also be appreciated. Briefly, we need a camera for colour work, sensitive enough for epifluorescence, and let's say at least 4-5 megapixels. -- //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet = From histosci <@t> shentel.net Fri Jun 3 09:41:36 2005 From: histosci <@t> shentel.net (HSRL) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Re: Which coverslipper? In-Reply-To: <20050603031815.C366E213029C4DF1@email-cooking.com> Message-ID: <001601c5684a$5d28c9f0$0200a8c0@HSRLMAIN> We have a Sakura Glas Coverslipper. We love the unit- and best of all the customer service is phenomenal. We have the best Sakura Rep in the business- Anna Magallanez. Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org -----Original Message----- From: Cooking.com [mailto:Cooking.com@email-cooking.com] Sent: Friday, June 03, 2005 5:18 AM To: beth@hsrl.org Subject: 15% Off a Single Item - 3 Days Only _____ Cooking.com - Top Brands . Best Selection . 100% Satisfaction Guaranteed _____ Receive 15% OFF a Single Item* - Only 3 More Days! To redeem, simply use Coupon Code C95873 at checkout. Hurry! Offer ends Sunday, June 5th. Redeem this offer today and save! Click here to shop. *The 15% discount will apply to the item with the highest price. Excludes Merchant Partner products, All-Clad, Breville, Capresso, Emerilware, Global, Henckels, Kershaw, KitchenAid Stand Mixers, Krups, KuhnS Rikon, Musso, Saeco, Seattle's Best Coffee, Starbucks, Torrefazione, Viking, Wusthof, clearance items and gift certificates. Cannot be combined with any other offers or discounts, promotional gift certificates, or third party site offers including but not limited to rebate or miles programs. Cannot be applied to previous purchase. Terms are subject to change. Offer ends 6/5/05. _____ Visit Our Most Popular Departments New Items | Cookware | Cutlery | Small Appliances | Cook's Tools | Cookbook Shop Coffee & Tea | Clearance | Email your Wish List | Gift Certificates | Wedding Registry | Recipes _____ YOUR NEWSLETTER SUBSCRIPTION Note that you're subscribed as: beth@hsrl.org Please do not reply to this email. If you would like to send us an email, click here. To unsubscribe or make other changes to your newsletter account, click here. If you prefer to talk to someone about your subscription or any question you may have about Cooking.com, you may call us toll-free at 1-877-999-2433. 100% SATISFACTION AND SECURE ORDERING GUARANTEED. NO CREDIT CARD RISK. Visit our site to read details about our security and privacy policies. Thank you for letting us share our products, recipes, and our love for cooking. We look forward to your next visit! C2005 Cooking.com | 2850 Ocean Park Blvd., Suite 310, Santa Monica, CA 90405-6209 From Janet.Keeping <@t> cna.nl.ca Fri Jun 3 09:45:28 2005 From: Janet.Keeping <@t> cna.nl.ca (Keeping, Janet) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] (no subject) Message-ID: Hello I have been "lurking" on your list for a few days. There is a great exchange of information going on here. I have recently moved from the clinical lab to a college where I teach the Histotechnology part of a medical Lab program. I wonder if any of you have any tips on how I might provide my students with tissues which simulate renal and liver needle biopsies. Any tissues that we may have are of course from autopsy. I also wondered about simulating GI biopsies and curettings. If any of you have done this I would appreciate your advice. Janet Keeping Instructor Medical Laboratory Science Program College of the North Atlantic Prince Philip Drive Campus St. John's, NL, A1C 5P7 709-758-7657 tel 709-758-7634 fax Email: janet.keeping@cna.nl.ca From jqb7 <@t> cdc.gov Fri Jun 3 09:51:11 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Re: Which coverslipper? Message-ID: I don't know.....we have Sharon Wehman and she is absolutely great! Maybe there should be some sort of "Survival" show pitting them against one another to see who's best! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of HSRL Sent: Friday, June 03, 2005 10:42 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Re: Which coverslipper? We have a Sakura Glas Coverslipper. We love the unit- and best of all the customer service is phenomenal. We have the best Sakura Rep in the business- Anna Magallanez. Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org -----Original Message----- From: Cooking.com [mailto:Cooking.com@email-cooking.com] Sent: Friday, June 03, 2005 5:18 AM To: beth@hsrl.org Subject: 15% Off a Single Item - 3 Days Only _____ Cooking.com - Top Brands . Best Selection . 100% Satisfaction Guaranteed _____ Receive 15% OFF a Single Item* - Only 3 More Days! To redeem, simply use Coupon Code C95873 at checkout. Hurry! Offer ends Sunday, June 5th. Redeem this offer today and save! Click here to shop. *The 15% discount will apply to the item with the highest price. Excludes Merchant Partner products, All-Clad, Breville, Capresso, Emerilware, Global, Henckels, Kershaw, KitchenAid Stand Mixers, Krups, KuhnS Rikon, Musso, Saeco, Seattle's Best Coffee, Starbucks, Torrefazione, Viking, Wusthof, clearance items and gift certificates. Cannot be combined with any other offers or discounts, promotional gift certificates, or third party site offers including but not limited to rebate or miles programs. Cannot be applied to previous purchase. Terms are subject to change. Offer ends 6/5/05. _____ Visit Our Most Popular Departments New Items | Cookware | Cutlery | Small Appliances | Cook's Tools | Cookbook Shop Coffee & Tea | Clearance | Email your Wish List | Gift Certificates | Wedding Registry | Recipes _____ YOUR NEWSLETTER SUBSCRIPTION Note that you're subscribed as: beth@hsrl.org Please do not reply to this email. If you would like to send us an email, click here. To unsubscribe or make other changes to your newsletter account, click here. If you prefer to talk to someone about your subscription or any question you may have about Cooking.com, you may call us toll-free at 1-877-999-2433. 100% SATISFACTION AND SECURE ORDERING GUARANTEED. NO CREDIT CARD RISK. Visit our site to read details about our security and privacy policies. Thank you for letting us share our products, recipes, and our love for cooking. We look forward to your next visit! C2005 Cooking.com | 2850 Ocean Park Blvd., Suite 310, Santa Monica, CA 90405-6209 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ian.Ward <@t> nottingham.ac.uk Fri Jun 3 10:02:24 2005 From: Ian.Ward <@t> nottingham.ac.uk (Ian Ward) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Re: Histonet Digest, Vol 19, Issue 5 (Out of Office) Message-ID: I will be on annual leave untill Monday 20th June. For bookings or in an emergency please contact either Susan Anderson or Kelly Draper-Morgan on: susan.anderson@nottingham.ac.uk kelly.draper-morgan@nottingham.ac.uk This message has been checked for viruses but the contents of an attachment may still contain software viruses, which could damage your computer system: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation. From jdmd77 <@t> hotmail.com Fri Jun 3 10:12:11 2005 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] (no subject) In-Reply-To: Message-ID: Janet - It is relatively simple to contact your local butcher to acquire cow and lamb internal organs. Kidney, liver, heart and all gastrointestinal organs. To simulate the biopsies from these tissues, aquire large bore needles and perform biopsies, or use soft forceps to avulse tissue from the GI tract. Hope this is of assistance! Julia Dahl, MD. >From: "Keeping, Janet" >To: >Subject: [Histonet] (no subject) >Date: Fri, 3 Jun 2005 12:15:28 -0230 >MIME-Version: 1.0 >Received: from swlx162.swmed.edu ([199.165.152.162]) by mc4-f23.hotmail.com >with Microsoft SMTPSVC(6.0.3790.211); Fri, 3 Jun 2005 07:46:03 -0700 >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34)id 1DeDQh-0007jR-Sv; Fri, 03 Jun >2005 09:45:52 -0500 >Received: from [199.165.152.167] (helo=swlx167.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34) id 1DeDQQ-0007j9-Bffor >histonet@lists.utsouthwestern.edu; Fri, 03 Jun 2005 09:45:35 -0500 >Received: from [209.128.23.29] (helo=mailmarshal.cna.nl.ca)by >swlx167.swmed.edu with esmtp (Exim 4.44) id 1DeDQP-0001lv-KPfor >histonet@lists.utsouthwestern.edu; Fri, 03 Jun 2005 09:45:30 -0500 >Received: from exchange.cna.nl.ca (Not Verified[10.200.11.100]) >bymailmarshal.cna.nl.ca with NetIQ MailMarshal (v6, 0, 3, 8)id >; Fri, 03 Jun 2005 12:15:28 -0230 >X-Message-Info: gUeNUVfFqHBSeC/Efsp97RfDLd6IRxCzUe7NSdqqnPk= >X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 >Content-class: urn:content-classes:message >X-MS-Has-Attach: X-MS-TNEF-Correlator: Thread-Index: >AcVoS/BIHTitFYlbSiqgoIAd/1ANnA== >X-Scan-Signature: c0d0b691c340a303b666656f3482430c >X-Content-Filtered-By: Mailman/MimeDel 2.1.5 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >Precedence: list >List-Id: For the exchange of information pertaining to histotechnology >andrelated fields >List-Unsubscribe: >, > >List-Archive: >List-Post: >List-Help: >List-Subscribe: >, >Errors-To: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 690e9200c080ca55fd7a8d0fb0193eeb >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: X-Spam-Status: No, hits=0.0 required=5.5 tests=none >autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >Return-Path: histonet-bounces@lists.utsouthwestern.edu >X-OriginalArrivalTime: 03 Jun 2005 14:46:03.0889 (UTC) >FILETIME=[F77BEE10:01C5684A] > >Hello I have been "lurking" on your list for a few days. There is a >great exchange of information going on here. I have recently moved from >the clinical lab to a college where I teach the Histotechnology part of >a medical Lab program. I wonder if any of you have any tips on how I >might provide my students with tissues which simulate renal and liver >needle biopsies. Any tissues that we may have are of course from >autopsy. I also wondered about simulating GI biopsies and curettings. >If any of you have done this I would appreciate your advice. > > > >Janet Keeping > >Instructor > >Medical Laboratory Science Program > >College of the North Atlantic > >Prince Philip Drive Campus > >St. John's, NL, A1C 5P7 > >709-758-7657 tel > >709-758-7634 fax > >Email: janet.keeping@cna.nl.ca > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Fri Jun 3 10:47:32 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Ventana Benchmark cytology protocols Message-ID: The few cytology preparations (FNAs and wet preps) we have done here have been stained using the "non-paraffin, wet slide load" protocol format on our Benchmark XT, and we chose the best temperature and incubation time for each antisera tested. The FNA slides and preps were previously fixed with our routine cytology spray fix. As no formalin fixation is used, no antigen retrieval is necessary. The antisera dilution may have to be altered from that used for wax sections (if you do them) - you'll just have to see. You may have the luxury of being able to try different fixatives rather than using the one you use routinely, so you could try choosing ones that are best suited to the antigen you are trying to detect. Let me know how you get on! Jacqui Malam Royal Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From John.Thweatt <@t> med.va.gov Fri Jun 3 11:33:41 2005 From: John.Thweatt <@t> med.va.gov (Thweatt, John T) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Problem with cutting sections on Microtome Message-ID: <7A00E2723D55D311AC240000F831866802606F54@vhaamaexc1.amarillo.med.va.gov> Dear Fellow Histonetters, We are experiencing some trouble cutting just regular skin that we have not seen before. There has been no change in the processing or any other process that we are doing yet our Pathologist pointed out here this week that she is seeing shattering, wavy sections starting in our skin specimens and now it is showing up in our polyps as well. We have tried to change the angle of the blade, different blades, chilling and warming the block (the usual method to clear this up) with no avail. These are very new microtomes that we purchased last fall (Microm 355 S) fully automatic microtomes (2) and we are using Richard Allen blades and tried some Duraedge samples also. We should not have such problems with this new of equipment. Any suggestions or ideas? Anyone experience the same with the same microtome? and how did you resolve it? Thanks in advance. Tom Thweatt From Linke_Noelle <@t> Allergan.com Fri Jun 3 11:42:27 2005 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] CD4, CD8 and Bcells Message-ID: Hi everyone I'm looking for antibodies to CD4, CD8 and a B-cell marker (CD20 or CD45RA perhaps) that work on rat tissues (paraffin). Does anyone have any suggestions? Thank you! Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 From David.Muskett <@t> RLC.NHS.UK Fri Jun 3 11:42:36 2005 From: David.Muskett <@t> RLC.NHS.UK (Muskett David) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] GLUT-1 antibody[Scanned] Message-ID: <6AB79F0BA7C4914BA8DD8F5E0C21B9755A72EF@AHEXMAIL01.xalderhey.com> Dear all Can anybody explain to me why syncytiotrophoblasts are positive for Glut-1 expression. Thanks David David Muskett Chief Biomedical Scientist, Histology Royal Liverpool Children's NHS Trust Eaton Road Liverpool L12 2AP Internal telephone x3615 Telephone (0151) 293 3656 Fax (0151) 293 3617 http://nhsald02/histopathology <> From MadaryJ <@t> MedImmune.com Fri Jun 3 12:28:45 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Summer job at Medimmune, Location Message-ID: <746FDB897740814EA52BDDCB5ED1DDBCBB1590@medimmune4.medimmune.com> Sorry my ealrier posting was weak. We are located in Gaithersburg Maryland about 20 minutes outside of Wash D.C. The amount is not set yet but for a decent tech we can work off past salaries. We want a responsible person that can hit the lab without much extra training, just the nuances of this lab should be all required. The lab is simple on the paraffin side, and requires a person who can cut good frozens, doing IHC on those frozens using the DAKO autostainer. The staining is set up so the person will need to be very comfortable making dilutions and reagents from scratch. We rarely use commercial anything except buffers. Again, a retired person who was really good would be perfect. From clarissabush <@t> sbcglobal.net Fri Jun 3 12:53:37 2005 From: clarissabush <@t> sbcglobal.net (CM Bush) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Why do some paraffin sections of brain wrinkle on the water bath? Message-ID: <20050603175337.42908.qmail@web80302.mail.yahoo.com> Dear Histonet, Seeking some some basic info: Sometimes, when cutting 8 um sections of brain, mouse or human (in standard size cassettes), the section will either wrinkle or pucker once on the water bath (set at ~43C), and then other sections (from different blocks) are great and flatten out, very well, looking pretty much like the tissue did at the face of the block. The mouse brains seem to wrinkle more randomly, where human brain sections pucker in a more regular pattern-does this depend at all on how the piece of human brain was cut-in as far as which section of brain, or ratio of white matter to gray, contours of the brain, or dehydration issues, or..? For the mouse brains, some extra time on the bath seems to iron out the wrinkles pretty much. Same for the human brain but not completely. If I could draw a picture, once the section of human brain has dried on to the slide there are regular patterns of slight folds where the puckers where, arranged around the, I think, tracks of white matter. Like I said, sometimes these wrinkle and puckers take care of them selves while on the water bath or drying on the slide- I'd like to try to improve the sections, what I can do to fix this, or at least find out what is going on with this, just to know. Thank you so much for the help. CMB From DonnaWillis <@t> texashealth.org Fri Jun 3 13:06:10 2005 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Electron Microscopy Workload Message-ID: <4483B6EE0FA82A4EAED2B1E161E5E41147C858@ftwex02.txhealth.org> What is the average workload for a TEM technician? How many cases a year should 1 FTE perform? Thanks, Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx 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 sbreeden <@t> nmda.nmsu.edu Fri Jun 3 13:24:44 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Coverslipper Thanks Message-ID: Thanks to everyone who has sent their comments (+ and -) about coverslippers. I have tucked these comments into my little file-cabinet-like gray matter and will make my decision based in good part on these gems. Thanks for taking your time to help out! Long Live Histonet!! Sara A. Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From John.Thweatt <@t> med.va.gov Fri Jun 3 13:51:43 2005 From: John.Thweatt <@t> med.va.gov (Thweatt, John T) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Qualifications to work in Histology Message-ID: <7A00E2723D55D311AC240000F831866802606F55@vhaamaexc1.amarillo.med.va.gov> Dear Fellow Histonetters, Two posts in one day ought to be a record for me, but as demand dictates, I have returned with yet another question. This question may be directed more toward the Histology managers or histologists who know a little about regulations in the group as I once again as for your most valuable help. I am inquiring on behalf of a friend and fellow Histologist who at the moment does not have access to a computer, so pardon me if this isn't very clear as this is coming to me second hand. The question is: What are the individual state requirements for a histology technician that is not certified? Is there such regulation that exists within the states that would regulate hiring staff and what kind of requirements would a person fall under in order to be able to be hired to work in histology (non-certified). What does your state say about this issue? I would be interested to see what different states (or facilities) have to say about the uncertified technician route as it got a little harder now to get certified under ASCP. If anyone has any facility type documentation that gives you guidance as to who you can hire to work in histology, what kind of qualifications do you require? Does your state or facility eventually push for certification (surely)? Are there cities out there that lay the groundwork for what qualifications are necessary to work in our field? I do realize that this is a complex question that may not a simple answer. If someone out there has the expertise in this could give me some guidance on this topic it would be most appreciated. Thank you in advance. Sincerely, Tom Thweatt From EJones <@t> Ventanamed.com Fri Jun 3 14:03:54 2005 From: EJones <@t> Ventanamed.com (Emma Jones) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] ventana benchmark Message-ID: hello, I have just read your question on histonet. I would highly recommend you contact the Ventana Help desk and ask them directly for information on protocol. If they don't know, then some of the technical support staff may. As technical support staff, if I do not know a protocol, invariably I send a global email to my colleagues to ask for help, much the same as Histonet. Someone somewhere has info. So instead of struggling yourself, ask Ventana for help. regards Emma E Jones Technical Specialist UK & EIRE Ventana Medical Systems Tel: +44 (0) 7973 190163 E-mail: HYPERLINK "mailto:ejones@ventanamed.com"ejones@ventanamed.com Website:www.ventanamed.com -- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.322 / Virus Database: 267.5.2 - Release Date: 03/06/2005 From EJones <@t> Ventanamed.com Fri Jun 3 14:03:57 2005 From: EJones <@t> Ventanamed.com (Emma Jones) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] coverslippers Message-ID: Hi Sara, I would recommend you get in contact with a Ventana representative. Although known for automated immunohistochemical stainers, we have moved into the field of H&E/coverslippers. Maybe you would be interested in our new Symphony stainer. Stainer, oven and glass coverslipper combined. regards Emma Emma E Jones Technical Specialist UK & EIRE Ventana Medical Systems E-mail: HYPERLINK "mailto:ejones@ventanamed.com"ejones@ventanamed.com Website:www.ventanamed.com -- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.322 / Virus Database: 267.5.2 - Release Date: 03/06/2005 From BDUE <@t> PARTNERS.ORG Fri Jun 3 14:11:49 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Why do some paraffin sections of brain wrinkle on thewater bath? Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB5027A6A@PHSXMB7.partners.org> Hello Clarissa, I have some experience with human brain, and what you describe is what I've noticed routinely for several years. That's just what brain does. Thicker paraffin sections (10-15um) have more wavy puckering problems than do thin sections (5um). Larger sections also have more problems. I cut autopsy tissues up to 1x1.5in and giant sections up to 4x6in. They often have variable puckering which seems related to white vs grey matter patterns in the tissue. Hotter water bath helps increase the flattening (47C), but also puts you at greater risk for exploding sections if the processing hasn't been perfect. The tissue has shrunk quite a bit during routine processing and I imagine the hot water bath makes it want to swell up to it's original size -- or more. The puckering is the result of differential swelling and the restrictions from the surrounding paraffin and other tissues. At 8um the paraffin border of the section can resist the heat of the water bath and the pressure of the expanding tissue, and prevent the tissue from spreading. Thin sections left on the water bath tend to end up with irregular outlines, somewhat reflecting the tissue shape and the placement of initial puckers. Blocks that have a wide margin constrict the tissue more than those with narrow margins. I have trimmed the margins from some sections on the bath and this allowed more even spreading. But of course excessive spreading will show artifacts microscopically. You could try thinner sections 7,6, etc until you find a new balance between signal strength and puckering problems. Or warmer water in the bath. I have experimented with floating sections on ethanols from 60-80% but wouldn't recommend it for routine work. There's both the fire hazard of warmed alcohols and alot of handling differences from normal water bath routines. However, it was impressive to see a large section (2x3in.) floating flat with zero swell and minimal spread -- not even collagen took up water from the ethanol bath. Along similar lines, you could also experiment with turning the water bath temp down -- alot. Cooler water won't swell the tissues as much. But you also have to have very good microtomy technique since you can't rely on using the water bath to fix the problems in the sections. It is possible to cut serial perfect sections of brain -- no chatter, no compression, no wrinkling in the tissue -- but it takes more effort, and more fresh blades. Puckers on the water bath don't necessarily have to translate into wrinkles on the slides. Several things seem to influence whether the puckers become wrinkles or simply go away with drying. Lots of my sections retain some gentle "waviness" when I pick them up, but once stained, the waves are gone. Differential swelling gives way to differential drying. In my experience, the exact things I do to dry the slides can make a difference between having flat, wavy, or lifting sections where originally there were puckers. Vertical draining at room temp for short time (< 30min) followed by overnight in a 37C oven is what works best for me. You could simply try varying your drying protocol. An old trick I was taught here helps sometimes, especially if there are large puckers. You wet a filter paper with 95-100% ETOH and place it on a flat surface. When you pick up the section, briefly let it drain, then lay it down on the filter paper section side down. Rub the back of the slide with firm pressure -- too much will make the section stick to the paper. Separating the slide from the filter paper generally goes smoothly -- surprising but true -- but takes a little practice. What you end up with is a section that has been both physically flattened against the slide and dehydrated (shrunk) at the same time. This is a technique for problem sections, not routine use (although for giant sections the time is worth the results). This trick doesn't get rid of real wrinkles, but it will tend to make them stick to the slide instead of falling off. It also will instantly turn large puckers into wrinkles -- squished. I hope some of this rambling helps, -brice Neuropathology Lab Brigham & Women's Hospital Boston -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of CM Bush Sent: Friday, June 03, 2005 1:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Why do some paraffin sections of brain wrinkle on thewater bath? Dear Histonet, Seeking some some basic info: Sometimes, when cutting 8 um sections of brain, mouse or human (in standard size cassettes), the section will either wrinkle or pucker once on the water bath (set at ~43C), and then other sections (from different blocks) are great and flatten out, very well, looking pretty much like the tissue did at the face of the block. The mouse brains seem to wrinkle more randomly, where human brain sections pucker in a more regular pattern-does this depend at all on how the piece of human brain was cut-in as far as which section of brain, or ratio of white matter to gray, contours of the brain, or dehydration issues, or..? For the mouse brains, some extra time on the bath seems to iron out the wrinkles pretty much. Same for the human brain but not completely. If I could draw a picture, once the section of human brain has dried on to the slide there are regular patterns of slight folds where the puckers where, arranged around the, I think, tracks of white matter. Like I said, sometimes these wrinkle and puckers take care of them selves while on the water bath or drying on the slide- I'd like to try to improve the sections, what I can do to fix this, or at least find out what is going on with this, just to know. Thank you so much for the help. CMB _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From slappycraw <@t> yahoo.com Fri Jun 3 14:17:01 2005 From: slappycraw <@t> yahoo.com (Larry Woody) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Qualifications to work in Histology Message-ID: <20050603191701.70027.qmail@web52604.mail.yahoo.com> Even though they dropped the high school diploma route to taking the exams I don't think it stops a facility from hiring a non-registered person for histology work, although this could vary from state to state and from facilty to facility. I personally think the exams are going to be on the honor system soon,(meaning you can take them from your home) and eventually phased out, but that is only one opinion. Larry --------------------------------- Discover Yahoo! Use Yahoo! to plan a weekend, have fun online & more. Check it out! From pruegg <@t> ihctech.net Fri Jun 3 14:32:08 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] reference for washing formalin fixed tissues Message-ID: <200506031932.j53JW5bm009022@chip.viawest.net> I have been asked to elaborate on my reference to washing tissues to reverse the effects of formalin fixation iliminating the possible need for AR. Here goes: I found the reference to Washing in a book called Introduction to Immunocytochemistry 3rd Edition J.M. Polak and S. Van Noorden pg. 24- 3.8.2 "The simplest form of reversing the effects of formalin is to wash the tissue well before processing, but this is not usually possible in histopathology laboratories, where rapid turnowver of specimens is required." I have not tested this extensively but have done a little, especially on over fixed samples, the data is yet to be confirmed but I am looking into it as we speak. I will keep you all posted. Patsy From ploykasek <@t> phenopath.com Fri Jun 3 14:41:46 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Aquaporin In-Reply-To: Message-ID: You might to read the recent article in Modern Pathology (2005) 18, 535-540: "Expression of aquaporins and PAX2......." We have recently begun investigating a polyclonal aquaporin 1 from chemicon. Which antibody you need depends on how you intend to use it. Patti Loykasek PhenoPath Laboratories Seattle, WA > Hello All > > I have been requested to be involved in a research study > demonstrating Aquaporin 1 (AQ1, AQP1) using IHC. To > date I have found 2 antibodies available from Chemicon. > My questions are: > 1. Are there other commercial suppliers of this Ab. > 2. If not, which of the 2 from Chemicon is the preferred > option. > 3. Do you realyy need to fix with paraformaldehyde as > stated in most methods, and why? > > i would appreciate any advice forwarded. > > regards > > > Tony Reilly > Chief Scientist > Anatomical Pathology > Northside Pathology > Prince Charles Hospital > Rode rd Chermside Q 4032 > Australia > Ph: 07 3350 8543 > Fax: 07 33508546 > tony_reilly@health.qld.gov.au > > > > > ****************************************************************************** > ***** > This email, including any attachments sent with it, is confidential and for > the sole use of the intended recipient(s). This confidentiality is not waived > or lost, if you receive it and you are not the intended recipient(s), or if it > is transmitted/received in error. > > Any unauthorised use, alteration, disclosure, distribution or review of this > email is prohibited. It may be subject to a statutory duty of confidentiality > if it relates to health service matters. > > If you are not the intended recipient(s), or if you have received this email > in error, you are asked to immediately notify the sender by telephone or by > return email. You should also delete this email and destroy any hard copies > produced. > ****************************************************************************** > ***** > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MadaryJ <@t> MedImmune.com Fri Jun 3 15:09:54 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Tissue Microarrays. Message-ID: <746FDB897740814EA52BDDCB5ED1DDBCBB1594@medimmune4.medimmune.com> What kind of response did you get on this? I have a pathologist who seems real hung up on contracting out TMA's to someone (actually in England, maybe you!). I am justm wondering from a routine standpoint and doing some pre-clinical work on rodents, what would be the benefit of us getting a system in place or contracting out this work? I mean I took a class in how to prepare them a few years back, bascially taking the specimens and putting them in spots on the block with the legend etc. I mean could one do this without fancy machinery, and what is the real use for this stuff? From MadaryJ <@t> MedImmune.com Fri Jun 3 15:27:59 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Tissue microarrays use? Message-ID: <746FDB897740814EA52BDDCB5ED1DDBC1E2E78@medimmune4.medimmune.com> Can someone explain to me what the big deal is on tissue microarrays? I understand that they are many perfect circles of known diseased or normal tissues than can be used as a control for various applications. What are the applications for this stuff on routine or research that would make it worth it for us to either do it ourselves or contract it out? Can it be done in house cheap? From Charlene.Henry <@t> STJUDE.ORG Fri Jun 3 15:43:13 2005 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Tissue microarrays use?. . Message-ID: <5CB39BCA5724F349BCB748675C6CA1A202C7572B@SJMEMXMB02.stjude.sjcrh.local> They are greatly instrumental in research. Working at a research facility, we were able to pay for the purchase of the instrument with only 1 research project. Example: A pathologist has a research project that he/she is doing say on Neuroblastoma. They have 150 blocks that they need a total of 10 IHC tests on each block. You take the 150 blocks and prepare 2 tissue micro array blocks and then run your 10 IHC test on the 2 TMA blocks. You have saved a great deal of money because you have 20 IHC tests instead of 1500 IHC tests that would have been needed without the tissue array blocks. At approximately $15 for each IHC test, you can see that $300 is much better than $22,500. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Friday, June 03, 2005 3:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue microarrays use?. . Can someone explain to me what the big deal is on tissue microarrays? I understand that they are many perfect circles of known diseased or normal tissues than can be used as a control for various applications. What are the applications for this stuff on routine or research that would make it worth it for us to either do it ourselves or contract it out? Can it be done in house cheap? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Jun 3 16:38:35 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Cutting undecalcified bone In-Reply-To: <20050602171759.81401.qmail@web30415.mail.mud.yahoo.com> Message-ID: <200506032138.j53LcWbm019799@chip.viawest.net> I have experience cutting frozen sections of whole rat tibias without decal (John Tarpley and I did this work together) which in my experience could only be done using the tape transfer technique. I use a D profile tungsten carbide knife(the bevel is only on one side unlike a C profile which has the same bevel on both sides, this is like the old triangle shaped glass knives) this knife requires a pretty straight cutting angle compared to the C profile which is more angled, thus the reference Liz had about me changing the knife angle. I have tried everything from heavily coated 6X to the lightest 1/2X coated slides. I have still to find a technique which will give me perfect sections of bone everytime. The problem for me is not getting the perfect section cut but getting it attached to the coated slide without bubbles under the tape and then removing the tape without leaving some of the section on the tape. This can be very frustrating and expensive to cut a beautiful section only to have it pull off with the tape, the expensive slide is lost and your beautiful section is useless. This is not only the case for undecalcified bone, I have been having the same old problem with cornea implants. Some things to try include: try to cut a thinner section like 4 microns (if your knife is really sharp this is not a problem), the thinner section will lay flatter on the coated slide. I do expose the UV for 2-3 times with about 10 secs. Between or the fuse will blow. Pull the tape off diagnally from corner to corner very slowly and smoothly. When appling the tape with the section to the coated slide "role the hell out of it" to borrow a phrase from Gayle, to make sure it gets really well attached to the slide before UV. I have had some luck with preparing the slide, exposing it to UV and then placing it on dryice for 5-10 min. before removing the tape, but again I never get every section in good shape and actually lately I have been getting more bad sections than good, which is driving me crazy. There are people up at CSU using this to cut horse carpal bone, I can give you their contact info if you like. I think I did better with bone than I am doing with cornea right now. Oh, for bone I try to cut as cold as possible, about -35 I have been cutting the softer tissue warmer -18---24, hey maybe that's my problem I should go back to the cold cold bone method, it can't hurt at this point. I snap freeze samples by surrounding them with OCT and either slowly lowering into liq. Nitrogen or isopentane and dryice. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen Peters M.D. Sent: Thursday, June 02, 2005 10:18 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Cutting undecalcified bone Nancy, I have experience cutting frozens on undecacified bone specimens for routine surg path cases using conventional technique but have no experience with the tape transfer system. I routinely use disposible low profile blades which in not optimum for bone. I imagine a stronger high profile blade or knife will work better. My success has varied with the hardness of the bone. Trebecular bone coming from older patients can cut fairly easily porvided the blade is sharp, and the section is taken with a fluid continuous motion. The result is a surprisingly intact section. The harder the bone ( typically the younger the patient ) there will be more damage to the blade in each pass. The sections will be streaked and splitting requiring one or more blade changes by time I get a resonable section. It is quite a juggling act. Cutting hard cortical bone is often very frustrating and very difficult to get any resonable section. From what you describe as bubbles, I am guessing you are getting sections of bone thicker than you are hoping for and as a result when you roll it out, the thicker bone pieces are protecting the rest of the section from rolling. Often the first section of any tissue will be considerably thicker than those that follow after a few fluid turns of the wheel. If you are not letting a few pass your first section may be considerably thicker and as a result will shatter more and create thicker trabeculae which may be leading to your problem. The equivalent using conventional technique will give me a section that my coverslip will not lie flat against the slide and my mounting medium will not spread. I look foreward to other peoples experience on this subject. Stephen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vitha <@t> mic.tamu.edu Fri Jun 3 16:51:03 2005 From: vitha <@t> mic.tamu.edu (Stanislav Vitha) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Attaching sections to slides (Cellulose nitrate?) Message-ID: <6.1.2.0.0.20050603161920.032f1100@mic.tamu.edu> Dear Histonetters, I have a user who is performing in situ hybridization on sections from plant floral tissue (Arabidopsis). The problem is that most sections are lost during the lengthy procedure. The tissue was embedded in a low-melting polyester wax (Steedman's wax) and 5-10 micron sections attached to either poly-L-lysine or silane-treated slides. This has worked great for immunos, but in situ hybridization seems too much for those sections to hold on. The usual tricks of drying the tissue after dewaxing have been applied, but without much effect. So we are contemplating using celloidin (cellulose nitrate) to coat the slides with dewaxed sections and thus prevent them from floating away. Do you think this is going to cause excessive background or other problems for in situ hybridization? The protocol I am planning to use is as follows: 1) De-wax the sections on slides as usual (in ethanol, the polyester wax is acohol-soluble) 2) Dip the slide in a solution of 50ml 100% ethanol, 49ml ether, 1g cellulose nitrate for few minutes 3) Lift the slide and hold in the air few seconds until it is just becinning to dry 4) place the slide in 80% ethanol to congeal the nitrocellulose 5) bring the slide to water or buffer through a graded ethanol series Thanks in advance for your advice Thank you. Stan Vitha Dr. Stanislav Vitha vitha@mic.tamu.edu Microscopy and Imaging Center Texas A&M University BSBW 119 College Station, TX 77843-2257 tel: 979-845-1129 (main desk) tel: 979-845-1607 (direct link) fax: 979-847-8933 From katri <@t> cogeco.ca Fri Jun 3 17:11:51 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Tissue microarrays use?. . References: <5CB39BCA5724F349BCB748675C6CA1A202C7572B@SJMEMXMB02.stjude.sjcrh.local> Message-ID: <009801c56889$3ecf7a00$6a9a9618@Katri> Hi Charlene, Knowing that so many tumors can give a very heterogenous staining patterns ( some areas strongly positive and others weak or even negative ), how do you overcome this problem with microarrays? I would have a hard time trusting the results with such small samples. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Henry, Charlene" To: "Madary, Joseph" ; Sent: Friday, June 03, 2005 4:43 PM Subject: RE: [Histonet] Tissue microarrays use?. . They are greatly instrumental in research. Working at a research facility, we were able to pay for the purchase of the instrument with only 1 research project. Example: A pathologist has a research project that he/she is doing say on Neuroblastoma. They have 150 blocks that they need a total of 10 IHC tests on each block. You take the 150 blocks and prepare 2 tissue micro array blocks and then run your 10 IHC test on the 2 TMA blocks. You have saved a great deal of money because you have 20 IHC tests instead of 1500 IHC tests that would have been needed without the tissue array blocks. At approximately $15 for each IHC test, you can see that $300 is much better than $22,500. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Friday, June 03, 2005 3:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue microarrays use?. . Can someone explain to me what the big deal is on tissue microarrays? I understand that they are many perfect circles of known diseased or normal tissues than can be used as a control for various applications. What are the applications for this stuff on routine or research that would make it worth it for us to either do it ourselves or contract it out? Can it be done in house cheap? _______________________________________________ Histonet mailing list Histonet@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 Luis.Chiriboga <@t> med.nyu.edu Fri Jun 3 17:24:47 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Tissue microarrays use?. . In-Reply-To: <5CB39BCA5724F349BCB748675C6CA1A202C7572B@SJMEMXMB02.stjude.sjcrh.local> Message-ID: Hi Joseph I have used commercial as well as built my own. There is plenty of literature discussing mostly the pros not to much on the drawbacks. I suggest you review some of the articles so that you can make an informed decision (I can email you some of the pdf files if you don't have access, email me offline). >From personal experience; financially, the technique can save you "boat loads" of money. From a technique perspective, having all the different tissues & controls in one block helps you normalize your technique. histopathologically, you are limiting what you see by arraying small cores. Scientifically, you need to be extremely careful how the data is analyzed and extrapolated, especially if your using clustering methods.... I hope this helps Luis ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Henry, Charlene Sent: Friday, June 03, 2005 4:43 PM To: Madary, Joseph; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Tissue microarrays use?. . They are greatly instrumental in research. Working at a research facility, we were able to pay for the purchase of the instrument with only 1 research project. Example: A pathologist has a research project that he/she is doing say on Neuroblastoma. They have 150 blocks that they need a total of 10 IHC tests on each block. You take the 150 blocks and prepare 2 tissue micro array blocks and then run your 10 IHC test on the 2 TMA blocks. You have saved a great deal of money because you have 20 IHC tests instead of 1500 IHC tests that would have been needed without the tissue array blocks. At approximately $15 for each IHC test, you can see that $300 is much better than $22,500. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Friday, June 03, 2005 3:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue microarrays use?. . Can someone explain to me what the big deal is on tissue microarrays? I understand that they are many perfect circles of known diseased or normal tissues than can be used as a control for various applications. What are the applications for this stuff on routine or research that would make it worth it for us to either do it ourselves or contract it out? Can it be done in house cheap? _______________________________________________ Histonet mailing list Histonet@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 jmy1967 <@t> comcast.net Fri Jun 3 18:22:12 2005 From: jmy1967 <@t> comcast.net (jmy1967@comcast.net) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Saturday work schedule Message-ID: <060320052322.28030.42A0E623000E795400006D7E2200734840C9C0C7CE970306@comcast.net> Dear Histonetters, We are considering adjusting our Saturday work schedule. Currently, two techs embed about 130 biopsy blocks and section and stain three slides for each. I'd like some information on how other high volume labs implement their Saturday schedule. Do you have a Saturday rotation? And if so, what's your work flow protocol, i.e. number of techs, type of specimens produced (biopsies and/or surgicals)? If your lab has two or more shifts, is the Saturday rotation equitable or does only day shift rotate on Saturdays? Many thanks for your responses! Jill From haldana <@t> unimoron.edu.ar Fri Jun 3 23:30:59 2005 From: haldana <@t> unimoron.edu.ar (Hernan Aldana) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Mouse anti-human androgen receptor Message-ID: <004201c568be$359ca360$9d25e818@b3w6zzmtb6juvbs> Dear Histonetters I need to work with Mouse anti-human androgen receptor from BD PharMingen applied in tissues of human testicles. Does anyone have any advices for me? Fixation, paraffin processing, antibody dilution, times, antigen recovering, etc. Thanks in advance Dr. Hern?n J. Aldana Marcos Secretario Acad?mico Facultad de Medicina. Universidad de Mor?n Machado 914. B1708JPD. Buenos Aires. Argentina e-mail alternativo hernanjavier@yahoo.com web:http://www.ht.org.ar/histologia/NUEVAS%20UNIDADES/home.htm From sulekhababy <@t> yahoo.co.uk Sat Jun 4 04:30:32 2005 From: sulekhababy <@t> yahoo.co.uk (Sulekha Ravi) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Shattering of sections. Message-ID: <20050604093033.75713.qmail@web25008.mail.ukl.yahoo.com> We had the same problem of microchattering of sections when we were using low profile disposable blades in our Leica automatic microtome. The height of this blade is only 8mm. and we had to use a small metallic rod like attachment at the bottom side of the blade to get the required height, to cut sections. We solved the problem of chattering, when we changed over to high profile disposable blade, which has a height of 14mm and there is no need of this attachment. --------------------------------- How much free photo storage do you get? Store your holiday snaps for FREE with Yahoo! Photos. Get Yahoo! Photos From lubbockcat <@t> hotmail.com Sat Jun 4 06:09:19 2005 From: lubbockcat <@t> hotmail.com (Terry Murphy) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Vision Bio Systems Message-ID: Does anyone on the histonet have any experience with Vision BioSystems. I am looking for any information from anyone who has dealt with the company and it's products, good or bad. Thank you Terry Murphy HTL (ASCP) _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From bmcmahill <@t> incytepathology.com Sat Jun 4 13:33:36 2005 From: bmcmahill <@t> incytepathology.com (Bonnie J. McMahill) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Qualifications to work in Histology Message-ID: Washington state does not have a requirement for licensure for histotechs - at this point. That is the only driving factor for qualified people filling these positions if a facility doesn't have a specific requirement. Our lab is a private reference lab and we have stated minimum qualifications for out positions that require HT or HTL's. We do have positions we can fill for "trainees" - which have requirements that match those in order for the individual to be certified after training for one year. Usually the trainees have a B.S. in Biology. Bonnie McMahill Histology Supervisor InCyte Pathology Spokane, WA > -----Original Message----- > From: Thweatt, John T [SMTP:John.Thweatt@med.va.gov] > Sent: Friday, June 03, 2005 11:52 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Qualifications to work in Histology > > Dear Fellow Histonetters, > Two posts in one day ought to be a record for me, but as demand > dictates, I have returned with yet another question. > This question may be directed more toward the Histology managers or > histologists who know a little about regulations in the group as I once > again as for your most valuable help. > I am inquiring on behalf of a friend and fellow Histologist who at the > moment does not have access to a computer, so pardon me if this isn't very > clear as this is coming to me second hand. > The question is: What are the individual state requirements for a > histology technician that is not certified? Is there such regulation that > exists within the states that would regulate hiring staff and what kind of > requirements would a person fall under in order to be able to be hired to > work in histology (non-certified). What does your state say about this > issue? > I would be interested to see what different states (or facilities) have > to say about the uncertified technician route as it got a little harder now > to get certified under ASCP. > If anyone has any facility type documentation that gives you guidance > as to who you can hire to work in histology, what kind of qualifications do > you require? Does your state or facility eventually push for certification > (surely)? Are there cities out there that lay the groundwork for what > qualifications are necessary to work in our field? > I do realize that this is a complex question that may not a simple > answer. If someone out there has the expertise in this could give me some > guidance on this topic it would be most appreciated. Thank you in advance. > > Sincerely, > > Tom Thweatt > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From billions <@t> public1.sz.js.cn Sat Jun 4 20:45:18 2005 From: billions <@t> public1.sz.js.cn (Sinoera Tech) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Re: BIOLOGICAL STAINS, DYES AND INDICATORS, DIAGNOSTICS Message-ID: <009601c56970$3d3711c0$a001a8c0@ming> Subject: Re: BIOLOGICAL STAINS, DYES AND INDICATORS, DIAGNOSTICS Dear Sirs, We manufacture following products for scientific research activities. BIOLOGICAL STAINS, DYES AND INDICATORS, DIAGNOSTICS Alcian Yellow Alcian Green Alcian Blue 8GX; Ingrain Blue 1; CI 74240 Alcian Blue, pyridine variant sulfobromophthalein sodium hydrate Thymolphthalein monophosphate magnesium salt Thymolphthalein monophosphate disodium salt o-Cresolphthalein monophosphate disodium salt o-Cresolphthalein monophosphate magnedium salt o-Cresolphthalein monophosphate di(cyclohexylammonium) salt other metal salts and other amine salts of o-Cresolphthalein monophosphoric acid phenolphthalein monophosphate bis(cyclohexylamine) salt 1-naphtholphthalein monophosphate derivatives Other phthalein monophosphoric acid and their salts Thymolphthalein complexone, and its metal salts sulfonated phenol(sulfo)phthalein series phthalein and fluorescein derivatives phthalein complexone dyes, and other phthalein, or complexone dyes 3',3'',5',5''-Tetrachlorophenol-3,4,5,6-tetrabromosulfophthalein Please contact us for custom manufacturing synthesis with greatful thanks. Kind Regards. - Minggeng Wang, Ph.D / President SUZHOU SINOERA CHEM CO., LTD. No.5508, 125 Binhe Road Suzhou New & Hi-Tech District 215011 China Fax: +86 512 68224995; +86 512 68258994 Tel: +86 1380 6217039 +86 512 68246939 E-mail: billions@public1.sz.js.cn From madbaza <@t> powerup.com.au Sun Jun 5 18:02:16 2005 From: madbaza <@t> powerup.com.au (Barry Madigan) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Vision Bio Systems In-Reply-To: Message-ID: <20050605230216.TTMS16131.smta05.mail.ozemail.net@WORKSTATION1> Hello Terry We have two of Visions Bond Immunostainers and a number of their Peloris tissue processors. Have had the Bonds for about 18 months now and the tissue processors almost a year. We have been very pleased with their service. Just send me an email on the info you require. Regards Barry Madigan Immunohistochemistry Queensland Health Pathology Services (QHPS) Royal Brisbane Hospital Australia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry Murphy Sent: Saturday, 4 June 2005 9:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Vision Bio Systems Does anyone on the histonet have any experience with Vision BioSystems. I am looking for any information from anyone who has dealt with the company and it's products, good or bad. Thank you Terry Murphy HTL (ASCP) _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From antje.marcantonio <@t> novartis.com Mon Jun 6 01:39:03 2005 From: antje.marcantonio <@t> novartis.com (antje.marcantonio@novartis.com) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] CD4, CD8 and Bcells Message-ID: Noelle, for B cells in rat FFPE sections we use the mouse anti-rat CD45R from Pharmingen. AR with citrate buffer needed. The antibody works well for us with a dilution of 1/5000 and overnight incubation. For CD4 (Serotec MCA55G) and CD8 (Serotec MCA48G) we use frozens. According to the datasheets these antibodies can be used as well for paraffin-embedded material after PLP fixation. Hope this helps somehow. Cheers, Antje Marcantonio Novartis Pharma AG BU Transplantation Research Basel, Switzerland Tel. +41 61 324 6730 antje.marcantonio@novartis.com From pengbw <@t> sjtu.edu.cn Mon Jun 6 03:19:45 2005 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] skeletal muscle regeneration marker? Message-ID: <20050606081945.4966611139CF@sjtu.edu.cn> Hi to all, Does anyone know if there is a marker for skeletal muscle regeneration, especialy for myoblast? Which company can I get it? Thanks in advance! Baowei Peng 1954 Huashan Road Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China Ph:86-21-62932108 E-mail:pengbw@sjtu.edu.cn From dsnider <@t> shrinenet.org Mon Jun 6 09:01:29 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Processing GMA Specimens Message-ID: Hello again everyone! You have all been very helpful and I need (yet again) to gather more information from everyone! As previously stated, I have little knowledge working with GMA specimens. It has been a learning experience for me! I process clinically engeineered skin samples. They are very small and thin. The program I use, (which was programmed into the RMC, EMP5160 when I took the job) is almost 5 hours long. The last 3 steps are 100% GMA preperation solution, each an hour long. After reviewing other protocols, this seems quite long for the samples I am working with. I would appreciate any protocols or suggestions you may have. You can email or fax them to me! Again, ALL help is greatly appreciated, and thank you in advance! Deanna Snider HT ASCP Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 513-872-6072 (fax) CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From tissuearray <@t> hotmail.com Mon Jun 6 09:55:34 2005 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Tissue Microarrays. In-Reply-To: <746FDB897740814EA52BDDCB5ED1DDBCBB1594@medimmune4.medimmune.com> Message-ID: Tissue Microarrays are not hard to do once you understand the process. I have several articles on constructing microarrays and I answer questions in email and phone often. You can go the my website and see a basic array created without using expensive instruments and other helpful information. arrayworkshop.com I hope this helps Thom Jensen >From: "Madary, Joseph" >To: >Subject: [Histonet] Tissue Microarrays. >Date: Fri, 3 Jun 2005 16:09:54 -0400 >MIME-Version: 1.0 >X-Originating-IP: [199.68.16.3] >Received: from swlx162.swmed.edu ([199.165.152.162]) by mc4-f27.hotmail.com with Microsoft SMTPSVC(6.0.3790.211); Fri, 3 Jun 2005 13:10:44 -0700 >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by swlx162.swmed.edu with esmtp (Exim 4.34)id 1DeIUi-0003Ni-UE; Fri, 03 Jun 2005 15:10:22 -0500 >Received: from [199.165.152.167] (helo=swlx167.swmed.edu)by swlx162.swmed.edu with esmtp (Exim 4.34) id 1DeIUR-0003NZ-GDfor histonet@lists.utsouthwestern.edu; Fri, 03 Jun 2005 15:10:04 -0500 >Received: from [216.82.255.51] (helo=mail64.messagelabs.com)by swlx167.swmed.edu with smtp (Exim 4.44) id 1DeIUP-0000hC-N5for histonet@lists.utsouthwestern.edu; Fri, 03 Jun 2005 15:09:59 -0500 >Received: (qmail 23381 invoked from network); 3 Jun 2005 20:09:56 -0000 >Received: from medimmune3.medimmune.com (HELO medimmune6.medimmune.com)(199.68.16.3) by server-5.tower-64.messagelabs.com with SMTP;3 Jun 2005 20:09:56 -0000 >Received: from medimmune4.medimmune.com ([10.10.100.4]) bymedimmune6.medimmune.com with Microsoft SMTPSVC(5.0.2195.6713); Fri, 3 Jun 2005 16:09:54 -0400 >X-Message-Info: gUeNUVfFqHAmHwmm5+Z7Ul7CSrCIdpLGNLZgBo+SnaE= >X-VirusChecked: Checked >X-Env-Sender: MadaryJ@MedImmune.com >X-Msg-Ref: server-5.tower-64.messagelabs.com!1117829395!47476931!1 >X-StarScan-Version: 5.4.15; banners=-,-,- >X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 >Content-Class: urn:content-classes:message >X-MS-Has-Attach: >X-MS-TNEF-Correlator: >Thread-Topic: [Histonet] Tissue Microarrays. >Thread-Index: AcVoeDUV7if1Nq7YSyyJ0nz2eO1vug== >X-OriginalArrivalTime: 03 Jun 2005 20:09:54.0870 (UTC)FILETIME=[35405560:01C56878] >X-Scan-Signature: 2f26e54c73378134ed276f0a1e9fcee8 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >Precedence: list >List-Id: For the exchange of information pertaining to histotechnology andrelated fields >List-Unsubscribe: , >List-Archive: >List-Post: >List-Help: >List-Subscribe: , >Errors-To: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: ef1ad7719a1154f9450b1c67eafa0a9b >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=5.5 tests=none autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >Return-Path: histonet-bounces@lists.utsouthwestern.edu > >What kind of response did you get on this? I have a pathologist who seems real hung up on contracting out TMA's to someone (actually in England, maybe you!). I am justm wondering from a routine standpoint and doing some pre-clinical work on rodents, what would be the benefit of us getting a system in place or contracting out this work? I mean I took a class in how to prepare them a few years back, bascially taking the specimens and putting them in spots on the block with the legend etc. I mean could one do this without fancy machinery, and what is the real use for this stuff? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From John.Thweatt <@t> med.va.gov Mon Jun 6 10:00:43 2005 From: John.Thweatt <@t> med.va.gov (Thweatt, John T) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] State requirements for HT Message-ID: <7A00E2723D55D311AC240000F831866802606F57@vhaamaexc1.amarillo.med.va.gov> Dear Fellow Histonetters, Does anyone know or heard of any "State" requirements for Histotechs to work in the lab setting without certification? Thanks in advance, Tom Thweatt From Charlene.Henry <@t> STJUDE.ORG Mon Jun 6 10:22:11 2005 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Tissue microarrays use?. . Message-ID: <5CB39BCA5724F349BCB748675C6CA1A202C7572C@SJMEMXMB02.stjude.sjcrh.local> Our pathologist review H&E slides of each block to be used in a tissue array and they determine which blocks are grouped together for each tissue array block. They also mark on the H&E slide the exact area they want punched. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: Katri Tuomala [mailto:katri@cogeco.ca] Sent: Friday, June 03, 2005 5:12 PM To: Henry, Charlene; Madary, Joseph; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Tissue microarrays use?. . Hi Charlene, Knowing that so many tumors can give a very heterogenous staining patterns ( some areas strongly positive and others weak or even negative ), how do you overcome this problem with microarrays? I would have a hard time trusting the results with such small samples. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Henry, Charlene" To: "Madary, Joseph" ; Sent: Friday, June 03, 2005 4:43 PM Subject: RE: [Histonet] Tissue microarrays use?. . They are greatly instrumental in research. Working at a research facility, we were able to pay for the purchase of the instrument with only 1 research project. Example: A pathologist has a research project that he/she is doing say on Neuroblastoma. They have 150 blocks that they need a total of 10 IHC tests on each block. You take the 150 blocks and prepare 2 tissue micro array blocks and then run your 10 IHC test on the 2 TMA blocks. You have saved a great deal of money because you have 20 IHC tests instead of 1500 IHC tests that would have been needed without the tissue array blocks. At approximately $15 for each IHC test, you can see that $300 is much better than $22,500. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Friday, June 03, 2005 3:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue microarrays use?. . Can someone explain to me what the big deal is on tissue microarrays? I understand that they are many perfect circles of known diseased or normal tissues than can be used as a control for various applications. What are the applications for this stuff on routine or research that would make it worth it for us to either do it ourselves or contract it out? Can it be done in house cheap? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Mon Jun 6 10:48:07 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:10 2005 Subject: FW: [Histonet] Tissue microarrays use?. . In-Reply-To: <009801c56889$3ecf7a00$6a9a9618@Katri> Message-ID: I think everyone has had very good comments about the use of microarrays, and I agree with all of them. I would like to add, that it is the usual practice to put more than 1 core from a block into an array. This somewhat addresses the problem with heterogeneity, but doesn't totally solve it. Just my 2 cents. Patti Loykasek PhenoPath Laboratories Seattle, WA ------ Forwarded Message From: "Katri Tuomala" Date: Fri, 3 Jun 2005 18:11:51 -0400 To: "Henry, Charlene" , "Madary, Joseph" , Subject: Re: [Histonet] Tissue microarrays use?. . Hi Charlene, Knowing that so many tumors can give a very heterogenous staining patterns ( some areas strongly positive and others weak or even negative ), how do you overcome this problem with microarrays? I would have a hard time trusting the results with such small samples. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Henry, Charlene" To: "Madary, Joseph" ; Sent: Friday, June 03, 2005 4:43 PM Subject: RE: [Histonet] Tissue microarrays use?. . They are greatly instrumental in research. Working at a research facility, we were able to pay for the purchase of the instrument with only 1 research project. Example: A pathologist has a research project that he/she is doing say on Neuroblastoma. They have 150 blocks that they need a total of 10 IHC tests on each block. You take the 150 blocks and prepare 2 tissue micro array blocks and then run your 10 IHC test on the 2 TMA blocks. You have saved a great deal of money because you have 20 IHC tests instead of 1500 IHC tests that would have been needed without the tissue array blocks. At approximately $15 for each IHC test, you can see that $300 is much better than $22,500. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Friday, June 03, 2005 3:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue microarrays use?. . Can someone explain to me what the big deal is on tissue microarrays? I understand that they are many perfect circles of known diseased or normal tissues than can be used as a control for various applications. What are the applications for this stuff on routine or research that would make it worth it for us to either do it ourselves or contract it out? Can it be done in house cheap? _______________________________________________ Histonet mailing list Histonet@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 Forwarded Message From Janet.Bonner <@t> FLHOSP.ORG Mon Jun 6 10:48:10 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Saturday work schedule Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42B3@fh2k093.fhmis.net> We have approximately the same workload and have two techs and a Lab Aide. One tech comes in around 1AM and embeds biopsies and as much Sat. work as possible, cuts the blocks and sp. stains if she has the time. The second Tech comes in around 3-4AM and cuts, does the special stains, and embeds the Monday work (we process all our tissue on Friday night and embed it Sat incase the Pathologist decides he wants a case slated for Monday). The Lab Aide comes in around 4-6AM and checks to make sure we have all the slated Sat. work, labels the slides, distributes the slides to the two Saturday Pathologists, and cleans up the stainer and processors when all the slides redistributed. The second tech is working until noon, and then we have an on-call person from noon until 4PM, and on Sunday on-call from 10AM to 4PM. We have a separate immuno dept. (- we do about 300,000 slides per year). Hope this helps - Janet --Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of jmy1967@comcast.net Sent: Friday, June 03, 2005 7:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Saturday work schedule Dear Histonetters, We are considering adjusting our Saturday work schedule. Currently, two techs embed about 130 biopsy blocks and section and stain three slides for each. I'd like some information on how other high volume labs implement their Saturday schedule. Do you have a Saturday rotation? And if so, what's your work flow protocol, i.e. number of techs, type of specimens produced (biopsies and/or surgicals)? If your lab has two or more shifts, is the Saturday rotation equitable or does only day shift rotate on Saturdays? Many thanks for your responses! Jill _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Mon Jun 6 11:04:58 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Saturday work schedule Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42B4@fh2k093.fhmis.net> I neglected the rest of your question! About the shift rotation. I'm the latest shift that applies to working Saturday - 10AM to 6:30PM. Obviously I have to take Fridays off to get up at 2AM on Saturday to make it by 3AM(I'm then the late Saturday person - until 12 noon) The next shift that does not apply to the Saturday rotation comes in at 3PM-11:30PM. The majority of our people work 10PM-6:30AM and 6AM-2:30PM. This shift covers early Saturdays and has Mondays off. (By "shift", we actually have our own hours skewed throughout the day, ie- I'm the only 10AMer.) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of jmy1967@comcast.net Sent: Friday, June 03, 2005 7:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Saturday work schedule Dear Histonetters, We are considering adjusting our Saturday work schedule. Currently, two techs embed about 130 biopsy blocks and section and stain three slides for each. I'd like some information on how other high volume labs implement their Saturday schedule. Do you have a Saturday rotation? And if so, what's your work flow protocol, i.e. number of techs, type of specimens produced (biopsies and/or surgicals)? If your lab has two or more shifts, is the Saturday rotation equitable or does only day shift rotate on Saturdays? Many thanks for your responses! Jill _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Frederick.Fifield <@t> sunhealth.org Mon Jun 6 11:11:09 2005 From: Frederick.Fifield <@t> sunhealth.org (Frederick.Fifield@sunhealth.org) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Qualifications to work in Histology Message-ID: Hello Tom I'm in Arizona and as far as I know there are no state regulations regarding the certification of histotechs, nor does my facility have any requirements in that regard although we give preference to those who are certified in those situations when we have a certified and non-certified applicant for an opening. Most labs where I have worked have non-certified techs. At the lab that I currently manage we have two certified techs and two non-certified techs. The non-certified techs are in the process of getting their certification. Most facilities in Phoenix would prefer that their techs are certified and certified techs will usually be hired before non-certified ones. Since there is no state requirement many non-certified techs in Arizona have never felt it necessary to challenge the exam because in the long run it has no effect on their pay or their ability to be hired. Our non-certified techs receive the same pay that a certified tech would receive. I have questioned this with our HR department and was told that wages are based on the ability to do the job and not of certification. If and when my non-certified techs become HT's they will not receive any additional pay for being certified. As long as non-certified techs have the ability to be hired and receive the same wages as certified techs this situation is not like to change soon in this area. I hope this helps to answer your questions. Fred Fifield BS, HT (ASCP) Pathology Section Manager Sun Health - Boswell Memorial Hospital (623) 876-5338 Dear Fellow Histonetters, Two posts in one day ought to be a record for me, but as demand dictates, I have returned with yet another question. This question may be directed more toward the Histology managers or histologists who know a little about regulations in the group as I once again as for your most valuable help. I am inquiring on behalf of a friend and fellow Histologist who at the moment does not have access to a computer, so pardon me if this isn't very clear as this is coming to me second hand. The question is: What are the individual state requirements for a histology technician that is not certified? Is there such regulation that exists within the states that would regulate hiring staff and what kind of requirements would a person fall under in order to be able to be hired to work in histology (non-certified). What does your state say about this issue? I would be interested to see what different states (or facilities) have to say about the uncertified technician route as it got a little harder now to get certified under ASCP. If anyone has any facility type documentation that gives you guidance as to who you can hire to work in histology, what kind of qualifications do you require? Does your state or facility eventually push for certification (surely)? Are there cities out there that lay the groundwork for what qualifications are necessary to work in our field? I do realize that this is a complex question that may not a simple answer. If someone out there has the expertise in this could give me some guidance on this topic it would be most appreciated. Thank you in advance. Sincerely, Tom Thweatt ******************************************************************************* The information contained in this transmission may be legally privileged and/or confidential information. Any dissemination, distribution or copying of this transmission by anyone other than the intended recipient is strictly prohibited. If you receive this in error, please inform the sender immediately and remove any record of this message. ******************************************************************************* For more information about Sun Health, visit us at: www.sunhealth.org From info <@t> instrumedics.com Mon Jun 6 11:34:39 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:25:10 2005 Subject: [Histonet] Cutting undecalcified bone References: <200506032138.j53LcWbm019799@chip.viawest.net> Message-ID: <008201c56ab5$ad674be0$6401a8c0@INSTRUMEDICS22> Patsy, I know from experience that many bone labs with the CryoJane system have difficulty in getting "perfect" sections and transferring them fully intact. However, many of them seem to find a technique for success. Dr. Rowe and his colleagues at UConn have been very successful with tape-transfer in cutting decalcified and undecalcified bone. I believe he has given a workshop(s) on his method. Perhaps you can contact Dr. Rowe's lab. Bernice Instrumedics ----- Original Message ----- From: "Patsy Ruegg" To: "'Stephen Peters M.D.'" ; Sent: Friday, June 03, 2005 5:38 PM Subject: RE: [Histonet] Cutting undecalcified bone >I have experience cutting frozen sections of whole rat tibias without decal > (John Tarpley and I did this work together) which in my experience could > only be done using the tape transfer technique. I use a D profile > tungsten > carbide knife(the bevel is only on one side unlike a C profile which has > the > same bevel on both sides, this is like the old triangle shaped glass > knives) > this knife requires a pretty straight cutting angle compared to the C > profile which is more angled, thus the reference Liz had about me changing > the knife angle. I have tried everything from heavily coated 6X to the > lightest 1/2X coated slides. I have still to find a technique which will > give me perfect sections of bone everytime. The problem for me is not > getting the perfect section cut but getting it attached to the coated > slide > without bubbles under the tape and then removing the tape without leaving > some of the section on the tape. This can be very frustrating and > expensive > to cut a beautiful section only to have it pull off with the tape, the > expensive slide is lost and your beautiful section is useless. > This is not only the case for undecalcified bone, I have been having the > same old problem with cornea implants. Some things to try include: try > to > cut a thinner section like 4 microns (if your knife is really sharp this > is > not a problem), the thinner section will lay flatter on the coated slide. > I > do expose the UV for 2-3 times with about 10 secs. Between or the fuse > will > blow. Pull the tape off diagnally from corner to corner very slowly and > smoothly. When appling the tape with the section to the coated slide > "role > the hell out of it" to borrow a phrase from Gayle, to make sure it gets > really well attached to the slide before UV. I have had some luck with > preparing the slide, exposing it to UV and then placing it on dryice for > 5-10 min. before removing the tape, but again I never get every section in > good shape and actually lately I have been getting more bad sections than > good, which is driving me crazy. There are people up at CSU using this to > cut horse carpal bone, I can give you their contact info if you like. I > think I did better with bone than I am doing with cornea right now. > Oh, for bone I try to cut as cold as possible, about -35 I have been > cutting > the softer tissue warmer -18---24, hey maybe that's my problem I should go > back to the cold cold bone method, it can't hurt at this point. > I snap freeze samples by surrounding them with OCT and either slowly > lowering into liq. Nitrogen or isopentane and dryice. > Patsy > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen > Peters M.D. > Sent: Thursday, June 02, 2005 10:18 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Cutting undecalcified bone > > Nancy, > > I have experience cutting frozens on undecacified bone specimens for > routine > surg path cases using conventional technique but have no experience with > the > tape transfer system. > I routinely use disposible low profile blades which in not optimum for > bone. I imagine a stronger high profile blade or knife will work better. > My > success has varied with the hardness of the bone. Trebecular bone coming > from older patients can cut fairly easily porvided the blade is sharp, and > the section is taken with a fluid continuous motion. The result is a > surprisingly intact section. The harder the bone ( typically the younger > the > patient ) there will be more damage to the blade in each pass. The > sections > will be streaked and splitting requiring one or more blade changes by time > I > get a resonable section. It is quite a juggling act. > Cutting hard cortical bone is often very frustrating and very difficult to > get any resonable section. From what you describe as bubbles, I am > guessing > you are getting sections of bone thicker than you are hoping for and as a > result when you roll it out, the thicker bone pieces are protecting the > rest > of the section from rolling. Often the first section of any tissue will be > considerably thicker than those that follow after a few fluid turns of the > wheel. If you are not letting a few pass your first section may be > considerably thicker and as a result will shatter more and create thicker > trabeculae which may be leading to your problem. > The equivalent using conventional technique will give me a section that my > coverslip will not lie flat against the slide and my mounting medium will > not spread. > I look foreward to other peoples experience on this subject. > > Stephen > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From DEllenburg2 <@t> stfrancishealth.org Mon Jun 6 13:07:18 2005 From: DEllenburg2 <@t> stfrancishealth.org (DEllenburg2@stfrancishealth.org) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] State requirements for an HT Message-ID: <0A502E8156DAA4468CB8979B2717755520613F@bssmsx5501.intranet.stfrancishealth.com> As far as I know we do not have any State regulations requiring an individual to be certified to work in the histology dept. However, there is a difference in the pay grade for certified and non-certified techs. We currently have one certified and 2 non-certified techs. Hope this helps. Debbie Ellenburg Supervisor Bon Secours St. Francis Health System 864-255-1582 -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: None To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 19, Issue 9 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: Vision Bio Systems (Barry Madigan) 2. Re: CD4, CD8 and Bcells (antje.marcantonio@novartis.com) 3. skeletal muscle regeneration marker? (Baowei Peng) 4. Processing GMA Specimens (Snider, Deanna) 5. RE: Tissue Microarrays. (Thom Jensen) 6. State requirements for HT (Thweatt, John T) 7. RE: Tissue microarrays use?. . (Henry, Charlene) 8. FW: [Histonet] Tissue microarrays use?. . (Patti Loykasek) 9. RE: Saturday work schedule (Bonner, Janet) 10. RE: Saturday work schedule (Bonner, Janet) 11. Qualifications to work in Histology (Frederick.Fifield@sunhealth.org) 12. Re: Cutting undecalcified bone (Instrumedics) ---------------------------------------------------------------------- Message: 1 Date: Mon, 6 Jun 2005 09:02:16 +1000 From: "Barry Madigan" Subject: RE: [Histonet] Vision Bio Systems To: "'Terry Murphy'" , Message-ID: <20050605230216.TTMS16131.smta05.mail.ozemail.net@WORKSTATION1> Content-Type: text/plain; charset="us-ascii" Hello Terry We have two of Visions Bond Immunostainers and a number of their Peloris tissue processors. Have had the Bonds for about 18 months now and the tissue processors almost a year. We have been very pleased with their service. Just send me an email on the info you require. Regards Barry Madigan Immunohistochemistry Queensland Health Pathology Services (QHPS) Royal Brisbane Hospital Australia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry Murphy Sent: Saturday, 4 June 2005 9:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Vision Bio Systems Does anyone on the histonet have any experience with Vision BioSystems. I am looking for any information from anyone who has dealt with the company and it's products, good or bad. Thank you Terry Murphy HTL (ASCP) _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Mon, 6 Jun 2005 08:39:03 +0200 From: antje.marcantonio@novartis.com Subject: Re: [Histonet] CD4, CD8 and Bcells To: "Linke_Noelle" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Noelle, for B cells in rat FFPE sections we use the mouse anti-rat CD45R from Pharmingen. AR with citrate buffer needed. The antibody works well for us with a dilution of 1/5000 and overnight incubation. For CD4 (Serotec MCA55G) and CD8 (Serotec MCA48G) we use frozens. According to the datasheets these antibodies can be used as well for paraffin-embedded material after PLP fixation. Hope this helps somehow. Cheers, Antje Marcantonio Novartis Pharma AG BU Transplantation Research Basel, Switzerland Tel. +41 61 324 6730 antje.marcantonio@novartis.com ------------------------------ Message: 3 Date: Mon, 6 Jun 2005 16:19:45 +0800 (BEIST) From: Baowei Peng Subject: [Histonet] skeletal muscle regeneration marker? To: Histonet@lists.utsouthwestern.edu Message-ID: <20050606081945.4966611139CF@sjtu.edu.cn> Content-Type: text/plain; charset="gb2312" Hi to all, Does anyone know if there is a marker for skeletal muscle regeneration, especialy for myoblast? Which company can I get it? Thanks in advance! Baowei Peng 1954 Huashan Road Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China Ph:86-21-62932108 E-mail:pengbw@sjtu.edu.cn ------------------------------ Message: 4 Date: Mon, 6 Jun 2005 10:01:29 -0400 From: "Snider, Deanna" Subject: [Histonet] Processing GMA Specimens To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="ISO-8859-1" Hello again everyone! You have all been very helpful and I need (yet again) to gather more information from everyone! As previously stated, I have little knowledge working with GMA specimens. It has been a learning experience for me! I process clinically engeineered skin samples. They are very small and thin. The program I use, (which was programmed into the RMC, EMP5160 when I took the job) is almost 5 hours long. The last 3 steps are 100% GMA preperation solution, each an hour long. After reviewing other protocols, this seems quite long for the samples I am working with. I would appreciate any protocols or suggestions you may have. You can email or fax them to me! Again, ALL help is greatly appreciated, and thank you in advance! Deanna Snider HT ASCP Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 513-872-6072 (fax) CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. ------------------------------ Message: 5 Date: Mon, 06 Jun 2005 14:55:34 +0000 From: "Thom Jensen" Subject: RE: [Histonet] Tissue Microarrays. To: MadaryJ@MedImmune.com, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format="flowed" Tissue Microarrays are not hard to do once you understand the process. I have several articles on constructing microarrays and I answer questions in email and phone often. You can go the my website and see a basic array created without using expensive instruments and other helpful information. arrayworkshop.com I hope this helps Thom Jensen >From: "Madary, Joseph" >To: >Subject: [Histonet] Tissue Microarrays. >Date: Fri, 3 Jun 2005 16:09:54 -0400 >MIME-Version: 1.0 >X-Originating-IP: [199.68.16.3] >Received: from swlx162.swmed.edu ([199.165.152.162]) by mc4-f27.hotmail.com with Microsoft SMTPSVC(6.0.3790.211); Fri, 3 Jun 2005 13:10:44 -0700 >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by swlx162.swmed.edu with esmtp (Exim 4.34)id 1DeIUi-0003Ni-UE; Fri, 03 Jun 2005 15:10:22 -0500 >Received: from [199.165.152.167] (helo=swlx167.swmed.edu)by swlx162.swmed.edu with esmtp (Exim 4.34) id 1DeIUR-0003NZ-GDfor histonet@lists.utsouthwestern.edu; Fri, 03 Jun 2005 15:10:04 -0500 >Received: from [216.82.255.51] (helo=mail64.messagelabs.com)by swlx167.swmed.edu with smtp (Exim 4.44) id 1DeIUP-0000hC-N5for histonet@lists.utsouthwestern.edu; Fri, 03 Jun 2005 15:09:59 -0500 >Received: (qmail 23381 invoked from network); 3 Jun 2005 20:09:56 -0000 >Received: from medimmune3.medimmune.com (HELO medimmune6.medimmune.com)(199.68.16.3) by server-5.tower-64.messagelabs.com with SMTP;3 Jun 2005 20:09:56 -0000 >Received: from medimmune4.medimmune.com ([10.10.100.4]) bymedimmune6.medimmune.com with Microsoft SMTPSVC(5.0.2195.6713); Fri, 3 Jun 2005 16:09:54 -0400 >X-Message-Info: gUeNUVfFqHAmHwmm5+Z7Ul7CSrCIdpLGNLZgBo+SnaE= >X-VirusChecked: Checked >X-Env-Sender: MadaryJ@MedImmune.com >X-Msg-Ref: server-5.tower-64.messagelabs.com!1117829395!47476931!1 >X-StarScan-Version: 5.4.15; banners=-,-,- >X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 >Content-Class: urn:content-classes:message >X-MS-Has-Attach: >X-MS-TNEF-Correlator: >Thread-Topic: [Histonet] Tissue Microarrays. >Thread-Index: AcVoeDUV7if1Nq7YSyyJ0nz2eO1vug== >X-OriginalArrivalTime: 03 Jun 2005 20:09:54.0870 (UTC)FILETIME=[35405560:01C56878] >X-Scan-Signature: 2f26e54c73378134ed276f0a1e9fcee8 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >Precedence: list >List-Id: For the exchange of information pertaining to histotechnology andrelated fields >List-Unsubscribe: , >List-Archive: >List-Post: >List-Help: >List-Subscribe: , >Errors-To: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: ef1ad7719a1154f9450b1c67eafa0a9b >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=5.5 tests=none autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >Return-Path: histonet-bounces@lists.utsouthwestern.edu > >What kind of response did you get on this? I have a pathologist who seems real hung up on contracting out TMA's to someone (actually in England, maybe you!). I am justm wondering from a routine standpoint and doing some pre-clinical work on rodents, what would be the benefit of us getting a system in place or contracting out this work? I mean I took a class in how to prepare them a few years back, bascially taking the specimens and putting them in spots on the block with the legend etc. I mean could one do this without fancy machinery, and what is the real use for this stuff? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 6 Jun 2005 08:00:43 -0700 From: "Thweatt, John T" Subject: [Histonet] State requirements for HT To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <7A00E2723D55D311AC240000F831866802606F57@vhaamaexc1.amarillo.med.va.gov> Content-Type: text/plain; charset="iso-8859-1" Dear Fellow Histonetters, Does anyone know or heard of any "State" requirements for Histotechs to work in the lab setting without certification? Thanks in advance, Tom Thweatt ------------------------------ Message: 7 Date: Mon, 6 Jun 2005 10:22:11 -0500 From: "Henry, Charlene" Subject: RE: [Histonet] Tissue microarrays use?. . To: "Katri Tuomala" , "Madary, Joseph" , Message-ID: <5CB39BCA5724F349BCB748675C6CA1A202C7572C@SJMEMXMB02.stjude.sjcrh.local> Content-Type: text/plain; charset="us-ascii" Our pathologist review H&E slides of each block to be used in a tissue array and they determine which blocks are grouped together for each tissue array block. They also mark on the H&E slide the exact area they want punched. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: Katri Tuomala [mailto:katri@cogeco.ca] Sent: Friday, June 03, 2005 5:12 PM To: Henry, Charlene; Madary, Joseph; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Tissue microarrays use?. . Hi Charlene, Knowing that so many tumors can give a very heterogenous staining patterns ( some areas strongly positive and others weak or even negative ), how do you overcome this problem with microarrays? I would have a hard time trusting the results with such small samples. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Henry, Charlene" To: "Madary, Joseph" ; Sent: Friday, June 03, 2005 4:43 PM Subject: RE: [Histonet] Tissue microarrays use?. . They are greatly instrumental in research. Working at a research facility, we were able to pay for the purchase of the instrument with only 1 research project. Example: A pathologist has a research project that he/she is doing say on Neuroblastoma. They have 150 blocks that they need a total of 10 IHC tests on each block. You take the 150 blocks and prepare 2 tissue micro array blocks and then run your 10 IHC test on the 2 TMA blocks. You have saved a great deal of money because you have 20 IHC tests instead of 1500 IHC tests that would have been needed without the tissue array blocks. At approximately $15 for each IHC test, you can see that $300 is much better than $22,500. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Friday, June 03, 2005 3:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue microarrays use?. . Can someone explain to me what the big deal is on tissue microarrays? I understand that they are many perfect circles of known diseased or normal tissues than can be used as a control for various applications. What are the applications for this stuff on routine or research that would make it worth it for us to either do it ourselves or contract it out? Can it be done in house cheap? _______________________________________________ Histonet mailing list Histonet@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: 8 Date: Mon, 06 Jun 2005 08:48:07 -0700 From: Patti Loykasek Subject: FW: [Histonet] Tissue microarrays use?. . To: histonet Message-ID: Content-Type: text/plain; charset="US-ASCII" I think everyone has had very good comments about the use of microarrays, and I agree with all of them. I would like to add, that it is the usual practice to put more than 1 core from a block into an array. This somewhat addresses the problem with heterogeneity, but doesn't totally solve it. Just my 2 cents. Patti Loykasek PhenoPath Laboratories Seattle, WA ------ Forwarded Message From: "Katri Tuomala" Date: Fri, 3 Jun 2005 18:11:51 -0400 To: "Henry, Charlene" , "Madary, Joseph" , Subject: Re: [Histonet] Tissue microarrays use?. . Hi Charlene, Knowing that so many tumors can give a very heterogenous staining patterns ( some areas strongly positive and others weak or even negative ), how do you overcome this problem with microarrays? I would have a hard time trusting the results with such small samples. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Henry, Charlene" To: "Madary, Joseph" ; Sent: Friday, June 03, 2005 4:43 PM Subject: RE: [Histonet] Tissue microarrays use?. . They are greatly instrumental in research. Working at a research facility, we were able to pay for the purchase of the instrument with only 1 research project. Example: A pathologist has a research project that he/she is doing say on Neuroblastoma. They have 150 blocks that they need a total of 10 IHC tests on each block. You take the 150 blocks and prepare 2 tissue micro array blocks and then run your 10 IHC test on the 2 TMA blocks. You have saved a great deal of money because you have 20 IHC tests instead of 1500 IHC tests that would have been needed without the tissue array blocks. At approximately $15 for each IHC test, you can see that $300 is much better than $22,500. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Friday, June 03, 2005 3:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue microarrays use?. . Can someone explain to me what the big deal is on tissue microarrays? I understand that they are many perfect circles of known diseased or normal tissues than can be used as a control for various applications. What are the applications for this stuff on routine or research that would make it worth it for us to either do it ourselves or contract it out? Can it be done in house cheap? _______________________________________________ Histonet mailing list Histonet@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 Forwarded Message ------------------------------ Message: 9 Date: Mon, 6 Jun 2005 11:48:10 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] Saturday work schedule To: "'jmy1967@comcast.net'" , histonet@lists.utsouthwestern.edu Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42B3@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 We have approximately the same workload and have two techs and a Lab Aide. One tech comes in around 1AM and embeds biopsies and as much Sat. work as possible, cuts the blocks and sp. stains if she has the time. The second Tech comes in around 3-4AM and cuts, does the special stains, and embeds the Monday work (we process all our tissue on Friday night and embed it Sat incase the Pathologist decides he wants a case slated for Monday). The Lab Aide comes in around 4-6AM and checks to make sure we have all the slated Sat. work, labels the slides, distributes the slides to the two Saturday Pathologists, and cleans up the stainer and processors when all the slides redistributed. The second tech is working until noon, and then we have an on-call person from noon until 4PM, and on Sunday on-call from 10AM to 4PM. We have a separate immuno dept. (- we do about 300,000 slides per year). Hope this helps - Janet --Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of jmy1967@comcast.net Sent: Friday, June 03, 2005 7:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Saturday work schedule Dear Histonetters, We are considering adjusting our Saturday work schedule. Currently, two techs embed about 130 biopsy blocks and section and stain three slides for each. I'd like some information on how other high volume labs implement their Saturday schedule. Do you have a Saturday rotation? And if so, what's your work flow protocol, i.e. number of techs, type of specimens produced (biopsies and/or surgicals)? If your lab has two or more shifts, is the Saturday rotation equitable or does only day shift rotate on Saturdays? Many thanks for your responses! Jill _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Mon, 6 Jun 2005 12:04:58 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] Saturday work schedule To: "'jmy1967@comcast.net'" , histonet@lists.utsouthwestern.edu Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42B4@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 I neglected the rest of your question! About the shift rotation. I'm the latest shift that applies to working Saturday - 10AM to 6:30PM. Obviously I have to take Fridays off to get up at 2AM on Saturday to make it by 3AM(I'm then the late Saturday person - until 12 noon) The next shift that does not apply to the Saturday rotation comes in at 3PM-11:30PM. The majority of our people work 10PM-6:30AM and 6AM-2:30PM. This shift covers early Saturdays and has Mondays off. (By "shift", we actually have our own hours skewed throughout the day, ie- I'm the only 10AMer.) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of jmy1967@comcast.net Sent: Friday, June 03, 2005 7:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Saturday work schedule Dear Histonetters, We are considering adjusting our Saturday work schedule. Currently, two techs embed about 130 biopsy blocks and section and stain three slides for each. I'd like some information on how other high volume labs implement their Saturday schedule. Do you have a Saturday rotation? And if so, what's your work flow protocol, i.e. number of techs, type of specimens produced (biopsies and/or surgicals)? If your lab has two or more shifts, is the Saturday rotation equitable or does only day shift rotate on Saturdays? Many thanks for your responses! Jill _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Mon, 6 Jun 2005 09:11:09 -0700 From: Frederick.Fifield@sunhealth.org Subject: [Histonet] Qualifications to work in Histology To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hello Tom I'm in Arizona and as far as I know there are no state regulations regarding the certification of histotechs, nor does my facility have any requirements in that regard although we give preference to those who are certified in those situations when we have a certified and non-certified applicant for an opening. Most labs where I have worked have non-certified techs. At the lab that I currently manage we have two certified techs and two non-certified techs. The non-certified techs are in the process of getting their certification. Most facilities in Phoenix would prefer that their techs are certified and certified techs will usually be hired before non-certified ones. Since there is no state requirement many non-certified techs in Arizona have never felt it necessary to challenge the exam because in the long run it has no effect on their pay or their ability to be hired. Our non-certified techs receive the same pay that a certified tech would receive. I have questioned this with our HR department and was told that wages are based on the ability to do the job and not of certification. If and when my non-certified techs become HT's they will not receive any additional pay for being certified. As long as non-certified techs have the ability to be hired and receive the same wages as certified techs this situation is not like to change soon in this area. I hope this helps to answer your questions. Fred Fifield BS, HT (ASCP) Pathology Section Manager Sun Health - Boswell Memorial Hospital (623) 876-5338 Dear Fellow Histonetters, Two posts in one day ought to be a record for me, but as demand dictates, I have returned with yet another question. This question may be directed more toward the Histology managers or histologists who know a little about regulations in the group as I once again as for your most valuable help. I am inquiring on behalf of a friend and fellow Histologist who at the moment does not have access to a computer, so pardon me if this isn't very clear as this is coming to me second hand. The question is: What are the individual state requirements for a histology technician that is not certified? Is there such regulation that exists within the states that would regulate hiring staff and what kind of requirements would a person fall under in order to be able to be hired to work in histology (non-certified). What does your state say about this issue? I would be interested to see what different states (or facilities) have to say about the uncertified technician route as it got a little harder now to get certified under ASCP. If anyone has any facility type documentation that gives you guidance as to who you can hire to work in histology, what kind of qualifications do you require? Does your state or facility eventually push for certification (surely)? Are there cities out there that lay the groundwork for what qualifications are necessary to work in our field? I do realize that this is a complex question that may not a simple answer. If someone out there has the expertise in this could give me some guidance on this topic it would be most appreciated. Thank you in advance. Sincerely, Tom Thweatt **************************************************************************** *** The information contained in this transmission may be legally privileged and/or confidential information. Any dissemination, distribution or copying of this transmission by anyone other than the intended recipient is strictly prohibited. If you receive this in error, please inform the sender immediately and remove any record of this message. **************************************************************************** *** For more information about Sun Health, visit us at: www.sunhealth.org ------------------------------ Message: 12 Date: Mon, 6 Jun 2005 12:34:39 -0400 From: "Instrumedics" Subject: Re: [Histonet] Cutting undecalcified bone To: "Patsy Ruegg" Cc: HistoNet Server Message-ID: <008201c56ab5$ad674be0$6401a8c0@INSTRUMEDICS22> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Patsy, I know from experience that many bone labs with the CryoJane system have difficulty in getting "perfect" sections and transferring them fully intact. However, many of them seem to find a technique for success. Dr. Rowe and his colleagues at UConn have been very successful with tape-transfer in cutting decalcified and undecalcified bone. I believe he has given a workshop(s) on his method. Perhaps you can contact Dr. Rowe's lab. Bernice Instrumedics ----- Original Message ----- From: "Patsy Ruegg" To: "'Stephen Peters M.D.'" ; Sent: Friday, June 03, 2005 5:38 PM Subject: RE: [Histonet] Cutting undecalcified bone >I have experience cutting frozen sections of whole rat tibias without decal > (John Tarpley and I did this work together) which in my experience could > only be done using the tape transfer technique. I use a D profile > tungsten > carbide knife(the bevel is only on one side unlike a C profile which has > the > same bevel on both sides, this is like the old triangle shaped glass > knives) > this knife requires a pretty straight cutting angle compared to the C > profile which is more angled, thus the reference Liz had about me changing > the knife angle. I have tried everything from heavily coated 6X to the > lightest 1/2X coated slides. I have still to find a technique which will > give me perfect sections of bone everytime. The problem for me is not > getting the perfect section cut but getting it attached to the coated > slide > without bubbles under the tape and then removing the tape without leaving > some of the section on the tape. This can be very frustrating and > expensive > to cut a beautiful section only to have it pull off with the tape, the > expensive slide is lost and your beautiful section is useless. > This is not only the case for undecalcified bone, I have been having the > same old problem with cornea implants. Some things to try include: try > to > cut a thinner section like 4 microns (if your knife is really sharp this > is > not a problem), the thinner section will lay flatter on the coated slide. > I > do expose the UV for 2-3 times with about 10 secs. Between or the fuse > will > blow. Pull the tape off diagnally from corner to corner very slowly and > smoothly. When appling the tape with the section to the coated slide > "role > the hell out of it" to borrow a phrase from Gayle, to make sure it gets > really well attached to the slide before UV. I have had some luck with > preparing the slide, exposing it to UV and then placing it on dryice for > 5-10 min. before removing the tape, but again I never get every section in > good shape and actually lately I have been getting more bad sections than > good, which is driving me crazy. There are people up at CSU using this to > cut horse carpal bone, I can give you their contact info if you like. I > think I did better with bone than I am doing with cornea right now. > Oh, for bone I try to cut as cold as possible, about -35 I have been > cutting > the softer tissue warmer -18---24, hey maybe that's my problem I should go > back to the cold cold bone method, it can't hurt at this point. > I snap freeze samples by surrounding them with OCT and either slowly > lowering into liq. Nitrogen or isopentane and dryice. > Patsy > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen > Peters M.D. > Sent: Thursday, June 02, 2005 10:18 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Cutting undecalcified bone > > Nancy, > > I have experience cutting frozens on undecacified bone specimens for > routine > surg path cases using conventional technique but have no experience with > the > tape transfer system. > I routinely use disposible low profile blades which in not optimum for > bone. I imagine a stronger high profile blade or knife will work better. > My > success has varied with the hardness of the bone. Trebecular bone coming > from older patients can cut fairly easily porvided the blade is sharp, and > the section is taken with a fluid continuous motion. The result is a > surprisingly intact section. The harder the bone ( typically the younger > the > patient ) there will be more damage to the blade in each pass. The > sections > will be streaked and splitting requiring one or more blade changes by time > I > get a resonable section. It is quite a juggling act. > Cutting hard cortical bone is often very frustrating and very difficult to > get any resonable section. From what you describe as bubbles, I am > guessing > you are getting sections of bone thicker than you are hoping for and as a > result when you roll it out, the thicker bone pieces are protecting the > rest > of the section from rolling. Often the first section of any tissue will be > considerably thicker than those that follow after a few fluid turns of the > wheel. If you are not letting a few pass your first section may be > considerably thicker and as a result will shatter more and create thicker > trabeculae which may be leading to your problem. > The equivalent using conventional technique will give me a section that my > coverslip will not lie flat against the slide and my mounting medium will > not spread. > I look foreward to other peoples experience on this subject. > > Stephen > _______________________________________________ > Histonet mailing list > Histonet@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 19, Issue 9 *************************************** The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at (864) 255-1000, and permanently delete the original e-mail, attachment(s), and any copies. From algranth <@t> u.arizona.edu Mon Jun 6 13:17:22 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] question on stains for plant histology Message-ID: <4.3.2.7.2.20050606102906.0b9b1d60@algranth.inbox.email.arizona.edu> Good Monday morning! I'm asked to do several stains on plant tissue this week and I have the protocols from Ruzin's book but have a few questions. A Safranin and Hematoxylin stain calls for differentiation in acidified 50% ETOH. Can somebody tell me what acid do you use to do this and do you just pH until the ETOH is acidic? The stain also calls for slightly acidified DI (doesn't say but I'm assuming DI water) to wash the slides after staining in Hematoxylin. Again - do I just add a few drops of acid - like glacial acetic to the DI water? Last - if anybody has ever done Flemming's Triple Stain can you contact me? 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 sinbody <@t> kumc.edu Mon Jun 6 13:33:41 2005 From: sinbody <@t> kumc.edu (Sarah Inbody) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] microglia in macaques Message-ID: Hi all-- I was wondering if anyone had done staining for microglia in macaque brains. I have an activation marker (LN3), but I would like a stain that's not dependent upon activation. I have tried the RCA-1 lectin (Vector), but it doesn't seem to be specific in macaque brains--all I get are endothelial cells and background. Thanks so much for your help! sarah inbody sinbody@kumc.edu From mcauliff <@t> umdnj.edu Mon Jun 6 13:50:17 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] microglia in macaques In-Reply-To: References: Message-ID: <42A49AE9.6050303@umdnj.edu> Hi Sarah: Try F4/80 or Mac1. I suspect a quick lit search on PubMed would turn up someone working on microglia in non-human primates. Geoff Sarah Inbody wrote: >Hi all-- > >I was wondering if anyone had done staining for microglia in macaque >brains. I have an activation marker (LN3), but I would like a stain >that's not dependent upon activation. I have tried the RCA-1 lectin >(Vector), but it doesn't seem to be specific in macaque brains--all I >get are endothelial cells and background. > >Thanks so much for your help! >sarah inbody >sinbody@kumc.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From doninn <@t> mail.nih.gov Mon Jun 6 15:19:23 2005 From: doninn <@t> mail.nih.gov (Donin, Nick (NIH/NCI)) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] LFB with Vector ABC kits Message-ID: <16A0583FB1644E4DB8C0A0265028B6FD02B4E74A@nihexchange13.nih.gov> I am trying to stain using LFB in combination with the Vector ABC Staining kits. For the Vector Kit staining, I am using a biotin-conjugated secondary antibody, which then conjugates with an avidin-peroxidase molecule which reacts with a DAB substrate. Does anyone know if these two methods are compatible? If so, is it preferable to do either one first? Additionally, my lab has run out of Lithium Carbonate for the LFB protocol, can I substitute with Lithium Chloride? I'm trying to stain formalin fixed, paraffin embedded mouse brain. Thanks for your help. Email me any questions or suggestions you have. Thanks. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 From liz <@t> premierlab.com Mon Jun 6 15:43:00 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] LFB with Vector ABC kits In-Reply-To: <16A0583FB1644E4DB8C0A0265028B6FD02B4E74A@nihexchange13.nih.gov> Message-ID: <000001c56ad8$5769f850$a7d48a80@AMY> Nick I have done Immunohistochemical staining with DAB as a chromagen and then performed an acid fast stain on top of it. It worked just fine. If you want to see an image there is on our web page. I would not think that staining with LFB after the immuno would be a problem as long as you use DAB for a chromagen. There is a nice article in Histologic (I don't know what issue, maybe Vinnie will know) which demonstrated a method for immunohistochemistry and PAS staining in kidney sections. You just need to go the sakurausa.com and search the older issues of that publication. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Donin, Nick (NIH/NCI) Sent: Monday, June 06, 2005 1:19 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] LFB with Vector ABC kits I am trying to stain using LFB in combination with the Vector ABC Staining kits. For the Vector Kit staining, I am using a biotin-conjugated secondary antibody, which then conjugates with an avidin-peroxidase molecule which reacts with a DAB substrate. Does anyone know if these two methods are compatible? If so, is it preferable to do either one first? Additionally, my lab has run out of Lithium Carbonate for the LFB protocol, can I substitute with Lithium Chloride? I'm trying to stain formalin fixed, paraffin embedded mouse brain. Thanks for your help. Email me any questions or suggestions you have. Thanks. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cmconway <@t> usgs.gov Mon Jun 6 16:18:04 2005 From: cmconway <@t> usgs.gov (Carla M Conway) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] stain needed to dye external pores Message-ID: Hello all, A colleague has some juvenile trout (4-8 inches long) that have been fixed in 10% NBF then stored in ethanol. He would like to count the mandibular pores located on the underside of the lower jaw. Can anyone suggest a stain or procedure which would make these external pores more visible? They would prefer to stain the area directly and not process/paraffin embed, etc.. Any suggestions will be greatly appreciated. Carla Conway Histologist Western Fisheries Research Center 6505 NE 65th St. Seattle, WA 98115 ph: 206-526-6282 ext 242 fax: 206-526-6654 cmconway@usgs.gov From peoshel <@t> wisc.edu Mon Jun 6 16:49:04 2005 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] stain needed to dye external pores In-Reply-To: References: Message-ID: Carla, I used to do this with sculpins from Lake Michigan. Mostly, though we didn't need to do anything to see the lateral line pores. But! it's not hard to make a cast of the canal and it's pores. Just take a blunt syringe needle (aka "90 deg tip") and inject any of several different things into the canal: Bateson's or Mercox methylmethacrylate, or the MMA or GMA resins work, but silicone bathtub chaulk also works. These all give the LL pores and impressions of the neuromasts within the canal. India ink can be used for a temporary fill. One problem with staining the LL canals is that the canals are usually filled with mucus, typically somewhat thicker than that covering the fish. One reason the injection methods work is that they flush out the mucus. But ... fixed in formalin and stored in EtOH ... this may well cause problems getting stuff into the canal. Phil >Hello all, > >A colleague has some juvenile trout (4-8 inches long) that have been fixed >in 10% NBF then stored in ethanol. He would like to count the mandibular >pores located on the underside of the lower jaw. Can anyone suggest a stain >or procedure which would make these external pores more visible? They would >prefer to stain the area directly and not process/paraffin embed, etc.. Any >suggestions will be greatly appreciated. > >Carla Conway > >Histologist >Western Fisheries Research Center >6505 NE 65th St. >Seattle, WA 98115 >ph: 206-526-6282 ext 242 >fax: 206-526-6654 >cmconway@usgs.gov > > > >_______________________________________________ >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 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm From anh2006 <@t> med.cornell.edu Mon Jun 6 16:54:27 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Need a chemist's opinion on formalin fixation reversal In-Reply-To: <200506031932.j53JW5bm009022@chip.viawest.net> References: <200506031932.j53JW5bm009022@chip.viawest.net> Message-ID: Can one of the chemists on board please address the issue of washing to reverse formalin fixation? (sorry if you have done so already but I must have missed it). This is a new concept to me and makes me very very nervous about some of my routines in the lab as often times I can only occasionally process and have samples in the fridge for sometime after fixation yet before processing ... yikes! Thank you thank you! Andrea >I have been asked to elaborate on my reference to washing tissues to reverse >the effects of formalin fixation iliminating the possible need for AR. >Here goes: > > >I found the reference to Washing in a book called Introduction to >Immunocytochemistry 3rd Edition J.M. Polak and S. Van Noorden pg. 24- 3.8.2 >"The simplest form of reversing the effects of formalin is to wash the >tissue well before processing, but this is not usually possible in >histopathology laboratories, where rapid turnowver of specimens is >required." > >I have not tested this extensively but have done a little, especially on >over fixed samples, the data is yet to be confirmed but I am looking into it >as we speak. I will keep you all posted. > >Patsy -- From lpwenk <@t> sbcglobal.net Mon Jun 6 16:59:56 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Tissue Microarrays. In-Reply-To: Message-ID: If it's any help, there will be an NSH teleconference on tissue microarrays on Wed. June 15, 2005, at 1 pm EDST. If interested, go to: http://www.nsh.org Click on Education Click on Teleconference Call IMMEDIATELY, as the handouts, slides, etc. have already been mailed. The following is the abstract. Tissue Microarrays Tissue microarray has emerged as a great breakthrough in the field of histotechnology. This technique allows for multiple patients to be reviewed on the same slide by a pathologist with the aid of a computer. Once the tissue microarray is constructed and sectioned, a wide range of staining techniques can be performed, such as IHC, IF, ISH, special stains and even QC controls for H&E stains. A practical histological approach and method will be discussed, to present the purpose, design, block selection, array construction and sectioning of tissue microarrays. Presenters: Wanda Jones, HT(ASCP) & Paul Billings, University of Alabama, Birmingham, AL (I am the NSH Teleconference Coordinator, but, no, I don't get paid to do this. It's volunteer.) Peggy Wenk William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Thom Jensen Sent: Monday, June 06, 2005 10:56 AM To: MadaryJ@MedImmune.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Tissue Microarrays. Tissue Microarrays are not hard to do once you understand the process. I have several articles on constructing microarrays and I answer questions in email and phone often. You can go the my website and see a basic array created without using expensive instruments and other helpful information. arrayworkshop.com I hope this helps Thom Jensen >From: "Madary, Joseph" >To: >Subject: [Histonet] Tissue Microarrays. >Date: Fri, 3 Jun 2005 16:09:54 -0400 >MIME-Version: 1.0 >X-Originating-IP: [199.68.16.3] >Received: from swlx162.swmed.edu ([199.165.152.162]) by mc4-f27.hotmail.com with Microsoft SMTPSVC(6.0.3790.211); Fri, 3 Jun 2005 13:10:44 -0700 >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by swlx162.swmed.edu with esmtp (Exim 4.34)id 1DeIUi-0003Ni-UE; Fri, 03 Jun 2005 15:10:22 -0500 >Received: from [199.165.152.167] (helo=swlx167.swmed.edu)by swlx162.swmed.edu with esmtp (Exim 4.34) id 1DeIUR-0003NZ-GDfor histonet@lists.utsouthwestern.edu; Fri, 03 Jun 2005 15:10:04 -0500 >Received: from [216.82.255.51] (helo=mail64.messagelabs.com)by swlx167.swmed.edu with smtp (Exim 4.44) id 1DeIUP-0000hC-N5for histonet@lists.utsouthwestern.edu; Fri, 03 Jun 2005 15:09:59 -0500 >Received: (qmail 23381 invoked from network); 3 Jun 2005 20:09:56 -0000 >Received: from medimmune3.medimmune.com (HELO medimmune6.medimmune.com)(199.68.16.3) by server-5.tower-64.messagelabs.com with SMTP;3 Jun 2005 20:09:56 -0000 >Received: from medimmune4.medimmune.com ([10.10.100.4]) bymedimmune6.medimmune.com with Microsoft SMTPSVC(5.0.2195.6713); Fri, 3 Jun 2005 16:09:54 -0400 >X-Message-Info: gUeNUVfFqHAmHwmm5+Z7Ul7CSrCIdpLGNLZgBo+SnaE= >X-VirusChecked: Checked >X-Env-Sender: MadaryJ@MedImmune.com >X-Msg-Ref: server-5.tower-64.messagelabs.com!1117829395!47476931!1 >X-StarScan-Version: 5.4.15; banners=-,-,- >X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 >Content-Class: urn:content-classes:message >X-MS-Has-Attach: >X-MS-TNEF-Correlator: >Thread-Topic: [Histonet] Tissue Microarrays. >Thread-Index: AcVoeDUV7if1Nq7YSyyJ0nz2eO1vug== >X-OriginalArrivalTime: 03 Jun 2005 20:09:54.0870 (UTC)FILETIME=[35405560:01C56878] >X-Scan-Signature: 2f26e54c73378134ed276f0a1e9fcee8 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >Precedence: list >List-Id: For the exchange of information pertaining to histotechnology andrelated fields >List-Unsubscribe: , >List-Archive: >List-Post: >List-Help: >List-Subscribe: , >Errors-To: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: ef1ad7719a1154f9450b1c67eafa0a9b >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=5.5 tests=none autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >Return-Path: histonet-bounces@lists.utsouthwestern.edu > >What kind of response did you get on this? I have a pathologist who seems real hung up on contracting out TMA's to someone (actually in England, maybe you!). I am justm wondering from a routine standpoint and doing some pre-clinical work on rodents, what would be the benefit of us getting a system in place or contracting out this work? I mean I took a class in how to prepare them a few years back, bascially taking the specimens and putting them in spots on the block with the legend etc. I mean could one do this without fancy machinery, and what is the real use for this stuff? > >_______________________________________________ >Histonet mailing list >Histonet@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 Luis.Chiriboga <@t> med.nyu.edu Mon Jun 6 11:11:56 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Tissue microarrays use?. . In-Reply-To: <5CB39BCA5724F349BCB748675C6CA1A202C7572C@SJMEMXMB02.stjude.sjcrh.local> Message-ID: Katri, your correct, "sampling" is one of the major issues of the technique. Most importantly is to design the microarray such that you have multiple cores from different areas of the same block. Design and adequate sampling are the key see the following references: Nocito, A. et al 2001: Microarrays of bladder cancer tissue are highly representative of proliferation index and histological grade. J Pathol 194(3) 349-57 Rubin, M.A. et al 2002: Tissue microarray sampling strategy for prostate cancer biomarker analysis. Am J Surg Path 26(3) 312-9 we routinely put 4 cores/block, and if indicated we have put as many as 8 from the same block. Even if the requires creating additional blocks, you still get the cost benefit from the reduced volume of samples... LC>>> ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Henry, Charlene Sent: Monday, June 06, 2005 11:22 AM To: Katri Tuomala; Madary, Joseph; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Tissue microarrays use?. . Our pathologist review H&E slides of each block to be used in a tissue array and they determine which blocks are grouped together for each tissue array block. They also mark on the H&E slide the exact area they want punched. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: Katri Tuomala [mailto:katri@cogeco.ca] Sent: Friday, June 03, 2005 5:12 PM To: Henry, Charlene; Madary, Joseph; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Tissue microarrays use?. . Hi Charlene, Knowing that so many tumors can give a very heterogenous staining patterns ( some areas strongly positive and others weak or even negative ), how do you overcome this problem with microarrays? I would have a hard time trusting the results with such small samples. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Henry, Charlene" To: "Madary, Joseph" ; Sent: Friday, June 03, 2005 4:43 PM Subject: RE: [Histonet] Tissue microarrays use?. . They are greatly instrumental in research. Working at a research facility, we were able to pay for the purchase of the instrument with only 1 research project. Example: A pathologist has a research project that he/she is doing say on Neuroblastoma. They have 150 blocks that they need a total of 10 IHC tests on each block. You take the 150 blocks and prepare 2 tissue micro array blocks and then run your 10 IHC test on the 2 TMA blocks. You have saved a great deal of money because you have 20 IHC tests instead of 1500 IHC tests that would have been needed without the tissue array blocks. At approximately $15 for each IHC test, you can see that $300 is much better than $22,500. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Friday, June 03, 2005 3:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue microarrays use?. . Can someone explain to me what the big deal is on tissue microarrays? I understand that they are many perfect circles of known diseased or normal tissues than can be used as a control for various applications. What are the applications for this stuff on routine or research that would make it worth it for us to either do it ourselves or contract it out? Can it be done in house cheap? _______________________________________________ Histonet mailing list Histonet@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 Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue Jun 7 02:36:42 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] CD 138 Message-ID: Has anyone tried to get CD 138:- clone 5F7 (Vector), clone B-B4 (Lab Vision) or clone MI 15 (Dako) to work on a Ventana Benchmark XT? Tried Vector's but no luck so far - it may just not like that particular clone. Don't want to purchase any more antisera until I know which one works well. Would appreciate your info. Jacqui Malam Royal Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue Jun 7 02:38:52 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Processing with vacuum and/or pressure Message-ID: Would tissue processor users please let me know whether it is believed best to use vacuum only or both vacuum and pressure when processing, especially with blocks of fatty tissues eg breast. We use two Shandon Pathcentres. I was recommended to use vacuum only but we do get breast blocks which still aren't processed in the centre despite using heat with the processing reagents. Thanks Jacqui Malam Royal Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From KPercival <@t> wyeth.com Tue Jun 7 06:32:39 2005 From: KPercival <@t> wyeth.com (Karen Percival) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Processing with vacuum and/or pressure Message-ID: Jacqui, The best process I found for fatty tissues such as breast was to add an additional absolute alcohol step and to increase the times on each (3 in total). The breast tissues process and cut wonderfully, and we rarely have unfixed tissues or tissues fallilng off of the slides even for IHC with HIER. Karen Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com >>> Malam Jacqueline 6/7/2005 3:38:52 AM >>> Would tissue processor users please let me know whether it is believed best to use vacuum only or both vacuum and pressure when processing, especially with blocks of fatty tissues eg breast. We use two Shandon Pathcentres. I was recommended to use vacuum only but we do get breast blocks which still aren't processed in the centre despite using heat with the processing reagents. Thanks Jacqui Malam Royal Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue Jun 7 06:59:40 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Processing Message-ID: Hi Karen Couldn't get through to your e-mail address to answer personally. We use a 70% - I hr, 90% - 1 hr and 3 100% IMS steps (11/2, 1, 11/2hrs each). Do you use just vacuum or pressure as well, and what time in each step? Cheers Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From mbecker <@t> pathlabinc.com Tue Jun 7 09:12:23 2005 From: mbecker <@t> pathlabinc.com (Michelle Becker) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Processing with mesh cassettes Message-ID: We recently started using mesh cassettes for small biopsies, also called micromesh, histoscreen or micro biopsy cassettes, depending on the manufacturer. The techs, as well the pathologists, love them. Unfortunately, it has caused reagent carryover in our tissue processors (VIP 3000 & Leica TP1050)after only one run. The first reagent level (formalin)decreases significantly; the 2nd reagent level(Prefer), as well the next 3 alcohol levels, have significant increases. The tissue processor used does not seem to make a difference. We run a 4 hour short program with vacuum for biopsies. Has anyone else had this problem and if so, how did you resolve it? I really hate to stop using something the techs and pathologists love and actually agree on! Thank-you. Michele Becker, HTL(ASCP) Histology Manager Laboratory Corporation of America Portsmouth, NH From funderwood <@t> mcohio.org Tue Jun 7 09:41:27 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Processing with mesh cassettes Message-ID: Hi Michelle, I've not attempted this. Perhaps if you exposed the cassettes to a solution to reduce the surface tension of the screens. Possibly putting a small amount of a detergent in the first formalin. Or dipping the cassettes in a detergent solution before placing them on the processor. Fred >>> "Michelle Becker" 06/07/05 10:12AM >>> We recently started using mesh cassettes for small biopsies, also called micromesh, histoscreen or micro biopsy cassettes, depending on the manufacturer. The techs, as well the pathologists, love them. Unfortunately, it has caused reagent carryover in our tissue processors (VIP 3000 & Leica TP1050)after only one run. The first reagent level (formalin)decreases significantly; the 2nd reagent level(Prefer), as well the next 3 alcohol levels, have significant increases. The tissue processor used does not seem to make a difference. We run a 4 hour short program with vacuum for biopsies. Has anyone else had this problem and if so, how did you resolve it? I really hate to stop using something the techs and pathologists love and actually agree on! Thank-you. Michele Becker, HTL(ASCP) Histology Manager Laboratory Corporation of America Portsmouth, NH _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Tue Jun 7 09:57:29 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] LFB with Vector ABC kits In-Reply-To: <16A0583FB1644E4DB8C0A0265028B6FD02B4E74A@nihexchange13.nih.gov> References: <16A0583FB1644E4DB8C0A0265028B6FD02B4E74A@nihexchange13.nih.gov> Message-ID: <42A5B5D9.8080005@umdnj.edu> Hi Nick: You may have to 'do the experiment' yourself to answer the stain compatibility question. Vector's "Intense" reagent makes DAB a very nice gold color, should look nice with LFB. I have had excellent results with "Intense". I suspect that it is a gold cholride-based intensifer. As for lithium carbonate, I as pretty sure that it is the carbonate (ie alkalinity) you want, not the lithium, so try something with one (more?) CO3 ion. Years ago one of the grad students in the anatomy dept. at U. of Louisville med school did some testing on lithium versus other alkalies, I don't remember exactly what she found. Geoff Donin, Nick (NIH/NCI) wrote: >I am trying to stain using LFB in combination with the Vector ABC Staining >kits. For the Vector Kit staining, I am using a biotin-conjugated secondary >antibody, which then conjugates with an avidin-peroxidase molecule which >reacts with a DAB substrate. Does anyone know if these two methods are >compatible? If so, is it preferable to do either one first? Additionally, >my lab has run out of Lithium Carbonate for the LFB protocol, can I >substitute with Lithium Chloride? I'm trying to stain formalin fixed, >paraffin embedded mouse brain. Thanks for your help. Email me any >questions or suggestions you have. Thanks. > > > >Nick Donin > >CRTA > >Neuro-Oncology Branch > >National Cancer Institute > >National Institutes of Health > >9000 Rockville Pike > >Building 35, Room 2B-203 > >Bethesda, MD 20892 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** 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 pmarcum <@t> vet.upenn.edu Tue Jun 7 10:02:08 2005 From: pmarcum <@t> vet.upenn.edu (pmarcum@vet.upenn.edu) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Processing with mesh cassettes In-Reply-To: References: Message-ID: <1118156528.42a5b6f05d321@imp.vet.upenn.edu> Be very careful adding detergent to your fixative or dipping cassettes in it. The result can dissolve the lipids in the cell walls and cause very serious problems with the microscopic examination. The cytoplasm can become very indistinct and difficult to understand in context of the tissue. The tissue processor manufacturers don't usually like detergent in the systems as it can cause foaming through out the plumbing with air pockets. Pam Marcum Quoting Fred Underwood : > Hi Michelle, > > I've not attempted this. Perhaps if you exposed the cassettes to a > solution to reduce the surface tension of the screens. Possibly putting > a small amount of a detergent in the first formalin. Or dipping the > cassettes in a detergent solution before placing them on the processor. > > Fred > > >>> "Michelle Becker" 06/07/05 10:12AM >>> > We recently started using mesh cassettes for small biopsies, also > called > micromesh, histoscreen or micro biopsy cassettes, depending on the > manufacturer. The techs, as well the pathologists, love them. > Unfortunately, it has caused reagent carryover in our tissue processors > (VIP > 3000 & Leica TP1050)after only one run. The first reagent level > (formalin)decreases significantly; the 2nd reagent level(Prefer), as > well > the next 3 alcohol levels, have significant increases. The tissue > processor > used does not seem to make a difference. We run a 4 hour short program > with > vacuum for biopsies. Has anyone else had this problem and if so, how > did you > resolve it? I really hate to stop using something the techs and > pathologists love and actually agree on! > Thank-you. > > Michele Becker, HTL(ASCP) > Histology Manager > Laboratory Corporation of America > Portsmouth, NH > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@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 Tue Jun 7 10:03:00 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] CD 138 Message-ID: I use Cell Marque's at 1:50 with standard CC1, 32min incubation. Their clone is B-B4. Hope this helps. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malam Jacqueline Sent: Tuesday, June 07, 2005 2:37 AM To: Histonet submissions (histonet@lists.utsouthwestern.edu) Subject: [Histonet] CD 138 Has anyone tried to get CD 138:- clone 5F7 (Vector), clone B-B4 (Lab Vision) or clone MI 15 (Dako) to work on a Ventana Benchmark XT? Tried Vector's but no luck so far - it may just not like that particular clone. Don't want to purchase any more antisera until I know which one works well. Would appreciate your info. Jacqui Malam Royal Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bwhitaker <@t> brownpathology.com Tue Jun 7 10:14:26 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Processing with mesh cassettes In-Reply-To: Message-ID: <000001c56b73$986b3120$3601a8c0@brownpathology.net> On our processor (Thermo's Excelsior), it was a very simple matter to increase the drain times in between each reagent. That did the trick for us. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michelle Becker Sent: Tuesday, June 07, 2005 9:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Processing with mesh cassettes We recently started using mesh cassettes for small biopsies, also called micromesh, histoscreen or micro biopsy cassettes, depending on the manufacturer. The techs, as well the pathologists, love them. Unfortunately, it has caused reagent carryover in our tissue processors (VIP 3000 & Leica TP1050)after only one run. The first reagent level (formalin)decreases significantly; the 2nd reagent level(Prefer), as well the next 3 alcohol levels, have significant increases. The tissue processor used does not seem to make a difference. We run a 4 hour short program with vacuum for biopsies. Has anyone else had this problem and if so, how did you resolve it? I really hate to stop using something the techs and pathologists love and actually agree on! Thank-you. Michele Becker, HTL(ASCP) Histology Manager Laboratory Corporation of America Portsmouth, NH _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DonnaWillis <@t> texashealth.org Tue Jun 7 10:39:55 2005 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:25:11 2005 Subject: FW: [Histonet] Electron Microscopy Workload Message-ID: <4483B6EE0FA82A4EAED2B1E161E5E41147C861@ftwex02.txhealth.org> Are there other EM labs performing TEM similarly to Tim's processing requiring their staff to produce more than 200 cases a year for 1 FTE? We are currently still doing things the way he did but on 95% kidney. 200 has been the target in my lab for 1FTE. I am wanting to put this in a graph for CAP. I know there are procedures that could be automated (ex. Processing) to allow workload increases but currently we are still do a lot of manual methods and that is what I would like to benchmark. Thanks, Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Friday, June 03, 2005 1:35 PM To: Willis, Donna Subject: RE: [Histonet] Electron Microscopy Workload Donna, I did em for 11 years and had about 200 cases per year. This was about 70 percent kidney and 30 percent tumor. I did all the processing, cutting, staining, initial screening, negative development and made enlargments. I also attended most of the kidney biopsies to get the tissue. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Willis, Donna Sent: Friday, June 03, 2005 11:06 AM To: HistoNet Subject: [Histonet] Electron Microscopy Workload What is the average workload for a TEM technician? How many cases a year should 1 FTE perform? Thanks, Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx 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. _______________________________________________ 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 jqb7 <@t> cdc.gov Tue Jun 7 10:30:53 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] job at CDC Message-ID: All: We have an opening in our lab here at CDC and will be posting the position in the near future. As soon as it is officially listed I will send out the information for formal application. If anyone would like to go ahead and send a CV for review that would be acceptable. The position is in is the Infectious Disease Pathology Activity lab in the Division of Viral and Rickettsial Diseases which is part of the National Center for Infectious Diseases. Basically we would like someone with a strong histology background and immuno. knowledge would be a big plus. The CVs can be mailed to: Lisa Harper, Health Scientist 1600 Clifton Rd. MailStop G-32 Atlanta, GA 30333 (404) 639-3131 Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 (404) 639-3590 From fbolognani <@t> salud.unm.edu Tue Jun 7 10:41:25 2005 From: fbolognani <@t> salud.unm.edu (Federico Bolognani) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] methacarm and luxol fast blue Message-ID: Does anybody knows if Luxol Fast Blue staining works in samples fixed with Methacarm? Thanks Federico Bolognani, MD, PhD Research Assistant Professor Department of Cell Biology and Physiology University of New Mexico MSC08 4750 1 University of New Mexico Albuquerque NM 87131-0001 USA Phone: (505) 272-4010 Fax: (505) 272-9105 From Janet.Bonner <@t> FLHosp.org Tue Jun 7 10:48:12 2005 From: Janet.Bonner <@t> FLHosp.org (Bonner, Janet) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Processing with mesh cassettes Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42BA@fh2k093.fhmis.net> Are your cassettes packed in too tightly or do they "stick" together during processing, and do you use an agitation in your program? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu; mbecker@pathlabinc.com Sent: 6/7/2005 10:41 AM Subject: Re: [Histonet] Processing with mesh cassettes Hi Michelle, I've not attempted this. Perhaps if you exposed the cassettes to a solution to reduce the surface tension of the screens. Possibly putting a small amount of a detergent in the first formalin. Or dipping the cassettes in a detergent solution before placing them on the processor. Fred >>> "Michelle Becker" 06/07/05 10:12AM >>> We recently started using mesh cassettes for small biopsies, also called micromesh, histoscreen or micro biopsy cassettes, depending on the manufacturer. The techs, as well the pathologists, love them. Unfortunately, it has caused reagent carryover in our tissue processors (VIP 3000 & Leica TP1050)after only one run. The first reagent level (formalin)decreases significantly; the 2nd reagent level(Prefer), as well the next 3 alcohol levels, have significant increases. The tissue processor used does not seem to make a difference. We run a 4 hour short program with vacuum for biopsies. Has anyone else had this problem and if so, how did you resolve it? I really hate to stop using something the techs and pathologists love and actually agree on! Thank-you. Michele Becker, HTL(ASCP) Histology Manager Laboratory Corporation of America Portsmouth, NH _______________________________________________ Histonet mailing list Histonet@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 dahmed <@t> mdanderson.org Tue Jun 7 10:59:59 2005 From: dahmed <@t> mdanderson.org (dahmed@mdanderson.org) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Formic Acid and Bone Marrow Biopsies Message-ID: For those labs using formic acid for decalcification of bone marrow biopsies, What is the concentration and for what length of time is it used? David S. Ahmed, HT(ASCP) Chief Histology Technician Department of Pathology M.D. Anderson Cancer Center From dpahisto <@t> yahoo.com Tue Jun 7 11:32:40 2005 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Cassette Labeler Message-ID: <20050607163240.64704.qmail@web33415.mail.mud.yahoo.com> Can anyone suggest a good cassette labeler that is "inexpensive" ? We are not looking for anything fancy - one line - prints numbers and letters. I am getting tired of the pens and pencils coming off in Penfix and Carnoy's. Cindy DuBois __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Luis.Chiriboga <@t> med.nyu.edu Tue Jun 7 13:20:40 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Need a chemist's opinion on formalin fixation reversal In-Reply-To: Message-ID: Hey Andrea I don't know if the exact chemical mechanism is known. Most likely, the hydroxymethyl adducts and early cross links (lysine and arginine predominately) are unstable and easily reversed by h20(and alcohol?) There was a really nice 2 part article published in JOH 2001 by Eltoum IE. et al (unfortunately I don't have those issue because if it's not nailed down, somebody steals it !) I believe Jerry Fredenburgh was a co-author (yup, he presented at NYSHS) so you may want to contact him......let me know L -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrea T. Hooper Sent: Monday, June 06, 2005 5:54 PM To: Patsy Ruegg; histonet@lists.utsouthwestern.edu Subject: [Histonet] Need a chemist's opinion on formalin fixation reversal Can one of the chemists on board please address the issue of washing to reverse formalin fixation? (sorry if you have done so already but I must have missed it). This is a new concept to me and makes me very very nervous about some of my routines in the lab as often times I can only occasionally process and have samples in the fridge for sometime after fixation yet before processing ... yikes! Thank you thank you! Andrea >I have been asked to elaborate on my reference to washing tissues to reverse >the effects of formalin fixation iliminating the possible need for AR. >Here goes: > > >I found the reference to Washing in a book called Introduction to >Immunocytochemistry 3rd Edition J.M. Polak and S. Van Noorden pg. 24- 3.8.2 >"The simplest form of reversing the effects of formalin is to wash the >tissue well before processing, but this is not usually possible in >histopathology laboratories, where rapid turnowver of specimens is >required." > >I have not tested this extensively but have done a little, especially on >over fixed samples, the data is yet to be confirmed but I am looking into it >as we speak. I will keep you all posted. > >Patsy -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BDUE <@t> PARTNERS.ORG Tue Jun 7 13:42:57 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Need a chemist's opinion on formalin fixation reversal Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB5027A76@PHSXMB7.partners.org> Hi, I have a related tidbit for you. I am not a chemist, but I just recently read deep on HIER for an ISH protocol we started. Several sources (DAKO manual I think, and a couple rigorous papers I found) suggested that Heat Induced Epitope Rerieval (HIER) on Formalin Fixed Paraffin Embedded (FFPE) tissue sections serves (in part) to accelerate what washing at room temp could in theory do alone. In other words, washing long enough can remove the bound formaldehyde and/or reverse other FFPE fixation effects. It was estimated that washing alone could take months to achieve what HIER does in 30-60min. Sorry I don't have references handy, but if you want them, bug me and I'll dig through my folders sometime. -brice Neuropathology Lab Brigham & Women's Hospital Boston "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Luis Chiriboga Sent: Tuesday, June 07, 2005 2:21 PM To: Andrea T. Hooper; Patsy Ruegg; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Need a chemist's opinion on formalin fixation reversal Hey Andrea I don't know if the exact chemical mechanism is known. Most likely, the hydroxymethyl adducts and early cross links (lysine and arginine predominately) are unstable and easily reversed by h20(and alcohol?) There was a really nice 2 part article published in JOH 2001 by Eltoum IE. et al (unfortunately I don't have those issue because if it's not nailed down, somebody steals it !) I believe Jerry Fredenburgh was a co-author (yup, he presented at NYSHS) so you may want to contact him......let me know L -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrea T. Hooper Sent: Monday, June 06, 2005 5:54 PM To: Patsy Ruegg; histonet@lists.utsouthwestern.edu Subject: [Histonet] Need a chemist's opinion on formalin fixation reversal Can one of the chemists on board please address the issue of washing to reverse formalin fixation? (sorry if you have done so already but I must have missed it). This is a new concept to me and makes me very very nervous about some of my routines in the lab as often times I can only occasionally process and have samples in the fridge for sometime after fixation yet before processing ... yikes! Thank you thank you! Andrea >I have been asked to elaborate on my reference to washing tissues to reverse >the effects of formalin fixation iliminating the possible need for AR. >Here goes: > > >I found the reference to Washing in a book called Introduction to >Immunocytochemistry 3rd Edition J.M. Polak and S. Van Noorden pg. 24- 3.8.2 >"The simplest form of reversing the effects of formalin is to wash the >tissue well before processing, but this is not usually possible in >histopathology laboratories, where rapid turnowver of specimens is >required." > >I have not tested this extensively but have done a little, especially on >over fixed samples, the data is yet to be confirmed but I am looking into it >as we speak. I will keep you all posted. > >Patsy -- _______________________________________________ Histonet mailing list Histonet@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 Charlene.Henry <@t> STJUDE.ORG Tue Jun 7 13:55:51 2005 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Cassette Labeler. . Message-ID: <5CB39BCA5724F349BCB748675C6CA1A202C7572D@SJMEMXMB02.stjude.sjcrh.local> Cindy, We have a Surgipath Millennium that we have been very pleased with and the price in 2003 was approximately $10,000. It is simple and very easy to use. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cindy DuBois Sent: Tuesday, June 07, 2005 11:33 AM To: Histonet Subject: [Histonet] Cassette Labeler. . Can anyone suggest a good cassette labeler that is "inexpensive" ? We are not looking for anything fancy - one line - prints numbers and letters. I am getting tired of the pens and pencils coming off in Penfix and Carnoy's. Cindy DuBois __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DonnaWillis <@t> texashealth.org Tue Jun 7 14:18:19 2005 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Electron Microscopy Workload Message-ID: <4483B6EE0FA82A4EAED2B1E161E5E41147C865@ftwex02.txhealth.org> Thanks Joan, I know that I am on the right wave length with 1 FTE and 200 cases. Donna -----Original Message----- From: Sempf, Joan M. [mailto:Joan.Sempf@med.va.gov] Sent: Tuesday, June 07, 2005 12:15 PM To: Willis, Donna Subject: RE: [Histonet] Electron Microscopy Workload Sorry, there are two full time employees here. Joan -----Original Message----- From: Willis, Donna [mailto:DonnaWillis@texashealth.org] Sent: Tuesday, June 07, 2005 11:58 AM To: Sempf, Joan M. Subject: RE: [Histonet] Electron Microscopy Workload Joan, When you say "WE", how many Full Time Employees do you have working in the EM area? Donna -----Original Message----- From: Sempf, Joan M. [mailto:Joan.Sempf@med.va.gov] Sent: Tuesday, June 07, 2005 11:36 AM To: Willis, Donna Subject: RE: [Histonet] Electron Microscopy Workload We here at the VA in Madison do about 350 specimens a year. 90% are kidney biopsies. We process, cut, stain, develop our own negatives and print our own pictures; maintaining a two day turn-around time. We also have a large research program with two graduate students working on immunogold projects. When we are not busy with clinical work we have research projects to work on. Joan Sempf VA Hospital Madison Wisconsin -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Willis, Donna Sent: Tuesday, June 07, 2005 10:40 AM To: HistoNet Subject: FW: [Histonet] Electron Microscopy Workload Are there other EM labs performing TEM similarly to Tim's processing requiring their staff to produce more than 200 cases a year for 1 FTE? We are currently still doing things the way he did but on 95% kidney. 200 has been the target in my lab for 1FTE. I am wanting to put this in a graph for CAP. I know there are procedures that could be automated (ex. Processing) to allow workload increases but currently we are still do a lot of manual methods and that is what I would like to benchmark. Thanks, Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Friday, June 03, 2005 1:35 PM To: Willis, Donna Subject: RE: [Histonet] Electron Microscopy Workload Donna, I did em for 11 years and had about 200 cases per year. This was about 70 percent kidney and 30 percent tumor. I did all the processing, cutting, staining, initial screening, negative development and made enlargments. I also attended most of the kidney biopsies to get the tissue. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Willis, Donna Sent: Friday, June 03, 2005 11:06 AM To: HistoNet Subject: [Histonet] Electron Microscopy Workload What is the average workload for a TEM technician? How many cases a year should 1 FTE perform? Thanks, Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx 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. _______________________________________________ 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. _______________________________________________ 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. 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 liz <@t> premierlab.com Tue Jun 7 14:41:09 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] goldners trichrome Message-ID: <000001c56b98$df0c6660$a7d48a80@AMY> Hello Everyone I have to perform a goldners trichrome. The two protocols that I have do not use bouins as a mordant. One does state that the trichrome stain benefits from picric acid fixation but also states that 10% NBF is fine. Is this correct or should I mordant in bouins prior to staining. I figured I would ask the histonet first before I run the stain with and without using bouins as a mordant. Any advice would be helpful. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From bohlmta <@t> auburn.edu Tue Jun 7 14:44:40 2005 From: bohlmta <@t> auburn.edu (Tiffany Bohlmann) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Lung prep Message-ID: Hello all. I'm sorry if this is a no brainer... I have searched and searched for the answer... including the histonet archives. If you air infuse a lung and fix in gluteraldehyde, will this wash out any edema that was in the alveoli? Will edema be washed out with ordinary purfusion fixation (I'd assume not since we do see it?)? If you did want to air purfuse a lung for fixation, what would you have to do to keep edema intact? Thank you for any suggestions you may have for me! Tiffany Bohlmann Auburn University Collage of Veterinary Medicine bohlmta@auburn.edu From Vickroy.Jim <@t> mhsil.com Tue Jun 7 15:46:21 2005 From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] p63 antibody Message-ID: I have just been told by a vendor that the developer of the clone for p63 has switched this to an RUO antibody instead of a IVD antibody which it was in the past. Obviously this becomes a hassle for diagnostic labs. Does anybody know of a vendor that has a different antibody clone for p63 that is an IVD antibody? Jim Vickroy Memorial Medical Center Springfield, Illinois. From Barb.Richmond <@t> mckennan.org Tue Jun 7 16:35:47 2005 From: Barb.Richmond <@t> mckennan.org (Barb Richmond) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Weighing specimens Message-ID: Does formalin decrease the weight of tissue? In surgery for breast reductions the tissue removed is weighing > 400 grams. After the addition of formalin the lab is getting a weight that is 20 grams less. The balance/scales used have been compared and they only have 2-3 gram diffrence. What do you think?? ____________________ 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 TJasper <@t> smdc.org Tue Jun 7 16:47:06 2005 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Weighing specimens Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E45175@harrier> Used formalin, particularly from breast cases will retain components from the tissue. We see these as the non-recyclable by-product after used formalin is recaptured. Perhaps this explains, at least partially, this phenomenon. Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Barb Richmond [mailto:Barb.Richmond@mckennan.org] Sent: Tuesday, June 07, 2005 4:36 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Weighing specimens Does formalin decrease the weight of tissue? In surgery for breast reductions the tissue removed is weighing > 400 grams. After the addition of formalin the lab is getting a weight that is 20 grams less. The balance/scales used have been compared and they only have 2-3 gram diffrence. What do you think?? ____________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. From peoshel <@t> wisc.edu Tue Jun 7 16:55:20 2005 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Weighing specimens In-Reply-To: <1DB04B57B04C5747B87C3B39F3E605AA02E45175@harrier> References: <1DB04B57B04C5747B87C3B39F3E605AA02E45175@harrier> Message-ID: Formalin fixation causes both weight loss and specimen shrinkage, mostly from extraction of tissue components. A 5% weight loss doesn't sound surprising. Maybe. How long have the specimens been in formalin, and are they weighed after removal from the formalin and blotting to soak up the excess formalin? Were they blotted before weighing in surgery? This sort of question comes up in the ecological (especially physiological ecology) literature all the time. Phil >Used formalin, particularly from breast cases will retain components from >the tissue. We see these as the non-recyclable by-product after used >formalin is recaptured. Perhaps this explains, at least partially, this >phenomenon. > >Thomas Jasper HT(ASCP)BAS >Anatomic Pathology Coordinator >SMDC Clinical Laboratory >Duluth, MN >tjasper@smdc.org > > >-----Original Message----- >From: Barb Richmond [mailto:Barb.Richmond@mckennan.org] >Sent: Tuesday, June 07, 2005 4:36 PM >To: 'histonet@lists.utsouthwestern.edu' >Subject: [Histonet] Weighing specimens > > >Does formalin decrease the weight of tissue? In surgery for breast >reductions the tissue removed is weighing > 400 grams. After the addition >of formalin the lab is getting a weight that is 20 grams less. The >balance/scales used have been compared and they only have 2-3 gram >diffrence. What do you think?? > >____________________ >Confidentiality Notice: This e-mail message, including any attachments, is >for the sole use of the intended recipient(s) and may contain confidential >and privileged information. Any unauthorized review, use, disclosure, or >distribution is prohibited. If you are not the intended recipient, please >contact the sender by reply e-mail and destroy all copies of the original >message. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >This e-mail communication and any attachments may contain >confidential and privileged information for the use of the >designated recipients named above. If you are not the intended >recipient, you are hereby notified that you have received this >communication in error and that any review, disclosure, >dissemination, distribution or copying of it or its contents is >prohibited. As required by federal and state laws, you need to hold >this information as privileged and confidential. If you have >received this communication in error, please notify the sender and >destroy all copies of this communication and any attachments. > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm From cwscouten <@t> myneurolab.com Tue Jun 7 17:08:48 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Weighing specimens Message-ID: <5784D843593D874C93E9BADCB87342AB44FA33@tpiserver03.Coretech-holdings.com> Intesting. In soft tissue, formalin shrinks the tissue by removing extracellular space, and crosslinking proteins in neighboring cells together. It makes sense that the fluid that had been in the extracellular space is squeezed out. The tissue is smaller, and should be lighter. 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 Barb Richmond Sent: Tuesday, June 07, 2005 4:36 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Weighing specimens Does formalin decrease the weight of tissue? In surgery for breast reductions the tissue removed is weighing > 400 grams. After the addition of formalin the lab is getting a weight that is 20 grams less. The balance/scales used have been compared and they only have 2-3 gram diffrence. What do you think?? ____________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Tue Jun 7 17:33:28 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Weighing specimens Message-ID: Is the standard for "medically necessary" breast reduction still a minimum of 500g (each) (i.e., what health insurance companies will pay for). I recall this being a huge (no pun) issue some time ago - where the surgeon said the specimen weighed 500g, but the pathology scale indicated less - thereby screwing the patient out of the insurance coverage. My gripe is the ongoing resentment of insurance companies determining what is medically neccessary, rather than the physicians. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist GPRD Cancer Research Abbott Laboratories, Abbott Park, IL Jackie.OConnor@abbott.com "Charles Scouten" Sent by: histonet-bounces@lists.utsouthwestern.edu 06/07/2005 05:08 PM To: "Barb Richmond" , cc: Subject: RE: [Histonet] Weighing specimens Intesting. In soft tissue, formalin shrinks the tissue by removing extracellular space, and crosslinking proteins in neighboring cells together. It makes sense that the fluid that had been in the extracellular space is squeezed out. The tissue is smaller, and should be lighter. 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 Barb Richmond Sent: Tuesday, June 07, 2005 4:36 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Weighing specimens Does formalin decrease the weight of tissue? In surgery for breast reductions the tissue removed is weighing > 400 grams. After the addition of formalin the lab is getting a weight that is 20 grams less. The balance/scales used have been compared and they only have 2-3 gram diffrence. What do you think?? ____________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sandy.Nelson <@t> tenethealth.com Tue Jun 7 17:42:59 2005 From: Sandy.Nelson <@t> tenethealth.com (Nelson, Sandy - PPH) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Posting for Histology List Serve Message-ID: Subject: Histology Management Position A 500 bed hospital adjacent to the Houston Medical Center has an opening for a full time Histology Supervisor. This candidate should have strong leadership skills and prior management experience is preferred. Interested? Call 713.527.5292. Sandra Nelson Director Laboratory Services Park Plaza Hospital Houston, Texas 713.527.5292 sandy.nelson@tenethealth.com ** This is not intended to be a legally binding or legally effective signature From Secholsb <@t> aol.com Tue Jun 7 19:34:02 2005 From: Secholsb <@t> aol.com (Secholsb@aol.com) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] MACCHIAVELLO STAIN Message-ID: <1df.3d8334e8.2fd796fa@aol.com> Has anybody done a macchiavello stain for chlamydia? If so, HELP.... I am unable to get my methylene blue to stain my background. I can leave the slides in methylene blue over night and it still will not stain. What could I possibly be doing incorrectly? Thanks From barbara.bublava <@t> meduniwien.ac.at Wed Jun 8 03:24:57 2005 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] goldners trichrome References: <000001c56b98$df0c6660$a7d48a80@AMY> Message-ID: <002f01c56c03$90702ee0$dd01a8c0@GERICHTS9XOZZ8> Hi Liz I do Goldner routinely and there is an improvement in colour after postfixation with bouin?s. Muscle for example tends to bee greenish without bouin. After posttfixation it turns out red. Every trichrome (except van Gieson) will profit from postfixation with Bouin?s. I postfix at room temperature for at least two hours but most often over night (i have no time pressure) Best would be to fix in bouin only (did try that with fozens) but in routine it is not always possible and it is said that ihc will not work after bouin and dna is rouined ... sorry for bad english and best wishes Barbara ----- Original Message ----- From: "Elizabeth Chlipala" To: "'histonet'" Sent: Tuesday, June 07, 2005 9:41 PM Subject: [Histonet] goldners trichrome > Hello Everyone > > I have to perform a goldners trichrome. The two protocols that I have > do not use bouins as a mordant. One does state that the trichrome stain > benefits from picric acid fixation but also states that 10% NBF is fine. > Is this correct or should I mordant in bouins prior to staining. I > figured I would ask the histonet first before I run the stain with and > without using bouins as a mordant. Any advice would be helpful. > > Thanks in advance. > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > Manager > Premier Laboratory, LLC > P.O. Box 18592 > Boulder, Colorado 80308 > Office: (303) 735-5001 > Fax: (303) 735-3540 > liz@premierlab.com > www.premierlab.com > > Ship to Address: > Premier Laboratory > University of Colorado > MCDB, Room A3B40 > Boulder, Colorado 80309 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jorge.tornero <@t> gmail.com Wed Jun 8 03:36:59 2005 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Technovit staining Message-ID: <8c964a7905060801367dbad84c@mail.gmail.com> Hi, do you know a good process for staining with H&E cuts of specimens embedded in technovit 7100 resin? We usually stain them manually, with a simple 7.5 minutes of harris hematoxilin bath, followed by rinsing int tap water for another 7.5 minutes (with lots of agitation, if not, blue patches remains even in the glass surface of the slide), 7.5 minutes in eosin and a final rinse in water for 5-7 minutes, but we have recently adquired an autostainer from leica and we eant to use a more complex protocol. ?do you have any good experience? Thank you From barbara.bublava <@t> meduniwien.ac.at Wed Jun 8 04:34:34 2005 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] MACCHIAVELLO STAIN References: <1df.3d8334e8.2fd796fa@aol.com> Message-ID: <005901c56c0d$47af1b80$dd01a8c0@GERICHTS9XOZZ8> Hi all I am also unable to get my methylene blue to work. Different solutions, different concentrations with and without acidic acid, with and without postfixation in bouin ... (trying to do a Masson) Whats going wrong? ----- Original Message ----- From: To: Sent: Wednesday, June 08, 2005 2:34 AM Subject: [Histonet] MACCHIAVELLO STAIN > Has anybody done a macchiavello stain for chlamydia? If so, HELP.... > I am unable to get my methylene blue to stain my background. I can leave > the slides in methylene blue over night and it still will not stain. What > could I possibly > be doing incorrectly? Thanks > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From barbara.bublava <@t> meduniwien.ac.at Wed Jun 8 04:37:39 2005 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] MACCHIAVELLO STAIN References: <1df.3d8334e8.2fd796fa@aol.com> Message-ID: <006201c56c0d$b62f8770$dd01a8c0@GERICHTS9XOZZ8> forgot to mention - all I produce is a layer above parts of the section thanks for your help Barbara ----- Original Message ----- From: To: Sent: Wednesday, June 08, 2005 2:34 AM Subject: [Histonet] MACCHIAVELLO STAIN > Has anybody done a macchiavello stain for chlamydia? If so, HELP.... > I am unable to get my methylene blue to stain my background. I can leave > the slides in methylene blue over night and it still will not stain. What > could I possibly > be doing incorrectly? Thanks > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From barbara.bublava <@t> meduniwien.ac.at Wed Jun 8 06:18:52 2005 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Is PTAH sensible to autolysis? Message-ID: <00af01c56c1b$da2e4770$dd01a8c0@GERICHTS9XOZZ8> Hi everyone Working in forensic medicine different stages of "autolysis" are a problem I have to deal with. Sometimes PTAH is beautiful, sometimes there is nearly no stain or some fibres are stained, some not or sometimes half the fibre is stained. Is it possible that "autolysis" is the problem? What is the maximum I can expect with this unsolvable problem? I postfix with Bouin over night at RT, do Kaliumpermanganate and oxalic acid 5 min each (or Lugo?s over night and sodiumthisulfate). I do rinse well with tap water and distilled water bevor I go to the staining solution. There is no staining at RT over night, 2 h at 52?C gives "best" results. PTAH is ripend with Kaliumpermanganate Thanks for help and patience barbara From JNocito <@t> Pathreflab.com Wed Jun 8 07:25:19 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] p63 antibody In-Reply-To: Message-ID: Jim, have you contacted Biocare Medical, 1-800-799-9499? 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 Vickroy, Jim Sent: Tuesday, June 07, 2005 3:46 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] p63 antibody I have just been told by a vendor that the developer of the clone for p63 has switched this to an RUO antibody instead of a IVD antibody which it was in the past. Obviously this becomes a hassle for diagnostic labs. Does anybody know of a vendor that has a different antibody clone for p63 that is an IVD antibody? Jim Vickroy Memorial Medical Center Springfield, Illinois. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joycejudge259 <@t> hotmail.com Wed Jun 8 07:43:17 2005 From: joycejudge259 <@t> hotmail.com (joyce judge) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Ventana Discovery vs. Ventana Benchmark Message-ID: I am currently a Ventana Benchmark customer and use the machine for IHC of commercial and novel antibody characterization. I am exploring the purchase of a Ventana Discovery and would like to hear from people that have experience with both. Any advice would be greatly appreciated. Joyce Judge From portera203 <@t> yahoo.com Wed Jun 8 08:42:34 2005 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Immuno Info Please Message-ID: <20050608134234.80978.qmail@web40914.mail.yahoo.com> I am looking for basic information on immunohisto. FFPE for monkey. If anyone has any information or insights they could relay to me I would be so thankful. Trying to get some things up and going here, just looking for general information about what kind of staining and systems people are using. Thanks in advance, Amy S.Porter, HT(ASCP), QIHC Michigan State University - Department of Physiology Division of Human Pathology Investigative Histopathology Laboratory 2201 Biomedical Physical Sciences Room 2133 East Lansing, MI 48824-3320 Ph: 517-355-6475 ext 1480/1481 Fax: 517-432-1368 portera@msu.edu --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. From Nancy.Lowen <@t> med.va.gov Wed Jun 8 08:45:56 2005 From: Nancy.Lowen <@t> med.va.gov (Lowen, Nancy) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Thanks for the help Message-ID: I would like to say thanks to all who replied to my questions about the Cryojane Tape Transfer system. I got a lot of good tips and I will certainly try them. Thanks again. Nancy From settembr <@t> umdnj.edu Wed Jun 8 08:50:22 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] p63 antibody Message-ID: Jim, I use p63 clone 4A4 from DakoCytomation Cat# M7247. The lot that I recieved from them is for in vitro diagnostic use. Call them to ask if their current lot is IVD also. Their Customer Service & Tech Service depts are fantastic. 1-800-235-5763 / 800-424-0021 respectively. If you have any other questions don't hesitate to ask me. Dana Settembre University Hospital - UMDNJ Newark, NJ >>> "Vickroy, Jim" 6/7/2005 4:46:21 PM >>> I have just been told by a vendor that the developer of the clone for p63 has switched this to an RUO antibody instead of a IVD antibody which it was in the past. Obviously this becomes a hassle for diagnostic labs. Does anybody know of a vendor that has a different antibody clone for p63 that is an IVD antibody? Jim Vickroy Memorial Medical Center Springfield, Illinois. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charlene.Henry <@t> STJUDE.ORG Wed Jun 8 10:15:33 2005 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Ventana Discovery vs. Ventana Benchmark. . Message-ID: <5CB39BCA5724F349BCB748675C6CA1A202C7572F@SJMEMXMB02.stjude.sjcrh.local> We currently have both instruments and the Benchmark is used for clinical and the Discovery for research. The benefit of using the Discovery: it is a more open system allowing freedom to program both IHC, FISH, and ISH protocols using your own probes, antibodies, secondary antibodies, and reagents. We even have the TUNEL assay programmed on our Discovery. Hope this helps. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of joyce judge Sent: Wednesday, June 08, 2005 7:43 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Ventana Discovery vs. Ventana Benchmark. . I am currently a Ventana Benchmark customer and use the machine for IHC of commercial and novel antibody characterization. I am exploring the purchase of a Ventana Discovery and would like to hear from people that have experience with both. Any advice would be greatly appreciated. Joyce Judge _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Wed Jun 8 10:30:05 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:11 2005 Subject: AW: [Histonet] MACCHIAVELLO STAIN In-Reply-To: <1df.3d8334e8.2fd796fa@aol.com> Message-ID: I have learnd that acid solution inhibits the staining of methylenblue. We use it after decolorisation the Ziehl-Neelson with 3% HCl-alcohol and wash the slides very well. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Secholsb@aol.com Gesendet: Mittwoch, 08. Juni 2005 02:34 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] MACCHIAVELLO STAIN Has anybody done a macchiavello stain for chlamydia? If so, HELP.... I am unable to get my methylene blue to stain my background. I can leave the slides in methylene blue over night and it still will not stain. What could I possibly be doing incorrectly? Thanks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sagitalcuts <@t> yahoo.com Wed Jun 8 11:07:46 2005 From: sagitalcuts <@t> yahoo.com (Yolanda Maldonado) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Any Entomologist out there? Message-ID: <20050608160746.65770.qmail@web53501.mail.yahoo.com> Dear Histonet members: I've found a couple of ticks at my apartment and I am a bit worried. Any entomologist out there can recognize it? I have posted a couple of photos on the server (named tick 1 and tick 2) I observed it under the microscope and I can tell you it has 6 legs, 2 antennae and some sort of fangs (dots on pic1), and it doesn't like the light!! Thanks! Patricia Maldonado HT (ASCP) --------------------------------- Discover Yahoo! Have fun online with music videos, cool games, IM & more. Check it out! From gentras <@t> vetmed.auburn.edu Wed Jun 8 11:20:14 2005 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Adipocyte staining Message-ID: <6.0.1.1.0.20050608111452.01b9f848@mailhost.vetmed.auburn.edu> Hello, please is anyone out there familiar with the Oil Red O protocol for adipocytes which requires differentiation in 1% Dextrin? A colleague of mine needs to know if the Dextrin has to made in H2O or Isopropanol ? The Oil Red O is made in Isopropanol. Your prompt replies will be much appreciated. Atoska From pruegg <@t> ihctech.net Wed Jun 8 11:39:06 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Adipocyte staining In-Reply-To: <6.0.1.1.0.20050608111452.01b9f848@mailhost.vetmed.auburn.edu> Message-ID: <200506081639.j58Gd5t1026960@chip.viawest.net> I make my ORO in polyethyleen glycol and differentiate in the glycol. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Atoska S. Gentry Sent: Wednesday, June 08, 2005 9:20 AM To: Histonet Subject: [Histonet] Adipocyte staining Hello, please is anyone out there familiar with the Oil Red O protocol for adipocytes which requires differentiation in 1% Dextrin? A colleague of mine needs to know if the Dextrin has to made in H2O or Isopropanol ? The Oil Red O is made in Isopropanol. Your prompt replies will be much appreciated. Atoska _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Jun 8 11:46:45 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Technovit staining In-Reply-To: <8c964a7905060801367dbad84c@mail.gmail.com> Message-ID: <200506081646.j58Gkht1029314@chip.viawest.net> I use Gills 3 for 15 min. for GMA plastic sections H&E. It is very concentrated and does not require acid alcohol differention. I use aqueous eosin (0.2%) for 20-30 min. with a very quick dips in 95% ETOH to set the eosin and get it out of the background. I airdry and permount after 95% ETOH. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jorge Tornero Sent: Wednesday, June 08, 2005 1:37 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Technovit staining Hi, do you know a good process for staining with H&E cuts of specimens embedded in technovit 7100 resin? We usually stain them manually, with a simple 7.5 minutes of harris hematoxilin bath, followed by rinsing int tap water for another 7.5 minutes (with lots of agitation, if not, blue patches remains even in the glass surface of the slide), 7.5 minutes in eosin and a final rinse in water for 5-7 minutes, but we have recently adquired an autostainer from leica and we eant to use a more complex protocol. ?do you have any good experience? Thank you _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kbroomal <@t> NEMOURS.ORG Wed Jun 8 11:52:19 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Any Entomologist out there? Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824335@wlmmsx02.nemours.org> I looked, but the pictures aren't up yet. What part of the country/climate do you live in? Do you have pets that go outdoors? Why are you worried? Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Yolanda Maldonado Sent: Wednesday, June 08, 2005 12:08 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Any Entomologist out there? Dear Histonet members: I've found a couple of ticks at my apartment and I am a bit worried. Any entomologist out there can recognize it? I have posted a couple of photos on the server (named tick 1 and tick 2) I observed it under the microscope and I can tell you it has 6 legs, 2 antennae and some sort of fangs (dots on pic1), and it doesn't like the light!! Thanks! Patricia Maldonado HT (ASCP) --------------------------------- Discover Yahoo! Have fun online with music videos, cool games, IM & more. Check it out! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BDUE <@t> PARTNERS.ORG Wed Jun 8 11:57:12 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Adipocyte staining... ORO Churukian Style... Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB5027A79@PHSXMB7.partners.org> Hello Atoska, your protocol should explain the mixing. I use Churukian's ORO as printed in Bancroft 5th ed. The dextrin is a stabilizer of some sort, not a differentiator. The procedure is one step plus counterstain. For Churukian's procedure, the working solution is a combination of the ORO-isopropanol and the aqueous dextrin, so the literal answer to your questions is: mix the dextrin in water, then combine before use. Filtering or letting stand may be needed depending on which variant of the procedure you are following. In the variant I use (p. 208 in Bancroft 5ed.) the solutions are mixed and then remain stable for years -- until they stop working. I use this every week for muscle bxs and it never fails and is never fickle. -brice Neuropathology Lab Brigham & Women's Hospital Boston "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Atoska S. Gentry Sent: Wednesday, June 08, 2005 12:20 PM To: Histonet Subject: [Histonet] Adipocyte staining Hello, please is anyone out there familiar with the Oil Red O protocol for adipocytes which requires differentiation in 1% Dextrin? A colleague of mine needs to know if the Dextrin has to made in H2O or Isopropanol ? The Oil Red O is made in Isopropanol. Your prompt replies will be much appreciated. Atoska _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From schnegelsberg <@t> xgene.com Wed Jun 8 10:03:08 2005 From: schnegelsberg <@t> xgene.com (Birthe Schnegelsberg) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] anti human Laminin-5 antibody In-Reply-To: <6.0.1.1.0.20050608111452.01b9f848@mailhost.vetmed.auburn.edu> Message-ID: Hello. I am performing a fluorescent staining on cryosections from human skin with a monoclonal antibody from Chemicon for Laminin-5. I used concentrations from 1:400- very/too weak staining suggesting to be at expected location- to up to 1:80 -too high background, specific staining not detectable. Does anybody have experience with this antibody using cryosections? I did a 10min RT aceton fixation of the tissue, would anybody suggest a different fixation to improve the staining? Thanks, Birthe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Atoska S. Gentry Sent: Wednesday, June 08, 2005 11:20 AM To: Histonet Subject: [Histonet] Adipocyte staining Hello, please is anyone out there familiar with the Oil Red O protocol for adipocytes which requires differentiation in 1% Dextrin? A colleague of mine needs to know if the Dextrin has to made in H2O or Isopropanol ? The Oil Red O is made in Isopropanol. Your prompt replies will be much appreciated. Atoska _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JosefaNava <@t> texashealth.org Wed Jun 8 12:50:04 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Full time Histotech Opening Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944AB@phdex03.txhealth.org> A full time Histotech or eligible position is available at Presbyterian Hospital of Dallas, Texas. Interested candidates may contact Maureen Hoops (214-345-7795) for details and benefit information. 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 pex0220 <@t> yahoo.com.cn Wed Jun 8 15:08:06 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] anti-osterix antibody Message-ID: <20050608200806.94497.qmail@web15509.mail.cnb.yahoo.com> Hello, all, If you have some information about anti-osterix antibody from company or institute, I hope you can do me a favor! Thank you! Guofeng --------------------------------- DO YOU YAHOO!? ÑÅ»¢Ãâ·ÑGÓÊÏ䣭Öйú×îÇ¿Ãâ·Ñ·À¶¾·´À¬»ø³¬´óÓÊÏä From Charles.Embrey <@t> carle.com Wed Jun 8 15:13:22 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] looking for doug showers Message-ID: Sorry about going through the forum for this but I am trying to get hold of Doug Showers. Any contact info would be appreciated. Chuck Embrey From degaboh <@t> rice.edu Wed Jun 8 15:20:07 2005 From: degaboh <@t> rice.edu (ZP) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] brushes for cryotome anti-roll plate In-Reply-To: <20040731170552.23F671DB44@fungible8.mail.rice.edu> Message-ID: <20050608202006.843D71DB02@handler2.mail.rice.edu> Hello, I'm trying to order a couple of brushes that we use for cleaning the anti-roll plate and blade after a section has been taken in our cryotome. The previous brushes I used were worn down to a nub, and nobody knows where we got them from. Fisher doesn't have the right type at all. Ours were ~1.5 cm in length and had medium stiff bristles. Does anyone have any suggestions? Thank you! Zarana Patel degaboh@rice.edu From RizoC <@t> chi.osu.edu Wed Jun 8 15:20:32 2005 From: RizoC <@t> chi.osu.edu (Rizo, Christian) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] EM Specialist/ Technologist Position in Columbus, Ohio Message-ID: <979FF5962E234F45B06CF0DB7C1AABB201FEA785@chi2k3ms01.columbuschildrens.net> My name is Christian M. Rizo. I am the Manager of the Anatomic Pathology Lab at Children's Hospital in Columbus Ohio. The reason for this e-mail is that I wanted to inform you that we have an opening for an EM Technologist at our institution. There is a first shift (8:00 A.M. - 4:30 P.M.) Electron Microscopy Technologist position in the Anatomic Pathology Laboratory that is immediately available. The position is a full-time position, working Monday through Friday. Applicants must be a B.S. degree. Preference is given to candidates with some electron microscopy experience or M.S. degree candidate. This position is responsible for varied laboratory procedures, including specimen processing, electron microtomy, immunogold labeling and utilization of electron microscope. Duties may change depending on workload. Applicants should have moderate computer skills. Good customer relations, teamwork, dependability and communication are a must for successful candidates. Children's Hospital Columbus Founded by a determined group of women in 1892, Children's Hospital began as a local charity to serve a dozen very ill children. Throughout the following century, this tiny community-funded mission matured into a health care system that today spans the Midwest as one of its preferred providers of pediatric health care. Columbus Children's today is ranked as one of the nation's ten largest children's hospitals and pediatric research centers. Our hospital campus has nearly 700,000 patient visits every year. Located just minutes from downtown Columbus, Children's Hospital is one of the nation's most progressive and sophisticated health care institutions. Specialty areas include * Surgery * Heart transplant/cardiac care * Cancer * Trauma * Rehabilitation * Dialysis * Bone Marrow Transplant * Neurosciences * Research Children's is also the region's only pediatric Level 1 Trauma Center. With over 1.5-million square feet of interior space, the campus at Children's Hospital is vast. * The main hospital is home to the emergency department (one of the busiest pediatric emergency departments in the nation), surgery suites, interventional radiology and all inpatient units. * In August 2005, a new clinical expansion will be established which includes 14 additional state-of-the-art operating suites (which include intra-operative MRI and robotic technologies), new space for the Children's Heart Center to include catheterization laboratories and an additional 28-bed neonatal intensive care unit as well as integrated clinical laboratory services Thank you for your time. Please forward the e-mail to personnel who you think will benefit from it. 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 laurie.colbert <@t> huntingtonhospital.com Wed Jun 8 15:23:08 2005 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Sialidase digestion Message-ID: <0BE6ADFAE4E7E04496BF21ABD34662801D0D42@EXCHANGE1.huntingtonhospital.com> Has anyone ever done (or heard of) a colloidal iron stain with sialidase digestion? Our pathologists have a mucinous skin tumor and are trying to differentiate between a primary breast CA and a primary eyelid tumor. It may be run in conjunction with hyaluronidase digestion. Any references or procedures?? Laurie Colbert Huntington Hospital (626) 397-8620 From liz <@t> premierlab.com Wed Jun 8 15:27:31 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] brushes for cryotome anti-roll plate In-Reply-To: <20050608202006.843D71DB02@handler2.mail.rice.edu> Message-ID: <000001c56c68$8357b8d0$a7d48a80@AMY> I would try an art supply store. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of ZP Sent: Wednesday, June 08, 2005 1:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] brushes for cryotome anti-roll plate Hello, I'm trying to order a couple of brushes that we use for cleaning the anti-roll plate and blade after a section has been taken in our cryotome. The previous brushes I used were worn down to a nub, and nobody knows where we got them from. Fisher doesn't have the right type at all. Ours were ~1.5 cm in length and had medium stiff bristles. Does anyone have any suggestions? Thank you! Zarana Patel degaboh@rice.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From julien_lambreydesouza <@t> uqar.qc.ca Wed Jun 8 15:30:01 2005 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien Lambrey de Souza) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Alizarin S mindbender question Message-ID: <5d6b9372e9fa482318505b69ad373136@uqar.qc.ca> Hello histonetters, In our lab we do routine staining of fish specimens by the Clear and stain technique of Dingerkus and Uhler 1977, and some of the existing modifications with very nice results. But in the last few days, this technique has turned sour.... Something is happening to the fish when they are left in the Alizarin red S solution made up with 0.5% KOH. The solution has been made fresh several times but gives the same result each time. The fish seem to contract (muscular?) so much that the caudal becomes deformed (S shaped), the fin rays curl up and detach and the cranial elements also seem very fragile. My first thought was that the KOH solution was too harsh, but then I realized that the same KOH (0,5%) is used with these fish in the bleaching solution without harming the fish (apart from normal bleaching). The protocol is summarized as follows: 1) dehydration in EtOH ---- the fish turn out fine 2) stain in alcian blue solution (EtOH+acetic acid+alcain blue 8GN)---- fish turn out fine 3) neutralization in borax ---- fish are still fine 4) bleach in H2O2 solution (H2O2 3%+ KOH 0,5%) ---- fish still very nice 5) trypsin digestion (borax+H2O+tryspin) ------ although trypsin has cleared the specimen of its "meat" the fish is still very nice and fin rays are intact. 6) Alizarin red S solution (KOH 0,5%+alizarin S to turn solution deep purple) ----- almost on contact with the solution, the fish rinkles, rays curl up and detach... the specimen becomes useless. (The solution used to color the bones red and the specimen was then cleared of excess red by KOH bath). I have tried changing borax and KOH solutions with no improvement. Could it be that Alizarin S has gone bad? Is this even possible? We have had very high temperatures in the lab lately (30 degrees celsius), could this have influenced the solutions? I am puzzled. Also, we keep our trypsin in the -20?C freezer. The freezer has had a problem and freezed to -40?C for a day. Could this have altered the trypsin in a way that specimen exposition to this trypsin is incompatible with a later exposition to alizarin? (I am out of good ideas, so I'm trying absurd ideas....). If anyone is willing to take a shot, I will accept all suggestions because I am high and dry. Thanks for any help. Julien De souza, Evolutionary biology, UQAR, Quebec. From Jackie.O'Connor <@t> abbott.com Wed Jun 8 15:35:43 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] brushes for cryotome anti-roll plate Message-ID: Paint store. Art supply store. "ZP" Sent by: histonet-bounces@lists.utsouthwestern.edu 06/08/2005 03:20 PM Please respond to degaboh To: cc: Subject: [Histonet] brushes for cryotome anti-roll plate Hello, I'm trying to order a couple of brushes that we use for cleaning the anti-roll plate and blade after a section has been taken in our cryotome. The previous brushes I used were worn down to a nub, and nobody knows where we got them from. Fisher doesn't have the right type at all. Ours were ~1.5 cm in length and had medium stiff bristles. Does anyone have any suggestions? Thank you! Zarana Patel degaboh@rice.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed Jun 8 16:21:48 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Sialidase digestion In-Reply-To: <0BE6ADFAE4E7E04496BF21ABD34662801D0D42@EXCHANGE1.huntingtonhospital.com> References: <0BE6ADFAE4E7E04496BF21ABD34662801D0D42@EXCHANGE1.huntingtonhospital.com> Message-ID: <42A7616C.2070006@umdnj.edu> Hi Laurie: I have not done CI with sialidase but I have used hyaluronaidase many years ago. Keep in mind that there are two kinds of hyaluronidase, Bovine testicular and Streptococcyl, I think the latter is more specific. You need a book on mucin/connective tissue histochemistry, a lot has been done using gut mucins as models but I forget the details. See a histochemistry text for the correct path to follow. Geoff Laurie Colbert wrote: >Has anyone ever done (or heard of) a colloidal iron stain with sialidase digestion? Our pathologists have a mucinous skin tumor and are trying to differentiate between a primary breast CA and a primary eyelid tumor. It may be run in conjunction with hyaluronidase digestion. Any references or procedures?? > >Laurie Colbert >Huntington Hospital >(626) 397-8620 > >_______________________________________________ >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 emry <@t> u.washington.edu Wed Jun 8 16:34:00 2005 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] waterbath Message-ID: Do any of you use tap water for the waterbath or do you have to use distilled water? Thanks, Trisha U of WA, Seattle From lhartson <@t> trudeauinstitute.org Wed Jun 8 16:50:13 2005 From: lhartson <@t> trudeauinstitute.org (Louise Hartson) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] waterbath Message-ID: RO water >>> "Trisha Emry" 06/08/05 5:34 PM >>> Do any of you use tap water for the waterbath or do you have to use distilled water? Thanks, Trisha U of WA, Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Wed Jun 8 16:56:35 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] brushes for cryotome anti-roll plate Message-ID: Or a craft store...ie Michaels or JoAnns Robyn OHSU From Janet.Bonner <@t> FLHOSP.ORG Wed Jun 8 17:03:00 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] brushes for cryotome anti-roll plate Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42BF@fh2k093.fhmis.net> ...OFFICE DEPOT!!!!..... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Robyn Vazquez Sent: Wednesday, June 08, 2005 5:57 PM To: histonet@lists.utsouthwestern.edu; liz@premierlab.com; degaboh@rice.edu Subject: RE: [Histonet] brushes for cryotome anti-roll plate Or a craft store...ie Michaels or JoAnns Robyn OHSU _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Wed Jun 8 17:07:54 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] brushes for cryotome anti-roll plate Message-ID: <4.3.2.7.2.20050608150128.00cd7f08@algranth.inbox.email.arizona.edu> Years ago I saw some labs using Crayola brushes to clean up the microtomes and cryostat. I started using them and get them at the office supply store. I also saw them once in Walgreens. They are called Crayola "So Big" No. 208. Very inexpensive just don't use them with xylene since the handles are plastic! Speaking of brushes - sometimes I like to brush down sections while doing frozens and fabric painting brushes work well for this. I get mine at JoAnn Fabrics and Crafts. They are made by Loew Cornell and a pack of them is just a few bucks. Andi Paint store. Art supply store. "ZP" Sent by: histonet-bounces@lists.utsouthwestern.edu 06/08/2005 03:20 PM Please respond to degaboh To: cc: Subject: [Histonet] brushes for cryotome anti-roll plate Hello, I'm trying to order a couple of brushes that we use for cleaning the anti-roll plate and blade after a section has been taken in our cryotome. The previous brushes I used were worn down to a nub, and nobody knows where we got them from. Fisher doesn't have the right type at all. Ours were ~1.5 cm in length and had medium stiff bristles. Does anyone have any suggestions? Thank you! Zarana Patel degaboh@rice.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 runyanc <@t> mail.nih.gov Wed Jun 8 17:25:06 2005 From: runyanc <@t> mail.nih.gov (Runyan, Caroline (NIH/NIMH)) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Thionin staining method Message-ID: <5F9DE1C25708B04EAD634A1AE3D911300B8DB562@nihexchange20.nih.gov> I am trying to re-stain brain sections with thionin (they had been stained four years ago initially but were originally very light and have now faded beyond recognition). I popped off the coverslips, and then left the sections to sit in xylenes for about five days, at which point the dpx used for coverslipping appeared to have been removed from the tissue. Then, I re-hydrated and incubated in thionin for fifteen minutes-the thionin did not stick to the tissue at all. As soon as I drained the slides in water, all color was removed. I then went back and looked at the protocol that was apparently used to stain the tissue originally, and it looks as though the tissue was not de-fatted prior to thionin. The slides went from 70% EtOH (no water or 50%) through to 100%, into thionin, and then straight to 95%, 100%, xylene. Is tissue's current lack of thionin reactivity due to the fact that the original stain had been done so long ago, or because of the procedure originally used? Thanks. From katri <@t> cogeco.ca Wed Jun 8 18:07:54 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] waterbath References: Message-ID: <002101c56c7e$e71158c0$6a9a9618@Katri> Hi Trisha, We used to use tap water, but with the advent of immuno and charged slides, we have changed totally to distilled water and no additives. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Trisha Emry" To: "histo" Sent: Wednesday, June 08, 2005 5:34 PM Subject: [Histonet] waterbath > Do any of you use tap water for the waterbath or do you have to use > distilled water? > > Thanks, > Trisha > U of WA, Seattle > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histo20 <@t> hotmail.com Wed Jun 8 18:41:30 2005 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Histotech trainee Message-ID: Hello everyone! I have a cytoprep tech in our institution who has expressed a real desire to become a histotech. I have a position open, but have never "trained" a non-tech. Does anyone have training checklists- timeframes, etc. for this type of situation that you might be able to share? Any help would be most appreciated! Paula Wilder St. Joseph Medical Center Towson, MD 21204 From barbara.bublava <@t> meduniwien.ac.at Thu Jun 9 01:32:54 2005 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] waterbath References: Message-ID: <002901c56cbd$112c1e90$dd01a8c0@GERICHTS9XOZZ8> I also use tapwater and there is no problem. I even use it with slides silanised by myself. That works for special stains very well. In my last job I also did immunos as well but thats years ago and some things (HIER) changed in immunoprocessing. Greetings barbara ----- Original Message ----- From: "Trisha Emry" To: "histo" Sent: Wednesday, June 08, 2005 11:34 PM Subject: [Histonet] waterbath > Do any of you use tap water for the waterbath or do you have to use > distilled water? > > Thanks, > Trisha > U of WA, Seattle > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kcboswell <@t> grandecom.net Thu Jun 9 03:09:08 2005 From: kcboswell <@t> grandecom.net (Kevin and Chantel Bosewell) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] tap water on stainer Message-ID: <000601c56cca$87cd9d00$35069b18@Rascal> I was wondering what water you guys are running on your stainers, tap or deionized? Currently we are using tap but I was wondering if deionized would make a crisper stain? Thank you, Chantel Boswell HT (ASCP) Waco, Tx From bliven.laura <@t> marshfieldclinic.org Thu Jun 9 08:38:07 2005 From: bliven.laura <@t> marshfieldclinic.org (bliven.laura@marshfieldclinic.org) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Properdin Antibody Message-ID: <3986a01c56cf8$7821afd0$8e0110ac@mfldclinframe.org> I'm looking for an antibody source for Properdin FITC conjugated. Our current antibody has been discontinued and a new one flunked QC. Usage: Frozen kidney biopsies. Any company sources? Thanks much, Laura Bliven Marshfield Laboratories (715)387-7810 bliven.laura@marshfieldclinic.org From NMargaryan <@t> childrensmemorial.org Thu Jun 9 08:40:48 2005 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] tap water on stainer Message-ID: <63B8B599DE283148B92E83C78B32C15DB91E6F@cmhexbe2.childrensmemorial.org> I use dionized water. Tap has a lot of bleach in. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kevin and Chantel Bosewell Sent: Thursday, June 09, 2005 3:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] tap water on stainer I was wondering what water you guys are running on your stainers, tap or deionized? Currently we are using tap but I was wondering if deionized would make a crisper stain? Thank you, Chantel Boswell HT (ASCP) Waco, Tx _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The contents of this e-mail message and any attachments are private and confidential communications intended solely for the addressee(s) named in this message. If you are not the intended recipient of this message, please 1) immediately notify the sender by reply e-mail and then delete this message and its attachments and 2) do not read, use, distribute disclose or copy this message and/or any attachments. From vazquezr <@t> ohsu.edu Thu Jun 9 08:46:15 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] waterbath Message-ID: Use distilled, not tap... Robyn OHSU From asmith <@t> mail.barry.edu Thu Jun 9 08:48:15 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Sialidase digestion [long] Message-ID: <5D2189E74151CC42BEC02906BA8996322B90BF@exchsrv01.barrynet.barry.edu> Many years ago, I used sialidase and a control, followed by colloidal iron, to study primate salivary glands (A.A. Smith: Major salivary glands of the Philippine tarsier. Folia Primatologica 10: 113-130, 1969). I added 2 ml of reconstituted cholera filtrate (4000 microgram units of sialidase) to 41 ml of 0.05 M pH 5.5 acetate buffer (36 ml 0.05 ml sodium acetate adjusted to pH 5.5 with 0.1 M acetic acid plus 324 mg sodium chloride and 48 mg calcium chloride dehydrate). The control was the buffer without the cholera filtrate. I incubated for 24 hours at 37 degrees C. Audrea Vaughan (former graduate student, now teaching at Miami-Dade College) used sialidase and colloidal iron to study rhesus monkey salivary glands. She used 1.0 micromolar units of Clostridium perfringens sialidase (Sigma type V) in 40 ml of 0.1 M pH 5.5 acetate buffer. She incubated 24 hours at 37 C. Both of us used Mowry's colloidal iron ( R.W. Mowry: J. Clin. Invest. 7: 566-576, 1958). The procedure is given in Humason's ANIMAL TISSUE METHODS (pp. 301-303 in my copy of the 4th edition). I have also attached it here. Colloidal iron is a more sensitive detector of sialomucins than alcian blue or zirconyl hematoxylin. Audrea Vaughan found that sialidase worked well on tissues fixed in formalin, alcohol-formalin, or Carnoy's. It did not work on tissue fixed in Zenker's or Helly's fluid. Even the buffer removed the sialomucins from tissues fixed in Bouin's fluid. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert Sent: Wednesday, June 08, 2005 4:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sialidase digestion Has anyone ever done (or heard of) a colloidal iron stain with sialidase digestion? Our pathologists have a mucinous skin tumor and are trying to differentiate between a primary breast CA and a primary eyelid tumor. It may be run in conjunction with hyaluronidase digestion. Any references or procedures?? Laurie Colbert Huntington Hospital (626) 397-8620 _______________________________________________ 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 pmarcum <@t> vet.upenn.edu Thu Jun 9 09:05:00 2005 From: pmarcum <@t> vet.upenn.edu (Pamela A. Marcum) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] tap water on stainer In-Reply-To: <63B8B599DE283148B92E83C78B32C15DB91E6F@cmhexbe2.childrensmemorial.org> References: <63B8B599DE283148B92E83C78B32C15DB91E6F@cmhexbe2.childrensmemorial.org> Message-ID: <1118325900.42a84c8c32f12@imp.vet.upenn.edu> Hi, Water has always been an issue and it depends on where you live as to what is best. I have lived in areas where the pH of tap water was in the 8.0> range and other areas where you can smell the chlorine or the pH was <6.0. DI water is great if you have the option and if you don't you should find out the pH and try to work with it by understanding the chemistry of the tap water and how it will effect your stains. Some areas tap water is just fine and may even be better than DI if no one is changing the filters routinely. Best Regards, Pamela A. Marcum, Histology Manager University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 W. Street Road Kennett Square, PA 19348-1692 Phone: 610-925-6278 e-mail: pmarcum@vet.upenn.edu Quoting "Margaryan, Naira" : > I use dionized water. Tap has a lot of bleach in. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kevin > and Chantel Bosewell > Sent: Thursday, June 09, 2005 3:09 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] tap water on stainer > > I was wondering what water you guys are running on your stainers, tap > or deionized? Currently we are using tap but I was wondering if > deionized would make a crisper stain? > Thank you, > Chantel Boswell HT (ASCP) > Waco, Tx > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > The contents of this e-mail message and any attachments are private and > confidential communications intended solely for the addressee(s) named in > this message. If you are not the intended recipient of this message, please > 1) immediately notify the sender by reply e-mail and then delete this message > and its attachments and 2) do not read, use, distribute disclose or copy this > message and/or any attachments. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From mcauliff <@t> umdnj.edu Thu Jun 9 09:21:49 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:11 2005 Subject: [Histonet] Thionin staining method In-Reply-To: <5F9DE1C25708B04EAD634A1AE3D911300B8DB562@nihexchange20.nih.gov> References: <5F9DE1C25708B04EAD634A1AE3D911300B8DB562@nihexchange20.nih.gov> Message-ID: <42A8507D.9050804@umdnj.edu> Hi Caroline: John Kiernan is might be the best person to ask about this but for now......... Lack of de-fatting should not be the cause of your problems, at least not in my experience. There was considerable de-fatting in the graded ethanols, there is experimental evidence for this. Did you put some acetic acid in your thionin? If the stain is not acidic it will wash right out. If all else fails, try an oxidation with 0.25% potassium permanganate for a few minutes followed by bleaching the ugly brown color with 2% oxalic acid, then wash and restain with thionin. Geoff Runyan, Caroline (NIH/NIMH) wrote: >I am trying to re-stain brain sections with thionin (they had been stained >four years ago initially but were originally very light and have now faded >beyond recognition). I popped off the coverslips, and then left the >sections to sit in xylenes for about five days, at which point the dpx used >for coverslipping appeared to have been removed from the tissue. Then, I >re-hydrated and incubated in thionin for fifteen minutes-the thionin did not >stick to the tissue at all. As soon as I drained the slides in water, all >color was removed. I then went back and looked at the protocol that was >apparently used to stain the tissue originally, and it looks as though the >tissue was not de-fatted prior to thionin. The slides went from 70% EtOH >(no water or 50%) through to 100%, into thionin, and then straight to 95%, >100%, xylene. > >Is tissue's current lack of thionin reactivity due to the fact that the >original stain had been done so long ago, or because of the procedure >originally used? Thanks. >_______________________________________________ >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 Gregor.Majdic <@t> vf.uni-lj.si Thu Jun 9 09:35:56 2005 From: Gregor.Majdic <@t> vf.uni-lj.si (Majdic Gregor) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Vibroslice Message-ID: Hi everybody, I am new to this, but I have a question for anybody who uses Vibroslice from Campden instruments. I bought their Vibroslice MA752 exactly a year ago. For last few weeks we had problems with cutting brains (tissue was sheared) and I could feel very slight up and down movement of the nose (blade holder). After talking with people at Campden instruments, I decided to sent it back for repairs and today they contacted me telling that this is usual wear (initially they said due to years of use and after I told them the machine is only one year old they corrected themselves it could be one year use) on the bearings and I will have to do this repair regularly. I was pretty upset about that as they charge 150 GBP for repair (and I have to pay the postage), and they never mentioned to me when I was buying it that it will need a regular change of bearings. As I am not at all happy with that I would like to check if anybody else has the same machine and same problems with it - I would really like to hear it. Thanks to everybody, Gregor ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Possunt quia posse videntur ----------------------------------------------------------------- Gregor Majdic Head of Center for Animal Genomics Veterinary faculty, University of Ljubljana Gerbiceva 60 SI-1000 Ljubljana, Slovenia Phone: +386 1 4779210 Fax: +386 1 2832243 Email: gregor.majdic@vf.uni-lj.si From asmith <@t> mail.barry.edu Thu Jun 9 10:12:43 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] colloidal iron Message-ID: <5D2189E74151CC42BEC02906BA8996322B90C0@exchsrv01.barrynet.barry.edu> Since my attachment didn't work, I am sending my protocol for Mowry's colloidal iron as text. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 COLLOIDAL IRON Mowry, R.W. (1958) Improved procedure for the staining of acidic polysaccharides by Muller's colloidal (hydrous) ferric oxide and its combination with the Feulgen and periodic acid-Schiff reactions. J. Clin. Invest. -7-: 566-576. Stock "solution" (actually a colloidal suspension): 6.75 g FeCl3.6H2O 6 ml distilled water 0.4 ml conc. (37%) HCl Stir until dissolved. Bring 250 ml distilled water to a boil in a 1 liter container. Add ferric chloride solution to the boiling water. Cool. Colloidal iron: 15 ml distilled water 5 ml glacial acetic acid 20 ml stock "solution" Ferrocyanic acid: 15 ml 2% potassium ferrocyanide trihydrate [0.3 g K4Fe(CN)6.H2O in 15 ml H2O] Make 30 minutes before use. 15 ml 2% solution of conc. (37%) HCl [0.3 ml 37% HCl plus 14.7 ml H2O] Mix solutions just before use. 1 N HCl: 27.5 ml H2O 2.5 ml conc. (37%) HCl Sulfurous acid: 0.5 g sodium pyrosulfite = "sodium metabisulfite" = Na2S2O5 100 ml distilled water 0.5 ml conc. (37%) HCl 1. Bring to water. 2. 12% acetic acid, 30 seconds. 3. Colloidal iron, 1 hour. 4. 12% acetic acid, 4 changes, 3 minutes each. 5. Ferrocyanic acid, 20 minutes. 6. Running tap water, 5 minutes. 7. 1 N HCl at 60oC, 10 minutes. 8. Running tap water, 5 minutes. 9. Distilled water, rinse. 10. Schiff's reagent, 10 minutes. 11. Sulfurous acid, 3 changes, 2 minutes each. 12. Running water, 5 minutes. 13. Dehydrate, clear, and mount. 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 sjchtascp <@t> yahoo.com Thu Jun 9 10:14:45 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] ID rat capillarys in muscle Message-ID: <20050609151445.42997.qmail@web90205.mail.scd.yahoo.com> Working with ischemic rat hindleg injected with VEGF. I've read some articles recommending capillary endothelial cell identification with CD 31, Factor VIII and RECA-1, if anyone has experiance with any of these, others or similar work I'd appreciate the contact. Steve --------------------------------- Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. From conniemoss <@t> relia.net Thu Jun 9 10:22:13 2005 From: conniemoss <@t> relia.net (Connie McManus) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Any Entomologist out there? In-Reply-To: <20050608160746.65770.qmail@web53501.mail.yahoo.com> References: <20050608160746.65770.qmail@web53501.mail.yahoo.com> Message-ID: <49564.208.186.240.165.1118330533.squirrel@email.relia.net> HI, Patricia With 6 legs, it's not a tick. Ticks are arachnids (the spider group) and have eight legs. I didn't go to the server where your pics are, but if you go to this website http://www.ent.iastate.edu/imagegal/ticks/ you can find a wealth of info about all the ticks you would ever encounter. If I were you, I would take your pictures to the entomology dept of the nearest university. Usually there is someone there who will ID them for free. Extension entomologists are also a good place to look. -- Connie McManus Mt Ogden Scientific Services -Providing full service electron microscopy for you 950 W Kershaw, Suite E Ogden UT 84401 toll-free: 877/311-6677 direct: 801/745-2583 cell: 435/757/2975 fax: 435/514-1781 conniemoss@relia.net www.mtogdensci.com ====== Yolanda Maldonado said: > Dear Histonet members: > I've found a couple of ticks at my apartment and I am a bit worried. > Any entomologist out there can recognize it? I have posted a couple of photos on the > server (named tick 1 and tick 2) I observed it under the microscope and I can tell you > it has 6 legs, 2 antennae and some sort of fangs (dots on pic1), and it doesn't like the > light!! > Thanks! > Patricia Maldonado HT (ASCP) > > > > > --------------------------------- > Discover Yahoo! > Have fun online with music videos, cool games, IM & more. Check it out! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From dsnider <@t> shrinenet.org Thu Jun 9 10:35:33 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Reusing GMA Reagents Message-ID: Hello again histonetters. As I have said in the past, I am just learning all about GMA's. So far things are working out pretty well. Everyones advice has really helped. My question now is this, can the reagents used in processing be reused? I would assume it may be okay with the alcohols, but what about the actual GMA solutions? So far I am making fresh each time. (just to be on the safe side). But thought someone out there would know definantly. Thanks in advance, Deanna Snider HT ASCP Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From eshields <@t> bhset.org Thu Jun 9 10:42:42 2005 From: eshields <@t> bhset.org (E Sharon Shields) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Smooth Muscle Ysosin Heavy Chain Message-ID: Is any oune aout there in Histoland doing SM-MHC on the Ventana Benchmark? Would you care to share your antibody source and protocol? Thank you, Sharon Shields Baptist Hospital of East Tennessee 865 549-4351 The information in this e-mail message, including any attachments, may contain confidential and privileged information that is protected by law. It is intended for the sole use of the recipient named above. If you are not the intended recipient or the agent responsible for delivering it to the intended recipient, you are hereby notified that any unauthorized review, use, dissemination or copying is strictly prohibited. If you have received this electronic mail transmission in error please notify us immediately at bellis@bhset.org and delete any copies from your system. <<<>>> From rcharles <@t> state.pa.us Thu Jun 9 10:44:49 2005 From: rcharles <@t> state.pa.us (Charles, Roger) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] posting pictures Message-ID: <12E4E17FEF6EBE4BAE95BEB3CDCB570066D8D4@enhbgpri04.pa.lcl> When someone says they posted pictures on the Histonet sever where to you find them? I have looked over the Histonet page and see nothing about pictures. Roger Charles Microbiologist Pennsylvania Veterinary Laboratory 2305 N Cameron St Harrisburg, PA 17110 717-787-8808 From cgfields <@t> lexhealth.org Thu Jun 9 10:52:40 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] posting pictures Message-ID: I did the same looking for the bugs Yolanda posted. There is nothing about pictures. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC -----Original Message----- From: Charles, Roger [mailto:rcharles@state.pa.us] Sent: Thursday, June 09, 2005 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] posting pictures When someone says they posted pictures on the Histonet sever where to you find them? I have looked over the Histonet page and see nothing about pictures. Roger Charles Microbiologist Pennsylvania Veterinary Laboratory 2305 N Cameron St Harrisburg, PA 17110 717-787-8808 _______________________________________________ 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 kbroomal <@t> NEMOURS.ORG Thu Jun 9 11:00:37 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] posting pictures Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824339@wlmmsx02.nemours.org> The directions for posting pictures indicate that it could take 24 hours for the pictures to be posted. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Carole Fields Sent: Thursday, June 09, 2005 11:53 AM To: 'Charles, Roger' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] posting pictures I did the same looking for the bugs Yolanda posted. There is nothing about pictures. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC -----Original Message----- From: Charles, Roger [mailto:rcharles@state.pa.us] Sent: Thursday, June 09, 2005 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] posting pictures When someone says they posted pictures on the Histonet sever where to you find them? I have looked over the Histonet page and see nothing about pictures. Roger Charles Microbiologist Pennsylvania Veterinary Laboratory 2305 N Cameron St Harrisburg, PA 17110 717-787-8808 _______________________________________________ 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rocan <@t> mac.com Thu Jun 9 11:49:41 2005 From: rocan <@t> mac.com (Rocan) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] ID rat capillarys in muscle In-Reply-To: <20050609151445.42997.qmail@web90205.mail.scd.yahoo.com> References: <20050609151445.42997.qmail@web90205.mail.scd.yahoo.com> Message-ID: <421d80358bd0932d822d7887f04a0d33@mac.com> Steve, I would strongly suggest that you look up the papers from Donald M. McDonald (UCSF). Search on medline for author "McDonald DM". There absolutely no other group in the world better for vessel staining in vivo. I do recommend you do it in vivo before euthanasia. We have developed all our methods his papers. In brief: you inject the animal with a very small amount (less than 1ug), of griffonia isolectin B4 conjugated to a fluorochrome or biotin. Then you must perfuse the animal with 0.1% FRESH payaformaldehyde in (PBS). The perfusion done at 120 mmHg via left ventricle; the right atrium is punctured for drainage. It works like a charm! This procedure will have all the vessels lumens stained with fluorescence and the perfusion prevents vascular collapse. They have modified their techniques so it is important to start with his newest papers and go back to older ones as needed for details. In your case it would be great because it would also tell you if the vessels had flow. VEGF can create hemangioma-like networks that are not perfused properly . Contact me directly if you need any details. Good luck ----- Dr.Rocio Sierra-Honigmann Director Engineered Wound Repair Laboratory Cedars Sinai Research Institute 310-423-1882 Honigmannr@cshs.org On Jun 9, 2005, at 8:14 AM, Steven Coakley wrote: > Working with ischemic rat hindleg injected with VEGF. I've read some > articles recommending capillary endothelial cell identification with > CD 31, Factor VIII and RECA-1, if anyone has experiance with any of > these, others or similar work I'd appreciate the contact. > > Steve > > > --------------------------------- > Do you Yahoo!? > Yahoo! Mail - Helps protect you from nasty viruses. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dmikita <@t> wmcnet.org Thu Jun 9 12:16:31 2005 From: dmikita <@t> wmcnet.org (Daryl Mikita) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] PAPs on the GLX linear slide stainer? Message-ID: Hello, Does anyone know if you can do the cytology PAP stain on the GLX linear slide stainer? If so, does anyone have the protocol for it. We are going to be doing cytology in house, and we have two GLX slide stainers that are not in use. If possible we would like to use one for the PAP stain. Thanks, Daryl Mikita, HT(ASCP)cm Daryl A. Mikita, HT(ASCP)cm Wyoming Medical Center Anatomical Pathology 1233 E. 2nd St. Casper, WY 82601 From Jacquie.Farnsworth <@t> cls.ab.ca Thu Jun 9 12:21:06 2005 From: Jacquie.Farnsworth <@t> cls.ab.ca (Jacquie.Farnsworth@cls.ab.ca) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Mercurochrome in IBF--replacement? Message-ID: <4BC300747AF87A48BCDF8E48BC2885CECCD254@mail1.calgary.com> I've read many articles in the archives about replacing mercurochrome with eosin, etc. We are looking for a dye solution to use in IBF fixative for our prostate biopsies. (In order to see the tissue easier at embedding). We are currently trying Mrs. Stewart's bluing solution. With this, the pathologists have noticed a blue halo around the sections. Generally it does not interfere with diagnosis, but am wondering if there's something better out there? Thank you in advance, Jacquie Jacqueline Farnsworth Anatomic Pathology Tech III Calgary Laboratory Services Jacquie.Farnsworth@CLS.ab.ca PH: (403) 944-1578 Pager: 212-8223 #1933 CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. From KConway <@t> cmcc.ca Thu Jun 9 12:41:44 2005 From: KConway <@t> cmcc.ca (Kevin Conway) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Nerve biopsy - muscular dystrophy Message-ID: Hi everyone, I was wondering if you or your friendly neighbourhood pathologist might have access to (and be willing to share) sections or blocks of nerve biopsies from patients with muscular dystrophy. Southern Ontario is preferred but I'm sure we could work out a shipping arrangement. I'm trying to find a control tissue containing Renaut bodies, if you have any other suggestions I'd be happy to hear them. Thanks! Kevin Kevin Conway, PhD Assistant Professor Department of Anatomy Canadian Memorial Chiropractic College 416-482-2340 x.258 kconway@cmcc.ca From akbitting <@t> geisinger.edu Thu Jun 9 14:19:15 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Bile stain Message-ID: Does anyone have a good bile stain? Ours turns a drab olive green and our Dr. would like to see a "bright emerald green". Thanks in advance for your help. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From LuckG <@t> empirehealth.org Thu Jun 9 14:39:35 2005 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Mercurochrome in IBF--replacement? Message-ID: Jacquie, We use the orange dye that comes with the set of Davidson's tissue dyes. To set the ink better we soak our small biopsies briefly (after we wrap them in lens paper) in Bouin's solution. Although this is a bit messy it is very effective, has improved the visualizing for both embedding and cutting and the pathologists are happy as well. Good luck, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org www.deaconessmedicalcenter.org -----Original Message----- From: Jacquie.Farnsworth@cls.ab.ca [mailto:Jacquie.Farnsworth@cls.ab.ca] Sent: Thursday, June 09, 2005 10:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mercurochrome in IBF--replacement? I've read many articles in the archives about replacing mercurochrome with eosin, etc. We are looking for a dye solution to use in IBF fixative for our prostate biopsies. (In order to see the tissue easier at embedding). We are currently trying Mrs. Stewart's bluing solution. With this, the pathologists have noticed a blue halo around the sections. Generally it does not interfere with diagnosis, but am wondering if there's something better out there? Thank you in advance, Jacquie Jacqueline Farnsworth Anatomic Pathology Tech III Calgary Laboratory Services Jacquie.Farnsworth@CLS.ab.ca PH: (403) 944-1578 Pager: 212-8223 #1933 CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sjohnso616 <@t> aol.com Thu Jun 9 15:14:44 2005 From: Sjohnso616 <@t> aol.com (Sjohnso616@aol.com) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] (no subject) Message-ID: Histotechnologist ? Surgical Gross Lab in large hospital. Florida License and ASCP certification. Frozen section and grossing experience preferred. Daytime shifts. Excellent compensation and benefits including medical, dental, pension, generous paid time off. Near the beaches in beautiful Sarasota, Florida. EOE/DFWP Call (941) 362-8914 or Send resume to: e-mail: hr@sarapath.com Fax: (941) 362-8906 Sarasota Pathology _www.sarapath.com_ (http://www.sarapath.com/) From PMonfils <@t> Lifespan.org Thu Jun 9 15:55:08 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] New subscriber & VIP 1000 reservoirs Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171753D@lsexch.lsmaster.lifespan.org> Greetings from a new subscriber. I came across this list while searching online for something I need in my own laboratory, so I'll take this opportunity to mention that need. But I am also very happy to find such a list, the existence of which I was previously unaware of, and hope to continue participating in the future. I'm located in Rhode Island, U.S.A., and have been in histology for 35 years, about half clinical and half research-related. What I am looking for is six replacement solvent reservoirs for a Tissue-Tek VIP 1000 tissue processor (floor model). If anyone knows where these can be purchased - or if you have some you no longer need - I would appreciate hearing from you. Regards, Paul M. From dellav <@t> musc.edu Thu Jun 9 16:08:20 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Mercurochrome in IBF--replacement? Message-ID: Many laboratories will place a small quantity of eosin dye powder or eosin stain solution in the last alcohol on their processors. the specimen will be readily visible at the embedding station and the eosin is removed duirng staining (when you blue your hematoxylin) so your H&Es will not look differently. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> 06/09/05 01:21PM >>> I've read many articles in the archives about replacing mercurochrome with eosin, etc. We are looking for a dye solution to use in IBF fixative for our prostate biopsies. (In order to see the tissue easier at embedding). We are currently trying Mrs. Stewart's bluing solution. With this, the pathologists have noticed a blue halo around the sections. Generally it does not interfere with diagnosis, but am wondering if there's something better out there? Thank you in advance, Jacquie Jacqueline Farnsworth Anatomic Pathology Tech III Calgary Laboratory Services Jacquie.Farnsworth@CLS.ab.ca PH: (403) 944-1578 Pager: 212-8223 #1933 CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mmcgraw <@t> seattlecca.org Thu Jun 9 16:58:39 2005 From: mmcgraw <@t> seattlecca.org (McGraw, Medea J) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] BEST FIXATIVE FOR BRAIN TISSUE Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948033A89F3@wala01.seattlecca.org> I am looking for information on fixing brain tissue to give the best detail of tissue elements. I have read that Ammonium Bromide is currently using for fixing brain tissue (helps detail the astrocytes). My question is: can you post fix a brain that has already been fixed in 10% NBF with Ammonium Bromide or do you have to put the fresh tissue directly into the Ammonium Bromide? Any advice would be greatly appreciated. Medea J. McGraw , B.S. HT (ASCP) SCCA Pathology Department Desk: 206-288-1358 Main Lab: 206-288-1355 Email: mmcgraw@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 zacharyweil <@t> yahoo.com Thu Jun 9 18:16:30 2005 From: zacharyweil <@t> yahoo.com (Zach Weil) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] (no subject) Message-ID: <20050609231630.40449.qmail@web50604.mail.yahoo.com> I am trying to bring the animal model of MS (EAE) into our lab. In reading the literature it seems as if the best way to analyze the nervous tissue would be staining with H&E, Luxol fast blue and Bielschowsky. I want to use both brain and spinal cord tissue. However, I am unsure of how to proceed in several areas. 1) It is very difficult for me to remove the spinal cord from the animal without perfusing it first. However, in the literature most people remove the cord and drop it in formalin, is there any reason why perfusion would not work? 2) The other question is paraffin embedding vs. frozen and OCT. I would have to have the tissue embedded at a commercial lab so I would prefer to use frozen sections, but I can?t tell if that would interfere with these stains. Thanks so much, Zach Zachary Weil Graduate Research Associate Departments of Psychology and Neuroscience The Ohio State University 614-538-9540 fax (614) 451-3116 http://www.psy.ohio-state.edu/nelson/zach_weil.htm __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Sjohnso616 <@t> aol.com Thu Jun 9 19:50:12 2005 From: Sjohnso616 <@t> aol.com (Sjohnso616@aol.com) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Seeking histologist-fl Message-ID: <1ff.35a11e1.2fda3dc4@aol.com> Histotechnologist ? Surgical Gross Lab in large hospital. Florida License and ASCP certification. Frozen section and grossing experience preferred. Daytime shifts. Excellent compensation and benefits including medical, dental, pension, generous paid time off. Near the beaches in beautiful Sarasota, Florida. EOE/DFWP Call (941) 362-8914 or Send resume to: e-mail: hr@sarapath.com Fax: (941) 362-8906 Sarasota Pathology _www.sarapath.com_ (http://www.sarapath.com/) From jim.manavis <@t> imvs.sa.gov.au Fri Jun 10 01:57:09 2005 From: jim.manavis <@t> imvs.sa.gov.au (Jim Manavis) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Combined congo red & Biel Message-ID: <000601c56d89$9f1e7e10$636c140a@itp36533> Has anyone come across a combined congo red and Bielchowsky method. Cheers Jim From Jacqueline.Malam <@t> rli.mbht.nhs.uk Fri Jun 10 03:46:51 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Smooth muscle myosin Message-ID: Hi Sharon We have a Benchmark XT and use Dako's M3558 at 1/100 on a room temp incubation time of 88 mins with mild CC1 retrieval. This protocol replaced our standard CC1 retrieval with 60 mins incubation to reduce a morphology problem. Hope this helps Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From hts <@t> gator.net Fri Jun 10 06:47:30 2005 From: hts <@t> gator.net (hts@gator.net) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] CPT codes and pricing histology work Message-ID: <50584.65.81.70.231.1118404050.squirrel@webmail.gru.net> Hi Everyone! We are a contract lab who's main focus has been animal tissue prep and research projects. Because of our TAT and quality slides we have attracted a human pathologist who has become our Medical Director and now we are branching out into human diagnostic tissue. Please help! Though half of my career has been in human tissue hospitals, I am now separated from that by about 4 years and have no idea what to charge for human diagnostic tissue. Please hit the reply button and fill me in or let me know where to find the pricing that private labs and hospitals charge for the various histology techniques. Thanks so much for all your help in advance. Beverly Robinson HT(ASCP) Owner/Lab Manager Histology Tech Services "A CLIA and GLP Compliant Lab" Gainesville, FL Ph: 352-338-0045 Fx: 352-338-0027 From ajennings <@t> unmc.edu Fri Jun 10 08:23:32 2005 From: ajennings <@t> unmc.edu (Anita Jennings) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] processor input Message-ID: I have read up on some of the opinions about processors, I know there were more, but my searches are coming up thin and I need input asap since it is the end of the fiscal year for us I am in the market for a tissue processor, I want the Shandon/Thermo, mostly because I have a great rep, but I am willing to believe all the companies have reps like him, the admins aka powers that be and have the $$$ want some specs on other units and I would be interested to know who can compete with Thermo on what they offer....I am looking at the pathcentre I would wait to see what is out there when I go to NSH C/S but my present processor is acting up and I have to replace it soon thanks From ryaskovich <@t> dir.nidcr.nih.gov Fri Jun 10 09:11:54 2005 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] processor input Message-ID: <8F3AB322628548428A992EFB0E80F5D315D63188@nihexchange8.nih.gov> Anita, I love the Sakura VIP's they never go down. They are like Toyota's. Ruth Yaskovich N.I.H. N.I.D.C.R. Pain and Neurosensory Mechanism Branch > ---------- > From: Anita Jennings > Sent: Friday, June 10, 2005 8:23 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] processor input > > > > > > > I have read up on some of the opinions about processors, I know there were > more, but my searches are coming up thin and I need input asap since it is > the end of the fiscal year for us > I am in the market for a tissue processor, I want the Shandon/Thermo, > mostly because I have a great rep, but I am willing to believe all the > companies have reps like him, the admins aka powers that be and have the > $$$ want some specs on other units and I would be interested to know who > can compete with Thermo on what they offer....I am looking at the > pathcentre > I would wait to see what is out there when I go to NSH C/S but my present > processor is acting up and I have to replace it soon > > > thanks > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From cwscouten <@t> myneurolab.com Fri Jun 10 09:24:40 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] posting pictures Message-ID: <5784D843593D874C93E9BADCB87342AB44FA48@tpiserver03.Coretech-holdings.com> The histonet is an email listserve. No pictures may be attached to the mail to avoid problems with viruses. You may reference a picture, and mention its file name, in a histonet email. Then, the actual picture with that name is sent to pictures@histonet.org. Allow as much as 24 hours after sending the picuture for it to be posted to http://www.histonet.org/. If a picture has been referenced on the histonet, it may be seen at http://www.histonet.org/ within a day, and will remain there as an archive. To see the picture, go the website, and click "view listserve images" in the menu on the right. Scroll down through time to the date of the histonet refererence. 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 Carole Fields Sent: Thursday, June 09, 2005 10:53 AM To: 'Charles, Roger' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] posting pictures I did the same looking for the bugs Yolanda posted. There is nothing about pictures. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC -----Original Message----- From: Charles, Roger [mailto:rcharles@state.pa.us] Sent: Thursday, June 09, 2005 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] posting pictures When someone says they posted pictures on the Histonet sever where to you find them? I have looked over the Histonet page and see nothing about pictures. Roger Charles Microbiologist Pennsylvania Veterinary Laboratory 2305 N Cameron St Harrisburg, PA 17110 717-787-8808 _______________________________________________ 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From B427297 <@t> aol.com Fri Jun 10 09:30:40 2005 From: B427297 <@t> aol.com (B427297@aol.com) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Cadaver Search Dog Training Message-ID: <158.5296baeb.2fdafe10@aol.com> Would someone from the Austin/San Antonio area contact me regarding cadavers? Thanks. Jackie O'Connor From pex0220 <@t> yahoo.com.cn Fri Jun 10 16:25:05 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] antigen retrieval in bone paraffin sections Message-ID: <20050610212505.64592.qmail@web15502.mail.cnb.yahoo.com> Hello, all, Do you have some good suggestions about antigen retrieval in bone paraffin sections? Thank you! Guofeng __________________________________________________ ¸Ï¿ì×¢²áÑÅ»¢³¬´óÈÝÁ¿Ãâ·ÑÓÊÏä? http://cn.mail.yahoo.com From liz <@t> premierlab.com Fri Jun 10 16:42:01 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] antigen retrieval in bone paraffin sections In-Reply-To: <20050610212505.64592.qmail@web15502.mail.cnb.yahoo.com> Message-ID: <000201c56e05$3c16dbb0$a7d48a80@AMY> Guofeng I frequently use the ready to use proteinase K from DakoCytomation on formalin fixed, formic acid decalcified bone sections. I get good results using proteinase K with antibodies such as F4/80 in mouse joints, ED-1 in rat joints. IL-6 in both mouse and rat joints, mouse neutrophils, factor H, BrdU and Factor VIII. The other retrieval method I use is BioGenex's DeCal. I use that for CD45R in mice. I try to stay away from HEIR. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com =20 Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pex Sent: Friday, June 10, 2005 2:25 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] antigen retrieval in bone paraffin sections Hello, all,=20 Do you have some good suggestions about antigen retrieval in bone paraffin sections?=20 Thank you!=20 =20 Guofeng __________________________________________________ =B8=CF=BF=EC=D7=A2=B2=E1=D1=C5=BB=A2=B3=AC=B4=F3=C8=DD=C1=BF=C3=E2=B7=D1=D3= =CA=CF=E4? http://cn.mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Tracey.Lenek <@t> CLS.ab.ca Fri Jun 10 17:36:12 2005 From: Tracey.Lenek <@t> CLS.ab.ca (Tracey.Lenek@CLS.ab.ca) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] IVIEW DAB detection kits Message-ID: <4BC300747AF87A48BCDF8E48BC2885CE010E0F10@mail1.calgary.com> Hi, We are a high volume IHC department and use Ventana's systems exclusively. Over the past 6 months we have noticed inconsistent staining and batch to batch variation with different lot #'s of detection kits. Very difficult to maintain consistency with a large number of primary antibodies that we offer. Has anyone else experienced a similar problem? Thanks Tracey CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. From Salvacion.DeLovino <@t> cshs.org Sat Jun 11 12:43:21 2005 From: Salvacion.DeLovino <@t> cshs.org (DeLovino, Salvacion S.) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Electric Forceps Message-ID: Does anyone know where we could get a UL approved electric or heated forceps used in embedding? It does away with flames or forceps warmer. We're interested to get one except that we heard that it's not UL approved. We have strict regulations in California so that everything electric need to be UL approved. I hope we could find one that's UL approved. We've had problems with contamination and I think this may be the answer. Thanks! Salve Delovino Cedars-Sinai Medical Center Los Angeles, California IMPORTANT WARNING: This message is intended for the use of the person or entity to which it is addressed and may contain information that is privileged and confidential, the disclosure of which is governed by applicable law. If the reader of this message is not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this information is STRICTLY PROHIBITED. If you have received this message in error, please notify us immediately by calling (310) 423-6428 and destroy the related message. Thank You for your cooperation. From scoop <@t> mail.nih.gov Sat Jun 11 16:17:52 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] erythroid precursor marker for bone marrow, IF Message-ID: Dear Histonetters, I'm trying to find an antibody to use for a marker for erythroid precursors in mouse bone marrow. I'm able to use TER-119 in FFPE tissue using DAB but when I do immunofluorescence there's too much background fluorescence. I managed to quench the background fluorescence by fixing in methacarn and then leaving the sections in 3% H2O2/50% methanol for several hours, but then TER-119 wouldn't stain the sections (other abs worked so I must have damaged the antigen). I'm thinking about trying cd235a/glycoophorin A either clone HIR2 or JC159. Any suggestions? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From lpwenk <@t> sbcglobal.net Sun Jun 12 07:01:51 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Bile stain In-Reply-To: Message-ID: Depending upon the amount of bilirubin in the specimen, the shades of green may range from the drab olive to the brilliant emerald. So this may not be a staining problem. It just may be the state of the bile in the specimen. That said, I'll include our procedure, which is from the Freida Carson book. Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 - - - - - - BILE - FOUCHET PREPARED BY: Peggy A. Wenk, BS, HTL(ASCP)SLS ADOPTED BY:_____________________________________ DATE:__________________________ SUPERSEDES: PURPOSE: Bile and lipofuchsin appear as brown-yellow pigments. This stain demonstrates bilirubin (bile pigment) in tissue sections. Red blood cells, when destroyed, release their hemoglobin. This hemoglobin is broken down into globin (protein) and heme (iron). When the iron is removed from the heme portion, the residue is known as biliverdin. The biliverdin is transported to the liver where it is reduced to bilirubin, and then converted into bile, where it passes into the gall-bladder, then into the duodenum. In the cases of biliary obstruction or extensive liver damage, biliverdin, bilirubin and bile may accumulate in the liver. PRINCIPLE: Trichloroacetic acid and ferric chloride will oxidize the bilirubin to biliverdin, to produce a green color. The van Gieson is the counterstain. FIXATION: Any well fixed tissue, except Zenker. 10% neutral buffered formalin preferred. TECHNIQUE: Cut routine paraffin sections at 5 um. CONTROL: Section of tissue with bile QUALITY CONTROL: 1. Make Fouchet reagent just before use and filter. EQUIPMENT: Balance, Erlenmeyer flasks, graduated cylinders, magnetic stirrer, non-forceps. CAUTION: Follow standard safety procedures when preparing stains. TRICHLOROACETIC ACID is an acid. Add slowly to water. May cause severe skin and eye burns. May be irritating to eyes and respiratory system. FERRIC CHLORIDE is a corrosive. May cause skin and eye burns. Can be irritating to respiratory tract. ACID FUCHSIN is an irritant. PICRIC ACID is toxic, highly reactive (4 - NFPA) and an extreme fire hazard (4 - NFPA). Keep picric acid moist at all times. If dry around top of jar, wash off dry particles with running water before opening. Store in an explosion proof jar. REAGENTS: 10% FERRIC CHLORIDE Ferric chloride (FeCl3?6H2O) 10.0 g Distilled water 100.0 mL OR 29% Stock ferric chloride 3.5 mL Distilled water 6.5 mL Stir together with magnetic stirrer. Store at room temperature. Stable for 1 month. FOUCHET REAGENT Trichloroacetic acid (CCl3COOH) 12.5 g Distilled water 50.0 mL 10% ferric chloride 5.0 mL JUST BEFORE USE, carefully add trichloroacetic acid to distilled water. Stir until dissolved. Add 10% ferric chloride. Filter before use. 1% ACID FUCHSIN Acid fuchsin (CI 42685) 1.0 g Distilled water 100.0 mL Dissolve together. Store at room temperature. Stable for months. SATURATED AQUEOUS PICRIC ACID Picric acid (2,4,6-(NO2)3C6H2OH) Distilled water 100.0 mL Add picric acid, a gram at a time, to the distilled water, and stir using a magnetic stirrer. When dissolved, add another gram and stir. Continue to do this until no more picric acid will dissolve into the distilled water. Store at room temperature. Stable for months. CAUTION See above warning of picric acid. REAGENTS: continued VAN GIESON COUNTERSTAIN 1% acid fuchsin 5.0 mL Saturated aqueous picric acid 100.0 mL Mix together. Store at room temperature. Stable for 4-6 months. May be reused until weak. PROCEDURE - Fouchet: 1. Deparaffinize and hydrate sections through graded alcohol to distilled water. 2. Place slides in freshly filtered Fouchet reagent 5 minutes 3. Rinse in distilled water, 3-5 changes 5-10 seconds each 4. Stain in van Gieson counterstain 1 minute 5. Place slides directly into 95% alcohol, 2 changes 5-10 seconds each 6. Dehydrate through absolute alcohols and clear in xylene. 7. Coverslip using a synthetic mounting media. RESULTS: Bile or bilirubin shades of green Collagen red Cytoplasm, red blood cells yellow PROCEDURAL NOTES: 1. Depending upon the concentration of bilirubin, the colors may range from olive green to emerald green. 2. After the van Gieson counterstain, go directly to 95% alcohol. Placing the slides in water may decrease the amount of counterstain. REFERENCES: Bancroft JD, Stevens A: Theory and Practice of Histological Techniques, 3rd ed. New York, NY, Churchill Livingstone, 1990. * Carson FL: Histotechnology: A Self-Instructional Text, Chicago, IL, ASCP Press, 1990. Sheehan DC, Hrapchak BB: Theory and Practice of Histotechnology, 2nd edition. Columbus, Ohio, Battelle Press, 1980. Vacca L: laboratory Manual of Histochemistry. New York, NY, Raven Press, 1985. * Source of procedure. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Thursday, June 09, 2005 3:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bile stain Does anyone have a good bile stain? Ours turns a drab olive green and our Dr. would like to see a "bright emerald green". Thanks in advance for your help. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emelinepg <@t> yahoo.com.br Sun Jun 12 09:40:34 2005 From: emelinepg <@t> yahoo.com.br (Emeline) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] fish scales decalcification Message-ID: <20050612144035.10923.qmail@web30915.mail.mud.yahoo.com> Hi, I am from Brazil, and I just subscribed this list, please forgive me if my English isn't good! I'm trying to study the skin of a flatfish, but something is wrong. I tried to decalcify the scales with EDTA, but it was very slow and didn't work. Then I used Davidson's fixative for 2 weeks and more, and it worked, but when I tried to section it, the tissues scattered in the histological bath, and I lost the material. I found that lower temperatures of the bath were less dangerous to the tissues, but still damages them. I don't know what to do to preserve the structure of the entire skin, with all of its layers. I didn't tried nitric acid, formic acid or anyone else. Maybe the problem is in the fixative/decalcifier, or maybe it is in the sectioning, I really don't know. Have anyone worked with this kind of material? I just want some tips that might help. Thanks, Emeline --------------------------------- Yahoo! Mail: agora com 1GB de espa?o gr?tis. Abra sua conta! From MGomez <@t> ameripath.com Sun Jun 12 20:50:20 2005 From: MGomez <@t> ameripath.com (Gomez, Milton) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] nail Message-ID: Dear Histonetters: Do you have a consistent procedure for cutting nails? I am using ammonia to soak the tissue and PLL coated slides but the nail tissue falls after performing some special and immuno stains. Thanks, Milton From jstaruk <@t> masshistology.com Sun Jun 12 21:28:36 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] nail In-Reply-To: Message-ID: <4404j6$10knqmt@mxip20a.cluster1.charter.net> We adhere nails to slides using diluted (5% or so) yellow wood glue. Haven't lost a nail since! Jim ____________________ Jim Staruk www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gomez, Milton Sent: Sunday, June 12, 2005 9:50 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] nail Dear Histonetters: Do you have a consistent procedure for cutting nails? I am using ammonia to soak the tissue and PLL coated slides but the nail tissue falls after performing some special and immuno stains. Thanks, Milton _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From themagoos <@t> rushmore.com Sun Jun 12 21:32:26 2005 From: themagoos <@t> rushmore.com (Jason and Heather) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] nail References: Message-ID: <000801c56fc0$26701210$b3fc9fd1@magoo> We use Nair to soften the nail and use wood glue in the water bath for an adhesive. Jason McGough HT(ASCP) Clinical Laboratory of the Black Hills From ROrr <@t> enh.org Mon Jun 13 07:25:47 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Ventana Iview kits, Message-ID: Hi Tracey, I use Ventana Iview kits as well, and have not noticed the variability you are describing. Please ask your Ventana Rep for contacts of other users in your area and ask them if they are experiencing the same issues. There may be other problems providing these symptoms.. Are you getting these results in ALL antibodies? Is it only antibodies getting a specific pretreatment? CC1 as opposed to Protease... Have you changed your control tissue or how that control tissue and the patient tissue has been handled before it got to you? Are you cutting and storing your control tissue ahead of time? Most control tissues can last a long time on the slide once they have been cut, but if you have tissue that has been cut and stored on the slide for a long time, you MAY want to look at that, a long time could be 6 months to a year...I know a lot of labs cut and store their tissue for extended periods of time and have no problems! Since you are experiencing inconsistencies, I suggest checking out this aspect... And while you're at it, there's always the WATER!!! It's summertime and water changes, maybe check out the water supply, the filtration process, bring in water from an outside source to make up your EZ prep and Reaction Buffer.... Hope this helps... Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 19, Issue 17 **************************************** From TJJ <@t> Stowers-Institute.org Mon Jun 13 09:49:22 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Destabilizing Technovit 9100 NEW monomer Message-ID: Hi all, I have the Technovit 9100 NEW kit and in the instructions they recommend destabilizing the basic solution (monomer) using aluminum oxide in a chromatography column. Has anybody done this and can you give me the benefit of your experience? It looks as though I'll need to buy a column, and I'm not sure what size or setup configuration to buy (diameter, fritted?). Thanks for any help you can offer! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From kosmicdog <@t> hotmail.com Mon Jun 13 09:57:55 2005 From: kosmicdog <@t> hotmail.com (J m) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] IHC artifact Message-ID: I have posted a picture of an IHC artifact on the image server (artifact1.jpg). This problem has turned up on some immunos done in my lab. Can anyone identify the source or has anyone had this problem before? Thanks for any help. Jason. From galinadeyneko <@t> yahoo.com Mon Jun 13 10:53:38 2005 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] goldners trichrome Message-ID: <20050613155338.68891.qmail@web33112.mail.mud.yahoo.com> Dear Colleagues , I also do a lot of routine Masson Trichrome on murine hearts and have been interesting with this method. Could you please share with me some information: protocol , targets for this staining etc. or where I can find information. I would like to find something more sensitive for fine diffuse fibrosis. Thank you. Galina Deyneko Novartis, Cambridge MA 617- 871-7613 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From PMonfils <@t> Lifespan.org Mon Jun 13 11:24:36 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] nail Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717541@lsexch.lsmaster.lifespan.org> In sectioning nails, I use a combination of several procedures: - a couple of drops of Nonidet P-40 in the water on the cold plate (I use Tissue-Tek cold plates, stored in the -20 C freezer when not in use). Soak for 15-30 minutes before sectioning. (A couple of drops of liquid dish detergent - not dishwasher detergent - could substitute). - 10 to 20 ml PVA (polyvinyl acetate) in the water bath. PVA is quite viscous and does not easily dissolve into the water bath. To get around this, I place a flask with 300 ml water on the hotplate stirrer at 70 C; then while stirring, pour the PVA into the flask. Let it stir for a couple of minutes, then pour the contents of the flask into the water bath and stir it in manually. Rinse out the flask with hot water immediately because once the PVA dries it is difficult to remove. The down side of using PVA is that it makes the water bath "milky", so the floating sections are more difficult to see. But it does hold difficult tissues on the slides very securely, and it causes a minimum of background staining with either immunohistochemical or standard histochemical procedures. Glues like "Elmer's Glue-All" and "Sobo Glue" are good sources of PVA, and are cheaper then purchasing PVA from a chemical company. - a warmer than usual flotation bath, 70 C, to better flatten the sections. This is warm enough to melt the paraffin, so you cannot place multiple pieces of tissue in one block; but that usually isn't a good idea anyway with such tough to section tissues. Allow the sections to float for a few minutes if necessary, until they appear flat. - flattening and initial adherence of sections on an 80 C slide warmer for 5 minutes, after allowing slides to drain vertically for a couple of minutes. Followed by standard drying in a 60 C drying oven for at least an hour. Paul M. From Sandra.Eyton-Jones <@t> CLS.ab.ca Mon Jun 13 11:30:44 2005 From: Sandra.Eyton-Jones <@t> CLS.ab.ca (Sandra.Eyton-Jones@CLS.ab.ca) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Automated Special Stains System Message-ID: <4BC300747AF87A48BCDF8E48BC2885CE013495D8@mail1.calgary.com> Hi, does anyone know of an automated special stain system, other than Ventana. We would like to demo a few systems. Thank you, Sandy Sandra Eyton-Jones Tech III Phone (403) 770-3588 Fax (403) 770-3778 Pager 212-8223 #0540 Diagnostic and Scientific Center (DSC) sandra.eyton-jones@cls.ab.ca CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. From JWEEMS <@t> sjha.org Mon Jun 13 11:41:44 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Automated Special Stains System Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45FD5@sjhaexc02.sjha.org> DAKO has a good one - the Artisan. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sandra.Eyton-Jones@CLS.ab.ca Sent: Monday, June 13, 2005 12:31 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated Special Stains System Hi, does anyone know of an automated special stain system, other than Ventana. We would like to demo a few systems. Thank you, Sandy Sandra Eyton-Jones Tech III Phone (403) 770-3588 Fax (403) 770-3778 Pager 212-8223 #0540 Diagnostic and Scientific Center (DSC) sandra.eyton-jones@cls.ab.ca CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** 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 tsai0058 <@t> umn.edu Mon Jun 13 12:30:18 2005 From: tsai0058 <@t> umn.edu (tsai0058) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Trouble Adhering Post-Fixed Frozen Cartilage to Slides... Message-ID: <200506131730.j5DHUIEm027656@qix.software.umn.edu> Hello, I'm trying to adhere frozen intervertebral discs to slides for H&E staining as well as immunohistochemistry, and am having a hard time getting the tissues to stick to the slides throughout the process. The tissues were obtained fresh, then fixed for 24 hours in Lanas solution (4% paraformaldehyde, 14% picric acid in phosphate buffer solution), and then stored in a cryopreservative for approximately 1 week, after which they were quick frozen in liquid nitrogen. Does anyone have any protocols that they use for post-fixed frozen cartilage? Also, I have a colleague who is working with paraffin embedded Temporal Mandibular Discs (connective tissue, synomium, muscle, cartilage), and was trying to do immunohistochemistry, and he was having some difficulty getting the tissue to stick to the slides as well. Does anyone have any thoughts regarding this problem? Thanks for your input! Phil Tsai University of Minnesota From bsylinda <@t> aol.com Mon Jun 13 12:39:19 2005 From: bsylinda <@t> aol.com (bsylinda@aol.com) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Automated Special Stains System In-Reply-To: <4BC300747AF87A48BCDF8E48BC2885CE013495D8@mail1.calgary.com> References: <4BC300747AF87A48BCDF8E48BC2885CE013495D8@mail1.calgary.com> Message-ID: <8C73E55F63B39AB-C3C-106DD@MBLK-M31.sysops.aol.com> Biogenix has a ihc/special stain stainer. Sylinda -----Original Message----- From: Sandra.Eyton-Jones@CLS.ab.ca To: Histonet@lists.utsouthwestern.edu Sent: Mon, 13 Jun 2005 10:30:44 -0600 Subject: [Histonet] Automated Special Stains System Hi, does anyone know of an automated special stain system, other than Ventana. We would like to demo a few systems. Thank you, Sandy Sandra Eyton-Jones Tech III Phone (403) 770-3588 Fax (403) 770-3778 Pager 212-8223 #0540 Diagnostic and Scientific Center (DSC) sandra.eyton-jones@cls.ab.ca CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Mon Jun 13 13:35:31 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Trouble Adhering Post-Fixed Frozen Cartilage to Slides... In-Reply-To: <200506131730.j5DHUIEm027656@qix.software.umn.edu> References: <200506131730.j5DHUIEm027656@qix.software.umn.edu> Message-ID: <42ADD1F3.1010700@umdnj.edu> Many years ago (1985?) there was a paper in Stain Technology, now known as Biotechnique and Histochemistry, called "hot melt corner point method" in which the authors used hot melt glue to hold down sections of vertebral bodies (or intervertebral disks). The glue was applied at the corners of the sections but I think the sections were embedded in celloidin. Also, you might want to try staining the sections free-floating in watch glasses (or a similar shallow container) and only mounting the sections when staining is finished. Geoff tsai0058 wrote: >Hello, > > I'm trying to adhere frozen intervertebral discs to slides for H&E >staining as well as immunohistochemistry, and am having a hard time getting >the tissues to stick to the slides throughout the process. The tissues >were obtained fresh, then fixed for 24 hours in Lanas solution (4% >paraformaldehyde, 14% picric acid in phosphate buffer solution), and then >stored in a cryopreservative for approximately 1 week, after which they >were quick frozen in liquid nitrogen. Does anyone have any protocols that >they use for post-fixed frozen cartilage? > > Also, I have a colleague who is working with paraffin embedded Temporal >Mandibular Discs (connective tissue, synomium, muscle, cartilage), and was >trying to do immunohistochemistry, and he was having some difficulty >getting the tissue to stick to the slides as well. Does anyone have any >thoughts regarding this problem? > > Thanks for your input! > >Phil Tsai >University of Minnesota > > > > >_______________________________________________ >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 jane.moose <@t> newberryhospital.org Mon Jun 13 16:52:42 2005 From: jane.moose <@t> newberryhospital.org (Jane Moose) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] melanin renoval Message-ID: <00cb01c57062$39d16fa0$18041f0a@LAB2> Does anyone have a procedure to bleach unstained slides (from FFPE tissue) in order to remove melanin in order to stain with DAB for PECAM? Jane Moose LIS Coordinator/Histology Newberry County Memorial Hospital Newberry, SC 803-405-7129 ************************************************************************************ This 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 message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. From celebrej <@t> HHSC.CA Mon Jun 13 13:59:05 2005 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Controls used for Muscle biopsies Message-ID: <3AADFB88753AD31189C100902786B91C14E6DFFC@hch_nt_exchange.hhsc.ca> Hello Histonetters... This is a question for all those labs out there that do enzyme histochemistry on muscle biopsies. Does anyone run controls along with there test muscle biopsies? If your honest answer is yes (not like mine was), what do you use? Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca The Cornerstone of Care Campaign for Hamilton Health Sciences needs your support. Please, visit hamiltonhealthsciences.ca today and help make something great even greater. This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From skouko <@t> po-box.mcgill.ca Mon Jun 13 14:15:52 2005 From: skouko <@t> po-box.mcgill.ca (skouko@po-box.mcgill.ca) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] deparaffinization won't work! Message-ID: <1118690152.42addb68e7e50@www.webmail.mcgill.ca> Dear Histoneters, I am trying to do an FMRP immuno on brain specimens that were fixed and embedded in paraffin. Unfortunately I couldn't get staining. So I did a Nissl and I don't get staining with that as well so I am guessing my tissue isn't fully deparaffinized. I have tried everything from: -Preheating the tissue on a hot plate for 2 hours. -Spraying the sections with ParaGuard (a paraffin repellant used to remove paraffin stains) after heating the sections. My protocol for deparaffinization is currently: -2 times 30 min in Xylene -10 minutes in 100% Ethanol -5 minutes in 95% Ethanol -5 minutes in 70% Ehtanol -5 minutes in 50% Ethanol -2 times 5 minute washes in 0.05% PBST -2 rinses in dH2O. Then I do an antigen retrieval step with Vector antigen Retrieval kit and then my immuno. Result = NO STAINING! Any help would be greatly appreciated. Best, Sophia Koukoui MBNS, Psychology McGill University Montreal Canada. From liz <@t> premierlab.com Mon Jun 13 14:58:28 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] melanin renoval In-Reply-To: <00cb01c57062$39d16fa0$18041f0a@LAB2> Message-ID: <000801c57052$449c4b90$a7d48a80@AMY> Jane Rather than perform a melanin bleach why don't you switch to a alkaline phosphatase detection system and use fast red as your chromagen. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jane Moose Sent: Monday, June 13, 2005 2:53 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] melanin renoval Does anyone have a procedure to bleach unstained slides (from FFPE tissue) in order to remove melanin in order to stain with DAB for PECAM? Jane Moose LIS Coordinator/Histology Newberry County Memorial Hospital Newberry, SC 803-405-7129 ************************************************************************ ************ This 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 message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Mon Jun 13 15:04:59 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] melanin renoval Message-ID: Or how about a red or purple DAB replacement? Biocare has a couple of permanent chromogens that can be used for HRP. I plan to do exactly this for some melanin-loaded xenografts. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist GPRD Cancer Research Abbott Laboratories, Abbott Park, IL Jackie.OConnor@abbott.com "Elizabeth Chlipala" Sent by: histonet-bounces@lists.utsouthwestern.edu 06/13/2005 02:58 PM To: "'Jane Moose'" , cc: Subject: RE: [Histonet] melanin renoval Jane Rather than perform a melanin bleach why don't you switch to a alkaline phosphatase detection system and use fast red as your chromagen. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jane Moose Sent: Monday, June 13, 2005 2:53 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] melanin renoval Does anyone have a procedure to bleach unstained slides (from FFPE tissue) in order to remove melanin in order to stain with DAB for PECAM? Jane Moose LIS Coordinator/Histology Newberry County Memorial Hospital Newberry, SC 803-405-7129 ************************************************************************ ************ This 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 message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Mon Jun 13 16:32:47 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] CPT coding question - 88360 Message-ID: For those of you performing CPT coding I have question. Is your technical charge (Part A) for 88360 the same or higher than that for 88342? In my opinion, it should be the same, but our hospital is charging a great deal more for this service. I think the change in RVU's should be on the interpretation side (Part B - Pathologist), not on the technical side. Thank you! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From Rcartun <@t> harthosp.org Mon Jun 13 16:34:31 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Tonsils - submission for pathology Message-ID: What guidelines (patient age) do people use for submitting tonsils for pathologic examination? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From deb.vaneyck <@t> phci.org Mon Jun 13 16:46:19 2005 From: deb.vaneyck <@t> phci.org (Van Eyck, Deb) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] polyoma virus Message-ID: Hi all, A question for any of you out there doing polyoma virus on urine cyto specs----I am working up Nova Castra's JC/BVK immuno stain for acetone fixed cell smears. (Works well on thin prep material with a 20n minute cold acetone post fix). In your experiences using this or other immuno procedures are you getting staining in most of the decoy cells as well as cytoplasmic staining in infected cells and free virus particles? My results are very clean and I am seeing some total nuclear staining in some of the decoy cells, but not all, some nuclear ring like staining, some cytoplasmic inclusion like staining and some clumps of what I am thinking are free virus particles? Any one out there using SV-40 for cyto specs and polyoma virus? If so would you please share your protocol with me? thanks very much. This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. From jdmd77 <@t> hotmail.com Mon Jun 13 16:50:23 2005 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Tonsils - submission for pathology In-Reply-To: Message-ID: The age range for each of the three institutions that I have been in was between 12 and 16 for routine tonsillectomy to get histologic examination... and of course, any masses, necrosis or something unusual (like a staple - apparently swallowed at age 10 and with a nifty foreign body reaction). Julia >From: "Richard Cartun" >To: >Subject: [Histonet] Tonsils - submission for pathology >Date: Mon, 13 Jun 2005 17:34:31 -0400 >MIME-Version: 1.0 >Received: from swlx162.swmed.edu ([199.165.152.162]) by mc7-f36.hotmail.com >with Microsoft SMTPSVC(6.0.3790.211); Mon, 13 Jun 2005 14:35:52 -0700 >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34)id 1Dhwaj-0001tB-5f; Mon, 13 Jun >2005 16:35:38 -0500 >Received: from [199.165.152.167] (helo=swlx167.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34) id 1DhwaS-0001sE-DDfor >histonet@lists.utsouthwestern.edu; Mon, 13 Jun 2005 16:35:21 -0500 >Received: from [199.231.28.53] (helo=hcnwgwds01.hh.chs)by swlx167.swmed.edu >with esmtp (Exim 4.44) id 1DhwaS-00057Y-1sfor >histonet@lists.utsouthwestern.edu; Mon, 13 Jun 2005 16:35:16 -0500 >Received: from HHCC-MTA by hcnwgwds01.hh.chswith Novell_GroupWise; Mon, 13 >Jun 2005 17:35:05 -0400 >X-Message-Info: gUeNUVfFqHBxE7JvKVytr/xaii1VcEUVvnYcKZJdG3A= >X-Mailer: Novell GroupWise Internet Agent 6.5.4 X-Scan-Signature: >ab3f4c55490e4d672805895add812bc5 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >Precedence: list >List-Id: For the exchange of information pertaining to histotechnology >andrelated fields >List-Unsubscribe: >, > >List-Archive: >List-Post: >List-Help: >List-Subscribe: >, >Errors-To: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 7bf0d7e947a05afa907d3090d45e1f8d >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: X-Spam-Status: No, hits=0.0 required=5.5 tests=none >autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >Return-Path: histonet-bounces@lists.utsouthwestern.edu >X-OriginalArrivalTime: 13 Jun 2005 21:35:52.0956 (UTC) >FILETIME=[DFD74BC0:01C5705F] > >What guidelines (patient age) do people use for submitting tonsils for >pathologic examination? Thank you. > >Richard > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 >(860) 545-1596 >(860) 545-0174 Fax > > > > >----------------------------------------------- >Confidentiality Notice > >This e-mail message, including any attachments, is for the sole use of the >intended recipient(s) and may contain confidential or proprietary >information which is legally privileged. Any unauthorized review, use, >disclosure, or distribution is prohibited. If you are not the intended >recipient, please promptly contact the sender by reply e-mail and destroy >all copies of the original message. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jessgrocki <@t> yahoo.com Mon Jun 13 20:53:32 2005 From: jessgrocki <@t> yahoo.com (Jessica Piche) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Controls used for Muscle biopsies In-Reply-To: <3AADFB88753AD31189C100902786B91C14E6DFFC@hch_nt_exchange.hhsc.ca> Message-ID: <20050614015332.1766.qmail@web81706.mail.yahoo.com> Hi Julia, The lab where I did muscle biopsy stains never used controls, but you really should. There are a bunch of techs on the histonet that helped me when I needed information so I'm sure they'll see your question and help you too! I'd like to learn what to use as controls as well. Thanks for asking! Jessica Piche-Grocki, HT Celebre Julia wrote: Hello Histonetters... This is a question for all those labs out there that do enzyme histochemistry on muscle biopsies. Does anyone run controls along with there test muscle biopsies? If your honest answer is yes (not like mine was), what do you use? Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca The Cornerstone of Care Campaign for Hamilton Health Sciences needs your support. Please, visit hamiltonhealthsciences.ca today and help make something great even greater. This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue Jun 14 05:17:06 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Re: nails Message-ID: Try processing as routine tissue, block out the nail so you are sectioning it along the vertical or diagonal (assuming it has been cut transversely), soften in Baker's fluid for half hour (10% glycerine in 70% alcohol) and mount onto gelatin/chrome alum-coated slides (0.1% chrome alum in 1% gelatin). Haven't tried immunos on nails - best of luck! Jacqui Royal Infirmary Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From cfockler <@t> mail1.vcu.edu Tue Jun 14 06:23:03 2005 From: cfockler <@t> mail1.vcu.edu (cfockler@mail1.vcu.edu) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Controls used for Muscle biopsies In-Reply-To: <20050614015332.1766.qmail@web81706.mail.yahoo.com> Message-ID: <200506141123.HAA08359@despina.vcu.edu> ------------------- > Hi Julia, > The lab where I did muscle biopsy stains never used controls, but you really should. There are a bunch of techs on the histonet that helped me when I needed information so I'm sure they'll see your question and help you too! I'd like to learn what to use as controls as well. Thanks for asking! > > Jessica Piche-Grocki, HT > > Celebre Julia wrote: > Hello Histonetters... > This is a question for all those labs out there that do enzyme > histochemistry on muscle biopsies. > Does anyone run controls along with there test muscle biopsies? If your > honest answer is yes (not like mine was), what do you use? > > Julia Celebre MLT > Anatomic Pathology > Hamilton General Hospital > 905-527-0271 ext46145 > email: celebrej@hhsc.ca > > > The Cornerstone of Care Campaign for Hamilton Health Sciences needs your > support. Please, visit hamiltonhealthsciences.ca today and help make > something great even greater. > > This information is directed in confidence solely to the person named above > and may not otherwise be distributed, copied or disclosed. Therefore, this > information should be considered strictly confidential. If you have > received this email in error, please notify the sender immediately via a > return email for further direction. Thank you for your assistance. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > We have experimented using controls. We get a lot of research, and have used fresh rat muscle (quad muscle) and the controls have worked beautifully. The only problem is the tissue must be fresh and you can only store cut controls for so long (in a -80 deep freezer) so it can be difficult to keep up with. Candyce Fockler Histotechnician Anatomic Pathology From lwhite <@t> lakeridgehealth.on.ca Tue Jun 14 06:46:42 2005 From: lwhite <@t> lakeridgehealth.on.ca (White, Lori) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] RE: IVIEW DAB detection kits Message-ID: Hi Tracey, I am assuming from your email address that you are from Canada? I have been experiencing significant background staining and variability for the last couple of weeks. I have been working with the Ventana tech support and discovered there are other "unhappy" accounts in Canada related to a specific master lot. We seemed to have resolved the problem after receiving some new kits with a different lot #. I am still cautious however and advise you to follow up with your Ventana rep. Good luck! Lori -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Saturday, June 11, 2005 1:01 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 19, Issue 17 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. antigen retrieval in bone paraffin sections (pex) 2. RE: antigen retrieval in bone paraffin sections (Elizabeth Chlipala) 3. IVIEW DAB detection kits (Tracey.Lenek@CLS.ab.ca) ---------------------------------------------------------------------- Message: 1 Date: Sat, 11 Jun 2005 05:25:05 +0800 (CST) From: pex Subject: [Histonet] antigen retrieval in bone paraffin sections To: Histonet@lists.utsouthwestern.edu Message-ID: <20050610212505.64592.qmail@web15502.mail.cnb.yahoo.com> Content-Type: text/plain; charset=gb2312 Hello, all, Do you have some good suggestions about antigen retrieval in bone paraffin sections? Thank you! Guofeng __________________________________________________ ????????????????????????????? http://cn.mail.yahoo.com ------------------------------ Message: 2 Date: Fri, 10 Jun 2005 15:42:01 -0600 From: "Elizabeth Chlipala" Subject: RE: [Histonet] antigen retrieval in bone paraffin sections To: "'pex'" , Message-ID: <000201c56e05$3c16dbb0$a7d48a80@AMY> Content-Type: text/plain; charset="gb2312" Guofeng I frequently use the ready to use proteinase K from DakoCytomation on formalin fixed, formic acid decalcified bone sections. I get good results using proteinase K with antibodies such as F4/80 in mouse joints, ED-1 in rat joints. IL-6 in both mouse and rat joints, mouse neutrophils, factor H, BrdU and Factor VIII. The other retrieval method I use is BioGenex's DeCal. I use that for CD45R in mice. I try to stay away from HEIR. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pex Sent: Friday, June 10, 2005 2:25 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] antigen retrieval in bone paraffin sections Hello, all, Do you have some good suggestions about antigen retrieval in bone paraffin sections? Thank you! Guofeng __________________________________________________ ????????????????????????????? http://cn.mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Fri, 10 Jun 2005 16:36:12 -0600 From: Subject: [Histonet] IVIEW DAB detection kits To: Message-ID: <4BC300747AF87A48BCDF8E48BC2885CE010E0F10@mail1.calgary.com> Content-Type: text/plain; charset="iso-8859-1" Hi, We are a high volume IHC department and use Ventana's systems exclusively. Over the past 6 months we have noticed inconsistent staining and batch to batch variation with different lot #'s of detection kits. Very difficult to maintain consistency with a large number of primary antibodies that we offer. Has anyone else experienced a similar problem? Thanks Tracey CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 19, Issue 17 **************************************** From GDawson <@t> dynacaremilwaukee.com Tue Jun 14 07:25:44 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] melanin renoval Message-ID: Jane, I've found that a melanin bleach hurts IHC staining. Have you considered using something like Nova Red from Vector as a good DAB substitute that yields red staining? This red chromagen contrasts much better with the melanin in the tissue and I've found I don't have to modify any of my usual procedures to achieve good staining. It is also permanent and does not wash out in alcohols like say an alc. phos. red. Let me know if you need the catalogue #. Good Luck, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: Jane Moose [mailto:jane.moose@newberryhospital.org] Sent: Monday, June 13, 2005 3:53 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] melanin renoval Does anyone have a procedure to bleach unstained slides (from FFPE tissue) in order to remove melanin in order to stain with DAB for PECAM? Jane Moose LIS Coordinator/Histology Newberry County Memorial Hospital Newberry, SC 803-405-7129 **************************************************************************** ******** This 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 message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ryakay <@t> shands.ufl.edu Tue Jun 14 09:04:17 2005 From: ryakay <@t> shands.ufl.edu (Kaye Ryan) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] decaling of bone marrow specimens for subsequent IHC staining Message-ID: Hi Everyone, Could you please share your choice of decal solutions for bone marrow specimens that will need IHC staining? Thanks for your time, Kaye Ryan Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 From Emily.Wiesner <@t> medecine.unige.ch Tue Jun 14 09:36:14 2005 From: Emily.Wiesner <@t> medecine.unige.ch (Emily Jane Wiesner-Camm) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Cryostat sections with small bubbles Message-ID: <42AEEB5E.3070104@medecine.unige.ch> Hi All, I was wondering whether anyone can let me know why small bubbles appear underneath my rat brain sections when I cut them on a cryostat (at -35) when they appear perfectly flat before placing them on a hotplate. How they can be avoided? Thanks, Emily From sjchtascp <@t> yahoo.com Tue Jun 14 09:49:02 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] iron hematoxylins Message-ID: <20050614144903.12252.qmail@web90202.mail.scd.yahoo.com> Does anyone know if using a bluing agent after weigerts iron hematoxylin will achieve the same results a lengthy tap water rince? Steve --------------------------------- Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. From chiggerson <@t> memhosp.com Tue Jun 14 10:04:21 2005 From: chiggerson <@t> memhosp.com (chiggerson@memhosp.com) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Histotech Trainee Pay Ranges Message-ID: Hi everyone, I am hoping that some of you are willing to share pay scale information. I am mostly interested in the hourly pay rate range for histotechnician/histotechnologist trainee positions and how those compare to the histotechnician and histotechnologist pay rate ranges. Our trainee pay range seems a bit out of whack. When an employee passes the BOR exam they essentially double their salary when they move to a tech scale. I would like to raise the pay scale for our trainees, and need to gather some info from those willing to share. It would be helpful to include the histotech and histotechnologist rates as well so I can see the overlap or lack of between the trainee scale and the tech scales. Thanks, Cindy Cindy Higgerson HTL(ASCP)CM Surgical Pathology Supervisor Memorial Hospital chiggerson@memhosp.com office: 618-257-5227 fax: 618-257-6868 From pruegg <@t> ihctech.net Tue Jun 14 10:33:14 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] cryo base molds Message-ID: <200506141533.j5EFX4t1027731@chip.viawest.net> I am looking for where to buy the ?Tissue Tek thin plastic base molds we use for freezing tissue. I have cut them so that they fit inside a mega cassette. I cannot find these anywhere anymore. I hope my description is clear. Patsy From DonnaWillis <@t> texashealth.org Tue Jun 14 11:08:49 2005 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] decaling of bone marrow specimens for subsequent IHCstaining Message-ID: <4483B6EE0FA82A4EAED2B1E161E5E41147C871@ftwex02.txhealth.org> Kaye, We use IED (Ion-Exchange Decal) from Biocare. 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 Kaye Ryan Sent: Tuesday, June 14, 2005 9:04 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] decaling of bone marrow specimens for subsequent IHCstaining Hi Everyone, Could you please share your choice of decal solutions for bone marrow specimens that will need IHC staining? Thanks for your time, Kaye Ryan Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 _______________________________________________ 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 POWELL_SA <@t> Mercer.edu Tue Jun 14 11:17:56 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] cryo base molds In-Reply-To: <200506141533.j5EFX4t1027731@chip.viawest.net> Message-ID: <01LPG8VC2HCC8WXF45@Macon2.Mercer.edu> Hi Patsy, If you are talking about the 4557 Tissue Tek Cryomolds, I got mine from EMS, the number on the box is 197555-081, but I ordered these some time ago, 4 or 5 years maybe. Shirley Powell -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Tuesday, June 14, 2005 10:33 AM To: 'Histonet' Subject: [Histonet] cryo base molds I am looking for where to buy the ?Tissue Tek thin plastic base molds we use for freezing tissue. I have cut them so that they fit inside a mega cassette. I cannot find these anywhere anymore. I hope my description is clear. Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Tue Jun 14 12:36:45 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] melanin renoval In-Reply-To: References: Message-ID: Along those same lines ... for staining CD31/PECAM in the melanoma samples I work with I use True Blue chromagen and it works beautifully! Andrea At 7:25 AM -0500 6/14/05, Dawson, Glen wrote: >Jane, > >I've found that a melanin bleach hurts IHC staining. Have you considered >using something like Nova Red from Vector as a good DAB substitute that >yields red staining? This red chromagen contrasts much better with the >melanin in the tissue and I've found I don't have to modify any of my usual >procedures to achieve good staining. It is also permanent and does not wash >out in alcohols like say an alc. phos. red. Let me know if you need the >catalogue #. > >Good Luck, > >Glen Dawson BS, HT & QIHC (ASCP) >IHC Manager >Milwaukee, WI -- From anh2006 <@t> med.cornell.edu Tue Jun 14 12:43:07 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] melanin renoval In-Reply-To: References: Message-ID: Sorry, I forgot to mention the True Blue is from KPL. Andrea At 1:36 PM -0400 6/14/05, Andrea Hooper wrote: >Along those same lines ... for staining CD31/PECAM in the melanoma >samples I work with I use True Blue chromagen and it works >beautifully! > -- From jlf <@t> jax.org Tue Jun 14 13:04:30 2005 From: jlf <@t> jax.org (Judi Ford) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] stains on mouse embryos Message-ID: <6.0.3.0.0.20050614140232.01b4c880@aretha.jax.org> Hi All, I have someone who wants to locate the gonadal cavity in a 10 day old mouse embryo. Is there a stain that may help to locate the area? Thanks, Judi Ford The Jackson Laboratory Bar Harbor, ME From info <@t> instrumedics.com Tue Jun 14 14:07:19 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] frozen sections of bone not sticking to slide Message-ID: <006e01c57114$4eba8b60$6401a8c0@INSTRUMEDICS22> Phil, I think you might be interested in the CryoJane Tape-Transfer system which is used by many labs cutting bone and cartilage. It installs in your existing cryostat Please visit our web site www.instrumedics.com and click on frozen sections on the home page. Also click on "gallery" to see photomicrographs of CryoJane prepared sections. Bernice Instrumedics From petepath <@t> yahoo.com Tue Jun 14 15:37:56 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Tonsils - submission for pathology Message-ID: <20050614203757.44709.qmail@web30413.mail.mud.yahoo.com> HI Richard, We submit all our tonsils. A few years back I brought a list of specimen types to our medical board that we felt could be grossed only or not sent to pathology. Our medical board wanted everything sent to path except normal placentas. They wanted all tonsils examined. Sounds like a bit of over kill but demonstrates how ideas can vary from institution to institution. Another comment on "insignificant specimens". We do a lot of gastric bypass surgery. We found out that the surgeons were throwing the anastomotic rings in the can. We asked if they would send them to pathology. To our surprise we started finding quite a few helicobacters gastritis cases. Maybe one in five or so. So make sure they are not tossing these. I have also turned up a few cancers in the macromastia specimens and a handful of occult mets in hernia sacs. Sometimes those "insignificant specimens" are not so insignificant. Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From ploykasek <@t> phenopath.com Tue Jun 14 17:34:53 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Job opening Message-ID: I would like to announce an opening at PhenoPath Laboratories in Seattle, WA. This position is in our contract research department. This position involves research immunohistochemistry projects at the bench level. Will need to provide oversight, training, evaluation, and prioritization of work for 3+ technologists; communicate with clients and manager in planning, executing and documenting projects. Quality control, safety, and preventive maintenance oversight also required. Bachelor?s of Science with a background in immunohistochemistry and project management; strong administrative, organizational, and interpersonal skills; laboratory supervision and experience with GLP projects desired; histology experience a plus. For more information, visit www.phenopath.com. Thank you. Patti Loykasek PhenoPath Laboratories From lpwenk <@t> sbcglobal.net Tue Jun 14 19:07:25 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] iron hematoxylins In-Reply-To: <20050614144903.12252.qmail@web90202.mail.scd.yahoo.com> Message-ID: I've never blued after a Weigert iron hematoxylin stain. Nuclei are either black, or they aren't. If they are not, then the iron hematoxylin is old (more than a couple days old), and putting the tissue into an alkali solution won't bring the color back. I'm curious, why are you bluing? What is the purpose? Maybe then I can help. Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Tuesday, June 14, 2005 10:49 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] iron hematoxylins Does anyone know if using a bluing agent after weigerts iron hematoxylin will achieve the same results a lengthy tap water rince? Steve --------------------------------- Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From llewllew <@t> shaw.ca Tue Jun 14 20:29:02 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] iron hematoxylins References: <0II300576N5ZN000@l-daemon> Message-ID: <000a01c57149$9cff5e60$7e034246@yourlk4rlmsu> I have blued Weigert's iron hematoxylin in the past, although I didn't do it as a routine. I wondered if the acid in the Weigert's solution affected the colour and I tried it to find out. Blueing does deepen the colour, although not as significantly as with alum hematoxylin. However, the acid solutions I used afterwards in Masson's trichrome undid it and the end result was the same. It is useful if the iron hematoxylin is used alone without a counterstain, perhaps on smears for amoebae, but otherwise not. Other iron hematoxylin stains of the Heidenhain type do not need blueing. The mordant does that. Bryan Llewellyn ----- Original Message ----- From: "Lee & Peggy Wenk" To: "'Steven Coakley'" ; Sent: Tuesday, June 14, 2005 5:07 PM Subject: RE: [Histonet] iron hematoxylins > I've never blued after a Weigert iron hematoxylin stain. Nuclei are either > black, or they aren't. If they are not, then the iron hematoxylin is old > (more than a couple days old), and putting the tissue into an alkali > solution won't bring the color back. > > I'm curious, why are you bluing? What is the purpose? Maybe then I can > help. > > Peggy Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven > Coakley > Sent: Tuesday, June 14, 2005 10:49 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] iron hematoxylins > > Does anyone know if using a bluing agent after weigerts iron hematoxylin > will achieve the same results a lengthy tap water rince? > > Steve > > > --------------------------------- > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > _______________________________________________ > Histonet mailing list > Histonet@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 katri <@t> cogeco.ca Tue Jun 14 20:39:51 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Cryostat sections with small bubbles References: <42AEEB5E.3070104@medecine.unige.ch> Message-ID: <004001c5714b$1fbb1f50$6a9a9618@Katri> Hi Emily, I have never heard of hot plating cryostat sections. You may be boiling the moisture under the section forming bubbles or do you air dry them well first? Also -35 C seems too cold for brain sections, you may be getting some cutting artifacts. I'm sure there'll be others with helpful information for you. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Emily Jane Wiesner-Camm" To: Sent: Tuesday, June 14, 2005 10:36 AM Subject: [Histonet] Cryostat sections with small bubbles > Hi All, > I was wondering whether anyone can let me know why small bubbles appear > underneath my rat brain sections when I cut them on a cryostat (at -35) > when they appear perfectly flat before placing them on a hotplate. How > they can be avoided? > Thanks, > Emily > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Jun 15 01:40:57 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] iron hematoxylins[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2AA@elht-exch1.xelht.nhs.uk> Confused???? Isn't haematin like litmus and changes colour in acid and alkali solutions? Doesn't the mordant act as a 'key' to the proteins and haematin causing the dye to 'bind'? So why, if the iron haematoxylin is in an acid 'environment' does it not require 'blueing'? -----Original Message----- From: Bryan Llewellyn [mailto:llewllew@shaw.ca] Sent: 15 June 2005 02:29 To: Histonet Subject: Re: [Histonet] iron hematoxylins[Scanned] I have blued Weigert's iron hematoxylin in the past, although I didn't do it as a routine. I wondered if the acid in the Weigert's solution affected the colour and I tried it to find out. Blueing does deepen the colour, although not as significantly as with alum hematoxylin. However, the acid solutions I used afterwards in Masson's trichrome undid it and the end result was the same. It is useful if the iron hematoxylin is used alone without a counterstain, perhaps on smears for amoebae, but otherwise not. Other iron hematoxylin stains of the Heidenhain type do not need blueing. The mordant does that. Bryan Llewellyn ----- Original Message ----- From: "Lee & Peggy Wenk" To: "'Steven Coakley'" ; Sent: Tuesday, June 14, 2005 5:07 PM Subject: RE: [Histonet] iron hematoxylins > I've never blued after a Weigert iron hematoxylin stain. Nuclei are either > black, or they aren't. If they are not, then the iron hematoxylin is old > (more than a couple days old), and putting the tissue into an alkali > solution won't bring the color back. > > I'm curious, why are you bluing? What is the purpose? Maybe then I can > help. > > Peggy Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven > Coakley > Sent: Tuesday, June 14, 2005 10:49 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] iron hematoxylins > > Does anyone know if using a bluing agent after weigerts iron hematoxylin > will achieve the same results a lengthy tap water rince? > > Steve > > > --------------------------------- > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > _______________________________________________ > Histonet mailing list > Histonet@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 Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Jun 15 02:42:20 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Melanin pigment Message-ID: I have a paper which compares two bleaching techniques which you might find useful: British Journal of Biomed. Science 1999; 56: 188-193. It examines the effect of the two methods on subsequent immunostaining with their pros and cons on various antibodies. If you don't want to change your immuno method and can't get hold of the info, let me know and I will summarize it for you. Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From BMolinari <@t> heart.thi.tmc.edu Wed Jun 15 05:47:39 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] cryomolds Message-ID: Hi all, Does anyone know if there are cryomolds that are deeper than 5mm? I have some mouse brains to cut and the researcher wants to keep them intact and not bisect them, any suggestions? Thanks, Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston, Texas 77030 832-355-6524 832-344-6812 (fax) From petepath <@t> yahoo.com Wed Jun 15 06:24:48 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] cryomolds Message-ID: <20050615112449.26577.qmail@web30410.mail.mud.yahoo.com> Hi Betsy, I custom make my my well bars and chucks to any depth and size. Visit http://pathologyinnovations.com/index.html to learn about these techniques. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From JWEEMS <@t> sjha.org Wed Jun 15 07:08:26 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] CPT modifier - 59 Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA46032@sjhaexc02.sjha.org> Billing questions again!!! I'm looing into the modifier 59, "Distinct Procedural Service". How many of you use it for additional tests performed on AP specimens, e.g. sentinel nodes that require immunos on more than one block of the same specimen? Thanks! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From llewllew <@t> shaw.ca Wed Jun 15 07:57:00 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] iron hematoxylins[Scanned] References: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2AA@elht-exch1.xelht.nhs.uk> Message-ID: <000601c571a9$b8bbbc10$7e034246@yourlk4rlmsu> Iron hematoxylins of the Heidenhain type do not use acids and usually no counterstain is applied. That type uses three bath staining. The first is mordanting in a ferric compound, usually ferric ammonium sulphate, the second is in the hematoxylin, and the third usually returns to the mordant to differentiate. Acids are not usually involved, and the third step leaves the tissue black stained. An exception is some modifications which replace the mordant with an acid, but these are not commonly used. Extensive washing is recommended with them, but to remove traces of mordant rather than to alkalise, although that may happen depending on the pH of the tap water. Bryan Llewellyn ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Bryan Llewellyn" ; "Histonet" Sent: Tuesday, June 14, 2005 11:40 PM Subject: RE: [Histonet] iron hematoxylins[Scanned] Confused???? Isn't haematin like litmus and changes colour in acid and alkali solutions? Doesn't the mordant act as a 'key' to the proteins and haematin causing the dye to 'bind'? So why, if the iron haematoxylin is in an acid 'environment' does it not require 'blueing'? -----Original Message----- From: Bryan Llewellyn [mailto:llewllew@shaw.ca] Sent: 15 June 2005 02:29 To: Histonet Subject: Re: [Histonet] iron hematoxylins[Scanned] I have blued Weigert's iron hematoxylin in the past, although I didn't do it as a routine. I wondered if the acid in the Weigert's solution affected the colour and I tried it to find out. Blueing does deepen the colour, although not as significantly as with alum hematoxylin. However, the acid solutions I used afterwards in Masson's trichrome undid it and the end result was the same. It is useful if the iron hematoxylin is used alone without a counterstain, perhaps on smears for amoebae, but otherwise not. Other iron hematoxylin stains of the Heidenhain type do not need blueing. The mordant does that. Bryan Llewellyn ----- Original Message ----- From: "Lee & Peggy Wenk" To: "'Steven Coakley'" ; Sent: Tuesday, June 14, 2005 5:07 PM Subject: RE: [Histonet] iron hematoxylins > I've never blued after a Weigert iron hematoxylin stain. Nuclei are either > black, or they aren't. If they are not, then the iron hematoxylin is old > (more than a couple days old), and putting the tissue into an alkali > solution won't bring the color back. > > I'm curious, why are you bluing? What is the purpose? Maybe then I can > help. > > Peggy Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven > Coakley > Sent: Tuesday, June 14, 2005 10:49 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] iron hematoxylins > > Does anyone know if using a bluing agent after weigerts iron hematoxylin > will achieve the same results a lengthy tap water rince? > > Steve > > > --------------------------------- > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Jun 15 08:52:49 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] iron hematoxylins[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD442@elht-exch1.xelht.nhs.uk> Heidenhain's uses sequence iron staining, as does Benda, Masson-Regaud and Mallory. Heidenhain's differentiates in an iron alum solution, Mallory in Ferric Chloride, Benda in acetic acid and Masson-Regaud in alcoholic picric acid. This depends on mordanting with an aqueous ferric salt solution, staining with haematoxylin until black, then differentiating with an acid or a ferric salt until you visualise cytologic detail. Progressive or regressive iron haematoxylin staining (Janssens or Weigerts) are similar to alum haematoxylin staining. Regressive staining depends on an optimal amount of iron for dense staining (eg for the demonstration of myelin), whilst progressive staining is made more selective for nuclei by addition of acid or by an excess of ferric salt. You see we are talking about different uses for iron haematoxylin, now I wish I knew how ICC worked!!! -----Original Message----- From: Bryan Llewellyn [mailto:llewllew@shaw.ca] Sent: 15 June 2005 13:57 To: Histonet Subject: Re: [Histonet] iron hematoxylins[Scanned] Iron hematoxylins of the Heidenhain type do not use acids and usually no counterstain is applied. That type uses three bath staining. The first is mordanting in a ferric compound, usually ferric ammonium sulphate, the second is in the hematoxylin, and the third usually returns to the mordant to differentiate. Acids are not usually involved, and the third step leaves the tissue black stained. An exception is some modifications which replace the mordant with an acid, but these are not commonly used. Extensive washing is recommended with them, but to remove traces of mordant rather than to alkalise, although that may happen depending on the pH of the tap water. Bryan Llewellyn ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" To: "Bryan Llewellyn" ; "Histonet" Sent: Tuesday, June 14, 2005 11:40 PM Subject: RE: [Histonet] iron hematoxylins[Scanned] Confused???? Isn't haematin like litmus and changes colour in acid and alkali solutions? Doesn't the mordant act as a 'key' to the proteins and haematin causing the dye to 'bind'? So why, if the iron haematoxylin is in an acid 'environment' does it not require 'blueing'? -----Original Message----- From: Bryan Llewellyn [mailto:llewllew@shaw.ca] Sent: 15 June 2005 02:29 To: Histonet Subject: Re: [Histonet] iron hematoxylins[Scanned] I have blued Weigert's iron hematoxylin in the past, although I didn't do it as a routine. I wondered if the acid in the Weigert's solution affected the colour and I tried it to find out. Blueing does deepen the colour, although not as significantly as with alum hematoxylin. However, the acid solutions I used afterwards in Masson's trichrome undid it and the end result was the same. It is useful if the iron hematoxylin is used alone without a counterstain, perhaps on smears for amoebae, but otherwise not. Other iron hematoxylin stains of the Heidenhain type do not need blueing. The mordant does that. Bryan Llewellyn ----- Original Message ----- From: "Lee & Peggy Wenk" To: "'Steven Coakley'" ; Sent: Tuesday, June 14, 2005 5:07 PM Subject: RE: [Histonet] iron hematoxylins > I've never blued after a Weigert iron hematoxylin stain. Nuclei are either > black, or they aren't. If they are not, then the iron hematoxylin is old > (more than a couple days old), and putting the tissue into an alkali > solution won't bring the color back. > > I'm curious, why are you bluing? What is the purpose? Maybe then I can > help. > > Peggy Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven > Coakley > Sent: Tuesday, June 14, 2005 10:49 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] iron hematoxylins > > Does anyone know if using a bluing agent after weigerts iron hematoxylin > will achieve the same results a lengthy tap water rince? > > Steve > > > --------------------------------- > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > _______________________________________________ > Histonet mailing list > Histonet@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 POWELL_SA <@t> Mercer.edu Wed Jun 15 09:55:22 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] cryomolds In-Reply-To: Message-ID: <01LPHKAB24C48WXJC3@Macon2.Mercer.edu> Try some of the molds sold for plastic embedding which are deeper and then transfer to your cryostat chucks. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Molinari, Betsy Sent: Wednesday, June 15, 2005 5:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cryomolds Hi all, Does anyone know if there are cryomolds that are deeper than 5mm? I have some mouse brains to cut and the researcher wants to keep them intact and not bisect them, any suggestions? Thanks, Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston, Texas 77030 832-355-6524 832-344-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sue.Kapoor <@t> uhsi.org Wed Jun 15 10:19:22 2005 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] alcohol/room size Message-ID: <61E9F2400F53D5119CFC00508B44E33B02E9E12D@khmcexch.uhsi.org> Hi, can anyone tell me or direct me to information of what amount of alchol is allowed in a room by the square footage? Thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From carolb <@t> mail.phys.mcw.edu Wed Jun 15 10:26:34 2005 From: carolb <@t> mail.phys.mcw.edu (Carol Bobrowitz) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] ergonomic table Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A969B@thor.phys.mcw.edu> I am searching for a source/vendors for an ergonomic table. A table with adjustable heights. It will be used for microtome use. The company we did order one from ignored to inform us they are no longer in business. Any leads would be greatly appreciated. Thanks in advance and Thank You All on the Histonet for all the help you have given me over the years. Carol Ann Bobrowitz cbobrowi@mcw.edu From Kemlo.Rogerson <@t> elht.nhs.uk Wed Jun 15 10:36:12 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] alcohol/room size[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2BA@elht-exch1.xelht.nhs.uk> Doesn't work like that, you need a detector to record the levels; there may be fire regs for thaT but that will be local -----Original Message----- From: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: 15 June 2005 16:19 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] alcohol/room size[Scanned] Hi, can anyone tell me or direct me to information of what amount of alchol is allowed in a room by the square footage? Thanks 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 kspencer <@t> utmem.edu Wed Jun 15 10:49:37 2005 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:25:12 2005 Subject: [Histonet] Cryostat sections with small bubbles In-Reply-To: <004001c5714b$1fbb1f50$6a9a9618@Katri> References: <42AEEB5E.3070104@medecine.unige.ch> <004001c5714b$1fbb1f50$6a9a9618@Katri> Message-ID: <6c8cb6f9da57811fb4731d13a09c4d2d@utmem.edu> Hi Emily, I cut rat brain sections at -18 to -20 and often have to warm the block face with my finger before each section. I also have better luck if I cut sections at 10u. These are fresh frozen brains. If I cut them at 20u I get bubbles under the sections. I keep them in the cryostat, and then fix the sections in cold fix, buffer wash, water wash, then air dry them in the hood. When I cut rat brain that has been perfused, I cut 20u floating sections. They never adhere well to slides, even if I use a hot plate. I always have bubbles under the section. That is why in this case we use floating sections. I hope this is helpful. Kathleen Spencer On Jun 14, 2005, at 8:39 PM, Katri Tuomala wrote: > Hi Emily, > I have never heard of hot plating cryostat sections. You may be > boiling the moisture under the section forming bubbles or do you air > dry them well first? Also -35 C seems too cold for brain sections, you > may be getting some cutting artifacts. > I'm sure there'll be others with helpful information for you. > Katri > > Katri Tuomala > Hamilton, Ontario, Canada > ----- Original Message ----- From: "Emily Jane Wiesner-Camm" > > To: > Sent: Tuesday, June 14, 2005 10:36 AM > Subject: [Histonet] Cryostat sections with small bubbles > > >> Hi All, >> I was wondering whether anyone can let me know why small bubbles >> appear underneath my rat brain sections when I cut them on a cryostat >> (at -35) when they appear perfectly flat before placing them on a >> hotplate. How they can be avoided? >> Thanks, >> Emily >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@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 livieira <@t> ualg.pt Wed Jun 15 10:52:15 2005 From: livieira <@t> ualg.pt (Lina Vieira) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Fish eggs Message-ID: <007b01c571c2$34016d30$2914100a@labhistologia> Helo Everyone, Can you give me some advise about fish eggs? I find a method to break/ destroy the fish eggs capsule. The method should be easy for aplication in many fish eggs and have no effect in the embryo. Tanks in advance LIna Vieira From Tbarnhart <@t> primecare.org Wed Jun 15 11:15:57 2005 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] alcohol/room size Message-ID: <1779904B5E82D511914C00D0B793339205BFD962@exchangent> I you check the CAP General Laboratory checklist, item GEN.72050 it gives the amount of flammable liquid that can be stored. You can store up to 1 gallon of Class I, II and IIIA liquids outside a fire-resistant cabinet for each 100 square feet of space. Up to 2 gallons if stored in safety cans or safety cabinets. If larger volumes are to be stored, they will be governed by you state or local fire laws and regulations. This could involve having the liquids stored in a building that has a blast wall along with other things such as a fire suppression system. You would have to check with your local fire marshal. Hope this helps..good luck. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND -----Original Message----- From: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: Wednesday, June 15, 2005 10:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] alcohol/room size Hi, can anyone tell me or direct me to information of what amount of alchol is allowed in a room by the square footage? Thanks 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 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 BDUE <@t> PARTNERS.ORG Wed Jun 15 11:53:50 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Cryostat sections with small bubbles Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB501027054@PHSXMB7.partners.org> Hello Emily, I have two or three tricks you might try, and a comment. The "bubbles" are probably similar to what I get whe picking up 10um human muscle. The tissue "jumps" to the slide in a semi-random fashion no matter how carefully you arrange the pick-up. So the "bubbles" are actually regions that get encircled before flattening out fully on the slide. They are really wrinkles that get "trapped" because the tissue can't spread once it hits the slide. Imagine trying to toss a wet blanket so it lands perfectly flat. 1.) try picking up on cold slides and then warming the back of the slide with your finger and air dry for a few minutes. If your sections are cold and flat, you may be able to get them to lay flat on a cold slide. The cold slide won't "grab" the section like a warm slide. But you do need to warm everything up to get proper adhesion. You can handle thick FS with forceps, especially if there is an OCT border. By controlling how fast you warm the slide, you can to some extent control how the section flattens. 2.) perfused tissue seems ok for you, so fixation is ok. Try floating your 20um fresh frozen sections on a small cup of cold PFA or cold formalin. You could put the fixative bath in your cryostat. Let them float flat like on a paraffin water bath. Pick up on charged or coated slides and air dry. Be careful about floaters. Don't float too long or the fixative will crosslink over the charged sites which you need for adhesion. (This is part of the reason that your perfused sections don't stick well.) 3.) Bancroft (5ed p. 103) lists a great section adhesive that I have used with thick fixed frozens. It should work with your 20um perfused floating sections. I use it modified, as follows. Make A) 1% gelatin in warm water and B) 2% formaldehyde (I use 37% formaldehyde, but you could probably adjust for PFA or NBF). Combine A and B 50:50 shortly before use. The gelatin slowly crosslinks and the soln is only usable for a few hours at room temp. To use, wet the surface of a charged slide with the soln -- use a swab or sponge or something. You only need a film of the adhesive, not a dripping wet slide. You pick up your FS on the wet slide. This takes a little practice to keep the crystat stage from getting covered. The sections spread flat on the film of adhesive. Then air dry the slide for 5-20 min. until visibly well dry -- a 37C oven is ok too. The sections will not fall off. I have done 20um fixed frozens of rat spinal cord (incl. cauda equina) and done silver stains and H&Es and LFBs, and not a single fiber ever fell off. The only down side is that the gelatin around the sections will pick up stains. Good Luck, -brice Neuropathology Lab Brigham & Women's Hospital Boston "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kathleen Spencer Sent: Wednesday, June 15, 2005 11:50 AM To: Katri Tuomala Cc: histonet@lists.utsouthwestern.edu; Emily Jane Wiesner-Camm Subject: Re: [Histonet] Cryostat sections with small bubbles Hi Emily, I cut rat brain sections at -18 to -20 and often have to warm the block face with my finger before each section. I also have better luck if I cut sections at 10u. These are fresh frozen brains. If I cut them at 20u I get bubbles under the sections. I keep them in the cryostat, and then fix the sections in cold fix, buffer wash, water wash, then air dry them in the hood. When I cut rat brain that has been perfused, I cut 20u floating sections. They never adhere well to slides, even if I use a hot plate. I always have bubbles under the section. That is why in this case we use floating sections. I hope this is helpful. Kathleen Spencer On Jun 14, 2005, at 8:39 PM, Katri Tuomala wrote: > Hi Emily, > I have never heard of hot plating cryostat sections. You may be > boiling the moisture under the section forming bubbles or do you air > dry them well first? Also -35 C seems too cold for brain sections, you > may be getting some cutting artifacts. > I'm sure there'll be others with helpful information for you. > Katri > > Katri Tuomala > Hamilton, Ontario, Canada > ----- Original Message ----- From: "Emily Jane Wiesner-Camm" > > To: > Sent: Tuesday, June 14, 2005 10:36 AM > Subject: [Histonet] Cryostat sections with small bubbles > > >> Hi All, >> I was wondering whether anyone can let me know why small bubbles >> appear underneath my rat brain sections when I cut them on a cryostat >> (at -35) when they appear perfectly flat before placing them on a >> hotplate. How they can be avoided? >> Thanks, >> Emily >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Wed Jun 15 12:48:41 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Stupid question about Biocare Romulin red Message-ID: I've never used this product before - can anyone tell me if it's supposed to appear red after it's been prepared? I expected to see some color - but it's clear as a bell. I'm not using it until I get an answer. Help! From dharclerode <@t> macropore.com Wed Jun 15 12:59:02 2005 From: dharclerode <@t> macropore.com (Donna Harclerode) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Deeper molds for mouse brains was cryomolds Message-ID: Hi Betsy I use PEEL-A-WAY molds made by POLYSCIENCES INC (available from VWR) for my bigger specimens. I do not use them for brains. The problem with trying to embed a brain in any sort of orientation in a mold is something I have never solved. When I put the brain in OCT it distorts. Rat and mouse brains cut very nicely (much better for me) if they are frozen by themselves. The 2 ways I do it is to make a foil boat and set the brain in and put it on dry ice with or without isopentane(slabs of dry ice are much better than pellets because they have a flat surface) for a minute or 2 (rats are more on the 2 minute side) till the top is frozen. The only thing with this is you get a flat spot on the bottom of the brain. These brains went for RNA ISH and we did 70 a week so we needed speed. The other way that takes a bit more effort is to hold the brain by the spinal cord and lower it into the isopentane in a beaker in dry ice 10-20 seconds depending on temp. One PI I worked with had a specific temperature range for the isopentane, but I do not remember that now. If you freeze it too long, you can get cracks or even multiple pieces if you leave it in way too long. After the brain is frozen (less is better than over freezing) I put the brains in conical tubes on dry ice till I get them to the -80 freezer. I have not had problems with ice artifact with either of these methods. For OCT embedded "regular" tissue either of these 2 methods have given me ice artifact either due to the warmer temperatures or increased time because of the OCT. I always use isopentane and liquid nitrogen for tissue other than brain Maybe the high lipid content in the brain makes the ice artifact less of a problem. To cut these I put a bit of OCT on the chuck and put the brain in the position I wanted. I usually cut the rostral end for coronal sections to make a flat spot for the chuck. One other tip we did was to have a small container of OCT and quickly dip the frozen mounted brain and refreeze the block. I made a hollow of dry ice in the container, dipped the brain and refroze the brain as quickly as possible. Good luck Donna Harclerode, HT, (ASCP), HTL, QIHC Immunohistochemist MacroPore Biosurgery, Inc 6740 Top Gun St. San Diego, CA 92121 858-458-0900 xt 322 dharclerode@macropore.com Subject: RE: [Histonet] cryomolds To: " Try some of the molds sold for plastic embedding which are deeper and then transfer to your cryostat chucks. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Molinari, Betsy Sent: Wednesday, June 15, 2005 5:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cryomolds Hi all, Does anyone know if there are cryomolds that are deeper than 5mm? I have some mouse brains to cut and the researcher wants to keep them intact and not bisect them, any suggestions? Thanks, Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston, Texas 77030 832-355-6524 832-344-6812 (fax) _______________________________________________ Histonet mailing list From sjchtascp <@t> yahoo.com Wed Jun 15 13:02:40 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Re: delayed processing Message-ID: <20050615180241.1268.qmail@web90205.mail.scd.yahoo.com> Steven Coakley wrote:Hello everyone, :-) <-------- "me again" Just started working in research so I am encountering many new things here. With animal, mainly mouse/rat muscle & liver tissue ( no fatty 1/4 pounders here), I've learned to cut back on the temps and time in the processing reagents. Heres my question, it been 3 weeks since our last processing cycle on our VIP. The reagents do not appear to have evaporated any. Should I have any concerns, especially of the absolute alcohols absorbing enough "moisture" to effect processing? Steve --------------------------------- Discover Yahoo! Have fun online with music videos, cool games, IM & more. Check it out! --------------------------------- Discover Yahoo! Get on-the-go sports scores, stock quotes, news & more. Check it out! From pzeitlow <@t> bbpllab.com Wed Jun 15 14:22:15 2005 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] UNSUBSCRIBE Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A47D64B@bbplsrv1.bbpl> Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. From BDUE <@t> PARTNERS.ORG Wed Jun 15 14:44:44 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:13 2005 Subject: OCT... RE: [Histonet] Deeper molds for mouse brains was cryomolds Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB501027055@PHSXMB7.partners.org> Hi Donna, I primarily work with human muscle bxs, which are sensitive to conditions that produce ice artifacts. My cold OCT trick may apply to rodent brains. I keep a bottle of OCT in a regular 4C refrigerator. I use it to embed troublesome muscles *after* they have been snap frozen in isopentane. No ice artifacts show up except maybe the outermost muscle fiber or two -- often none at all. After applying the cold OCT, I use either a freeze spray or dry ice or a -80C freezer to finish the job. For brains you could try snap freezing specimen and post-embedding like I do muscles. Or you could try using the cold OCT to fill your molds before freezing. This moves the specimen + OCT temp 15-20C south of RT, which means the isopentane has much less work to do to cross the freezing point. I don't freeze whole OCT blocks often, but I have noticed that cold OCT has fewer cracking problems than RT OCT when rapidly frozen. -brice Neuropathology Lab Brigham & Women's Hospital Boston "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Donna Harclerode Sent: Wednesday, June 15, 2005 1:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Deeper molds for mouse brains was cryomolds Hi Betsy I use PEEL-A-WAY molds made by POLYSCIENCES INC (available from VWR) for my bigger specimens. I do not use them for brains. The problem with trying to embed a brain in any sort of orientation in a mold is something I have never solved. When I put the brain in OCT it distorts. Rat and mouse brains cut very nicely (much better for me) if they are frozen by themselves. The 2 ways I do it is to make a foil boat and set the brain in and put it on dry ice with or without isopentane(slabs of dry ice are much better than pellets because they have a flat surface) for a minute or 2 (rats are more on the 2 minute side) till the top is frozen. The only thing with this is you get a flat spot on the bottom of the brain. These brains went for RNA ISH and we did 70 a week so we needed speed. The other way that takes a bit more effort is to hold the brain by the spinal cord and lower it into the isopentane in a beaker in dry ice 10-20 seconds depending on temp. One PI I worked with had a specific temperature range for the isopentane, but I do not remember that now. If you freeze it too long, you can get cracks or even multiple pieces if you leave it in way too long. After the brain is frozen (less is better than over freezing) I put the brains in conical tubes on dry ice till I get them to the -80 freezer. I have not had problems with ice artifact with either of these methods. For OCT embedded "regular" tissue either of these 2 methods have given me ice artifact either due to the warmer temperatures or increased time because of the OCT. I always use isopentane and liquid nitrogen for tissue other than brain Maybe the high lipid content in the brain makes the ice artifact less of a problem. To cut these I put a bit of OCT on the chuck and put the brain in the position I wanted. I usually cut the rostral end for coronal sections to make a flat spot for the chuck. One other tip we did was to have a small container of OCT and quickly dip the frozen mounted brain and refreeze the block. I made a hollow of dry ice in the container, dipped the brain and refroze the brain as quickly as possible. Good luck Donna Harclerode, HT, (ASCP), HTL, QIHC Immunohistochemist MacroPore Biosurgery, Inc 6740 Top Gun St. San Diego, CA 92121 858-458-0900 xt 322 dharclerode@macropore.com Subject: RE: [Histonet] cryomolds To: " Try some of the molds sold for plastic embedding which are deeper and then transfer to your cryostat chucks. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Molinari, Betsy Sent: Wednesday, June 15, 2005 5:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cryomolds Hi all, Does anyone know if there are cryomolds that are deeper than 5mm? I have some mouse brains to cut and the researcher wants to keep them intact and not bisect them, any suggestions? Thanks, Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston, Texas 77030 832-355-6524 832-344-6812 (fax) _______________________________________________ Histonet mailing list _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From froyer <@t> bitstream.net Wed Jun 15 15:54:40 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] alcohol/room size In-Reply-To: <61E9F2400F53D5119CFC00508B44E33B02E9E12D@khmcexch.uhsi.org> References: <61E9F2400F53D5119CFC00508B44E33B02E9E12D@khmcexch.uhsi.org> Message-ID: <42B09590.40700@bitstream.net> Just a guess... As much as you can drink without falling down? ~ Ford Ford Royer, MT(ASCP) Midwest Science & Biocenter Minneapolis, MN 800-745-4869 Kapoor, Sue wrote: >Hi, >can anyone tell me or direct me to information of what amount of alchol is >allowed in a room by the square footage? >Thanks 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 bwhitaker <@t> brownpathology.com Wed Jun 15 15:57:27 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] (no subject) Message-ID: <000801c571ec$d670e2b0$3601a8c0@brownpathology.net> Hi All, Our billing is set up to automatically charge an 88302 on tonsil/adenoid for patients < 14 years and an 88304 on > 14 years. Is this standard? Most lists I've seen from other institutions bill an 88304, with no age differentiation. I'd appreciate hearing what you all do. Thanks, Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 From petepath <@t> yahoo.com Wed Jun 15 16:24:33 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] tonsil cpt Message-ID: <20050615212434.35924.qmail@web30403.mail.mud.yahoo.com> Bonnie, All our tonsils are all billed 88304. The only age related CPT I can think of is foreskin. Newborn foreskin is 88302. Other than newborn is 88304. From BDUE <@t> PARTNERS.ORG Wed Jun 15 16:34:37 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:13 2005 Subject: Freeze Techniques RE: OCT... RE: [Histonet] Deeper molds for mouse brains was cryomolds Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB501027056@PHSXMB7.partners.org> Hi Donna, you're welcome. For the hearts, why not perfuse at RT, put in fridge for 5-10+ min to cool whole heart before embedding in cold OCT and snap freezing. The main trick is to minimize the temperature difference between the tissue + OCT and the actual freezing point. If you can get the center of the whole "block" cooler, then the snap freeze runs quicker. Chilled blocks can be larger since there isn't so much "thermal distance" to the center. Undiluted OCT doesn't adhere to tissues if it is colder than about 4C (refrig), so there are practical limits to how cold you can make the OCT if you apply *after* freezing. But if you apply the OCT *before chilling*, there should be no limits. Especially if your tissue can stand in OCT for a few minutes while chilling. Chill then freeze. Isopentane: I typically do what Frieda Carson does and let the isopentane freeze solid. Then I remove from the LN2 and break it up into a slurry as it warms up in the metal beaker. I find that a liquidy-slush does a better job extracting heat. Be sure to agitate the block in the isopentane to keep it in contact with the colder "ice-cubes". If you don't agitate then the isopentane local to the block warms up, even though elsewhere it is still good and cold. I freeze muscle bxs up to 2x2x4cm (that's centimeters) this way with almost no ice artifacts even in the dead center. Lastly, if blocks crack, the cracks can be repaired with cold OCT. I find it very hard to judge when to remove the tissue from the isopentane slurry -- you can't *see* when the center is frozen. Contrary to what others say, I find it much better to "over freeze" and deal with the cracks that happen. That's microtomy. I get much more consistant freeze quality this way. If you pull too early then ice artifacts will be forming. Uncracked blocks are easier to cut, but the morphology often suffers. Thanks for the paper ref -- I do some work on mouse eyes occasionally. -brice "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: Donna Harclerode [mailto:dharclerode@macropore.com] Sent: Wednesday, June 15, 2005 4:42 PM To: Due, Brice Subject: RE: OCT... RE: [Histonet] Deeper molds for mouse brains was cryomolds Hi Brice I do not do brains anymore, but the cold OCT idea may work for the hearts I am freezing in blocks that sometime I get artifact in the muscle. The whole hearts with OCT perfused freeze pretty good mostly. The problem with chilling the OCT for those would be that it would be too thick to perfuse. (but will check it out- I already dilute the OCT) I will play with that for future hearts. Muscle is one of the most difficult tissues I know so if things work for you they should work for pretty much anything. There is a paper in an old JOH where they mix OCT and aqua mount half and half for retina of eyes. Kent Keyser was a PI I worked for years ago that figured that out for more holding power for delicate retina. Thanks for the new ideas! Donna -----Original Message----- From: Due, Brice [mailto:BDUE@PARTNERS.ORG] Sent: Wednesday, June 15, 2005 12:45 PM To: Donna Harclerode; histonet@lists.utsouthwestern.edu Subject: OCT... RE: [Histonet] Deeper molds for mouse brains was cryomolds Hi Donna, I primarily work with human muscle bxs, which are sensitive to conditions that produce ice artifacts. My cold OCT trick may apply to rodent brains. I keep a bottle of OCT in a regular 4C refrigerator. I use it to embed troublesome muscles *after* they have been snap frozen in isopentane. No ice artifacts show up except maybe the outermost muscle fiber or two -- often none at all. After applying the cold OCT, I use either a freeze spray or dry ice or a -80C freezer to finish the job. For brains you could try snap freezing specimen and post-embedding like I do muscles. Or you could try using the cold OCT to fill your molds before freezing. This moves the specimen + OCT temp 15-20C south of RT, which means the isopentane has much less work to do to cross the freezing point. I don't freeze whole OCT blocks often, but I have noticed that cold OCT has fewer cracking problems than RT OCT when rapidly frozen. -brice Neuropathology Lab Brigham & Women's Hospital Boston "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Donna Harclerode Sent: Wednesday, June 15, 2005 1:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Deeper molds for mouse brains was cryomolds Hi Betsy I use PEEL-A-WAY molds made by POLYSCIENCES INC (available from VWR) for my bigger specimens. I do not use them for brains. The problem with trying to embed a brain in any sort of orientation in a mold is something I have never solved. When I put the brain in OCT it distorts. Rat and mouse brains cut very nicely (much better for me) if they are frozen by themselves. The 2 ways I do it is to make a foil boat and set the brain in and put it on dry ice with or without isopentane(slabs of dry ice are much better than pellets because they have a flat surface) for a minute or 2 (rats are more on the 2 minute side) till the top is frozen. The only thing with this is you get a flat spot on the bottom of the brain. These brains went for RNA ISH and we did 70 a week so we needed speed. The other way that takes a bit more effort is to hold the brain by the spinal cord and lower it into the isopentane in a beaker in dry ice 10-20 seconds depending on temp. One PI I worked with had a specific temperature range for the isopentane, but I do not remember that now. If you freeze it too long, you can get cracks or even multiple pieces if you leave it in way too long. After the brain is frozen (less is better than over freezing) I put the brains in conical tubes on dry ice till I get them to the -80 freezer. I have not had problems with ice artifact with either of these methods. For OCT embedded "regular" tissue either of these 2 methods have given me ice artifact either due to the warmer temperatures or increased time because of the OCT. I always use isopentane and liquid nitrogen for tissue other than brain Maybe the high lipid content in the brain makes the ice artifact less of a problem. To cut these I put a bit of OCT on the chuck and put the brain in the position I wanted. I usually cut the rostral end for coronal sections to make a flat spot for the chuck. One other tip we did was to have a small container of OCT and quickly dip the frozen mounted brain and refreeze the block. I made a hollow of dry ice in the container, dipped the brain and refroze the brain as quickly as possible. Good luck Donna Harclerode, HT, (ASCP), HTL, QIHC Immunohistochemist MacroPore Biosurgery, Inc 6740 Top Gun St. San Diego, CA 92121 858-458-0900 xt 322 dharclerode@macropore.com Subject: RE: [Histonet] cryomolds To: " Try some of the molds sold for plastic embedding which are deeper and then transfer to your cryostat chucks. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Molinari, Betsy Sent: Wednesday, June 15, 2005 5:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cryomolds Hi all, Does anyone know if there are cryomolds that are deeper than 5mm? I have some mouse brains to cut and the researcher wants to keep them intact and not bisect them, any suggestions? Thanks, Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston, Texas 77030 832-355-6524 832-344-6812 (fax) _______________________________________________ Histonet mailing list _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Wed Jun 15 16:47:05 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] alcohol/room size Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42CB@fh2k093.fhmis.net> It must be late in the day....@:) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 6/15/2005 4:54 PM Subject: Re: [Histonet] alcohol/room size Just a guess... As much as you can drink without falling down? ~ Ford Ford Royer, MT(ASCP) Midwest Science & Biocenter Minneapolis, MN 800-745-4869 Kapoor, Sue wrote: >Hi, >can anyone tell me or direct me to information of what amount of alchol is >allowed in a room by the square footage? >Thanks 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 > > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From POWELL_SA <@t> Mercer.edu Wed Jun 15 17:10:55 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] alcohol/room size In-Reply-To: <07AB60D5D7B9754EBF56F360F98D083DEB42CB@fh2k093.fhmis.net> Message-ID: <01LPHZHCJ0C48WXNJP@Macon2.Mercer.edu> Does that mean it is time to test your theory? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Wednesday, June 15, 2005 4:47 PM To: 'Ford Royer '; 'histonet-bounces@lists.utsouthwestern.edu '; 'histonet@lists.utsouthwestern.edu ' Subject: RE: [Histonet] alcohol/room size It must be late in the day....@:) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 6/15/2005 4:54 PM Subject: Re: [Histonet] alcohol/room size Just a guess... As much as you can drink without falling down? ~ Ford Ford Royer, MT(ASCP) Midwest Science & Biocenter Minneapolis, MN 800-745-4869 Kapoor, Sue wrote: >Hi, >can anyone tell me or direct me to information of what amount of alchol is >allowed in a room by the square footage? >Thanks 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 > > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Wed Jun 15 17:22:01 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] BCIP/NBT ready-to-use Message-ID: We are currently using a substrate system which has two separate components for BCIP and NBT, and then have to combine them just before use. I'm looking into alternate systems and have seen several vendors who offer what appears to be ready-to-use substrate. Is anybody using them and can you comment on stability with regard to expiration date, length of use, and lack of problems after the slides have been stained (I recall a recent thread regarding horrible overstaining after the slides had been coverslipped). Thanks muchly! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From cindipqr <@t> wowway.com Wed Jun 15 18:50:01 2005 From: cindipqr <@t> wowway.com (cindipqr@wowway.com) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Cryostat sections with small bubbles In-Reply-To: <004001c5714b$1fbb1f50$6a9a9618@Katri> References: <42AEEB5E.3070104@medecine.unige.ch> <004001c5714b$1fbb1f50$6a9a9618@Katri> Message-ID: <20050615224841.M28346@wowway.com> I agree, -35 seems way too cold. I cut rat brains around -18 ---------- Original Message ----------- From: "Katri Tuomala" To: "Emily Jane Wiesner-Camm" , Sent: Tue, 14 Jun 2005 21:39:51 -0400 Subject: Re: [Histonet] Cryostat sections with small bubbles > Hi Emily, > I have never heard of hot plating cryostat sections. You may be > boiling the moisture under the section forming bubbles or do you air > dry them well first? Also -35 C seems too cold for brain sections, > you may be getting some cutting artifacts. I'm sure there'll be > others with helpful information for you. Katri > > Katri Tuomala > Hamilton, Ontario, Canada > ----- Original Message ----- > From: "Emily Jane Wiesner-Camm" > To: > Sent: Tuesday, June 14, 2005 10:36 AM > Subject: [Histonet] Cryostat sections with small bubbles > > > Hi All, > > I was wondering whether anyone can let me know why small bubbles appear > > underneath my rat brain sections when I cut them on a cryostat (at -35) > > when they appear perfectly flat before placing them on a hotplate. How > > they can be avoided? > > Thanks, > > Emily > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------- End of Original Message ------- From kccatunda <@t> uol.com.br Wed Jun 15 20:20:59 2005 From: kccatunda <@t> uol.com.br (Katia Cristina Catunda) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Expiration date References: Message-ID: <000901c57211$a7ab5080$a279fea9@privatexx> What should we do with antibodies and reagents after their indicated expiration dates? Theorectically they shouldn't be estable after the expiration date but we do know that most of them still works perfectly... should we just throw them out and not use them anymore, wouldn't this be a way for the manufacturers manipulate their sellings? Need some opinions... Tks Katia From Gregor.Majdic <@t> vf.uni-lj.si Thu Jun 16 01:33:02 2005 From: Gregor.Majdic <@t> vf.uni-lj.si (Majdic Gregor) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] BCIP/NBT ready-to-use Message-ID: Hi, We are using Roche BCIP/NBT readyy to usee tablets for DIG - In situ hybridisation on both brain and testes and I think they work fine. If you have other conditions optimized, tehy give very little background. Gregor ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Possunt quia posse videntur ----------------------------------------------------------------- Gregor Majdic Head of Center for Animal Genomics Veterinary faculty, University of Ljubljana Gerbiceva 60 SI-1000 Ljubljana, Slovenia Phone: +386 1 4779210 Fax: +386 1 2832243 Email: gregor.majdic@vf.uni-lj.si ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Johnson, Teri Sent: ?et 16.6.2005 0:22 To: Histonet Subject: [Histonet] BCIP/NBT ready-to-use We are currently using a substrate system which has two separate components for BCIP and NBT, and then have to combine them just before use. I'm looking into alternate systems and have seen several vendors who offer what appears to be ready-to-use substrate. Is anybody using them and can you comment on stability with regard to expiration date, length of use, and lack of problems after the slides have been stained (I recall a recent thread regarding horrible overstaining after the slides had been coverslipped). Thanks muchly! 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 kwuny <@t> email.cs.nsw.gov.au Thu Jun 16 02:28:13 2005 From: kwuny <@t> email.cs.nsw.gov.au (Young Kwun) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] anti-mouse Cripto Antibody Message-ID: <200506161723505.SM01556@crgcsls814> Dear Histonetters, Does anyone know Cripto antibody which works on FFPE tissue sections? Has anyone tried R&D Systems' monoclonal Cripto antibody? Any info would be appreciated. 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 From Kemlo.Rogerson <@t> elht.nhs.uk Thu Jun 16 02:35:37 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Expiration date[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2BD@elht-exch1.xelht.nhs.uk> Depends on your SOPs. If the controls work Ok and you've stored them OK, you should be..um.... OK. But a Quality Management System should exclude anything that is out of date, including me. -----Original Message----- From: Katia Cristina Catunda [mailto:kccatunda@uol.com.br] Sent: 16 June 2005 02:21 To: histonet Subject: [Histonet] Expiration date[Scanned] What should we do with antibodies and reagents after their indicated expiration dates? Theorectically they shouldn't be estable after the expiration date but we do know that most of them still works perfectly... should we just throw them out and not use them anymore, wouldn't this be a way for the manufacturers manipulate their sellings? Need some opinions... Tks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Thu Jun 16 03:41:52 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Stain and pos control for copper Message-ID: I wonder if anyone with experience of copper stains can help me. We very rarely get asked to demonstrate copper and I know of only one positive case of Wilson's disease in 15 years which we were able to use as a pos control. We have just had another request. There was barely any control tissue left and both the test and control was neg. when stained with rhodanine. Two questions - is rhodanine the best stain, and what can we use for a pos. control? All ideas gratefully received! Jacqui Malam Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From abright <@t> brightinstruments.com Thu Jun 16 05:11:38 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Cryostat sections with small bubbles Message-ID: Emily, Basically you need to section brain at -8 to -12?C as colder temperatures will cause what you are getting plus cracking. However to achieve good quality sections at these temperatures you will need to maintain the knife and anti-roll temperatures at -20?C or lower to allow the section to slide in between the knife and the anti-roll plate unimpeded. Which is what Kathleen is doing when she warms the block face with her finger, a technique that went out with the Ark and should not be used as firstly you could have an infection issue and secondly it is not a very scientific approach in this day and age when there are devices fitted to some cryostats such as independent specimen temperature control to perform this task with full and repeatable control and of most importance, ease. 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: Kathleen Spencer [mailto:kspencer@utmem.edu] Sent: 15 June 2005 16:50 To: Katri Tuomala Cc: histonet@lists.utsouthwestern.edu; Emily Jane Wiesner-Camm Subject: Re: [Histonet] Cryostat sections with small bubbles Hi Emily, I cut rat brain sections at -18 to -20 and often have to warm the block face with my finger before each section. I also have better luck if I cut sections at 10u. These are fresh frozen brains. If I cut them at 20u I get bubbles under the sections. I keep them in the cryostat, and then fix the sections in cold fix, buffer wash, water wash, then air dry them in the hood. When I cut rat brain that has been perfused, I cut 20u floating sections. They never adhere well to slides, even if I use a hot plate. I always have bubbles under the section. That is why in this case we use floating sections. I hope this is helpful. Kathleen Spencer On Jun 14, 2005, at 8:39 PM, Katri Tuomala wrote: > Hi Emily, > I have never heard of hot plating cryostat sections. You may be > boiling the moisture under the section forming bubbles or do you air > dry them well first? Also -35 C seems too cold for brain sections, you > may be getting some cutting artifacts. > I'm sure there'll be others with helpful information for you. > Katri > > Katri Tuomala > Hamilton, Ontario, Canada > ----- Original Message ----- From: "Emily Jane Wiesner-Camm" > > To: > Sent: Tuesday, June 14, 2005 10:36 AM > Subject: [Histonet] Cryostat sections with small bubbles > > >> Hi All, >> I was wondering whether anyone can let me know why small bubbles >> appear underneath my rat brain sections when I cut them on a cryostat >> (at -35) when they appear perfectly flat before placing them on a >> hotplate. How they can be avoided? >> Thanks, >> Emily >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@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 livieira <@t> ualg.pt Thu Jun 16 06:05:22 2005 From: livieira <@t> ualg.pt (Lina Vieira) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Fish eggs References: Message-ID: <000901c57263$530a2630$2914100a@labhistologia> Hi Helen, Yes, but we want, if possible, measure the lenght of fresh fish embryos (without processing in GMA or paraffin). Lina Vieira Algarve University Faro Portugal ----- Original Message ----- From: "Helen Wimer" To: Sent: Thursday, June 16, 2005 11:45 AM Subject: Re: [Histonet] Fish eggs > Hi Lina, Do you access to or have you tried sectioning them in GMA? > > Helen F. Wimer > wimerh@si.edu > Smithsonian Institution MSC > NW Washington , DC 20560 > (301) 238-1180 > > Please note new phone number! > >>> "Lina Vieira" 06/15/05 11:52 AM >>> > Helo Everyone, > > Can you give me some advise about fish eggs? > I find a method to break/ destroy the fish eggs capsule. The method should > be easy for aplication in many fish eggs and have no effect in the embryo. > > Tanks in advance > > LIna Vieira > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From JNocito <@t> Pathreflab.com Thu Jun 16 07:08:31 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] (no subject) In-Reply-To: <000801c571ec$d670e2b0$3601a8c0@brownpathology.net> Message-ID: Bonnie, all our tonsil cases are 88304s regardless of age 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 Bonnie Whitaker Sent: Wednesday, June 15, 2005 3:57 PM To: 'histonet' Subject: [Histonet] (no subject) Hi All, Our billing is set up to automatically charge an 88302 on tonsil/adenoid for patients < 14 years and an 88304 on > 14 years. Is this standard? Most lists I've seen from other institutions bill an 88304, with no age differentiation. I'd appreciate hearing what you all do. Thanks, Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Thu Jun 16 07:47:29 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Expiration date Message-ID: Katia, CAP regulations do not allow for the use of expired antibodies and reagents so we try to purchase in volumes that will be used up by their expiry date. Also, our inhouse QA/QC personnel tend to go by the book as well so we don't have much choice in the matter. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Katia Cristina Catunda Sent: Wednesday, June 15, 2005 8:21 PM To: histonet Subject: [Histonet] Expiration date What should we do with antibodies and reagents after their indicated expiration dates? Theorectically they shouldn't be estable after the expiration date but we do know that most of them still works perfectly... should we just throw them out and not use them anymore, wouldn't this be a way for the manufacturers manipulate their sellings? Need some opinions... Tks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Thu Jun 16 08:15:14 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Stain and pos control for copper Message-ID: We've had always used the rhodanine stain for copper with good results. The control slides we get from Newcomer Supply. www.newcommersupply.com or 1-800-383-7799. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malam Jacqueline Sent: Thursday, June 16, 2005 3:42 AM To: Histonet submissions (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Stain and pos control for copper I wonder if anyone with experience of copper stains can help me. We very rarely get asked to demonstrate copper and I know of only one positive case of Wilson's disease in 15 years which we were able to use as a pos control. We have just had another request. There was barely any control tissue left and both the test and control was neg. when stained with rhodanine. Two questions - is rhodanine the best stain, and what can we use for a pos. control? All ideas gratefully received! Jacqui Malam Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Thu Jun 16 09:38:02 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Stain and pos control for copper Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42CC@fh2k093.fhmis.net> The Rhodamine stain is what we use (usually about once every 18 months), along with bought controls that we buy 10 at a time from Histology Control Systems. -Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet submissions (histonet@lists.utsouthwestern.edu) Sent: 6/16/2005 4:41 AM Subject: [Histonet] Stain and pos control for copper I wonder if anyone with experience of copper stains can help me. We very rarely get asked to demonstrate copper and I know of only one positive case of Wilson's disease in 15 years which we were able to use as a pos control. We have just had another request. There was barely any control tissue left and both the test and control was neg. when stained with rhodanine. Two questions - is rhodanine the best stain, and what can we use for a pos. control? All ideas gratefully received! Jacqui Malam Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacquie.Farnsworth <@t> cls.ab.ca Thu Jun 16 09:52:18 2005 From: Jacquie.Farnsworth <@t> cls.ab.ca (Jacquie.Farnsworth@cls.ab.ca) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Schiff's reagent in microwave? Message-ID: <4BC300747AF87A48BCDF8E48BC2885CECCD33D@mail1.calgary.com> Hi Histonetters, Does anyone have some thoughts on microwaving Schiff's reagent? We currently microwave Schiff's in our plastics laboratory (for PAS stain on our renal biopsies). We take the heated reagent out of the microwave, and place it in a fumehood immediately and remove slides. One of our techs is still having problems with the fumes. R95 masks are available, but I am wondering if anyone has any thoughts? Suggestions? Does anyone else microwave Schiff's reagent? Thank you in advance, Jacquie Jacqueline Farnsworth Anatomic Pathology Tech III Calgary Laboratory Services Jacquie.Farnsworth@CLS.ab.ca PH: (403) 944-1578 Pager: 212-8223 #1933 CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. From Dorothy.L.Webb <@t> HealthPartners.Com Thu Jun 16 09:52:51 2005 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Brain fixative Message-ID: <0E394B648E5284478A6CCB78E5AFDA2718C368@hpes1.HealthPartners.int> Does anyone fix their brain tissue from autopsies in anything other than 10% buffered formalin? The PA I am working with seems to feel they used a 4% solution of lightly buffered formalin from Mallincroft. Can anyone help me with this, as I am only used to using 10% formalin, as we use for all of our routine histology fixation. Any ideas would be greatly appreciated and thanks ahead of time to all of my fellow histonetters..what a great group you are!! +______________________________________+ ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From ryakay <@t> shands.ufl.edu Thu Jun 16 09:58:17 2005 From: ryakay <@t> shands.ufl.edu (Kaye Ryan) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] thanks Message-ID: Thanks to everyone who responded to my inquiry concerning decal solutions for bone marrows. Kaye Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 From Kemlo.Rogerson <@t> elht.nhs.uk Thu Jun 16 10:00:08 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Brain fixative[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2D1@elht-exch1.xelht.nhs.uk> Aren't they the same? We've had this debate; 4% formaldehyde is 10% formalin. 'Lightly buffered' is that like lightly salted? -----Original Message----- From: Webb, Dorothy L [mailto:Dorothy.L.Webb@HealthPartners.Com] Sent: 16 June 2005 15:53 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Brain fixative[Scanned] Does anyone fix their brain tissue from autopsies in anything other than 10% buffered formalin? The PA I am working with seems to feel they used a 4% solution of lightly buffered formalin from Mallincroft. Can anyone help me with this, as I am only used to using 10% formalin, as we use for all of our routine histology fixation. Any ideas would be greatly appreciated and thanks ahead of time to all of my fellow histonetters..what a great group you are!! +______________________________________+ ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Thu Jun 16 10:00:51 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Processing of research/mouse tissues (was delayed processing) Message-ID: Steve, welcome to research! It is a different world here. My advice would be to buy a hydrometer, and test your alcohols. When your last 100% starts to show some water, rotate it up to the position just past the 95% alcohol and make the later changes fresh. Also invest in the animal processing manual from the NSH. You'll find some very helpful schedules in there for all kinds of animal tissues. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From kspencer <@t> utmem.edu Thu Jun 16 10:05:24 2005 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Cryostat sections with small bubbles In-Reply-To: References: Message-ID: <0ff76615670407b429351b22aca29125@utmem.edu> Yes Alan, but when I put my finger on the block face to warm it, there is a kimwipe or gauze between my finger and the tissue. It works really well, so I guess the Ark is still around. As the owner of a company that sells cryostats, you need to be aware that not everyone has or can obtain such devices. Many of us, especially in research, work with old and crippled equipment, thus we need all the help we can get from our fellow technicians. Kathleen Spencer, HT ASCP Lab Manager/LCM Supervisor UTHSC On Jun 16, 2005, at 5:11 AM, Alan Bright wrote: > Emily, > > Basically you need to section brain at -8 to -12?C as colder > temperatures will cause what you are getting plus cracking. However to > achieve good quality sections at these temperatures you will need to > maintain the knife and anti-roll temperatures at -20?C or lower to > allow the section to slide in between the knife and the anti-roll > plate unimpeded. Which is what Kathleen is doing when she warms the > block face with her finger, a technique that went out with the Ark and > should not be used as firstly you could have an infection issue and > secondly it is not a very scientific approach in this day and age when > there are devices fitted to some cryostats such as independent > specimen temperature control to perform this task with full and > repeatable control and of most importance, ease. > > 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: Kathleen Spencer [mailto:kspencer@utmem.edu] > Sent: 15 June 2005 16:50 > To: Katri Tuomala > Cc: histonet@lists.utsouthwestern.edu; Emily Jane Wiesner-Camm > Subject: Re: [Histonet] Cryostat sections with small bubbles > > > Hi Emily, > > I cut rat brain sections at -18 to -20 and often have to warm the block > face with my finger before each section. I also have better luck if I > cut sections at 10u. These are fresh frozen brains. If I cut them at > 20u I get bubbles under the sections. I keep them in the cryostat, and > then fix the sections in cold fix, buffer wash, water wash, then air > dry them in the hood. > When I cut rat brain that has been perfused, I cut 20u floating > sections. They never adhere well to slides, even if I use a hot plate. > I always have bubbles under the section. That is why in this case we > use floating sections. > > I hope this is helpful. > > Kathleen Spencer > > > On Jun 14, 2005, at 8:39 PM, Katri Tuomala wrote: > >> Hi Emily, >> I have never heard of hot plating cryostat sections. You may be >> boiling the moisture under the section forming bubbles or do you air >> dry them well first? Also -35 C seems too cold for brain sections, you >> may be getting some cutting artifacts. >> I'm sure there'll be others with helpful information for you. >> Katri >> >> Katri Tuomala >> Hamilton, Ontario, Canada >> ----- Original Message ----- From: "Emily Jane Wiesner-Camm" >> >> To: >> Sent: Tuesday, June 14, 2005 10:36 AM >> Subject: [Histonet] Cryostat sections with small bubbles >> >> >>> Hi All, >>> I was wondering whether anyone can let me know why small bubbles >>> appear underneath my rat brain sections when I cut them on a cryostat >>> (at -35) when they appear perfectly flat before placing them on a >>> hotplate. How they can be avoided? >>> Thanks, >>> Emily >>> >>> >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@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 andrew.macduff <@t> ed.ac.uk Thu Jun 16 10:31:22 2005 From: andrew.macduff <@t> ed.ac.uk (andrew.macduff@ed.ac.uk) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Negatives for immunos using polyclonal antibody Message-ID: <1118935882.42b19b4a89a49@staffmail.ed.ac.uk> Dear all A quick question- when doing IHC with a polyclonal Ab is a serum negative sufficient since there can be no isotype specific control Ab? Thanks for the help Andrew Andrew MacDuff Clinical Research Fellow MRC Centre for Inflammation Research Medical School Edinburgh University From Janet.Bonner <@t> FLHOSP.ORG Thu Jun 16 10:53:56 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Schiff's reagent in microwave? Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42CD@fh2k093.fhmis.net> We did this microwaving of the Schiff reagent for awhile, but the step is only 15 minutes at room temperature so it wasn't worth using the microwave for this application. The Schiff Reagent last longer, too!! -Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jacquie.Farnsworth@cls.ab.ca Sent: Thursday, June 16, 2005 10:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Schiff's reagent in microwave? Hi Histonetters, Does anyone have some thoughts on microwaving Schiff's reagent? We currently microwave Schiff's in our plastics laboratory (for PAS stain on our renal biopsies). We take the heated reagent out of the microwave, and place it in a fumehood immediately and remove slides. One of our techs is still having problems with the fumes. R95 masks are available, but I am wondering if anyone has any thoughts? Suggestions? Does anyone else microwave Schiff's reagent? Thank you in advance, Jacquie Jacqueline Farnsworth Anatomic Pathology Tech III Calgary Laboratory Services Jacquie.Farnsworth@CLS.ab.ca PH: (403) 944-1578 Pager: 212-8223 #1933 CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.Eyres <@t> sanofi-aventis.com Thu Jun 16 10:49:39 2005 From: Stephen.Eyres <@t> sanofi-aventis.com (Stephen.Eyres@sanofi-aventis.com) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Microwave use in Pharmaceutical/CRO histo labs Message-ID: Hi All, Does anyone working in pharmaceutical/CRO's have any experience using microwave tissue processing? I'd be interested in your experiences, especially bad ones!! Cheers Steve ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- From dholmes <@t> anatomy.umsmed.edu Thu Jun 16 11:04:04 2005 From: dholmes <@t> anatomy.umsmed.edu (Dianne Holmes) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Required CEU's Message-ID: I have heard that there is a required number of CEU's for HT's to get within each year to maintain their certification. Is this so and if it is - how many? From NSEARCY <@t> swmail.sw.org Thu Jun 16 11:15:06 2005 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Muscle Referrals @ Baylor - Houston Message-ID: Does anyone know the contact person for the above? 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:;Anatomic Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD From dellav <@t> musc.edu Thu Jun 16 11:01:24 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] One of our own in need ! Please Read Message-ID: I am taking the liberty of sharing a correspondence I've had with a fellow Histotech who is facing some very difficult circumstances involving her baby's serious illness. I know that histotechs have big hearts and I would just like to make it possible for you to be aware of Lisa's circumstances. I am also hopeful that some of you may be able to offer her additional advice on where she might obtain financial support. you will need to scroll to the very bottom to see Lisa's first correspondence with me. I apologize in advance to anyone who may object to what may appear as fundraising on the list. I am simply making Lisa's circumstances known to you on her behalf. also, please forgive the lengthy message. her original message to me had a photo of her child attached but I know the list does not allow for attachments so it has been removed. I imagine that you can obtain the photo if you wish directly from Lisa. I have removed her home phone number from her original message at her request. It is circumstances like hers that remind me of what is most important in life . I'm sure that your prayers and good wishes would be most welcomed sincerely Vinnie Della Speranza >>> "lisa barbarisi" 06/16/05 01:56AM >>> Hello The address that I gave you for the Funds for Lisa was incomplete I corrected it below. Thanks again, Lisa ----- Original Message ----- From: "lisa barbarisi" To: "Vinnie Della Speranza" Sent: Wednesday, June 15, 2005 9:54 PM Subject: Re: Fw: Fund raising at the national convention > Dear Vinnie, > > Thank you for your response. I did not think of posting on the Histonet. > It sounds like a good idea. I would however like to keep my home phone > number out of the letter and please do add the web site > www.alysa-anne.com it is now up and is just wonderful. I was told that > it is still being worked on as they are going to add a guest book for > people to sign in on. Also, I would appreciate it if you mentioned the > "Funds for Lisa" account #44699 any donations may be sent to: > > Funds for Lisa Account 44699 > Research Administration > Mayo Clinic Scottsdale 13400 East Shea Blvd. > Scottsdale, AZ 85259 > > All donations are tax deductible and a donation receipt will be sent as > long as the name and address are included either on the check or the > envelope. > > I do appreciate all of the information that you sent there are some that I > have not called and I will do so. It is so hard to take the time needed to > research and call people so it is wonderful when others offer a hand. > Thank you for your wonderful suggestion. > > Please forward what you post on the Histonet and the link. I have trouble > keeping track of things so having them in a e-mail helps me keep track. I > appreciate your help with this. > > Thank you so much for your help I look forward to hearing from you again. > > Lisa & Alysa Anne > > > ----- Original Message ----- > From: "Vinnie Della Speranza" > To: ; ; > ; ; ; > ; ; > ; ; ; > ; ; ; > ; > Cc: ; > Sent: Monday, June 13, 2005 12:41 PM > Subject: Re: Fw: Fund raising at the national convention > > > Dear Lisa, > I cannot tell you how sad I am to learn of your circumstances and of > course your baby's illness. I have no doubt that this is a very difficult > time for you and I imagine that many could not begin to imagine the > challenges you are facing as a result. I know that you have been a member > of NSH for the last few years and everyone understands your lack of recent > participation. I will pray that yours and Alysa's circumstances begin to > improve. > > to be quite frank, I am struggling with figuring out how we might help > you. I would probably have to consult with the society's attorney due to > our non-profit status. fund raising of any sort could create unforeseen > problems for the society. given this concern, I don't believe that the > society could sanction or conduct formal fundraising on your behalf. > > I do not know if you have considered placing your letter on Histonet. I > would be happy to do this for you if you indicate that you would like me > to do so. The Histonet list reaches histotechs from around the globe and > is an independent group not constrained by tax and corporate laws. I don't > speak for that group but I've found that histotechs have big hearts and > at the least someone more knowledgeable than I may be a source of valuable > information that might ease your circumstances. I hope you will consider > this avenue and as I've said, I would be happy to post your letter to me > on that list. > > our Executive Director, Carrie Diamond, has pulled some information from > the American Cancer Society website that addresses avenues for financial > assistance for circumstances like yours. I apologize if you are already > aware of this information but here it is: > > from American Cancer Society website. > Where can families get help financially? > > Most families find it difficult to turn to others or to agencies and > funds for financial help. Families generally take pride in being > self-sufficient and providing for their own needs. The extra expenses of > a child's cancer may be for the first time with problems with money. > They should remember that their problems in such a situation are > temporary and not unique. In the future, they may be the ones in a > position to offer financial help to others. > > There are several possible sources of help for families in financial need: > > * income assistance for low-income families through Supplemental > Security Income (SSI) benefits > * income assistance for needy families from the Aid to Families with > Dependent Children (AFDC) program > * help with travel, meals and lodging from public and private programs > * assistance with basic living costs (rent, mortgage, insurance > premiums, utilities, telephone) from public and private programs > * help from church, civic, social, and fraternal groups in the > community > * general help from special funds in the medical center or community > * assistance from targeted fundraising for an individual patient or > family. > > Organizations > > American Cancer Society, Inc. 800-227-2345 > Candlelighters Childhood Cancer Foundation 800-366-2223 > Corporate Angel Network, Inc. 866-328-1313 > National Children's Cancer Society 800-532-6459 > National Marrow Donor Program 800-627-7692 > National Patient Air Transport Hotline 800-296-1217 > Ronald McDonald House Charities 630-623-7048 > The Leukemia & Lymphoma Society 800-955-4572 > > Organizations (Government): > > U.S. Department of Labor (COBRA information) > Pension and Welfare Benefits Administration 800-998-7542 (Publications > only) 202-219-8776 > Americans with Disabilities Act, Civil Right Division, Disability Rights > Section 800-514-0301 > Hill-Burton Program 800-638-0742 > Internal Revenue Service (publications) 800-829-3676 800-829-1040 > (Assistance and Information) > National Cancer Institute 800-422-6237 > Social Security Administration (SSI information) 800-772-1213 > > Other Resources: > > American Brain Tumor Association 800-886-2282 > American Kidney Fund 800-638-8299 > Bone Marrow Transplant Information Network 888-597-7674 > Brain Tumor Foundation for Children, Inc. 770-458-5554 > Children's Organ Transplant Association, Inc. 800-366-2682 > Cancervive 310-203-9232 > Curesearch National Childhood Cancer Foundation 800-458-6223 > Lymphoma Research Foundation of America 800-500-9976 > National Association of Hospital Hospitality Houses, Inc. 800-542-9730 > National Association of Personal Financial Advisors 800-366-2732 > National Bone Marrow Transplant Link 800-546-5268 > National Brain Tumor Foundation 800-934-2873 > National Children's Cancer Society 800-532-6459 > National Coalition for Cancer Survivorship 877-622-7937 > National Foundation for Credit Counseling 800-388-2227 > National Information Center for Children and > Youth with Disabilities 800-695-0285 > National Insurance Consumer Helpline 800-942-4242 > National Organization for Rare Disorders, Inc. 800-999-6673 > National Self-Help Clearinghouse 212-817-1822 > National Transplant Assistance Fund 800-642-8399 > Oley Foundation, Inc. (nutritional information) 800-776-6539 > Patient Advocate Foundation 800-532-5274 > > Other Organizations (Government): > > National Cancer Institute (NCI) > Cancer Information Service 800-422-6237 > Centers for Disease Control and Prevention 800-311-3435 > Department of Health and Human Services 877-696-6775 > Medicare Hotline 800-633-4227 > National Health Information Center 800-336-4797 > Veterans Benefits Administration 800-827-1000 > U.S. Department of Veterans Affairs > > I hope that you will let us know when the website you are planning is up > and running so it will be easier for people to offer donations. > > God bless you both. > sincerely > Vinnie Della Speranza > President, National Society for Histotechnology > > >>>> "lisa barbarisi" 06/10/05 02:37AM >>> > Please read. I was not sure who to send this to so I tried to include > everyone. > > Thank you in advance for your time. Prayers are very welcome also. > > Lisa A. Barbarisi HT(ASCP) > > ----- Original Message ----- > From: lisa barbarisi > To: histo@nsh.org > Cc: bali02@cox.net > Sent: Thursday, June 09, 2005 11:17 PM > Subject: Fund raising at the national convention > > > Hello All, > > I am writing to ask for help in the form of a fundraiser at the national > convention. I am an HT( ASCP) certified Histotech who has worked in the > field of histotechnology for over eighteen years. I worked as the > coordinator of the histology research facility at the Mayo Clinic I am > currently on unpaid leave. > > After almost three years of fertility treatments and > invitro-fertilizations I spent all of my savings and took out a second > mortgage on my home and I was finally blessed to become pregnant. I gave > birth to a beautiful baby girl on August 30th 2004. > > I am a single first time mom with one income of which is zero at the > moment since I am on an unpaid leave from my current position at the Mayo > Clinic research facility as a Histotech. I am looking for financial help. > My beautiful baby girl Alysa Anne was diagnosed with stage four > neuroblastoma on March 25th 2004 she was not yet seven months old. She has > had surgery to try and remove the primary tumor with no luck. She is > currently undergoing chemotherapy to try and shrink the primary tumor > which is located on her right adrenal gland and there are tumors present > in her liver (to many to count), bone marrow, her bones base of her skull, > chest bone, around both of her eyes and her pathology is unfavorable. I > have been told by her oncologist that this is going to be a long hard road > for both my daughter and I. Between the chemotherapy there are doctors > visits twice and sometimes three times a week to check her progress and > also her blood counts. She is on daily shots of Neupogen and multiple > medications. How is such a small body going to take so much? I just can't > believe this is happening. > > I'm sorry, I could go on, but you may not be interested in helping me and > I will understand. I am at the end of my rope. I know that many people may > ask for help, but I just had to try. My daughter is my life and I will do > whatever I have to to help her fight this horrible disease . I was unable > to attend the NSH convention last year in Canada as my daughter had > finally blessed me with her arrival (she was over a week late and after a > long hard labor I finally had to have a c-section). I will not be able to > attend this year either since my daughter is having chemotherapy she just > made it through her third round. I feel helpless watching the poison being > pumped inter her little body thru a port in her chest. > > I wonder if there is anyone who would like to take on this huge > undertaking of either a fundraiser or maybe just taking donations to the > "Funds for Lisa" a not for profit account set up by a co-worker who wanted > to help me. > > If anyone is interested I would be so happy. Please e-mail me at bali02@cox.net . There will be a > website that is not as far as I know up and running it will tell more > about me and my daughter. It will also have updates on her progress. Once > I get word that it is up and running I can e-mail the site. > > I would appreciate a response even if it is negative. > > I have to beat this MONSTER called NEUROBLASTOMA to save my daughter . > > Sincerely, > > Lisa A. Barbarisi > > Picture of Alysa Anne at six months old three weeks before diagnosis is > attached. > > I > From juan.gutierrez <@t> christushealth.org Thu Jun 16 11:26:11 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Schiff's reagent in microwave? Message-ID: Do you have room in the fume hood for the microwave? Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jacquie.Farnsworth@cls.ab.ca Sent: Thursday, June 16, 2005 9:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Schiff's reagent in microwave? Hi Histonetters, Does anyone have some thoughts on microwaving Schiff's reagent? We currently microwave Schiff's in our plastics laboratory (for PAS stain on our renal biopsies). We take the heated reagent out of the microwave, and place it in a fumehood immediately and remove slides. One of our techs is still having problems with the fumes. R95 masks are available, but I am wondering if anyone has any thoughts? Suggestions? Does anyone else microwave Schiff's reagent? Thank you in advance, Jacquie Jacqueline Farnsworth Anatomic Pathology Tech III Calgary Laboratory Services Jacquie.Farnsworth@CLS.ab.ca PH: (403) 944-1578 Pager: 212-8223 #1933 CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Thu Jun 16 11:45:10 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Brain fixative Message-ID: We use 20% formalin, and leave it in there for about a month. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Webb, Dorothy L Sent: Thursday, June 16, 2005 9:53 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Brain fixative Does anyone fix their brain tissue from autopsies in anything other than 10% buffered formalin? The PA I am working with seems to feel they used a 4% solution of lightly buffered formalin from Mallincroft. Can anyone help me with this, as I am only used to using 10% formalin, as we use for all of our routine histology fixation. Any ideas would be greatly appreciated and thanks ahead of time to all of my fellow histonetters..what a great group you are!! +______________________________________+ ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Edmondson <@t> christie-tr.nwest.nhs.uk Thu Jun 16 11:49:46 2005 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] BCIP/NBT ready-to-use Message-ID: DAKO do complete reagent in one, and the BCIP/NBT/INT that we use, its all used within date so can not help with long term stability. No ppt formed that we have seen Dave Christie Hosp Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Johnson, Teri Sent: 15 June 2005 23:22 To: Histonet Subject: [Histonet] BCIP/NBT ready-to-use We are currently using a substrate system which has two separate components for BCIP and NBT, and then have to combine them just before use. I'm looking into alternate systems and have seen several vendors who offer what appears to be ready-to-use substrate. Is anybody using them and can you comment on stability with regard to expiration date, length of use, and lack of problems after the slides have been stained (I recall a recent thread regarding horrible overstaining after the slides had been coverslipped). Thanks muchly! 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 mucram11 <@t> comcast.net Thu Jun 16 11:54:47 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Schiff's reagent in microwave? Message-ID: <061620051654.18290.42B1AED7000002C9000047722200748184CECE030E9D0C9A03@comcast.net> Technically we should all have the oven under the hood and it is not always possible if we need to use the hood for anything else. I know I was told to look into the cost of a laboratory microwave and thought the people I worked for at the time would have heart attack when they saw the quote. I used the Target MW as close to the hood as I could get it and that allowed me to move the Schiff's and anything else very quickly under the hood. I could hold my breathe and get it there. Otherwise I don't what would stop the fumes completely for you. In an ideal world we would all have large hoods and lab MWs for safety. Pam Marcum -------------- Original message -------------- > Do you have room in the fume hood for the microwave? > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > My opinions are my own and do not reflect those of my employer. Long live free > speech! > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Jacquie.Farnsworth@cls.ab.ca > Sent: Thursday, June 16, 2005 9:52 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Schiff's reagent in microwave? > > Hi Histonetters, > Does anyone have some thoughts on microwaving Schiff's reagent? We currently > microwave Schiff's in our plastics laboratory (for PAS stain on our renal > biopsies). We take the heated reagent out of the microwave, and place it in a > fumehood immediately and remove slides. One of our techs is still having > problems with the fumes. R95 masks are available, but I am wondering if anyone > has any thoughts? Suggestions? Does anyone else microwave Schiff's reagent? > > Thank you in advance, > Jacquie > > Jacqueline Farnsworth > Anatomic Pathology Tech III > Calgary Laboratory Services > Jacquie.Farnsworth@CLS.ab.ca > PH: (403) 944-1578 > Pager: 212-8223 #1933 > > > > > CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain > confidential, personal and/or privileged information intended for a specific > purpose and recipient. If you are not the intended recipient do not disclose, > copy, retain, distribute, use or modify any of the contents of this > transmission. If you received this transmission in error please notify me > immediately by return e-mail or telephone and destroy the entire transmission > and any copies produced. Thank you. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eca9 <@t> georgetown.edu Thu Jun 16 12:50:41 2005 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Ki-67 and PCNA Message-ID: <42B1BBF1.6020707@georgetown.edu> Hi Everyone, I am looking into staining with Ki-67 and PCNA. Which company do you prefer for these antibodies? Have you had any bad experiences with these antibodies for any particular provider? Greatly appreciate all your advice, Eva Andersson Georgetown Univetsity From petepath <@t> yahoo.com Thu Jun 16 13:20:05 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] re:One of our own in need ! Please Read Message-ID: <20050616182005.72992.qmail@web30408.mail.mud.yahoo.com> Here is a way anyone who wants to help Lisa. I am happy to offer 15% of the sale price on any Precision Cryoembedding System equiptment which is purchased through me with a reference to Funds for Lisa. As always I will send equiptment to demo which can be returned without obligation. I will carry this offer through the NSH meeting ( Booth 835). I do a lot of Pediatric Patholgy and I know what Lisa is going through. I apologize if this seems like a sleezy marketing ploy. In a way it is, but as I do not count on this buisness for income ( thank god) it is pretty much a charity anyway. Plus, anyone doing surg path who has not tried my toys might find that they are missing something. In any event it could help generate some help for a needy colleague. Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From Luis.Chiriboga <@t> med.nyu.edu Thu Jun 16 10:57:30 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Expiration date In-Reply-To: <000901c57211$a7ab5080$a279fea9@privatexx> Message-ID: Just a thought..... Perhaps people could donate expired reagents to a local histology program.....I don't know about the logistical or administrative issues that might be involved (there always are). Just as a side note.....I have antibodies (research) that are over 7 years old that still work perfectly fine. In fact, out of a ~400 hundred antibodies I have in stock, there are only a handful that truly "expire". The most unstable tend to be raw serum or ascities preparations with out preservatives, but this can be easily corrected to prevent deterioration. Commercial antibodies tend to be relatively stable if handled appropriately. L ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Katia Cristina Catunda Sent: Wednesday, June 15, 2005 9:21 PM To: histonet Subject: [Histonet] Expiration date What should we do with antibodies and reagents after their indicated expiration dates? Theorectically they shouldn't be estable after the expiration date but we do know that most of them still works perfectly... should we just throw them out and not use them anymore, wouldn't this be a way for the manufacturers manipulate their sellings? Need some opinions... Tks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From settembr <@t> umdnj.edu Thu Jun 16 13:55:23 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Ki-67 and PCNA Message-ID: Eva, For Ki-67 I use DakoCytomation Cat.#M7240 Works great on FFPE human tissue. Dana Settembre UMDNJ - University Hospital Newark, NJ >>> Eva C Andersson 6/16/2005 1:50:41 PM >>> Hi Everyone, I am looking into staining with Ki-67 and PCNA. Which company do you prefer for these antibodies? Have you had any bad experiences with these antibodies for any particular provider? Greatly appreciate all your advice, Eva Andersson Georgetown Univetsity _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kerry.l.crabb <@t> gsk.com Thu Jun 16 13:59:20 2005 From: kerry.l.crabb <@t> gsk.com (kerry.l.crabb@gsk.com) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] NSH S/C Posters Message-ID: The deadline to submit a Poster Session abstract for the NSH Symposium/Convention was June 15th. The deadline has been extended to July 1st. I want to encourage all to consider this method of sharing valuable information about the work you are doing. Please refer to the printed convention program for more information and the application form. You may also submit a Poster Session Application via the NSH web site (www.nshconvention.org). From the NSH Convention web site click on Poster Sessions. All the information you will need can be found there. If you have any questions or problems, contact the NSH office (301-262-6221). From anh2006 <@t> med.cornell.edu Thu Jun 16 14:12:49 2005 From: anh2006 <@t> med.cornell.edu (anh2006@med.cornell.edu) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Expiration date In-Reply-To: References: <000901c57211$a7ab5080$a279fea9@privatexx> Message-ID: <3954.140.251.146.2.1118949169.squirrel@webmail> I totally agree with you Luis, what a great idea! In fact I just used anti-CD31 MEC13.3 that was over 6 years old and the staining was fabulous :) Also, I find the ones that "expire" are the ones with lots of people using them and therefore have lots of dirty pipet tips dipping in or out or have constant temperature fluctuations from continuous (and sometimes careless) use. Andrea > Just a thought..... > Perhaps people could donate expired reagents to a local histology > program.....I don't know about the logistical or administrative issues > that > might be involved (there always are). > > Just as a side note.....I have antibodies (research) that are over 7 years > old that still work perfectly fine. In fact, out of a ~400 hundred > antibodies I have in stock, there are only a handful that truly "expire". > The most unstable tend to be raw serum or ascities preparations with out > preservatives, but this can be easily corrected to prevent deterioration. > Commercial antibodies tend to be relatively stable if handled > appropriately. > L > From rebecca.riesen <@t> dsilabs.com Thu Jun 16 14:24:20 2005 From: rebecca.riesen <@t> dsilabs.com (Riesen, Rebecca) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] PEL & STEL Message-ID: <05844092236E7749A1BBBCB1077C34A2DEA00E@dsi-ex01.gateway.dom> Hello HISTONETTERS: I have a new safety officer who wants me to just check formaldehyde and xylene levels for PEL & STEL with a "Room Monitor" instead of what I have always done, which is the individual personal badges. Are there any regulations concerning this? Is it ok to just monitor the "room"? If you do the room monitoring, do you need to monitor the individual with an individual badge for the initial level? Just curious as to what you folks are all doing out there. THANKS!!!! DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(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 ploykasek <@t> phenopath.com Thu Jun 16 14:27:36 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Expired reagents Message-ID: Donation is a wonderful idea. Some high schools are doing histo work, too. In regards to using expired reagents in the lab, I personally don't have a problem with it if your positive QC's are working, but (& this is a big but) CAP has a big problem with it. This is ANP.22432 on the CAP checklist & it is a phase II. If you are a CAP accredited lab, you can not use expired reagents no matter what. It makes me want to cry. To add more fuel to the flame, how are people handling the comment that goes with ANP.12425 that states you can use RUO reagents in "home brew" tests if you document that you searched for IVD or ASR class reagents & failed to find any. What do you think of that??? Doesn't seem impossible to do, just more paperwork/documentation. Patti Loykasek PhenoPath Laboratory Seattle, WA From emry <@t> u.washington.edu Thu Jun 16 16:54:06 2005 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] formic acid decal Message-ID: I need to speed up the decal process on some large bone samples. What is the highest precentage of formic acid that I can use without damaging the bones? I am using sodium formate with formic acid now. Thanks, Trisha Seattle From wwmn916 <@t> aol.com Thu Jun 16 16:56:51 2005 From: wwmn916 <@t> aol.com (wwmn916@aol.com) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Histotech position Message-ID: <8C740D56FA85185-B14-94B8@mblk-d45.sysops.aol.com> Hello histonetters, If anyone is interested in working with a very forward thinking and technology minded group of people and would like living in Sacramento, you are invited to "check us out". We are looking for an experienced histotechnician....much like many places.....but here at Path Logic, we have a lot for a good technician to explore employment opportunities for. To find out more please feel free to call 916-241-4518, fax 916-863-1498 or email dbrhkng@yahoo.com Deborah King Histology Supervisor Path Logic 3637 Mission Ave. Carmichael, CA 95608 From histology.bc <@t> shaw.ca Thu Jun 16 18:59:57 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Schiff's reagent in microwave? References: <4BC300747AF87A48BCDF8E48BC2885CECCD33D@mail1.calgary.com> Message-ID: <42B2127D.4050702@shaw.ca> Two points to make: 1. Microwaving Schiff's reagent will shorten its life and your operating costs will rise. 2. Unless you are using some really strange procedure, the time in Schiff reagent is only 10-15 minutes anyway. Think about it logically, are you really in that much of a rush? The health risks associated with hot Schiif fumes should be enough to disuade you from this procedure. Paul Bradbury Kamloops, BC Jacquie.Farnsworth@cls.ab.ca wrote: >Hi Histonetters, >Does anyone have some thoughts on microwaving Schiff's reagent? We currently microwave Schiff's in our plastics laboratory (for PAS stain on our renal biopsies). We take the heated reagent out of the microwave, and place it in a fumehood immediately and remove slides. One of our techs is still having problems with the fumes. R95 masks are available, but I am wondering if anyone has any thoughts? Suggestions? Does anyone else microwave Schiff's reagent? > >Thank you in advance, >Jacquie > >Jacqueline Farnsworth >Anatomic Pathology Tech III >Calgary Laboratory Services >Jacquie.Farnsworth@CLS.ab.ca >PH: (403) 944-1578 >Pager: 212-8223 #1933 > > > > >CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From AnthonyH <@t> chw.edu.au Thu Jun 16 19:00:50 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Stain and pos control for copper Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E2A3@simba.kids> Try a liver from primary biliary cirrhosis or even a cirrhotic liver. The copper will be seen around the edge of the hepatic nodules. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Malam Jacqueline [mailto:Jacqueline.Malam@rli.mbht.nhs.uk] Sent: Thursday, 16 June 2005 6:42 PM To: Histonet submissions (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Stain and pos control for copper I wonder if anyone with experience of copper stains can help me. We very rarely get asked to demonstrate copper and I know of only one positive case of Wilson's disease in 15 years which we were able to use as a pos control. We have just had another request. There was barely any control tissue left and both the test and control was neg. when stained with rhodanine. Two questions - is rhodanine the best stain, and what can we use for a pos. control? All ideas gratefully received! Jacqui Malam Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ 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 rschoon <@t> email.unc.edu Thu Jun 16 19:03:30 2005 From: rschoon <@t> email.unc.edu (Robert Schoonhoven) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] re:One of our own in need ! Please Read In-Reply-To: <20050616182005.72992.qmail@web30408.mail.mud.yahoo.com> References: <20050616182005.72992.qmail@web30408.mail.mud.yahoo.com> Message-ID: <42B21352.5050309@email.unc.edu> Stephen, Having been through the cancer gamut with both my wife and stepson (both are doing well), I hope that your offer will be taken advantage of. Robert Schoonhoven Stephen Peters M.D. wrote: >Here is a way anyone who wants to help Lisa. I am happy to offer 15% of the sale price on any Precision Cryoembedding System equiptment which is purchased through me with a reference to Funds for Lisa. As always I will send equiptment to demo which can be returned without obligation. I will carry this offer through the NSH meeting ( Booth 835). >I do a lot of Pediatric Patholgy and I know what Lisa is going through. >I apologize if this seems like a sleezy marketing ploy. In a way it is, but as I do not count on this buisness for income ( thank god) it is pretty much a charity anyway. Plus, anyone doing surg path who has not tried my toys might find that they are missing something. In any event it could help generate some help for a needy colleague. > >Stephen > > >Stephen Peters M.D. >Vice Chairman of Pathology >Hackensack University Medical Center >201 996 4836 > >Pathology Innovations, LLC >410 Old Mill Lane, >Wyckoff, NJ 07481 >201 847 7600 >www.pathologyinnovations.com > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From ajaivyas <@t> gmail.com Thu Jun 16 20:15:34 2005 From: ajaivyas <@t> gmail.com (Ajai Vyas) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Brain tissue with GFP: fixing or not Message-ID: <934be63705061618157452ab3c@mail.gmail.com> Greetings! I am trying to visualize some GFP +ve neurons in mouse. I have tried fresh freezing these by embedding in OCT and putting in slurry of alcohol+dry ice. I have problem with curling and tissue integrity when I section these brains in cryostat (-15 and -20 degree C). For the next lot I was wondering if I should attempt to 1) perfuse animals with fresh PFA, cryoprotect brains with sucrose and section or 2) try other methods of snap freezing suggested on histonet archives and/or try to play more with cryostat temperature Any suggestion and pointers will be greatly appreciated, Ajai From ssq5977 <@t> yahoo.com.cn Thu Jun 16 20:29:40 2005 From: ssq5977 <@t> yahoo.com.cn (=?gb2312?q?=CA=E7=C7=ED=20=C9=F2?=) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Is there anything better than OCT? Message-ID: <20050617012941.2507.qmail@web15602.mail.cnb.yahoo.com> Hi all: We are currently using OCT to embed renal biopsy tissue in cryostat mold. First of all ,we put a drop of OCT on the mold ,then we put the tissue on the top of the OCT and dip the mold into liquid nitrogen for 10 seconds. then the tissue is sent to cryosection. Does anyone know if there is anything better than OCT to embed the tissue? Thanks! Shuqiong Shen(ssq5977@yahoo.com.cn) Research Institute of Nephrology, Jinling Hospital 305 East Zhongshan Road, Nanjing Jiangsu Province,P.R of China Zip code: 210002 Tel: +86 25 85912177 --------------------------------- DO YOU YAHOO!? »¶Ó­Ê¹ÓÃÑÅ»¢³¬´óÈÝÁ¿Ãâ·ÑÓÊÏä From CrochiereSteve <@t> aol.com Thu Jun 16 20:57:15 2005 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] romulin red? Message-ID: <200.3cfba35.2fe387fb@aol.com> Do you mean Vulcan Fast Red? That is fairly clear but stains red. I thought Romulins were Blue? Anyway, try it to see if it works. I've had very good luck with Biocare's products Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 From JMacDonald <@t> mtsac.edu Thu Jun 16 23:36:17 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] romulin red? Message-ID: BioCare has a Romulin AEC. It is red. < To: histonet@lists.utsouthwestern.edu From: CrochiereSteve@aol.com< histonet-bounces@lists.utsouthwestern.edu Date: 06/16/2005 0 Subject: [Histonet] romulin red? Do you mean Vulcan Fast Red? That is fairly clear but stains red. I thought Romulins were Blue? Anyway, try it to see if it works. I've h Biocare's products Steven M. Crochiere, H Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 _______________ ______________________ 5F__ Histonet mailing list Histonet@lists.utsouthwes [1]http://lists.utsouthwestern.edu/mailman/listinfo/hi References 1. 3D"http://lists.utsouthwestern.edu/mailman/listinfo/his From c.m.vanderloos <@t> amc.uva.nl Fri Jun 17 02:45:54 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] RE: Stupid question about Biocare Romulin red Message-ID: Hi Jackie, I have tested that product for visualizing HRP activity too, but in my hands it wasn't red after preparation the full solution. There was a nice strong red/brown reaction product in the tissue section. The reaction product was not as red as AEC reaction for example. I have also tested Vector Nova Red and have the feeling that this product is identical with BioCare's Romulin Red. With both products your sections need to be mounted organically as the red reaction product will disappear in time after aqueous mounting. My preference goes to the Vector Nova Red. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 ---------------------------------------------------------------------- ----------------------------------------- Date: Wed, 15 Jun 2005 12:48:41 -0500 From: "Jackie M O'Connor" Subject: [Histonet] Stupid question about Biocare Romulin red To: histonet@lists.utsouthwestern.edu I've never used this product before - can anyone tell me if it's supposed to appear red after it's been prepared? I expected to see some color - but it's clear as a bell. I'm not using it until I get an answer. Help! From sakima <@t> bigpond.net.au Fri Jun 17 03:10:50 2005 From: sakima <@t> bigpond.net.au (Satoshi Akima) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Guava apoptosis assay suite Message-ID: Dear list members, does anybody have any experience yet with using this system to detect apoptosis? It's a new product but any opinions out there??? Toshi "...new Guava Apoptosis Assay Suite for detecting activated caspases. Guava Assay Suite uses a fluorescent caspase-specific inhibitor to identify individual apoptotic cells. The assays are optimized for 96- well plates so you can easily test multiple conditions and even screen clones or compounds using these assays. Guava Apoptosis Assay Suite simply adds to the expanding menu of assays that researchers can use for cancer research on the Guava systems. Since all the assays can be done on one platform, one scientist can characterize cellular reactions to multiple compounds in a single day (download the poster). All the Guava assays are optimized for the Guava microcytometry systems. Compact and light, these truly benchtop systems provide the power of flow cytometry analysis right in your own laboratory. The Guava EasyCyte is a five-parameter, three color system that can assay your samples directly from 96-well plates and is ideal for screening compounds, assessing tranfections or studying apoptosis pathways. The systems are easy to use and require minimal maintenance so you don?t have to have a dedicated technician for it. Learn more about Guava assays and the Guava EasyCyte." From abright <@t> brightinstruments.com Fri Jun 17 04:10:09 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Cryostat sections with small bubbles Message-ID: Dear Kathleen, Thank you for your reply. I do understand that Specimen Temperature Control may not be available to all cryostat users. Therefore I will set out a method that I have posted on Histonet previously to assist with sectioning problems on brain. You will see that this is a far more scientific method than warming up the face of a specimen with ones finger and will render repeatable good quality sections. Set the cryostat microtome chamber to -8n to -12?C, place containers made from aluminium kitchen foil around and in contact with the knife in areas that with not impede the specimens travel or the anti-roll plate, also do the same for where the anti-roll plate is parked away from the knife. Fill these containers with graduals of Solid C02. Allow 10 minutes for the solid C02 to cool the knife/ anti-roll plate to a lower temperature than the specimen, then start sectioning. 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: Kathleen Spencer [mailto:kspencer@utmem.edu] Sent: 16 June 2005 16:05 To: Alan Bright Cc: Emily Jane Wiesner-Camm; histonet@lists.utsouthwestern.edu; Katri Tuomala Subject: Re: [Histonet] Cryostat sections with small bubbles Yes Alan, but when I put my finger on the block face to warm it, there is a kimwipe or gauze between my finger and the tissue. It works really well, so I guess the Ark is still around. As the owner of a company that sells cryostats, you need to be aware that not everyone has or can obtain such devices. Many of us, especially in research, work with old and crippled equipment, thus we need all the help we can get from our fellow technicians. Kathleen Spencer, HT ASCP Lab Manager/LCM Supervisor UTHSC On Jun 16, 2005, at 5:11 AM, Alan Bright wrote: Emily, Basically you need to section brain at -8 to -12?C as colder temperatures will cause what you are getting plus cracking. However to achieve good quality sections at these temperatures you will need to maintain the knife and anti-roll temperatures at -20?C or lower to allow the section to slide in between the knife and the anti-roll plate unimpeded. Which is what Kathleen is doing when she warms the block face with her finger, a technique that went out with the Ark and should not be used as firstly you could have an infection issue and secondly it is not a very scientific approach in this day and age when there are devices fitted to some cryostats such as independent specimen temperature control to perform this task with full and repeatable control and of most importance, ease. 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: Kathleen Spencer [mailto:kspencer@utmem.edu] Sent: 15 June 2005 16:50 To: Katri Tuomala Cc: histonet@lists.utsouthwestern.edu; Emily Jane Wiesner-Camm Subject: Re: [Histonet] Cryostat sections with small bubbles Hi Emily, I cut rat brain sections at -18 to -20 and often have to warm the block face with my finger before each section. I also have better luck if I cut sections at 10u. These are fresh frozen brains. If I cut them at 20u I get bubbles under the sections. I keep them in the cryostat, and then fix the sections in cold fix, buffer wash, water wash, then air dry them in the hood. When I cut rat brain that has been perfused, I cut 20u floating sections. They never adhere well to slides, even if I use a hot plate. I always have bubbles under the section. That is why in this case we use floating sections. I hope this is helpful. Kathleen Spencer On Jun 14, 2005, at 8:39 PM, Katri Tuomala wrote: Hi Emily, I have never heard of hot plating cryostat sections. You may be boiling the moisture under the section forming bubbles or do you air dry them well first? Also -35 C seems too cold for brain sections, you may be getting some cutting artifacts. I'm sure there'll be others with helpful information for you. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Emily Jane Wiesner-Camm" To: Sent: Tuesday, June 14, 2005 10:36 AM Subject: [Histonet] Cryostat sections with small bubbles Hi All, I was wondering whether anyone can let me know why small bubbles appear underneath my rat brain sections when I cut them on a cryostat (at -35) when they appear perfectly flat before placing them on a hotplate. How they can be avoided? Thanks, Emily _______________________________________________ Histonet mailing list Histonet@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 pwg1 <@t> cdc.gov Fri Jun 17 05:45:31 2005 From: pwg1 <@t> cdc.gov (Greer, Patricia) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] knife sharpening Message-ID: Does anyone know of a company that sharpens microtome steel knives? A colleague here is have a problem with using disposable blades on his cryostat. The steel knife works well, but now needs sharpening. We long ago disposed of our knife sharpener and I seem to remember that there are companies that does microtome knife sharpening. Thanks, Pat Greer Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road Atlanta, GA 30333 From onep00 <@t> hotmail.com Fri Jun 17 05:58:01 2005 From: onep00 <@t> hotmail.com (Pam O'Neill) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] On Vacation Message-ID: Plesse hold mail from June 18 to June 27th. Thankyou Pam O'Neill _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar – get it now! http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/ From histology <@t> gradymem.org Fri Jun 17 06:56:24 2005 From: histology <@t> gradymem.org (histology@gradymem.org) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] xylene substitutes Message-ID: <160bb9c16095be.16095be160bb9c@onenet.net> Does anyone currently use xylene substitues in their H&E staining series? If so, which one have you found to clear best? Did you try several to come to that conclusion? We have tried several in the past and are currently using RA ClearRite. However, all of the sudden our pathologist thinks we can get better clearing. (I think he has been looking at xylene cleared slides from a reference lab and remembers how much better xylene clears.) If anyone has had wonderful results with any brand of clearing please let us know. Thanks, Angie Barnett, HTL(ASCP) Grady Memorial Hospital Chickasha, OK From Kemlo.Rogerson <@t> elht.nhs.uk Fri Jun 17 07:17:03 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] knife sharpening[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2E0@elht-exch1.xelht.nhs.uk> Diamond paste, a piece of wood and a knife back. Ah the good old days. The cut fingers, the visit to A&E for stitches, where did all that go? -----Original Message----- From: Greer, Patricia [mailto:pwg1@cdc.gov] Sent: 17 June 2005 11:46 To: Histonet (Histonet) Subject: [Histonet] knife sharpening[Scanned] Does anyone know of a company that sharpens microtome steel knives? A colleague here is have a problem with using disposable blades on his cryostat. The steel knife works well, but now needs sharpening. We long ago disposed of our knife sharpener and I seem to remember that there are companies that does microtome knife sharpening. Thanks, Pat Greer Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Don.Birgerson <@t> leica-microsystems.com Fri Jun 17 07:40:12 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] knife sharpening Message-ID: Hi Pat, There is a company close to Chicago that has been sharpening knives for years. Dorn and Hart Microedge Inc.,135 Home Ave., Villa Park, IL 60181.They have a web site www.dornandhart.com (630-530-2446). Good luck, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Greer, Patricia" To: "Histonet \(Histonet\)" Sent by: cc: (bcc: Don Birgerson/USDER/West/Leica) histonet-bounces@lists.utsouth Subject: [Histonet] knife sharpening western.edu 06/17/2005 05:45 AM Does anyone know of a company that sharpens microtome steel knives? A colleague here is have a problem with using disposable blades on his cryostat. The steel knife works well, but now needs sharpening. We long ago disposed of our knife sharpener and I seem to remember that there are companies that does microtome knife sharpening. Thanks, Pat Greer Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com From csn1x <@t> udcf.gla.ac.uk Fri Jun 17 08:28:13 2005 From: csn1x <@t> udcf.gla.ac.uk (Colin Nixon) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Species specific antibody marker Message-ID: <000801c57340$69fe6cb0$77e9d182@GLASGOWP7VFO15> I have received FFPE mouse tissue that has been treated with canine MDCK cells. It has been established that there are carcinoma cells present in the mice tissue. What I am trying to establish is whether the tumour has originated from a canine or murine origin. Does anyone know of any immunohistochemical markers that would be species specific for this task? Thanks, Colin From Barry.R.Rittman <@t> uth.tmc.edu Fri Jun 17 08:46:03 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Alizarin S mindbender question Message-ID: <566FB0B522443D43AF02D2ADBE35A6F001A9B64C@UTHEVS3.mail.uthouston.edu> Hi Julien Greetings from Houston where it is never cold. Come visit us sometime if you want to escape from the north. I have not worked with fish apart from ingesting several; however I have used an alizarin/ alcian blue technique several times to stain bone and cartilage for frogs and rodents. The technique that you use removes soft tissues and I am unclear as to why this is necessary. The techniques that I have used have retained the soft tissue and have made them permeable to clearing agents so that the details of bone and cartilage are very clear. My first thought on your technique is that the KOH is made up too strongly by mistake, can easily happen or that there is still trypsin present and acting (although the usual pH range for trypsin is 7 to 9). Both would explain the soft tissue maceration. I am not sure how long you use the trypsin or its concentration although it generally is fast acting and it is usually only active for about 3 hours. Trypsin can be sold with some gelatin in it as a stabilizing agent but not sure if this is the case here. The cloudiness may be a reaction between remnants of trypsin and the alizarin. Just some thoughts, not sure if these are correct or just BS. Good luck Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julien Lambrey de Souza Sent: Wednesday, June 08, 2005 3:30 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alizarin S mindbender question Hello histonetters, In our lab we do routine staining of fish specimens by the Clear and stain technique of Dingerkus and Uhler 1977, and some of the existing modifications with very nice results. But in the last few days, this technique has turned sour.... Something is happening to the fish when they are left in the Alizarin red S solution made up with 0.5% KOH. The solution has been made fresh several times but gives the same result each time. The fish seem to contract (muscular?) so much that the caudal becomes deformed (S shaped), the fin rays curl up and detach and the cranial elements also seem very fragile. My first thought was that the KOH solution was too harsh, but then I realized that the same KOH (0,5%) is used with these fish in the bleaching solution without harming the fish (apart from normal bleaching). The protocol is summarized as follows: 1) dehydration in EtOH ---- the fish turn out fine 2) stain in alcian blue solution (EtOH+acetic acid+alcain blue 8GN)---- fish turn out fine 3) neutralization in borax ---- fish are still fine 4) bleach in H2O2 solution (H2O2 3%+ KOH 0,5%) ---- fish still very nice 5) trypsin digestion (borax+H2O+tryspin) ------ although trypsin has cleared the specimen of its "meat" the fish is still very nice and fin rays are intact. 6) Alizarin red S solution (KOH 0,5%+alizarin S to turn solution deep purple) ----- almost on contact with the solution, the fish rinkles, rays curl up and detach... the specimen becomes useless. (The solution used to color the bones red and the specimen was then cleared of excess red by KOH bath). I have tried changing borax and KOH solutions with no improvement. Could it be that Alizarin S has gone bad? Is this even possible? We have had very high temperatures in the lab lately (30 degrees celsius), could this have influenced the solutions? I am puzzled. Also, we keep our trypsin in the -20?C freezer. The freezer has had a problem and freezed to -40?C for a day. Could this have altered the trypsin in a way that specimen exposition to this trypsin is incompatible with a later exposition to alizarin? (I am out of good ideas, so I'm trying absurd ideas....). If anyone is willing to take a shot, I will accept all suggestions because I am high and dry. Thanks for any help. Julien De souza, Evolutionary biology, UQAR, Quebec. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MDiCarlo <@t> KaleidaHealth.Org Fri Jun 17 08:54:51 2005 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] formic acid decal Message-ID: <139141F8BAF4A642A945ECC528511AF001F5ECE5@kalmb02.kaleidahealth.org> Trisha, I make my own 10% formic acid decal solution and use for large bone samples without any damage. And I get nice H&E staining. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Trisha Emry Sent: Thursday, June 16, 2005 17:54 To: histo Subject: [Histonet] formic acid decal I need to speed up the decal process on some large bone samples. What is the highest precentage of formic acid that I can use without damaging the bones? I am using sodium formate with formic acid now. Thanks, Trisha Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From POWELL_SA <@t> Mercer.edu Fri Jun 17 09:04:51 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] xylene substitutes In-Reply-To: <160bb9c16095be.16095be160bb9c@onenet.net> Message-ID: <01LPKB3AZ07K8WXLUG@Macon2.Mercer.edu> I have tried Formula 83 from CBG Biotech and it works quite well. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histology@gradymem.org Sent: Friday, June 17, 2005 6:56 AM To: Histonet Subject: [Histonet] xylene substitutes Does anyone currently use xylene substitues in their H&E staining series? If so, which one have you found to clear best? Did you try several to come to that conclusion? We have tried several in the past and are currently using RA ClearRite. However, all of the sudden our pathologist thinks we can get better clearing. (I think he has been looking at xylene cleared slides from a reference lab and remembers how much better xylene clears.) If anyone has had wonderful results with any brand of clearing please let us know. Thanks, Angie Barnett, HTL(ASCP) Grady Memorial Hospital Chickasha, OK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri Jun 17 09:08:17 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Species specific antibody marker Message-ID: Affinity BioReagents, (ABR) has a Mouse MAB (vimentin) which is specific for Mouse, Human and Rat. (MA3-745). You can use this to distinguish murine from canine cells, theoretically. I've used a human mitochondria marker to distinguish human from murine - everything that stains is human - the ones that don't stain are murine. This should work the same, I would hope. I haven't found a murine mitochondria marker so far. Jackie Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist GPRD Cancer Research Abbott Laboratories, Abbott Park, IL Jackie.OConnor@abbott.com "Colin Nixon" Sent by: histonet-bounces@lists.utsouthwestern.edu 06/17/2005 08:28 AM Please respond to Colin Nixon To: cc: Subject: [Histonet] Species specific antibody marker I have received FFPE mouse tissue that has been treated with canine MDCK cells. It has been established that there are carcinoma cells present in the mice tissue. What I am trying to establish is whether the tumour has originated from a canine or murine origin. Does anyone know of any immunohistochemical markers that would be species specific for this task? Thanks, Colin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BDUE <@t> PARTNERS.ORG Fri Jun 17 09:19:43 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] knife sharpening Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB50102705A@PHSXMB7.partners.org> You can also try C.L. Sturkey www.sturkey.com -- they are in PA. -brice "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Greer, Patricia Sent: Friday, June 17, 2005 6:46 AM To: Histonet (Histonet) Subject: [Histonet] knife sharpening Does anyone know of a company that sharpens microtome steel knives? A colleague here is have a problem with using disposable blades on his cryostat. The steel knife works well, but now needs sharpening. We long ago disposed of our knife sharpener and I seem to remember that there are companies that does microtome knife sharpening. Thanks, Pat Greer Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From POWELL_SA <@t> Mercer.edu Fri Jun 17 09:23:19 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Alizarin S mindbender question In-Reply-To: <566FB0B522443D43AF02D2ADBE35A6F001A9B64C@UTHEVS3.mail.uthouston.edu> Message-ID: <01LPKBQ7HYL88WXW7U@Macon2.Mercer.edu> Julien, I have performed the Dawson's technique (page 549 in CFA Culling third edition) on human embryos years ago using Alizarin to stain bone. I am not sure if this is suitable for what you are doing but if you contact me I will be happy to talk to you about what I encountered in the procedure. Your email address did not show up in the message. My email address is powell_sa@mercer.edu or you can call me. Shirley Powell Mercer University School of Medicine Macon, GA 478-301-2374 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R Sent: Friday, June 17, 2005 8:46 AM To: histonet Subject: RE: [Histonet] Alizarin S mindbender question Hi Julien Greetings from Houston where it is never cold. Come visit us sometime if you want to escape from the north. I have not worked with fish apart from ingesting several; however I have used an alizarin/ alcian blue technique several times to stain bone and cartilage for frogs and rodents. The technique that you use removes soft tissues and I am unclear as to why this is necessary. The techniques that I have used have retained the soft tissue and have made them permeable to clearing agents so that the details of bone and cartilage are very clear. My first thought on your technique is that the KOH is made up too strongly by mistake, can easily happen or that there is still trypsin present and acting (although the usual pH range for trypsin is 7 to 9). Both would explain the soft tissue maceration. I am not sure how long you use the trypsin or its concentration although it generally is fast acting and it is usually only active for about 3 hours. Trypsin can be sold with some gelatin in it as a stabilizing agent but not sure if this is the case here. The cloudiness may be a reaction between remnants of trypsin and the alizarin. Just some thoughts, not sure if these are correct or just BS. Good luck Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julien Lambrey de Souza Sent: Wednesday, June 08, 2005 3:30 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alizarin S mindbender question Hello histonetters, In our lab we do routine staining of fish specimens by the Clear and stain technique of Dingerkus and Uhler 1977, and some of the existing modifications with very nice results. But in the last few days, this technique has turned sour.... Something is happening to the fish when they are left in the Alizarin red S solution made up with 0.5% KOH. The solution has been made fresh several times but gives the same result each time. The fish seem to contract (muscular?) so much that the caudal becomes deformed (S shaped), the fin rays curl up and detach and the cranial elements also seem very fragile. My first thought was that the KOH solution was too harsh, but then I realized that the same KOH (0,5%) is used with these fish in the bleaching solution without harming the fish (apart from normal bleaching). The protocol is summarized as follows: 1) dehydration in EtOH ---- the fish turn out fine 2) stain in alcian blue solution (EtOH+acetic acid+alcain blue 8GN)---- fish turn out fine 3) neutralization in borax ---- fish are still fine 4) bleach in H2O2 solution (H2O2 3%+ KOH 0,5%) ---- fish still very nice 5) trypsin digestion (borax+H2O+tryspin) ------ although trypsin has cleared the specimen of its "meat" the fish is still very nice and fin rays are intact. 6) Alizarin red S solution (KOH 0,5%+alizarin S to turn solution deep purple) ----- almost on contact with the solution, the fish rinkles, rays curl up and detach... the specimen becomes useless. (The solution used to color the bones red and the specimen was then cleared of excess red by KOH bath). I have tried changing borax and KOH solutions with no improvement. Could it be that Alizarin S has gone bad? Is this even possible? We have had very high temperatures in the lab lately (30 degrees celsius), could this have influenced the solutions? I am puzzled. Also, we keep our trypsin in the -20?C freezer. The freezer has had a problem and freezed to -40?C for a day. Could this have altered the trypsin in a way that specimen exposition to this trypsin is incompatible with a later exposition to alizarin? (I am out of good ideas, so I'm trying absurd ideas....). If anyone is willing to take a shot, I will accept all suggestions because I am high and dry. Thanks for any help. Julien De souza, Evolutionary biology, UQAR, Quebec. _______________________________________________ Histonet mailing list Histonet@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 victoria.spoon <@t> bassett.org Fri Jun 17 09:25:47 2005 From: victoria.spoon <@t> bassett.org (Spoon, Victoria) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Histotechnician position available Message-ID: <052739589974CC44A7770DA22157C804099B5D@ex3.bassett.org> Bassett Healthcare in Cooperstown, New York has a part-time histotechnician position available immediately with the strong possibility of becoming full time within a year. Responsibilities include full range of histotechniques including embedding, sectioning, routine, special and immuno staining in a hospital based laboratory. If interested, contact: Victoria Spoon, Anatomic Pathology Manager Bassett Hospital Cooperstown, NY 13326 email: victoria.spoon@bassett.org (607)547-6357 NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by New York State, and Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient or have received this communication in error please contact the sender or email.security@bassett.org and destroy all copies of the original message. Thank you. From contact <@t> excaliburpathology.com Fri Jun 17 09:50:54 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Romulin red Message-ID: <20050617145054.85393.qmail@web50301.mail.yahoo.com> I thought Romulins looked like Vulcans, but were bad, hung out at the neutral zone and had the cloaking device. Maybe romulin red is like schiff's and is "cloaked" until used. ;) Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From Jackie.O'Connor <@t> abbott.com Fri Jun 17 09:57:18 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Romulin red Message-ID: Biocare is Star Trek oriented - kinda cool and fun - reminds me of doing business with Ben and Jerry's. (Hey, Hari - send me some ice cream). They actually have a "Decloaking Chamber" - their pressure cooker for HIER - as well as a Borg solution for HIER. Check out their website at www.Biocare.net if you haven't already. Nice people, too - I might add. I used the Romulin Red - and it worked just fine. I think, however, that a Methyl Green counterstain is best with it - kinda Christmas-sy. Jackie O' Paula Pierce Sent by: histonet-bounces@lists.utsouthwestern.edu 06/17/2005 09:50 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Romulin red I thought Romulins looked like Vulcans, but were bad, hung out at the neutral zone and had the cloaking device. Maybe romulin red is like schiff's and is "cloaked" until used. ;) Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Thomas.Crowell <@t> biogenidec.com Fri Jun 17 09:58:02 2005 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Rodent mast cell IHC In-Reply-To: Message-ID: Can anyone tell me if DakoCytomations C-kit CD117 polyclonal antibody crosses with mouse FFPE mast cells? Thanks Tom Crowell BiogenIdec Cambridge, MA From HACKERLAB <@t> aol.com Fri Jun 17 09:59:14 2005 From: HACKERLAB <@t> aol.com (HACKERLAB@aol.com) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Knife Sharpeneing Message-ID: <1ee.3dec0215.2fe43f42@aol.com> Hi Pat - I thought I'd pass another option your way - Hacker Instruments located in Winnsboro, South Carolina sharpens and reconditions knives as well. We also manufacture/distribute equipment and accessories used in Pathology/Histology - check out our web site, _www.hackerinstruments.com_ (http://www.hackerinstruments.com/) ; or give us a call; 803) 712-6100. Be Safe Shawnelle Shawnelle Powell Procurement/Purchasing Hacker Instruments 803) 712-6100 ext 20 From sjchtascp <@t> yahoo.com Fri Jun 17 10:07:04 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] elastic stain Message-ID: <20050617150704.1180.qmail@web90206.mail.scd.yahoo.com> Good morning everyone, Is there any elastic stain thats reliable for batch staining, that still give the domonstration of fine/course of a good VVG? Also has anyone the potassium permanganate/oxalic acid prior to the elastic working solution to achieve better staining of elastic fibers? Any opinions on whether using sodium thiosulfate or 95% alcohol is better for removing the iodine? Steve --------------------------------- Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. Learn more. From ploykasek <@t> phenopath.com Fri Jun 17 10:11:11 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Red chromogens Message-ID: Just to add a quick note, I like the Dako permanent red chromogen. It is for use with alkaline phosphatase detections. It is nice & bright, easy to use. Great for double labeling, too. Patti Loykasek PhenoPath Laboratories Seattle, WA From liz <@t> premierlab.com Fri Jun 17 10:17:28 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] formic acid decal In-Reply-To: Message-ID: <001401c5734f$adf76750$a7d48a80@AMY> Trisha I have used 20% formic acid made up in distilled water on sheep tibias and it worked just fine. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Trisha Emry Sent: Thursday, June 16, 2005 2:54 PM To: histo Subject: [Histonet] formic acid decal I need to speed up the decal process on some large bone samples. What is the highest precentage of formic acid that I can use without damaging the bones? I am using sodium formate with formic acid now. Thanks, Trisha Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Jun 17 11:28:46 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] elastic stain In-Reply-To: <20050617150704.1180.qmail@web90206.mail.scd.yahoo.com> Message-ID: <200506171628.j5HGSgt1005594@chip.viawest.net> I use Movat's Pentachrome as a batch stain for elastic fibers. I get the kit from Polyscientific and reuse many of the reagents. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Friday, June 17, 2005 8:07 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] elastic stain Good morning everyone, Is there any elastic stain thats reliable for batch staining, that still give the domonstration of fine/course of a good VVG? Also has anyone the potassium permanganate/oxalic acid prior to the elastic working solution to achieve better staining of elastic fibers? Any opinions on whether using sodium thiosulfate or 95% alcohol is better for removing the iodine? Steve --------------------------------- Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. Learn more. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Jun 17 11:29:34 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Rodent mast cell IHC In-Reply-To: Message-ID: <200506171629.j5HGTUt1005826@chip.viawest.net> Not in my experience. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Thomas Crowell Sent: Friday, June 17, 2005 7:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Rodent mast cell IHC Can anyone tell me if DakoCytomations C-kit CD117 polyclonal antibody crosses with mouse FFPE mast cells? Thanks Tom Crowell BiogenIdec Cambridge, MA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Fri Jun 17 12:04:30 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:13 2005 Subject: [Histonet] Xylene substitutes Message-ID: <000201c5735e$a0ff8eb0$1d2a14ac@wchsys.org> I use Clear-rite 3 and our pathologists have no problem with it. They all realize how bad Xylene is. I use H-2 Blue beads to help eliminate the water from our high humidity. ***** 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 mtitford <@t> aol.com Fri Jun 17 12:22:51 2005 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Formic acid decal Message-ID: <8C7417853058EC2-84-3706@mblk-r28.sysops.aol.com> Trisha in Seattle askes about formic acid decal. I trained in Great Britain and got used to using Gooding and Stewarts fluid which is formic acid based, and so I use it here in the colonies (!). Their formula is: Formic acid... 5 - 25 ml Formaldehyde (40%)... 5 ml Distilled water.. up to 100 ml. We have used this a long time. It works well and decals fast. We use the 25ml concentration. Of course if you leave the tissues in too long they look pretty tatty which I guess applies to all decalcifing fliuds except EDTA. Mike Titford USA Pathology Mobile AL USA From cekallsen <@t> ucdavis.edu Fri Jun 17 13:14:53 2005 From: cekallsen <@t> ucdavis.edu (Craig Kallsen) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Help, slide adhesive for wood Message-ID: <42B3131D.9080101@ucdavis.edu> I am having trouble getting wood specimens embedded in paraplast plus to stick to glass slides. The bark and phloem tissues stick fine, but the wood made up of the xylem does not. I have tried charges slides and a gelatin adhesive in the water bath. I am using ethanol based Safranin-O, Fast Green, Orange G and Iodine stains. Does anyone have an adhesive suggestion? From maria <@t> ski.org Fri Jun 17 13:36:52 2005 From: maria <@t> ski.org (Maria Mejia) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] knife sharpening References: Message-ID: <42B31844.703@ski.org> Over the last several years, I have used C.L. Sturkey, Inc. for reconditioning microtome knives of all sizes. For me, results have always been very good! Telephone 800-274-9446 They probably have a website too. Yours Maria Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Greer, Patricia wrote: >Does anyone know of a company that sharpens microtome steel knives? A >colleague here is have a problem with using disposable blades on his >cryostat. The steel knife works well, but now needs sharpening. We >long ago disposed of our knife sharpener and I seem to remember that >there are companies that does microtome knife sharpening. > >Thanks, > >Pat Greer >Centers for Disease Control and Prevention >Infectious Disease Pathology Activity >1600 Clifton Road >Atlanta, GA 30333 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Charles.Embrey <@t> carle.com Fri Jun 17 13:40:30 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:25:14 2005 Subject: FW: [Histonet] Xylene substitutes Message-ID: -----Original Message----- From: Charles.Embrey Sent: Friday, June 17, 2005 1:40 PM To: 'Joyce Cline' Subject: RE: [Histonet] Xylene substitutes Your quote "They all realize how bad Xylene is". Just how bad IS xylene? I have been using it for 25 years and am still alive and kicking. We recycle ours so we don't even have an environmental issue to deal with. I have never used any substitute that worked as well or better than xylene itself. Many claim to be almost as good as xylene and that is about as close as any get. As for me, I am sticking with what works. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Cline Sent: Friday, June 17, 2005 12:05 PM To: 'Histonet' Subject: [Histonet] Xylene substitutes I use Clear-rite 3 and our pathologists have no problem with it. They all realize how bad Xylene is. I use H-2 Blue beads to help eliminate the water from our high humidity. ***** 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 From BlazekL <@t> childrensdayton.org Fri Jun 17 13:51:02 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:25:14 2005 Subject: FW: [Histonet] Xylene substitutes Message-ID: Chuck.... you may be alive and kicking...... but..... from what I hear from reliable sources, leaves some questions. That "kicking" may be from to much saturation of xylene >>> "Charles.Embrey" 06/17/2005 2:40:30 PM >>> -----Original Message----- From: Charles.Embrey Sent: Friday, June 17, 2005 1:40 PM To: 'Joyce Cline' Subject: RE: [Histonet] Xylene substitutes Your quote "They all realize how bad Xylene is". Just how bad IS xylene? I have been using it for 25 years and am still alive and kicking. We recycle ours so we don't even have an environmental issue to deal with. I have never used any substitute that worked as well or better than xylene itself. Many claim to be almost as good as xylene and that is about as close as any get. As for me, I am sticking with what works. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Cline Sent: Friday, June 17, 2005 12:05 PM To: 'Histonet' Subject: [Histonet] Xylene substitutes I use Clear-rite 3 and our pathologists have no problem with it. They all realize how bad Xylene is. I use H-2 Blue beads to help eliminate the water from our high humidity. ***** 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From beckyjo <@t> email.unc.edu Fri Jun 17 14:08:11 2005 From: beckyjo <@t> email.unc.edu (Rebecca Jo) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Freezing Pig Eyes?? Message-ID: <20050617150811.1x7krtxxwsw4gcgs@webmail1.isis.unc.edu> Hello all, I will be obtaining some pig eyes in the near future and would like to know if anyone has a protocol for fixing and freezing pig eyes. I will be using the sections for in situ hybridization and immuno-staining. I've been working with frog eyes with some success but am finding it difficult to obtain really great sections. I usually make a slit in the eye, fix it in 4% paraformaldehyde overnight, and then put it through a series of 10%, 20%, and 30% sucrose. Then I snap freeze the tissue in isopentane cooled by liquid nitrogen. As stated before, I've had some success with this protocol but am unsure as to whether this will work with pig eyes. I've heard they can be tricky, especially with the retina becoming detached during the freezing process. If anyone has any tips for me, I would greatly appreciate it! Thanks. -- Rebecca E. Jo UNC-School of Medicine Department of Cell and Developmental Biology (919)843-9648 CB# 7090 From dellav <@t> musc.edu Fri Jun 17 14:08:54 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] PEL & STEL Message-ID: If you've already received replies on the list, I may have missed them so feel free to disregard. my understanding of the Formaldehyde standard is that it is NOT acceptable to measure only room air concentration. room sampling is not an accurate means for determining what your exposure was, since it is effected by air flow, room size and where in the room you are working. Personal vapor monitoring using a vapor monitor device positioned near the 'breathing zone', i.e. lab coat lapel, for example, is the method to be used. if your safety officer wants to measure both, no reason not to but if you expect to be doing only the one method of sampling, you must use the personal vapor monitoring device for sampling. When taking STEL measurements, be certain that you are performing those tasks likely to represent your highest exposure, such as pouring off formaldehyde waste or changing out your processor(s) Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Riesen, Rebecca" 06/16/05 03:24PM >>> Hello HISTONETTERS: I have a new safety officer who wants me to just check formaldehyde and xylene levels for PEL & STEL with a "Room Monitor" instead of what I have always done, which is the individual personal badges. Are there any regulations concerning this? Is it ok to just monitor the "room"? If you do the room monitoring, do you need to monitor the individual with an individual badge for the initial level? Just curious as to what you folks are all doing out there. THANKS!!!! DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Fri Jun 17 14:21:21 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Help, slide adhesive for wood Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42D2@fh2k093.fhmis.net> You could stain these sections by coverslipping with water and using an absorbant towel to exchange solutions (stains/dyes)by absorption and effusion. The slide could be cleared this way, but not very well. Try putting the fluids on the slide, let sit, and tip onto the towel to remove the solutions by osmosis. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 6/17/2005 2:14 PM Subject: [Histonet] Help, slide adhesive for wood I am having trouble getting wood specimens embedded in paraplast plus to stick to glass slides. The bark and phloem tissues stick fine, but the wood made up of the xylem does not. I have tried charges slides and a gelatin adhesive in the water bath. I am using ethanol based Safranin-O, Fast Green, Orange G and Iodine stains. Does anyone have an adhesive suggestion? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mplhisto <@t> aol.com Fri Jun 17 14:34:10 2005 From: mplhisto <@t> aol.com (mplhisto@aol.com) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Hirschsprung's and Disaccharidase In-Reply-To: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2AA@elht-exch1.xelht.nhs.uk> References: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2AA@elht-exch1.xelht.nhs.uk> Message-ID: <8C7418AAB0FEF6B-964-40F2@MBLK-M03.sysops.aol.com> I was wondering if any of you are performing Hirschspring's or Disaccharidase , in you laboratories? I would like to see procedures as well as specimen collection. Any information anyone can share would be greatly appreciated !! Thank You, Meredith Hale HT ( ASCP) Lead Histologist/ Lead IHC Tech Pathology Partners 8400 Esters Blvd. Suite 190 Irving, Texas 75063 214-277-8700 From Michael.Rice <@t> holy-cross.com Fri Jun 17 14:19:36 2005 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] RE: Histonet Digest, Vol 19, Issue 25 Message-ID: <3BC92F29BE821745AB15E04C98EE028DE3C270@HCH2KMAIL.holy-cross.com> I am sure the the Sturkey company who now makes disposables is still sharpening knives mike rice holy cross hosp Ft lauderdale -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Friday, June 17, 2005 1:07 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 19, Issue 25 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: formic acid decal (DiCarlo, Margaret) 2. RE: xylene substitutes (Shirley Powell) 3. Re: Species specific antibody marker (Jackie M O'Connor) 4. RE: knife sharpening (Due, Brice) 5. RE: Alizarin S mindbender question (Shirley Powell) 6. Histotechnician position available (Spoon, Victoria) 7. Romulin red (Paula Pierce) 8. Re: Romulin red (Jackie M O'Connor) 9. Rodent mast cell IHC (Thomas Crowell) 10. Knife Sharpeneing (HACKERLAB@aol.com) 11. elastic stain (Steven Coakley) 12. Red chromogens (Patti Loykasek) 13. RE: formic acid decal (Elizabeth Chlipala) 14. RE: elastic stain (Patsy Ruegg) 15. RE: Rodent mast cell IHC (Patsy Ruegg) ---------------------------------------------------------------------- Message: 1 Date: Fri, 17 Jun 2005 09:54:51 -0400 From: "DiCarlo, Margaret" Subject: RE: [Histonet] formic acid decal To: 'Trisha Emry' , histo Message-ID: <139141F8BAF4A642A945ECC528511AF001F5ECE5@kalmb02.kaleidahealth.org> Content-Type: text/plain; charset="iso-8859-1" Trisha, I make my own 10% formic acid decal solution and use for large bone samples without any damage. And I get nice H&E staining. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Trisha Emry Sent: Thursday, June 16, 2005 17:54 To: histo Subject: [Histonet] formic acid decal I need to speed up the decal process on some large bone samples. What is the highest precentage of formic acid that I can use without damaging the bones? I am using sodium formate with formic acid now. Thanks, Trisha Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. ------------------------------ Message: 2 Date: Fri, 17 Jun 2005 10:04:51 -0400 From: Shirley Powell Subject: RE: [Histonet] xylene substitutes To: histology@gradymem.org, 'Histonet' Message-ID: <01LPKB3AZ07K8WXLUG@Macon2.Mercer.edu> Content-Type: text/plain; charset=us-ascii I have tried Formula 83 from CBG Biotech and it works quite well. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histology@gradymem.org Sent: Friday, June 17, 2005 6:56 AM To: Histonet Subject: [Histonet] xylene substitutes Does anyone currently use xylene substitues in their H&E staining series? If so, which one have you found to clear best? Did you try several to come to that conclusion? We have tried several in the past and are currently using RA ClearRite. However, all of the sudden our pathologist thinks we can get better clearing. (I think he has been looking at xylene cleared slides from a reference lab and remembers how much better xylene clears.) If anyone has had wonderful results with any brand of clearing please let us know. Thanks, Angie Barnett, HTL(ASCP) Grady Memorial Hospital Chickasha, OK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Fri, 17 Jun 2005 09:08:17 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] Species specific antibody marker To: Colin Nixon Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Affinity BioReagents, (ABR) has a Mouse MAB (vimentin) which is specific for Mouse, Human and Rat. (MA3-745). You can use this to distinguish murine from canine cells, theoretically. I've used a human mitochondria marker to distinguish human from murine - everything that stains is human - the ones that don't stain are murine. This should work the same, I would hope. I haven't found a murine mitochondria marker so far. Jackie Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist GPRD Cancer Research Abbott Laboratories, Abbott Park, IL Jackie.OConnor@abbott.com "Colin Nixon" Sent by: histonet-bounces@lists.utsouthwestern.edu 06/17/2005 08:28 AM Please respond to Colin Nixon To: cc: Subject: [Histonet] Species specific antibody marker I have received FFPE mouse tissue that has been treated with canine MDCK cells. It has been established that there are carcinoma cells present in the mice tissue. What I am trying to establish is whether the tumour has originated from a canine or murine origin. Does anyone know of any immunohistochemical markers that would be species specific for this task? Thanks, Colin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Fri, 17 Jun 2005 10:19:43 -0400 From: "Due, Brice" Subject: RE: [Histonet] knife sharpening To: "Greer, Patricia" , "Histonet \(Histonet\)" Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB50102705A@PHSXMB7.partners.org> Content-Type: text/plain; charset="iso-8859-1" You can also try C.L. Sturkey www.sturkey.com -- they are in PA. -brice "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Greer, Patricia Sent: Friday, June 17, 2005 6:46 AM To: Histonet (Histonet) Subject: [Histonet] knife sharpening Does anyone know of a company that sharpens microtome steel knives? A colleague here is have a problem with using disposable blades on his cryostat. The steel knife works well, but now needs sharpening. We long ago disposed of our knife sharpener and I seem to remember that there are companies that does microtome knife sharpening. Thanks, Pat Greer Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Fri, 17 Jun 2005 10:23:19 -0400 From: Shirley Powell Subject: RE: [Histonet] Alizarin S mindbender question To: 'histonet' Message-ID: <01LPKBQ7HYL88WXW7U@Macon2.Mercer.edu> Content-Type: text/plain; charset=iso-8859-1 Julien, I have performed the Dawson's technique (page 549 in CFA Culling third edition) on human embryos years ago using Alizarin to stain bone. I am not sure if this is suitable for what you are doing but if you contact me I will be happy to talk to you about what I encountered in the procedure. Your email address did not show up in the message. My email address is powell_sa@mercer.edu or you can call me. Shirley Powell Mercer University School of Medicine Macon, GA 478-301-2374 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R Sent: Friday, June 17, 2005 8:46 AM To: histonet Subject: RE: [Histonet] Alizarin S mindbender question Hi Julien Greetings from Houston where it is never cold. Come visit us sometime if you want to escape from the north. I have not worked with fish apart from ingesting several; however I have used an alizarin/ alcian blue technique several times to stain bone and cartilage for frogs and rodents. The technique that you use removes soft tissues and I am unclear as to why this is necessary. The techniques that I have used have retained the soft tissue and have made them permeable to clearing agents so that the details of bone and cartilage are very clear. My first thought on your technique is that the KOH is made up too strongly by mistake, can easily happen or that there is still trypsin present and acting (although the usual pH range for trypsin is 7 to 9). Both would explain the soft tissue maceration. I am not sure how long you use the trypsin or its concentration although it generally is fast acting and it is usually only active for about 3 hours. Trypsin can be sold with some gelatin in it as a stabilizing agent but not sure if this is the case here. The cloudiness may be a reaction between remnants of trypsin and the alizarin. Just some thoughts, not sure if these are correct or just BS. Good luck Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julien Lambrey de Souza Sent: Wednesday, June 08, 2005 3:30 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alizarin S mindbender question Hello histonetters, In our lab we do routine staining of fish specimens by the Clear and stain technique of Dingerkus and Uhler 1977, and some of the existing modifications with very nice results. But in the last few days, this technique has turned sour.... Something is happening to the fish when they are left in the Alizarin red S solution made up with 0.5% KOH. The solution has been made fresh several times but gives the same result each time. The fish seem to contract (muscular?) so much that the caudal becomes deformed (S shaped), the fin rays curl up and detach and the cranial elements also seem very fragile. My first thought was that the KOH solution was too harsh, but then I realized that the same KOH (0,5%) is used with these fish in the bleaching solution without harming the fish (apart from normal bleaching). The protocol is summarized as follows: 1) dehydration in EtOH ---- the fish turn out fine 2) stain in alcian blue solution (EtOH+acetic acid+alcain blue 8GN)---- fish turn out fine 3) neutralization in borax ---- fish are still fine 4) bleach in H2O2 solution (H2O2 3%+ KOH 0,5%) ---- fish still very nice 5) trypsin digestion (borax+H2O+tryspin) ------ although trypsin has cleared the specimen of its "meat" the fish is still very nice and fin rays are intact. 6) Alizarin red S solution (KOH 0,5%+alizarin S to turn solution deep purple) ----- almost on contact with the solution, the fish rinkles, rays curl up and detach... the specimen becomes useless. (The solution used to color the bones red and the specimen was then cleared of excess red by KOH bath). I have tried changing borax and KOH solutions with no improvement. Could it be that Alizarin S has gone bad? Is this even possible? We have had very high temperatures in the lab lately (30 degrees celsius), could this have influenced the solutions? I am puzzled. Also, we keep our trypsin in the -20?C freezer. The freezer has had a problem and freezed to -40?C for a day. Could this have altered the trypsin in a way that specimen exposition to this trypsin is incompatible with a later exposition to alizarin? (I am out of good ideas, so I'm trying absurd ideas....). If anyone is willing to take a shot, I will accept all suggestions because I am high and dry. Thanks for any help. Julien De souza, Evolutionary biology, UQAR, Quebec. _______________________________________________ Histonet mailing list Histonet@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, 17 Jun 2005 10:25:47 -0400 From: "Spoon, Victoria" Subject: [Histonet] Histotechnician position available To: Message-ID: <052739589974CC44A7770DA22157C804099B5D@ex3.bassett.org> Content-Type: text/plain; charset="iso-8859-1" Bassett Healthcare in Cooperstown, New York has a part-time histotechnician position available immediately with the strong possibility of becoming full time within a year. Responsibilities include full range of histotechniques including embedding, sectioning, routine, special and immuno staining in a hospital based laboratory. If interested, contact: Victoria Spoon, Anatomic Pathology Manager Bassett Hospital Cooperstown, NY 13326 email: victoria.spoon@bassett.org (607)547-6357 NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by New York State, and Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient or have received this communication in error please contact the sender or email.security@bassett.org and destroy all copies of the original message. Thank you. ------------------------------ Message: 7 Date: Fri, 17 Jun 2005 07:50:54 -0700 (PDT) From: Paula Pierce Subject: [Histonet] Romulin red To: histonet@lists.utsouthwestern.edu Message-ID: <20050617145054.85393.qmail@web50301.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I thought Romulins looked like Vulcans, but were bad, hung out at the neutral zone and had the cloaking device. Maybe romulin red is like schiff's and is "cloaked" until used. ;) Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com ------------------------------ Message: 8 Date: Fri, 17 Jun 2005 09:57:18 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] Romulin red To: Paula Pierce Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Biocare is Star Trek oriented - kinda cool and fun - reminds me of doing business with Ben and Jerry's. (Hey, Hari - send me some ice cream). They actually have a "Decloaking Chamber" - their pressure cooker for HIER - as well as a Borg solution for HIER. Check out their website at www.Biocare.net if you haven't already. Nice people, too - I might add. I used the Romulin Red - and it worked just fine. I think, however, that a Methyl Green counterstain is best with it - kinda Christmas-sy. Jackie O' Paula Pierce Sent by: histonet-bounces@lists.utsouthwestern.edu 06/17/2005 09:50 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Romulin red I thought Romulins looked like Vulcans, but were bad, hung out at the neutral zone and had the cloaking device. Maybe romulin red is like schiff's and is "cloaked" until used. ;) Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Fri, 17 Jun 2005 10:58:02 -0400 From: Thomas Crowell Subject: [Histonet] Rodent mast cell IHC To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Can anyone tell me if DakoCytomations C-kit CD117 polyclonal antibody crosses with mouse FFPE mast cells? Thanks Tom Crowell BiogenIdec Cambridge, MA ------------------------------ Message: 10 Date: Fri, 17 Jun 2005 10:59:14 EDT From: HACKERLAB@aol.com Subject: [Histonet] Knife Sharpeneing To: pwg1@cdc.gov, Histonet@pathology.swmed.edu Message-ID: <1ee.3dec0215.2fe43f42@aol.com> Content-Type: text/plain; charset="US-ASCII" Hi Pat - I thought I'd pass another option your way - Hacker Instruments located in Winnsboro, South Carolina sharpens and reconditions knives as well. We also manufacture/distribute equipment and accessories used in Pathology/Histology - check out our web site, _www.hackerinstruments.com_ (http://www.hackerinstruments.com/) ; or give us a call; 803) 712-6100. Be Safe Shawnelle Shawnelle Powell Procurement/Purchasing Hacker Instruments 803) 712-6100 ext 20 ------------------------------ Message: 11 Date: Fri, 17 Jun 2005 08:07:04 -0700 (PDT) From: Steven Coakley Subject: [Histonet] elastic stain To: Histonet@lists.utsouthwestern.edu Message-ID: <20050617150704.1180.qmail@web90206.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Good morning everyone, Is there any elastic stain thats reliable for batch staining, that still give the domonstration of fine/course of a good VVG? Also has anyone the potassium permanganate/oxalic acid prior to the elastic working solution to achieve better staining of elastic fibers? Any opinions on whether using sodium thiosulfate or 95% alcohol is better for removing the iodine? Steve --------------------------------- Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. Learn more. ------------------------------ Message: 12 Date: Fri, 17 Jun 2005 08:11:11 -0700 From: Patti Loykasek Subject: [Histonet] Red chromogens To: histonet Message-ID: Content-Type: text/plain; charset="US-ASCII" Just to add a quick note, I like the Dako permanent red chromogen. It is for use with alkaline phosphatase detections. It is nice & bright, easy to use. Great for double labeling, too. Patti Loykasek PhenoPath Laboratories Seattle, WA ------------------------------ Message: 13 Date: Fri, 17 Jun 2005 09:17:28 -0600 From: "Elizabeth Chlipala" Subject: RE: [Histonet] formic acid decal To: "'Trisha Emry'" , "'histo'" Message-ID: <001401c5734f$adf76750$a7d48a80@AMY> Content-Type: text/plain; charset="us-ascii" Trisha I have used 20% formic acid made up in distilled water on sheep tibias and it worked just fine. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Trisha Emry Sent: Thursday, June 16, 2005 2:54 PM To: histo Subject: [Histonet] formic acid decal I need to speed up the decal process on some large bone samples. What is the highest precentage of formic acid that I can use without damaging the bones? I am using sodium formate with formic acid now. Thanks, Trisha Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Fri, 17 Jun 2005 10:28:46 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] elastic stain To: "'Steven Coakley'" , Message-ID: <200506171628.j5HGSgt1005594@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" I use Movat's Pentachrome as a batch stain for elastic fibers. I get the kit from Polyscientific and reuse many of the reagents. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Friday, June 17, 2005 8:07 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] elastic stain Good morning everyone, Is there any elastic stain thats reliable for batch staining, that still give the domonstration of fine/course of a good VVG? Also has anyone the potassium permanganate/oxalic acid prior to the elastic working solution to achieve better staining of elastic fibers? Any opinions on whether using sodium thiosulfate or 95% alcohol is better for removing the iodine? Steve --------------------------------- Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. Learn more. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Fri, 17 Jun 2005 10:29:34 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] Rodent mast cell IHC To: "'Thomas Crowell'" , Message-ID: <200506171629.j5HGTUt1005826@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Not in my experience. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Thomas Crowell Sent: Friday, June 17, 2005 7:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Rodent mast cell IHC Can anyone tell me if DakoCytomations C-kit CD117 polyclonal antibody crosses with mouse FFPE mast cells? Thanks Tom Crowell BiogenIdec Cambridge, MA _______________________________________________ Histonet mailing list Histonet@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 19, Issue 25 **************************************** ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From Charles.Embrey <@t> carle.com Fri Jun 17 14:47:00 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:25:14 2005 Subject: FW: FW: [Histonet] Xylene substitutes Message-ID: -----Original Message----- From: Charles.Embrey Sent: Friday, June 17, 2005 2:46 PM To: 'Linda Blazek' Subject: RE: FW: [Histonet] Xylene substitutes I did say kicking, not twitching. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Friday, June 17, 2005 1:51 PM To: Charles.Embrey; histonet@lists.utsouthwestern.edu Subject: Re: FW: [Histonet] Xylene substitutes Chuck.... you may be alive and kicking...... but..... from what I hear from reliable sources, leaves some questions. That "kicking" may be from to much saturation of xylene >>> "Charles.Embrey" 06/17/2005 2:40:30 PM >>> -----Original Message----- From: Charles.Embrey Sent: Friday, June 17, 2005 1:40 PM To: 'Joyce Cline' Subject: RE: [Histonet] Xylene substitutes Your quote "They all realize how bad Xylene is". Just how bad IS xylene? I have been using it for 25 years and am still alive and kicking. We recycle ours so we don't even have an environmental issue to deal with. I have never used any substitute that worked as well or better than xylene itself. Many claim to be almost as good as xylene and that is about as close as any get. As for me, I am sticking with what works. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Cline Sent: Friday, June 17, 2005 12:05 PM To: 'Histonet' Subject: [Histonet] Xylene substitutes I use Clear-rite 3 and our pathologists have no problem with it. They all realize how bad Xylene is. I use H-2 Blue beads to help eliminate the water from our high humidity. ***** 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 _______________________________________________ Histonet mailing list Histonet@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 debbiekeith <@t> cox.net Fri Jun 17 15:38:18 2005 From: debbiekeith <@t> cox.net (Debbie Keith) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] One of our own in need ! Please Read Message-ID: <5.2.0.9.0.20050617133749.020a2d30@pop.central.cox.net> Hi, I just wanted to let everyone know that we have updated Alysa's website to allow donations to be accepted via PayPal. I know through talking to Lisa that every single donation is needed and greatly appreciated. Her focus sincerely needs to be on the fight her daughter is fighting... not worrying about how she's going to pay her mortgage. In my first correspondence with Lisa she said something that has sort of echoed in my head ever since. Children... should NOT get cancer. they shouldn't. Debbie Keith HT(ASCP) From jcline <@t> wchsys.org Fri Jun 17 15:40:13 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] To Maria Mejia Message-ID: <000801c5737c$c3997440$1d2a14ac@wchsys.org> The H2 Blue beads are from American Master Tech Scientific, Inc 800-860-4073, they are in California. You use 10g, for every 500 mL xylene or substitute. ***** 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 BDUE <@t> PARTNERS.ORG Fri Jun 17 15:46:42 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Help, slide adhesive for wood Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB50102705C@PHSXMB7.partners.org> Not an adhesive, but along the same lines as the previous post, you could use what are called "capillary gap" slides. Same idea as using a coverslip. Cap-gap slides have extra frosted areas at the bottom corners, so when you double them up they maintain a gap of a known thickness unlike coverslips. Solns get sucked up into the gap and rinsing would not be so problematic as with coverslips. There are automated stainers based on cap-gap slides. You can place a cap-gap slide against a regular charged slide and the gap is roughly half the size. They cost about twice as much as charged slides, but for this purpose you could reuse them until the corner frostings get scraped off. To separate two slides, you must fully immerse the pair. Once submerged they come apart easily -- otherwise they stick together tightly. -brice "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonner, Janet Sent: Friday, June 17, 2005 3:21 PM To: 'Craig Kallsen '; 'histonet-bounces@lists.utsouthwestern.edu '; 'histonet@lists.utsouthwestern.edu ' Subject: RE: [Histonet] Help, slide adhesive for wood You could stain these sections by coverslipping with water and using an absorbant towel to exchange solutions (stains/dyes)by absorption and effusion. The slide could be cleared this way, but not very well. Try putting the fluids on the slide, let sit, and tip onto the towel to remove the solutions by osmosis. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 6/17/2005 2:14 PM Subject: [Histonet] Help, slide adhesive for wood I am having trouble getting wood specimens embedded in paraplast plus to stick to glass slides. The bark and phloem tissues stick fine, but the wood made up of the xylem does not. I have tried charges slides and a gelatin adhesive in the water bath. I am using ethanol based Safranin-O, Fast Green, Orange G and Iodine stains. Does anyone have an adhesive suggestion? _______________________________________________ Histonet mailing list Histonet@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 jcline <@t> wchsys.org Fri Jun 17 15:47:02 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] To Charles Embrey Message-ID: <000d01c5737d$b791a950$1d2a14ac@wchsys.org> There are 2 people in my lab that are sensitized to Xylene, my self also. Have you read the cautions on Xylene? NSH survey results? I have inspected labs that use Xylene and I still cannot be around it for very long without problems. ***** 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 debbiekeith <@t> cox.net Fri Jun 17 17:59:18 2005 From: debbiekeith <@t> cox.net (Debbie Keith) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] One of our own in need ! Please Read In-Reply-To: <200506172251.j5HMpArG015430@dbmail-mx1.orcon.net.nz> References: <5.2.0.9.0.20050617133749.020a2d30@pop.central.cox.net> Message-ID: <5.2.0.9.0.20050617155103.020a29d0@pop.central.cox.net> Darren... after sending the earlier message i realized i didn't include the link. www.alysa-anne.com her whole story is there... complete with pictures. i have a daughter that is 6 weeks younger than Alysa... this story hits too close to home. Everyone that i have told about Alysa is keeping her in their thoughts/prayers. we are all hoping for a miracle. (they HAPPEN!) Lisa is spending her time... focusing on keeping her Angel here on Earth. Debbie :) At 10:50 AM 6/18/05 +1200, you wrote: >Hi Debbie, >Could you please give me more information on Alysa. I have seen discussion >relating to her over the past week but don't know what the background is. >I am in NZ and as such may be "out of the loop". >As a father of two young children any mention of kiddies and cancer >immediately gets my attention. >A link to the website you mention could be useful too. > >Thanks >Darren James > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Debbie Keith >Sent: 18 June 2005 08:38 >To: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] One of our own in need ! Please Read > >Hi, > >I just wanted to let everyone know that we have updated Alysa's website to >allow donations to be accepted via PayPal. > >I know through talking to Lisa that every single donation is needed and >greatly appreciated. Her focus sincerely needs to be on the fight her >daughter is fighting... not worrying about how she's going to pay her >mortgage. > >In my first correspondence with Lisa she said something that has sort of >echoed in my head ever since. > >Children... should NOT get cancer. > >they shouldn't. > >Debbie Keith HT(ASCP) > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Fri Jun 17 18:18:09 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Microarray machines Message-ID: Greetings all, I need someone who has microarray equipment, experience, etc to contact me ASCP. Please call me at 210-231-8058 or email me back. Thanks Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX From histology.bc <@t> shaw.ca Fri Jun 17 19:39:46 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] elastic stain References: <20050617150704.1180.qmail@web90206.mail.scd.yahoo.com> Message-ID: <42B36D52.60203@shaw.ca> Hi Steve, One of the nicest, simplest stains for elastic fibres (large, small, and tiny) is orcein. The fibres are deep purple and no differentiation steps are involved. It's an ideal method for mass production. However, the purple of the elastin does not contrast well with the red of the van Gieson. Aldehyde fuchsin is also great for elastic fibres, does not require differentiation, and is reliable. Its contrast with van Gieson is not great though. A number of years ago, Keith Gordon and I wrote up a method in Bancroft's Theory and Practice of Histological Techniquers that used a dye called Dahlia, one of the triphenylmethane dyes related to the basic fuchsins. This method gave very good, reliable results, needed no differentiation, and stained the elastic fibres a very dark blue (almost black) that contrasted nicely with the van Gieson. The stain solution is essentially a varaint of aldehyde fuchsin, but uses dahlia in place of the fuchsin. Paul Bradbury Kamloops, BC, Canada Steven Coakley wrote: >Good morning everyone, > >Is there any elastic stain thats reliable for batch staining, that still give the domonstration of fine/course of a good VVG? Also has anyone the potassium permanganate/oxalic acid prior to the elastic working solution to achieve better staining of elastic fibers? Any opinions on whether using sodium thiosulfate or 95% alcohol is better for removing the iodine? > >Steve > > >--------------------------------- >Do you Yahoo!? > Yahoo! Mail - Find what you need with new enhanced search. Learn more. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From PKARLISCH <@t> psu.edu Fri Jun 17 19:39:58 2005 From: PKARLISCH <@t> psu.edu (Patricia Karlisch) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Re: Histonet Digest, Vol 19, Issue 26 (Out of office) Message-ID: I will be on vacation from June 21st through June 25th and will return on Monday June 28th. I will be in the process of moving from Danville to Palmyra. I can be reached at 570-271-0665 until June 22nd and hopefully will be connected to my new phone 717-832-2801 on June 24th. Tim Vickroy will be in charge for the week. If you have any questions please ask for Tim. Thanks From kccatunda <@t> uol.com.br Sat Jun 18 09:56:14 2005 From: kccatunda <@t> uol.com.br (Katia Cristina Catunda) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Expiration date and new EA question. References: <000901c57211$a7ab5080$a279fea9@privatexx> <3954.140.251.146.2.1118949169.squirrel@webmail> Message-ID: <008b01c57415$e04daf90$a279fea9@privatexx> Thanks for all your answers, we'll probably donnating the products to a resarch institution near here. Now we are having papanicolaou's EA problem over here... slides are getting too redish and my feelings tells me that our main problem our last eosin's batch, at the same time we are thying to addjust our EA recipe... Nowadays we use: eosin y, bismark, light green (main solutions dissolved in distilled water) absolute alcohol and some drops of phosphotungic acid and lithium carbonate. Trying to find new recipes I've found all sorts of them.. using vesuvine instead of bismark, using fast green instead of light green and dissolving main solutions in alcohol instead of distilled water. As we have been using this stain battery for more than 5 years and just know we thought we should better update it I am completely lost... What recipes do you use, is fast green better than light (read that it should be better fot stock solutions)? Thanks again, Katia From jnocito <@t> satx.rr.com Sat Jun 18 12:20:47 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:14 2005 Subject: Here I go again[Histonet] Expiration date References: <000901c57211$a7ab5080$a279fea9@privatexx> Message-ID: <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> Okay, I've been too quiet for too long. Here I go again. I am so tired of CAP and their ridiculous regulations. Every CAP inspection I fight this same question. When I was supervising the Immuno lab at AFIP, we froze concentrated antibodies at -70 for years. Antibodies that were frozen in 1080, were still viable in 1990. That means we were running immunos in 1990 with 1980 prices. What is so wrong about that? We also ran a known positive control with each batch. If the positive control worked, guess what? The antibody was still good. If the positive control did not work, we threw out that lot number and repeated it with another lot number. When we saw that there was a drop in the staining intensity, we tossed that lot and started another. My experience with antibodies is that they just don't go bad over night, they start staining with less intensity. I think that the CAP board members have stocks in the biochemical companies. Please don't get me wrong, I have many close friends that work in the biochemical side of this, but why would this drastic change in view? The FDA? I doubt it. Maybe I'm too one sides on this issue, but give me a break! If the positive control works in any other antibody, doesn't that mean the antibody is viable? I know companies have to put an expiration date on their products, but come on. If an antibody expires June 30, 2005, does it automatically go dead on July 1? As managers and supervisors, we are continuously bashed about saving money. This would be a great place to start, don't you think? That is all. Thank you Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Katia Cristina Catunda" To: "histonet" Sent: Wednesday, June 15, 2005 8:20 PM Subject: [Histonet] Expiration date > What should we do with antibodies and reagents after their indicated > expiration dates? Theorectically they shouldn't be estable after the > expiration date but we do know that most of them still works perfectly... > should we just throw them out and not use them anymore, wouldn't this be a > way for the manufacturers manipulate their sellings? Need some opinions... > > Tks > > Katia > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.7.8/22 - Release Date: 6/17/2005 > > From katri <@t> cogeco.ca Sat Jun 18 12:45:18 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:14 2005 Subject: Here I go again[Histonet] Expiration date References: <000901c57211$a7ab5080$a279fea9@privatexx> <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> Message-ID: <02b601c5742d$7e600720$6a9a9618@Katri> Hear, hear, Joe!!!!!! I'm in complete agreement with your points. This society has acquired a complete through away mentality...in the name of what? Certain decisions should be left up the professionals to make... ----- Original Message ----- From: "Joe Nocito" To: "Katia Cristina Catunda" ; "histonet" Sent: Saturday, June 18, 2005 1:20 PM Subject: Here I go again[Histonet] Expiration date > Okay, > I've been too quiet for too long. Here I go again. > > I am so tired of CAP and their ridiculous regulations. Every CAP > inspection I fight this same question. When I was supervising the Immuno > lab at AFIP, we froze concentrated antibodies at -70 for years. Antibodies > that were frozen in 1080, were still viable in 1990. That means we were > running immunos in 1990 with 1980 prices. What is so wrong about that? > We also ran a known positive control with each batch. If the positive > control worked, guess what? The antibody was still good. If the positive > control did not work, we threw out that lot number and repeated it with > another lot number. When we saw that there was a drop in the staining > intensity, we tossed that lot and started another. My experience with > antibodies is that they just don't go bad over night, they start staining > with less intensity. > I think that the CAP board members have stocks in the biochemical > companies. Please don't get me wrong, I have many close friends that work > in the biochemical side of this, but why would this drastic change in > view? The FDA? I doubt it. > Maybe I'm too one sides on this issue, but give me a break! If the > positive control works in any other antibody, doesn't that mean the > antibody is viable? > I know companies have to put an expiration date on their products, but > come on. If an antibody expires June 30, 2005, does it automatically go > dead on July 1? > As managers and supervisors, we are continuously bashed about saving > money. This would be a great place to start, don't you think? > That is all. Thank you > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > ----- Original Message ----- > From: "Katia Cristina Catunda" > To: "histonet" > Sent: Wednesday, June 15, 2005 8:20 PM > Subject: [Histonet] Expiration date > > >> What should we do with antibodies and reagents after their indicated >> expiration dates? Theorectically they shouldn't be estable after the >> expiration date but we do know that most of them still works perfectly... >> should we just throw them out and not use them anymore, wouldn't this be >> a way for the manufacturers manipulate their sellings? Need some >> opinions... >> >> Tks >> >> Katia >> >> >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> -- >> No virus found in this incoming message. >> Checked by AVG Anti-Virus. >> Version: 7.0.323 / Virus Database: 267.7.8/22 - Release Date: 6/17/2005 >> >> > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From stancelb <@t> msn.com Sat Jun 18 14:03:14 2005 From: stancelb <@t> msn.com (Barbara Stancel) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] RE: Xylene Substitutes Message-ID: I'm with Charles. We have been using xylene for 31 years. We have tried the xylene substitutes, but we and our pathologist prefer xylene. We recycle all the xylene to reduce cost of purchasing and cut hazardous waste. We have great ventilation in the lab. Our twice-a-year STEL monitoring has come back at way below the acceptable limits. If it ain't broke, don't fix it. Barbara Stancel USDA, FSIS, OPHS, EL, Pathology Athens, Georgia -----Original Message----- From: Charles.Embrey Sent: Friday, June 17, 2005 1:40 PM To: 'Joyce Cline' Subject: RE: [Histonet] Xylene substitutes Your quote "They all realize how bad Xylene is". Just how bad IS xylene? I have been using it for 25 years and am still alive and kicking. We recycle ours so we don't even have an environmental issue to deal with. I have never used any substitute that worked as well or better than xylene itself. Many claim to be almost as good as xylene and that is about as close as any get. As for me, I am sticking with what works. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Cline Sent: Friday, June 17, 2005 12:05 PM To: 'Histonet' Subject: [Histonet] Xylene substitutes I use Clear-rite 3 and our pathologists have no problem with it. They all realize how bad Xylene is. I use H-2 Blue beads to help eliminate the water from our high humidity. From Felixdustin <@t> aol.com Sat Jun 18 22:01:12 2005 From: Felixdustin <@t> aol.com (Felixdustin@aol.com) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] unsubscirbe Message-ID: <145.47894b81.2fe639f8@aol.com> Unsubscribe, please. From gu.lang <@t> gmx.at Sun Jun 19 10:57:32 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] VEGF - question Message-ID: Maybe a stupid question. Are endothel cells always stained positiv with vascular endothelial growth factor? I have got a tissue of a pyogenic granuloma as a positiv control from my pathologist. The endothel cells of the vessels are clear negativ, the surrounding area is positiv. Is this the expected result or is there any mistake? Thanks for your help. Gudrun Lang From beingmary53 <@t> sbcglobal.net Sun Jun 19 12:47:20 2005 From: beingmary53 <@t> sbcglobal.net (mary johnson) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Brain fixation In-Reply-To: <200506161659.j5GGxOje025248@ylpvm17.prodigy.net> Message-ID: Hi. Dorothy we fix our brain section in 20% formalin, and if your section is thin enough in two to three days they are ready for prosses oh yes the 20% for formalin is what we make up. Good luck mary Johnson HT ascp -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Thursday, June 16, 2005 11:59 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 19, Issue 23 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. Brain fixative (Webb, Dorothy L) 2. thanks (Kaye Ryan) 3. RE: Brain fixative[Scanned] (Rogerson Kemlo (ELHT) Pathology) 4. Processing of research/mouse tissues (was delayed processing) (Johnson, Teri) 5. Re: Cryostat sections with small bubbles (Kathleen Spencer) 6. Negatives for immunos using polyclonal antibody (andrew.macduff@ed.ac.uk) 7. RE: Schiff's reagent in microwave? (Bonner, Janet) 8. Microwave use in Pharmaceutical/CRO histo labs (Stephen.Eyres@sanofi-aventis.com) 9. Required CEU's (Dianne Holmes) 10. Muscle Referrals @ Baylor - Houston (Nita Searcy) 11. One of our own in need ! Please Read (Vinnie Della Speranza) 12. RE: Schiff's reagent in microwave? (GUTIERREZ, JUAN) 13. RE: Brain fixative (GUTIERREZ, JUAN) 14. RE: BCIP/NBT ready-to-use (Edmondson David (RBV) NHS Christie Tr) 15. RE: Schiff's reagent in microwave? (mucram11@comcast.net) ---------------------------------------------------------------------- Message: 1 Date: Thu, 16 Jun 2005 09:52:51 -0500 From: "Webb, Dorothy L" Subject: [Histonet] Brain fixative To: histonet@lists.utsouthwestern.edu Message-ID: <0E394B648E5284478A6CCB78E5AFDA2718C368@hpes1.HealthPartners.int> Content-Type: text/plain; charset="US-ASCII" Does anyone fix their brain tissue from autopsies in anything other than 10% buffered formalin? The PA I am working with seems to feel they used a 4% solution of lightly buffered formalin from Mallincroft. Can anyone help me with this, as I am only used to using 10% formalin, as we use for all of our routine histology fixation. Any ideas would be greatly appreciated and thanks ahead of time to all of my fellow histonetters..what a great group you are!! +______________________________________+ ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. ------------------------------ Message: 2 Date: Thu, 16 Jun 2005 10:58:17 -0400 From: "Kaye Ryan" Subject: [Histonet] thanks To: Message-ID: Content-Type: text/plain; charset=US-ASCII Thanks to everyone who responded to my inquiry concerning decal solutions for bone marrows. Kaye Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 ------------------------------ Message: 3 Date: Thu, 16 Jun 2005 16:00:08 +0100 From: "Rogerson Kemlo (ELHT) Pathology" Subject: RE: [Histonet] Brain fixative[Scanned] To: "Webb, Dorothy L" , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2D1@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" Aren't they the same? We've had this debate; 4% formaldehyde is 10% formalin. 'Lightly buffered' is that like lightly salted? -----Original Message----- From: Webb, Dorothy L [mailto:Dorothy.L.Webb@HealthPartners.Com] Sent: 16 June 2005 15:53 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Brain fixative[Scanned] Does anyone fix their brain tissue from autopsies in anything other than 10% buffered formalin? The PA I am working with seems to feel they used a 4% solution of lightly buffered formalin from Mallincroft. Can anyone help me with this, as I am only used to using 10% formalin, as we use for all of our routine histology fixation. Any ideas would be greatly appreciated and thanks ahead of time to all of my fellow histonetters..what a great group you are!! +______________________________________+ ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Thu, 16 Jun 2005 10:00:51 -0500 From: "Johnson, Teri" Subject: [Histonet] Processing of research/mouse tissues (was delayed processing) To: , "Histonet" Message-ID: Content-Type: text/plain; charset="US-ASCII" Steve, welcome to research! It is a different world here. My advice would be to buy a hydrometer, and test your alcohols. When your last 100% starts to show some water, rotate it up to the position just past the 95% alcohol and make the later changes fresh. Also invest in the animal processing manual from the NSH. You'll find some very helpful schedules in there for all kinds of animal tissues. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org ------------------------------ Message: 5 Date: Thu, 16 Jun 2005 10:05:24 -0500 From: Kathleen Spencer Subject: Re: [Histonet] Cryostat sections with small bubbles To: Alan Bright Cc: histonet@lists.utsouthwestern.edu, Katri Tuomala , Emily Jane Wiesner-Camm Message-ID: <0ff76615670407b429351b22aca29125@utmem.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Yes Alan, but when I put my finger on the block face to warm it, there is a kimwipe or gauze between my finger and the tissue. It works really well, so I guess the Ark is still around. As the owner of a company that sells cryostats, you need to be aware that not everyone has or can obtain such devices. Many of us, especially in research, work with old and crippled equipment, thus we need all the help we can get from our fellow technicians. Kathleen Spencer, HT ASCP Lab Manager/LCM Supervisor UTHSC On Jun 16, 2005, at 5:11 AM, Alan Bright wrote: > Emily, > > Basically you need to section brain at -8 to -12:C as colder > temperatures will cause what you are getting plus cracking. However to > achieve good quality sections at these temperatures you will need to > maintain the knife and anti-roll temperatures at -20:C or lower to > allow the section to slide in between the knife and the anti-roll > plate unimpeded. Which is what Kathleen is doing when she warms the > block face with her finger, a technique that went out with the Ark and > should not be used as firstly you could have an infection issue and > secondly it is not a very scientific approach in this day and age when > there are devices fitted to some cryostats such as independent > specimen temperature control to perform this task with full and > repeatable control and of most importance, ease. > > 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: Kathleen Spencer [mailto:kspencer@utmem.edu] > Sent: 15 June 2005 16:50 > To: Katri Tuomala > Cc: histonet@lists.utsouthwestern.edu; Emily Jane Wiesner-Camm > Subject: Re: [Histonet] Cryostat sections with small bubbles > > > Hi Emily, > > I cut rat brain sections at -18 to -20 and often have to warm the block > face with my finger before each section. I also have better luck if I > cut sections at 10u. These are fresh frozen brains. If I cut them at > 20u I get bubbles under the sections. I keep them in the cryostat, and > then fix the sections in cold fix, buffer wash, water wash, then air > dry them in the hood. > When I cut rat brain that has been perfused, I cut 20u floating > sections. They never adhere well to slides, even if I use a hot plate. > I always have bubbles under the section. That is why in this case we > use floating sections. > > I hope this is helpful. > > Kathleen Spencer > > > On Jun 14, 2005, at 8:39 PM, Katri Tuomala wrote: > >> Hi Emily, >> I have never heard of hot plating cryostat sections. You may be >> boiling the moisture under the section forming bubbles or do you air >> dry them well first? Also -35 C seems too cold for brain sections, you >> may be getting some cutting artifacts. >> I'm sure there'll be others with helpful information for you. >> Katri >> >> Katri Tuomala >> Hamilton, Ontario, Canada >> ----- Original Message ----- From: "Emily Jane Wiesner-Camm" >> >> To: >> Sent: Tuesday, June 14, 2005 10:36 AM >> Subject: [Histonet] Cryostat sections with small bubbles >> >> >>> Hi All, >>> I was wondering whether anyone can let me know why small bubbles >>> appear underneath my rat brain sections when I cut them on a cryostat >>> (at -35) when they appear perfectly flat before placing them on a >>> hotplate. How they can be avoided? >>> Thanks, >>> Emily >>> >>> >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Thu, 16 Jun 2005 16:31:22 +0100 From: andrew.macduff@ed.ac.uk Subject: [Histonet] Negatives for immunos using polyclonal antibody To: histonet@lists.utsouthwestern.edu Message-ID: <1118935882.42b19b4a89a49@staffmail.ed.ac.uk> Content-Type: text/plain; charset=ISO-8859-15 Dear all A quick question- when doing IHC with a polyclonal Ab is a serum negative sufficient since there can be no isotype specific control Ab? Thanks for the help Andrew Andrew MacDuff Clinical Research Fellow MRC Centre for Inflammation Research Medical School Edinburgh University ------------------------------ Message: 7 Date: Thu, 16 Jun 2005 11:53:56 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] Schiff's reagent in microwave? To: "'Jacquie.Farnsworth@cls.ab.ca'" , histonet@lists.utsouthwestern.edu Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42CD@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 We did this microwaving of the Schiff reagent for awhile, but the step is only 15 minutes at room temperature so it wasn't worth using the microwave for this application. The Schiff Reagent last longer, too!! -Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jacquie.Farnsworth@cls.ab.ca Sent: Thursday, June 16, 2005 10:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Schiff's reagent in microwave? Hi Histonetters, Does anyone have some thoughts on microwaving Schiff's reagent? We currently microwave Schiff's in our plastics laboratory (for PAS stain on our renal biopsies). We take the heated reagent out of the microwave, and place it in a fumehood immediately and remove slides. One of our techs is still having problems with the fumes. R95 masks are available, but I am wondering if anyone has any thoughts? Suggestions? Does anyone else microwave Schiff's reagent? Thank you in advance, Jacquie Jacqueline Farnsworth Anatomic Pathology Tech III Calgary Laboratory Services Jacquie.Farnsworth@CLS.ab.ca PH: (403) 944-1578 Pager: 212-8223 #1933 CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Thu, 16 Jun 2005 16:49:39 +0100 From: Stephen.Eyres@sanofi-aventis.com Subject: [Histonet] Microwave use in Pharmaceutical/CRO histo labs To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hi All, Does anyone working in pharmaceutical/CRO's have any experience using microwave tissue processing? I'd be interested in your experiences, especially bad ones!! Cheers Steve ---------------------------------------------------------- Le prisent message ainsi que ses iventuelles pihces jointes est exclusivement destini au(x) destinataire(s), personnes physiques ou morales, qu'il disigne. Il constitue de ce fait une correspondance ` caracthre privi et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser immidiatement l'expiditeur par retour de courrier ilectronique puis de le ditruire, ainsi que ses iventuelles pihces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- ------------------------------ Message: 9 Date: Thu, 16 Jun 2005 11:04:04 -0500 From: "Dianne Holmes" Subject: [Histonet] Required CEU's To: Message-ID: Content-Type: text/plain; charset=US-ASCII I have heard that there is a required number of CEU's for HT's to get within each year to maintain their certification. Is this so and if it is - how many? ------------------------------ Message: 10 Date: Thu, 16 Jun 2005 11:15:06 -0500 From: "Nita Searcy" Subject: [Histonet] Muscle Referrals @ Baylor - Houston To: Message-ID: Content-Type: text/plain; charset="us-ascii" Does anyone know the contact person for the above? 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:;Anatomic Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD ------------------------------ Message: 11 Date: Thu, 16 Jun 2005 12:01:24 -0400 From: "Vinnie Della Speranza" Subject: [Histonet] One of our own in need ! Please Read To: Cc: carrie@nsh.org, bali02@cox.net Message-ID: Content-Type: text/plain; charset=US-ASCII I am taking the liberty of sharing a correspondence I've had with a fellow Histotech who is facing some very difficult circumstances involving her baby's serious illness. I know that histotechs have big hearts and I would just like to make it possible for you to be aware of Lisa's circumstances. I am also hopeful that some of you may be able to offer her additional advice on where she might obtain financial support. you will need to scroll to the very bottom to see Lisa's first correspondence with me. I apologize in advance to anyone who may object to what may appear as fundraising on the list. I am simply making Lisa's circumstances known to you on her behalf. also, please forgive the lengthy message. her original message to me had a photo of her child attached but I know the list does not allow for attachments so it has been removed. I imagine that you can obtain the photo if you wish directly from Lisa. I have removed her home phone number from her original message at her request. It is circumstances like hers that remind me of what is most important in life . I'm sure that your prayers and good wishes would be most welcomed sincerely Vinnie Della Speranza >>> "lisa barbarisi" 06/16/05 01:56AM >>> Hello The address that I gave you for the Funds for Lisa was incomplete I corrected it below. Thanks again, Lisa ----- Original Message ----- From: "lisa barbarisi" To: "Vinnie Della Speranza" Sent: Wednesday, June 15, 2005 9:54 PM Subject: Re: Fw: Fund raising at the national convention > Dear Vinnie, > > Thank you for your response. I did not think of posting on the Histonet. > It sounds like a good idea. I would however like to keep my home phone > number out of the letter and please do add the web site > www.alysa-anne.com it is now up and is just wonderful. I was told that > it is still being worked on as they are going to add a guest book for > people to sign in on. Also, I would appreciate it if you mentioned the > "Funds for Lisa" account #44699 any donations may be sent to: > > Funds for Lisa Account 44699 > Research Administration > Mayo Clinic Scottsdale 13400 East Shea Blvd. > Scottsdale, AZ 85259 > > All donations are tax deductible and a donation receipt will be sent as > long as the name and address are included either on the check or the > envelope. > > I do appreciate all of the information that you sent there are some that I > have not called and I will do so. It is so hard to take the time needed to > research and call people so it is wonderful when others offer a hand. > Thank you for your wonderful suggestion. > > Please forward what you post on the Histonet and the link. I have trouble > keeping track of things so having them in a e-mail helps me keep track. I > appreciate your help with this. > > Thank you so much for your help I look forward to hearing from you again. > > Lisa & Alysa Anne > > > ----- Original Message ----- > From: "Vinnie Della Speranza" > To: ; ; > ; ; ; > ; ; > ; ; ; > ; ; ; > ; > Cc: ; > Sent: Monday, June 13, 2005 12:41 PM > Subject: Re: Fw: Fund raising at the national convention > > > Dear Lisa, > I cannot tell you how sad I am to learn of your circumstances and of > course your baby's illness. I have no doubt that this is a very difficult > time for you and I imagine that many could not begin to imagine the > challenges you are facing as a result. I know that you have been a member > of NSH for the last few years and everyone understands your lack of recent > participation. I will pray that yours and Alysa's circumstances begin to > improve. > > to be quite frank, I am struggling with figuring out how we might help > you. I would probably have to consult with the society's attorney due to > our non-profit status. fund raising of any sort could create unforeseen > problems for the society. given this concern, I don't believe that the > society could sanction or conduct formal fundraising on your behalf. > > I do not know if you have considered placing your letter on Histonet. I > would be happy to do this for you if you indicate that you would like me > to do so. The Histonet list reaches histotechs from around the globe and > is an independent group not constrained by tax and corporate laws. I don't > speak for that group but I've found that histotechs have big hearts and > at the least someone more knowledgeable than I may be a source of valuable > information that might ease your circumstances. I hope you will consider > this avenue and as I've said, I would be happy to post your letter to me > on that list. > > our Executive Director, Carrie Diamond, has pulled some information from > the American Cancer Society website that addresses avenues for financial > assistance for circumstances like yours. I apologize if you are already > aware of this information but here it is: > > from American Cancer Society website. > Where can families get help financially? > > Most families find it difficult to turn to others or to agencies and > funds for financial help. Families generally take pride in being > self-sufficient and providing for their own needs. The extra expenses of > a child's cancer may be for the first time with problems with money. > They should remember that their problems in such a situation are > temporary and not unique. In the future, they may be the ones in a > position to offer financial help to others. > > There are several possible sources of help for families in financial need: > > * income assistance for low-income families through Supplemental > Security Income (SSI) benefits > * income assistance for needy families from the Aid to Families with > Dependent Children (AFDC) program > * help with travel, meals and lodging from public and private programs > * assistance with basic living costs (rent, mortgage, insurance > premiums, utilities, telephone) from public and private programs > * help from church, civic, social, and fraternal groups in the > community > * general help from special funds in the medical center or community > * assistance from targeted fundraising for an individual patient or > family. > > Organizations > > American Cancer Society, Inc. 800-227-2345 > Candlelighters Childhood Cancer Foundation 800-366-2223 > Corporate Angel Network, Inc. 866-328-1313 > National Children's Cancer Society 800-532-6459 > National Marrow Donor Program 800-627-7692 > National Patient Air Transport Hotline 800-296-1217 > Ronald McDonald House Charities 630-623-7048 > The Leukemia & Lymphoma Society 800-955-4572 > > Organizations (Government): > > U.S. Department of Labor (COBRA information) > Pension and Welfare Benefits Administration 800-998-7542 (Publications > only) 202-219-8776 > Americans with Disabilities Act, Civil Right Division, Disability Rights > Section 800-514-0301 > Hill-Burton Program 800-638-0742 > Internal Revenue Service (publications) 800-829-3676 800-829-1040 > (Assistance and Information) > National Cancer Institute 800-422-6237 > Social Security Administration (SSI information) 800-772-1213 > > Other Resources: > > American Brain Tumor Association 800-886-2282 > American Kidney Fund 800-638-8299 > Bone Marrow Transplant Information Network 888-597-7674 > Brain Tumor Foundation for Children, Inc. 770-458-5554 > Children's Organ Transplant Association, Inc. 800-366-2682 > Cancervive 310-203-9232 > Curesearch National Childhood Cancer Foundation 800-458-6223 > Lymphoma Research Foundation of America 800-500-9976 > National Association of Hospital Hospitality Houses, Inc. 800-542-9730 > National Association of Personal Financial Advisors 800-366-2732 > National Bone Marrow Transplant Link 800-546-5268 > National Brain Tumor Foundation 800-934-2873 > National Children's Cancer Society 800-532-6459 > National Coalition for Cancer Survivorship 877-622-7937 > National Foundation for Credit Counseling 800-388-2227 > National Information Center for Children and > Youth with Disabilities 800-695-0285 > National Insurance Consumer Helpline 800-942-4242 > National Organization for Rare Disorders, Inc. 800-999-6673 > National Self-Help Clearinghouse 212-817-1822 > National Transplant Assistance Fund 800-642-8399 > Oley Foundation, Inc. (nutritional information) 800-776-6539 > Patient Advocate Foundation 800-532-5274 > > Other Organizations (Government): > > National Cancer Institute (NCI) > Cancer Information Service 800-422-6237 > Centers for Disease Control and Prevention 800-311-3435 > Department of Health and Human Services 877-696-6775 > Medicare Hotline 800-633-4227 > National Health Information Center 800-336-4797 > Veterans Benefits Administration 800-827-1000 > U.S. Department of Veterans Affairs > > I hope that you will let us know when the website you are planning is up > and running so it will be easier for people to offer donations. > > God bless you both. > sincerely > Vinnie Della Speranza > President, National Society for Histotechnology > > >>>> "lisa barbarisi" 06/10/05 02:37AM >>> > Please read. I was not sure who to send this to so I tried to include > everyone. > > Thank you in advance for your time. Prayers are very welcome also. > > Lisa A. Barbarisi HT(ASCP) > > ----- Original Message ----- > From: lisa barbarisi > To: histo@nsh.org > Cc: bali02@cox.net > Sent: Thursday, June 09, 2005 11:17 PM > Subject: Fund raising at the national convention > > > Hello All, > > I am writing to ask for help in the form of a fundraiser at the national > convention. I am an HT( ASCP) certified Histotech who has worked in the > field of histotechnology for over eighteen years. I worked as the > coordinator of the histology research facility at the Mayo Clinic I am > currently on unpaid leave. > > After almost three years of fertility treatments and > invitro-fertilizations I spent all of my savings and took out a second > mortgage on my home and I was finally blessed to become pregnant. I gave > birth to a beautiful baby girl on August 30th 2004. > > I am a single first time mom with one income of which is zero at the > moment since I am on an unpaid leave from my current position at the Mayo > Clinic research facility as a Histotech. I am looking for financial help. > My beautiful baby girl Alysa Anne was diagnosed with stage four > neuroblastoma on March 25th 2004 she was not yet seven months old. She has > had surgery to try and remove the primary tumor with no luck. She is > currently undergoing chemotherapy to try and shrink the primary tumor > which is located on her right adrenal gland and there are tumors present > in her liver (to many to count), bone marrow, her bones base of her skull, > chest bone, around both of her eyes and her pathology is unfavorable. I > have been told by her oncologist that this is going to be a long hard road > for both my daughter and I. Between the chemotherapy there are doctors > visits twice and sometimes three times a week to check her progress and > also her blood counts. She is on daily shots of Neupogen and multiple > medications. How is such a small body going to take so much? I just can't > believe this is happening. > > I'm sorry, I could go on, but you may not be interested in helping me and > I will understand. I am at the end of my rope. I know that many people may > ask for help, but I just had to try. My daughter is my life and I will do > whatever I have to to help her fight this horrible disease . I was unable > to attend the NSH convention last year in Canada as my daughter had > finally blessed me with her arrival (she was over a week late and after a > long hard labor I finally had to have a c-section). I will not be able to > attend this year either since my daughter is having chemotherapy she just > made it through her third round. I feel helpless watching the poison being > pumped inter her little body thru a port in her chest. > > I wonder if there is anyone who would like to take on this huge > undertaking of either a fundraiser or maybe just taking donations to the > "Funds for Lisa" a not for profit account set up by a co-worker who wanted > to help me. > > If anyone is interested I would be so happy. Please e-mail me at bali02@cox.net . There will be a > website that is not as far as I know up and running it will tell more > about me and my daughter. It will also have updates on her progress. Once > I get word that it is up and running I can e-mail the site. > > I would appreciate a response even if it is negative. > > I have to beat this MONSTER called NEUROBLASTOMA to save my daughter . > > Sincerely, > > Lisa A. Barbarisi > > Picture of Alysa Anne at six months old three weeks before diagnosis is > attached. > > I > ------------------------------ Message: 12 Date: Thu, 16 Jun 2005 11:26:11 -0500 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Schiff's reagent in microwave? To: , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Do you have room in the fume hood for the microwave? Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jacquie.Farnsworth@cls.ab.ca Sent: Thursday, June 16, 2005 9:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Schiff's reagent in microwave? Hi Histonetters, Does anyone have some thoughts on microwaving Schiff's reagent? We currently microwave Schiff's in our plastics laboratory (for PAS stain on our renal biopsies). We take the heated reagent out of the microwave, and place it in a fumehood immediately and remove slides. One of our techs is still having problems with the fumes. R95 masks are available, but I am wondering if anyone has any thoughts? Suggestions? Does anyone else microwave Schiff's reagent? Thank you in advance, Jacquie Jacqueline Farnsworth Anatomic Pathology Tech III Calgary Laboratory Services Jacquie.Farnsworth@CLS.ab.ca PH: (403) 944-1578 Pager: 212-8223 #1933 CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Thu, 16 Jun 2005 11:45:10 -0500 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Brain fixative To: "Webb, Dorothy L" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" We use 20% formalin, and leave it in there for about a month. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Webb, Dorothy L Sent: Thursday, June 16, 2005 9:53 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Brain fixative Does anyone fix their brain tissue from autopsies in anything other than 10% buffered formalin? The PA I am working with seems to feel they used a 4% solution of lightly buffered formalin from Mallincroft. Can anyone help me with this, as I am only used to using 10% formalin, as we use for all of our routine histology fixation. Any ideas would be greatly appreciated and thanks ahead of time to all of my fellow histonetters..what a great group you are!! +______________________________________+ ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. 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You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Thu, 16 Jun 2005 17:49:46 +0100 From: "Edmondson David \(RBV\) NHS Christie Tr" Subject: RE: [Histonet] BCIP/NBT ready-to-use To: "Johnson, Teri" Cc: "Histonet \(E-mail 2\)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" DAKO do complete reagent in one, and the BCIP/NBT/INT that we use, its all used within date so can not help with long term stability. No ppt formed that we have seen Dave Christie Hosp Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Johnson, Teri Sent: 15 June 2005 23:22 To: Histonet Subject: [Histonet] BCIP/NBT ready-to-use We are currently using a substrate system which has two separate components for BCIP and NBT, and then have to combine them just before use. I'm looking into alternate systems and have seen several vendors who offer what appears to be ready-to-use substrate. Is anybody using them and can you comment on stability with regard to expiration date, length of use, and lack of problems after the slides have been stained (I recall a recent thread regarding horrible overstaining after the slides had been coverslipped). Thanks muchly! 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 ------------------------------ Message: 15 Date: Thu, 16 Jun 2005 16:54:47 +0000 From: mucram11@comcast.net Subject: RE: [Histonet] Schiff's reagent in microwave? To: "GUTIERREZ, JUAN" , , Message-ID: <061620051654.18290.42B1AED7000002C9000047722200748184CECE030E9D0C9A03@comc ast.net> Content-Type: text/plain Technically we should all have the oven under the hood and it is not always possible if we need to use the hood for anything else. I know I was told to look into the cost of a laboratory microwave and thought the people I worked for at the time would have heart attack when they saw the quote. I used the Target MW as close to the hood as I could get it and that allowed me to move the Schiff's and anything else very quickly under the hood. I could hold my breathe and get it there. Otherwise I don't what would stop the fumes completely for you. In an ideal world we would all have large hoods and lab MWs for safety. Pam Marcum -------------- Original message -------------- > Do you have room in the fume hood for the microwave? > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > My opinions are my own and do not reflect those of my employer. Long live free > speech! > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Jacquie.Farnsworth@cls.ab.ca > Sent: Thursday, June 16, 2005 9:52 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Schiff's reagent in microwave? > > Hi Histonetters, > Does anyone have some thoughts on microwaving Schiff's reagent? We currently > microwave Schiff's in our plastics laboratory (for PAS stain on our renal > biopsies). We take the heated reagent out of the microwave, and place it in a > fumehood immediately and remove slides. One of our techs is still having > problems with the fumes. R95 masks are available, but I am wondering if anyone > has any thoughts? Suggestions? Does anyone else microwave Schiff's reagent? > > Thank you in advance, > Jacquie > > Jacqueline Farnsworth > Anatomic Pathology Tech III > Calgary Laboratory Services > Jacquie.Farnsworth@CLS.ab.ca > PH: (403) 944-1578 > Pager: 212-8223 #1933 > > > > > CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain > confidential, personal and/or privileged information intended for a specific > purpose and recipient. If you are not the intended recipient do not disclose, > copy, retain, distribute, use or modify any of the contents of this > transmission. If you received this transmission in error please notify me > immediately by return e-mail or telephone and destroy the entire transmission > and any copies produced. Thank you. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@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 19, Issue 23 **************************************** From amosbrooks <@t> earthlink.net Sun Jun 19 13:45:31 2005 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Expiration date In-Reply-To: <200506191300.1dK39P5zG3Nl34l0@mx-emperor.atl.sa.earthlink.net> References: <200506191300.1dK39P5zG3Nl34l0@mx-emperor.atl.sa.earthlink.net> Message-ID: <42B5BD4B.4060702@earthlink.net> Joe, A couple things come to mind here. Firstly, you must have one heck of a good freezer to maintain antibodies from 1080 to 1990. That predates the Magna Carta and the Battle of Hastings :-) (of course you meant 1980, I'm pulling your leg here) Secondly, It is worth noting that since immunohistochemistry is becoming so common these days one cannot assume everyone is as proficient at quality control as anyone else. So it follows that if an epitope becomes less intense in label over time it would not be surprising to find a tech that doesn't notice until it becomes problematic. This would also hold true for the less commonly used antibodies. We often have doctors request a new antibody and the confounded thing expires before he orders it a second time. Would I even notice if it was less intense than it was 2 years ago? I hope so but who knows? Granted this is fishing for a lowest common denominator, but that seems to be the way things are going these days. I agree that the CAP is a bit too obsessed with dates than with QC follow up and it becomes an expensive endeavor for labs to keep up with it. I think as long as the doctor reading the case is content with the signal on the test and control the CAP should leave well enough alone. But the lab should be responsible for making sure the antibody is reactive before using it. Let's face it we've all seen a case where there is one and only one slide with a lesion of interest on it. In a situation like that I'd like to know for sure the antibody is good the first time. Just some things to consider, Amos Brooks >Message: 1 >Date: Sat, 18 Jun 2005 12:20:47 -0500 >From: "Joe Nocito" >Subject: Here I go again[Histonet] Expiration date >To: "Katia Cristina Catunda" , "histonet" > >Message-ID: <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> >Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=response > >Okay, >I've been too quiet for too long. Here I go again. > >I am so tired of CAP and their ridiculous regulations. Every CAP inspection >I fight this same question. When I was supervising the Immuno lab at AFIP, >we froze concentrated antibodies at -70 for years. Antibodies that were >frozen in 1080, were still viable in 1990. That means we were running >immunos in 1990 with 1980 prices. What is so wrong about that? > We also ran a known positive control with each batch. If the positive >control worked, guess what? The antibody was still good. If the positive >control did not work, we threw out that lot number and repeated it with >another lot number. When we saw that there was a drop in the staining >intensity, we tossed that lot and started another. My experience with >antibodies is that they just don't go bad over night, they start staining >with less intensity. > I think that the CAP board members have stocks in the biochemical >companies. Please don't get me wrong, I have many close friends that work in >the biochemical side of this, but why would this drastic change in view? The >FDA? I doubt it. > Maybe I'm too one sides on this issue, but give me a break! If the >positive control works in any other antibody, doesn't that mean the antibody >is viable? > I know companies have to put an expiration date on their products, but >come on. If an antibody expires June 30, 2005, does it automatically go dead >on July 1? > As managers and supervisors, we are continuously bashed about saving >money. This would be a great place to start, don't you think? > That is all. Thank you > >Joe Nocito BS, HT(ASCP)QIHC >Histology Manager >Pathology Reference Lab >San Antonio, TX > From t-sherman <@t> comcast.net Sun Jun 19 15:41:27 2005 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:25:14 2005 Subject: Here I go again[Histonet] Expiration date (Joe Nocito) Message-ID: Joe, Regarding "If an antibody expires June 30, 2005, does it automatically go dead on July 1?" - according to the corporate legal teams and their liability concerns, of course. ;) But in all seriousness, you bring up a good point. I can understand a manufacturer needing to maintain QC and establish liability limits, not to mention bump potential premature boosts in sales from the dumping of viable product by enduser, but some modification should be considered. You mention a nice protocol of aliquoting, inventorying, and testing that seems entirely reasonable. Consider another lab that dumps lots according to date but that doesn't aliquot properly and lets an undiluted lot freeze and thaw such that it "expires" before the label date would indicate it has deteriorated. Would such a scenario be better? Obviously a more scaled weighting that considers the continual QC testing against positive and negative controls should be incorporated since that is the fundamental concern. If a lab can produce a series of time-dated QC test samples/slides associated with a particular lot/vial/aliquot that proves efficacy, then I should think that the Ab is still "good". Granted, the manufacturers should not be held responsible for extended usage beyond what they deem reasonable/optimal; however, thoughtless disposal of good product should not be a mandate by the governing histological agency either. $0.02 worth of validation. Regards, Todd Todd Sherman President HistoSoft Corporation www.histosoft.com Biology In A New Form (c) On Sun, 19 Jun 2005 12:00:23 -0500, wrote: > > Today's Topics: > > 1. Here I go again[Histonet] Expiration date (Joe Nocito) > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sat, 18 Jun 2005 12:20:47 -0500 > From: "Joe Nocito" > Subject: Here I go again[Histonet] Expiration date > To: "Katia Cristina Catunda" , "histonet" > > Message-ID: <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=response > > Okay, > I've been too quiet for too long. Here I go again. > > I am so tired of CAP and their ridiculous regulations. Every CAP > inspection > I fight this same question. When I was supervising the Immuno lab at > AFIP, > we froze concentrated antibodies at -70 for years. Antibodies that were > frozen in 1080, were still viable in 1990. That means we were running > immunos in 1990 with 1980 prices. What is so wrong about that? > We also ran a known positive control with each batch. If the positive > control worked, guess what? The antibody was still good. If the positive > control did not work, we threw out that lot number and repeated it with > another lot number. When we saw that there was a drop in the staining > intensity, we tossed that lot and started another. My experience with > antibodies is that they just don't go bad over night, they start staining > with less intensity. > I think that the CAP board members have stocks in the biochemical > companies. Please don't get me wrong, I have many close friends that > work in > the biochemical side of this, but why would this drastic change in view? > The > FDA? I doubt it. > Maybe I'm too one sides on this issue, but give me a break! If the > positive control works in any other antibody, doesn't that mean the > antibody > is viable? > I know companies have to put an expiration date on their products, > but > come on. If an antibody expires June 30, 2005, does it automatically go > dead > on July 1? > As managers and supervisors, we are continuously bashed about saving > money. This would be a great place to start, don't you think? > That is all. Thank you > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX From anh2006 <@t> med.cornell.edu Sun Jun 19 22:16:10 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] VEGF - question In-Reply-To: <200506191559.j5JFxIMM198616@smtp-in1.med.cornell.edu> References: <200506191559.j5JFxIMM198616@smtp-in1.med.cornell.edu> Message-ID: It is not unexpected that the endothelial cells should stain negative for VEGF. VEGF normally acts through a paracrine mechanism, whereby endothelial cells express the receptors for VEGF and the surrounding tissue (often times tumor cells) are those that secrete VEGF itself. VEGF can be present in a soluble form or as an ECM bound form so it is likely that VEGF immunostaining would appear "messy" as you might be detecting that which is soluble, ECM bound, or even cytoplasmic VEGF which hasn't yet been secreted from the cell. The only thing to consider is that many anti-VEGF antibodies are notorious for being non-specific so plan your experiments with careful controls. May I ask the source of your VEGF antibody and if you are staining human or mouse? Sincerely, Andrea At 5:57 PM +0200 6/19/05, Gudrun Lang wrote: >Maybe a stupid question. Are endothel cells always stained positiv with >vascular endothelial growth factor? > >I have got a tissue of a pyogenic granuloma as a positiv control from my >pathologist. The endothel cells of the vessels are clear negativ, the >surrounding area is positiv. > >Is this the expected result or is there any mistake? > > > >Thanks for your help. > >Gudrun Lang -- From Kemlo.Rogerson <@t> elht.nhs.uk Mon Jun 20 01:44:49 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] RE: Xylene Substitutes[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD44F@elht-exch1.xelht.nhs.uk> Acceptable limit????? Define an acceptable limit! There is no acceptable limit as far as I know, all exposure brings with it danger, but I concede there is a limit beneath which that danger is acceptable. But what are the unacceptable elements of this acceptability? Only 1 in a 1,000 women has a spontaneous abortions and I suppose that may be acceptable. Complete removal and replacement or extraction that results in zero exposure is acceptable, isn't it? -----Original Message----- From: Barbara Stancel [mailto:stancelb@msn.com] Sent: 18 June 2005 20:03 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Xylene Substitutes[Scanned] I'm with Charles. We have been using xylene for 31 years. We have tried the xylene substitutes, but we and our pathologist prefer xylene. We recycle all the xylene to reduce cost of purchasing and cut hazardous waste. We have great ventilation in the lab. Our twice-a-year STEL monitoring has come back at way below the acceptable limits. If it ain't broke, don't fix it. Barbara Stancel USDA, FSIS, OPHS, EL, Pathology Athens, Georgia -----Original Message----- From: Charles.Embrey Sent: Friday, June 17, 2005 1:40 PM To: 'Joyce Cline' Subject: RE: [Histonet] Xylene substitutes Your quote "They all realize how bad Xylene is". Just how bad IS xylene? I have been using it for 25 years and am still alive and kicking. We recycle ours so we don't even have an environmental issue to deal with. I have never used any substitute that worked as well or better than xylene itself. Many claim to be almost as good as xylene and that is about as close as any get. As for me, I am sticking with what works. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Cline Sent: Friday, June 17, 2005 12:05 PM To: 'Histonet' Subject: [Histonet] Xylene substitutes I use Clear-rite 3 and our pathologists have no problem with it. They all realize how bad Xylene is. I use H-2 Blue beads to help eliminate the water from our high humidity. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Jun 20 01:51:29 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Xylene substitutes[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2F1@elht-exch1.xelht.nhs.uk> Xylene makes you go 'see through' and I think it has benzene rings it. Sent a young Tech few years back to get some benzene rings from Pharmacy; he failed. -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: 17 June 2005 19:41 To: histonet@lists.utsouthwestern.edu Subject: FW: [Histonet] Xylene substitutes[Scanned] -----Original Message----- From: Charles.Embrey Sent: Friday, June 17, 2005 1:40 PM To: 'Joyce Cline' Subject: RE: [Histonet] Xylene substitutes Your quote "They all realize how bad Xylene is". Just how bad IS xylene? I have been using it for 25 years and am still alive and kicking. We recycle ours so we don't even have an environmental issue to deal with. I have never used any substitute that worked as well or better than xylene itself. Many claim to be almost as good as xylene and that is about as close as any get. As for me, I am sticking with what works. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Cline Sent: Friday, June 17, 2005 12:05 PM To: 'Histonet' Subject: [Histonet] Xylene substitutes I use Clear-rite 3 and our pathologists have no problem with it. They all realize how bad Xylene is. I use H-2 Blue beads to help eliminate the water from our high humidity. ***** 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From antje.marcantonio <@t> novartis.com Mon Jun 20 02:01:50 2005 From: antje.marcantonio <@t> novartis.com (antje.marcantonio@novartis.com) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] C3 antibody for RAT Message-ID: Hello Histonetters I badly need an antibody for C3 which reacts in rat tissue as the one I used in the past isn't available anymore (was from Biotrend) Any ideas ? I fully rely on YOU ! Thanks, Antje Marcantonio Novartis Pharma AG AT TX Research Basle, Switzerland Tel. +41 61 324 6730 antje.marcantonio@novartis.com From Stephen.Eyres <@t> sanofi-aventis.com Mon Jun 20 17:53:03 2005 From: Stephen.Eyres <@t> sanofi-aventis.com (Stephen.Eyres@sanofi-aventis.com) Date: Fri Sep 16 15:25:14 2005 Subject: Here I go again[Histonet] Expiration date (Joe Nocito) Message-ID: Hi, Our policy for ICC and special stains is that we use the positive control result as the standard marker. I commonly authorize for use special stain reagents that are past their expiry data. I work in a GLP environment and authorizing reagents for use that are beyond their expiry dates is acceptable from a GLP perspective if there is a stated scientific justification. Cheers Steve "Todd Sherman" To: histonet@lists.utsouthwestern.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: Re: Here I go again[Histonet] Expiration date (Joe Nocito) western.edu 19/06/2005 21:41 Joe, Regarding "If an antibody expires June 30, 2005, does it automatically go dead on July 1?" - according to the corporate legal teams and their liability concerns, of course. ;) But in all seriousness, you bring up a good point. I can understand a manufacturer needing to maintain QC and establish liability limits, not to mention bump potential premature boosts in sales from the dumping of viable product by enduser, but some modification should be considered. You mention a nice protocol of aliquoting, inventorying, and testing that seems entirely reasonable. Consider another lab that dumps lots according to date but that doesn't aliquot properly and lets an undiluted lot freeze and thaw such that it "expires" before the label date would indicate it has deteriorated. Would such a scenario be better? Obviously a more scaled weighting that considers the continual QC testing against positive and negative controls should be incorporated since that is the fundamental concern. If a lab can produce a series of time-dated QC test samples/slides associated with a particular lot/vial/aliquot that proves efficacy, then I should think that the Ab is still "good". Granted, the manufacturers should not be held responsible for extended usage beyond what they deem reasonable/optimal; however, thoughtless disposal of good product should not be a mandate by the governing histological agency either. $0.02 worth of validation. Regards, Todd Todd Sherman President HistoSoft Corporation www.histosoft.com Biology In A New Form (c) On Sun, 19 Jun 2005 12:00:23 -0500, wrote: > > Today's Topics: > > 1. Here I go again[Histonet] Expiration date (Joe Nocito) > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sat, 18 Jun 2005 12:20:47 -0500 > From: "Joe Nocito" > Subject: Here I go again[Histonet] Expiration date > To: "Katia Cristina Catunda" , "histonet" > > Message-ID: <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=response > > Okay, > I've been too quiet for too long. Here I go again. > > I am so tired of CAP and their ridiculous regulations. Every CAP > inspection > I fight this same question. When I was supervising the Immuno lab at > AFIP, > we froze concentrated antibodies at -70 for years. Antibodies that were > frozen in 1080, were still viable in 1990. That means we were running > immunos in 1990 with 1980 prices. What is so wrong about that? > We also ran a known positive control with each batch. If the positive > control worked, guess what? The antibody was still good. If the positive > control did not work, we threw out that lot number and repeated it with > another lot number. When we saw that there was a drop in the staining > intensity, we tossed that lot and started another. My experience with > antibodies is that they just don't go bad over night, they start staining > with less intensity. > I think that the CAP board members have stocks in the biochemical > companies. Please don't get me wrong, I have many close friends that > work in > the biochemical side of this, but why would this drastic change in view? > The > FDA? I doubt it. > Maybe I'm too one sides on this issue, but give me a break! If the > positive control works in any other antibody, doesn't that mean the > antibody > is viable? > I know companies have to put an expiration date on their products, > but > come on. If an antibody expires June 30, 2005, does it automatically go > dead > on July 1? > As managers and supervisors, we are continuously bashed about saving > money. This would be a great place to start, don't you think? > That is all. Thank you > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- From GDawson <@t> dynacaremilwaukee.com Mon Jun 20 17:53:03 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:25:14 2005 Subject: Here I go again[Histonet] Expiration date Message-ID: Joe, Agreed. Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: Joe Nocito [mailto:jnocito@satx.rr.com] Sent: Saturday, June 18, 2005 11:21 AM To: Katia Cristina Catunda; histonet Subject: Here I go again[Histonet] Expiration date Okay, I've been too quiet for too long. Here I go again. I am so tired of CAP and their ridiculous regulations. Every CAP inspection I fight this same question. When I was supervising the Immuno lab at AFIP, we froze concentrated antibodies at -70 for years. Antibodies that were frozen in 1080, were still viable in 1990. That means we were running immunos in 1990 with 1980 prices. What is so wrong about that? We also ran a known positive control with each batch. If the positive control worked, guess what? The antibody was still good. If the positive control did not work, we threw out that lot number and repeated it with another lot number. When we saw that there was a drop in the staining intensity, we tossed that lot and started another. My experience with antibodies is that they just don't go bad over night, they start staining with less intensity. I think that the CAP board members have stocks in the biochemical companies. Please don't get me wrong, I have many close friends that work in the biochemical side of this, but why would this drastic change in view? The FDA? I doubt it. Maybe I'm too one sides on this issue, but give me a break! If the positive control works in any other antibody, doesn't that mean the antibody is viable? I know companies have to put an expiration date on their products, but come on. If an antibody expires June 30, 2005, does it automatically go dead on July 1? As managers and supervisors, we are continuously bashed about saving money. This would be a great place to start, don't you think? That is all. Thank you Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Katia Cristina Catunda" To: "histonet" Sent: Wednesday, June 15, 2005 8:20 PM Subject: [Histonet] Expiration date > What should we do with antibodies and reagents after their indicated > expiration dates? Theorectically they shouldn't be estable after the > expiration date but we do know that most of them still works perfectly... > should we just throw them out and not use them anymore, wouldn't this be a > way for the manufacturers manipulate their sellings? Need some opinions... > > Tks > > Katia > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.7.8/22 - Release Date: 6/17/2005 > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Mon Jun 20 17:53:03 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Expiration date In-Reply-To: <42B5BD4B.4060702@earthlink.net> Message-ID: Amos, I understand. Many times we perform IHC on H & E slides because the lesion is so small, but there has to be a way to use expired immuno reagents as long as the QC is well monitored. We are throwing, literally, thousands of dollars down the drain each year. As for the freezer in 1080, my relatives came from the North Pole. Ice wasn't a problem. Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amos Brooks Sent: Sunday, June 19, 2005 1:46 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Expiration date Joe, A couple things come to mind here. Firstly, you must have one heck of a good freezer to maintain antibodies from 1080 to 1990. That predates the Magna Carta and the Battle of Hastings :-) (of course you meant 1980, I'm pulling your leg here) Secondly, It is worth noting that since immunohistochemistry is becoming so common these days one cannot assume everyone is as proficient at quality control as anyone else. So it follows that if an epitope becomes less intense in label over time it would not be surprising to find a tech that doesn't notice until it becomes problematic. This would also hold true for the less commonly used antibodies. We often have doctors request a new antibody and the confounded thing expires before he orders it a second time. Would I even notice if it was less intense than it was 2 years ago? I hope so but who knows? Granted this is fishing for a lowest common denominator, but that seems to be the way things are going these days. I agree that the CAP is a bit too obsessed with dates than with QC follow up and it becomes an expensive endeavor for labs to keep up with it. I think as long as the doctor reading the case is content with the signal on the test and control the CAP should leave well enough alone. But the lab should be responsible for making sure the antibody is reactive before using it. Let's face it we've all seen a case where there is one and only one slide with a lesion of interest on it. In a situation like that I'd like to know for sure the antibody is good the first time. Just some things to consider, Amos Brooks >Message: 1 >Date: Sat, 18 Jun 2005 12:20:47 -0500 >From: "Joe Nocito" >Subject: Here I go again[Histonet] Expiration date >To: "Katia Cristina Catunda" , "histonet" > >Message-ID: <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> >Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=response > >Okay, >I've been too quiet for too long. Here I go again. > >I am so tired of CAP and their ridiculous regulations. Every CAP inspection >I fight this same question. When I was supervising the Immuno lab at AFIP, >we froze concentrated antibodies at -70 for years. Antibodies that were >frozen in 1080, were still viable in 1990. That means we were running >immunos in 1990 with 1980 prices. What is so wrong about that? > We also ran a known positive control with each batch. If the positive >control worked, guess what? The antibody was still good. If the positive >control did not work, we threw out that lot number and repeated it with >another lot number. When we saw that there was a drop in the staining >intensity, we tossed that lot and started another. My experience with >antibodies is that they just don't go bad over night, they start staining >with less intensity. > I think that the CAP board members have stocks in the biochemical >companies. Please don't get me wrong, I have many close friends that work in >the biochemical side of this, but why would this drastic change in view? The >FDA? I doubt it. > Maybe I'm too one sides on this issue, but give me a break! If the >positive control works in any other antibody, doesn't that mean the antibody >is viable? > I know companies have to put an expiration date on their products, but >come on. If an antibody expires June 30, 2005, does it automatically go dead >on July 1? > As managers and supervisors, we are continuously bashed about saving >money. This would be a great place to start, don't you think? > That is all. Thank you > >Joe Nocito BS, HT(ASCP)QIHC >Histology Manager >Pathology Reference Lab >San Antonio, TX > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charles.Embrey <@t> carle.com Mon Jun 20 17:53:03 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] RE: Xylene Substitutes[Scanned] Message-ID: Kemlo, There is an acceptable limit to water ingestion. If you go past that it becomes toxic and even fatal. Sorry, I'm not going to stop drinking water just because at some point it becomes toxic. Many of these "warnings" in the lab boil down to junk science. I just can't take the "Henny Penny" approach to things and run around screaming "The sky is falling, the sky is falling". (My apologies to those that don't understand this analogy). Relax a little, life can be fun........................ Chuck Embrey -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rogerson Kemlo (ELHT) Pathology Sent: Monday, June 20, 2005 1:45 AM To: Barbara Stancel; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Xylene Substitutes[Scanned] Acceptable limit????? Define an acceptable limit! There is no acceptable limit as far as I know, all exposure brings with it danger, but I concede there is a limit beneath which that danger is acceptable. But what are the unacceptable elements of this acceptability? Only 1 in a 1,000 women has a spontaneous abortions and I suppose that may be acceptable. Complete removal and replacement or extraction that results in zero exposure is acceptable, isn't it? -----Original Message----- From: Barbara Stancel [mailto:stancelb@msn.com] Sent: 18 June 2005 20:03 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Xylene Substitutes[Scanned] I'm with Charles. We have been using xylene for 31 years. We have tried the xylene substitutes, but we and our pathologist prefer xylene. We recycle all the xylene to reduce cost of purchasing and cut hazardous waste. We have great ventilation in the lab. Our twice-a-year STEL monitoring has come back at way below the acceptable limits. If it ain't broke, don't fix it. Barbara Stancel USDA, FSIS, OPHS, EL, Pathology Athens, Georgia -----Original Message----- From: Charles.Embrey Sent: Friday, June 17, 2005 1:40 PM To: 'Joyce Cline' Subject: RE: [Histonet] Xylene substitutes Your quote "They all realize how bad Xylene is". Just how bad IS xylene? I have been using it for 25 years and am still alive and kicking. We recycle ours so we don't even have an environmental issue to deal with. I have never used any substitute that worked as well or better than xylene itself. Many claim to be almost as good as xylene and that is about as close as any get. As for me, I am sticking with what works. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Cline Sent: Friday, June 17, 2005 12:05 PM To: 'Histonet' Subject: [Histonet] Xylene substitutes I use Clear-rite 3 and our pathologists have no problem with it. They all realize how bad Xylene is. I use H-2 Blue beads to help eliminate the water from our high humidity. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Mon Jun 20 17:53:03 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:14 2005 Subject: Here I go again[Histonet] Expiration date In-Reply-To: <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> Message-ID: Joe - I totally agree with you. My only question is this: with the new CAP rule in place, have you been cited for using expired reagents? While I would be most concerned with proper QC, I know most inspectors go by the letter of the rule, and aren't open for debate. We got written up for having a box with an expired date on it - we were using it to store some excess non-expired reagent ( I know, exactly a smart thing to do). Patti Loykasek PhenoPath Laboratories Seattle, WA > Okay, > I've been too quiet for too long. Here I go again. > > I am so tired of CAP and their ridiculous regulations. Every CAP inspection > I fight this same question. When I was supervising the Immuno lab at AFIP, > we froze concentrated antibodies at -70 for years. Antibodies that were > frozen in 1080, were still viable in 1990. That means we were running > immunos in 1990 with 1980 prices. What is so wrong about that? > We also ran a known positive control with each batch. If the positive > control worked, guess what? The antibody was still good. If the positive > control did not work, we threw out that lot number and repeated it with > another lot number. When we saw that there was a drop in the staining > intensity, we tossed that lot and started another. My experience with > antibodies is that they just don't go bad over night, they start staining > with less intensity. > I think that the CAP board members have stocks in the biochemical > companies. Please don't get me wrong, I have many close friends that work in > the biochemical side of this, but why would this drastic change in view? The > FDA? I doubt it. > Maybe I'm too one sides on this issue, but give me a break! If the > positive control works in any other antibody, doesn't that mean the antibody > is viable? > I know companies have to put an expiration date on their products, but > come on. If an antibody expires June 30, 2005, does it automatically go dead > on July 1? > As managers and supervisors, we are continuously bashed about saving > money. This would be a great place to start, don't you think? > That is all. Thank you > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > ----- Original Message ----- > From: "Katia Cristina Catunda" > To: "histonet" > Sent: Wednesday, June 15, 2005 8:20 PM > Subject: [Histonet] Expiration date > > >> What should we do with antibodies and reagents after their indicated >> expiration dates? Theorectically they shouldn't be estable after the >> expiration date but we do know that most of them still works perfectly... >> should we just throw them out and not use them anymore, wouldn't this be a >> way for the manufacturers manipulate their sellings? Need some opinions... >> >> Tks >> >> Katia >> >> >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> -- >> No virus found in this incoming message. >> Checked by AVG Anti-Virus. >> Version: 7.0.323 / Virus Database: 267.7.8/22 - Release Date: 6/17/2005 >> >> > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Mon Jun 20 17:53:03 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] RE: Xylene Substitutes[Scanned] Message-ID: <566FB0B522443D43AF02D2ADBE35A6F001B0DDCA@UTHEVS3.mail.uthouston.edu> I must in general agree with Chuck. I have not used any clearing agents that I consider to be as good as xylenes. I think that we are now in an era where the pendulum has swung away from an almost total disregard for the use of chemicals to an era where we are sometimes overly cautious. This is not just with chemicals but many aspects in life. While it is critical to be very careful in the use of all chemicals and various practices (in the lab and at home)there is a point beyond which we would never work in a lab in case we absorbed some chemical or even leave the house in case we might get hit by a falling piece from an aircraft. When training in a lab we were taught to be aware of potential dangers from chemicals, equipment and especially trainees. Safe practices should not only be confined to the lab but should be a way of life for us all. After all we are exposed to enough pollution without our adding to it. If you cut your grass are you careful about not breathing exhaust fumes or getting gasoline on your hands? It seems at times as if we are trying to eliminate nature including us). Does this mean that we should not be using chemicals to clean ovens, to scrub sinks or to bleach clothes? I suggest that there is an acceptable limit to all these substances and practices but in order be realistic we always make ourselves fully aware of possible consequences of use. I make such decisions every day that I drive in Houston. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, June 20, 2005 8:24 AM To: Rogerson Kemlo (ELHT) Pathology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Xylene Substitutes[Scanned] Kemlo, There is an acceptable limit to water ingestion. If you go past that it becomes toxic and even fatal. Sorry, I'm not going to stop drinking water just because at some point it becomes toxic. Many of these "warnings" in the lab boil down to junk science. I just can't take the "Henny Penny" approach to things and run around screaming "The sky is falling, the sky is falling". (My apologies to those that don't understand this analogy). Relax a little, life can be fun........................ Chuck Embrey -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rogerson Kemlo (ELHT) Pathology Sent: Monday, June 20, 2005 1:45 AM To: Barbara Stancel; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Xylene Substitutes[Scanned] Acceptable limit????? Define an acceptable limit! There is no acceptable limit as far as I know, all exposure brings with it danger, but I concede there is a limit beneath which that danger is acceptable. But what are the unacceptable elements of this acceptability? Only 1 in a 1,000 women has a spontaneous abortions and I suppose that may be acceptable. Complete removal and replacement or extraction that results in zero exposure is acceptable, isn't it? -----Original Message----- From: Barbara Stancel [mailto:stancelb@msn.com] Sent: 18 June 2005 20:03 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Xylene Substitutes[Scanned] I'm with Charles. We have been using xylene for 31 years. We have tried the xylene substitutes, but we and our pathologist prefer xylene. We recycle all the xylene to reduce cost of purchasing and cut hazardous waste. We have great ventilation in the lab. Our twice-a-year STEL monitoring has come back at way below the acceptable limits. If it ain't broke, don't fix it. Barbara Stancel USDA, FSIS, OPHS, EL, Pathology Athens, Georgia -----Original Message----- From: Charles.Embrey Sent: Friday, June 17, 2005 1:40 PM To: 'Joyce Cline' Subject: RE: [Histonet] Xylene substitutes Your quote "They all realize how bad Xylene is". Just how bad IS xylene? I have been using it for 25 years and am still alive and kicking. We recycle ours so we don't even have an environmental issue to deal with. I have never used any substitute that worked as well or better than xylene itself. Many claim to be almost as good as xylene and that is about as close as any get. As for me, I am sticking with what works. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Cline Sent: Friday, June 17, 2005 12:05 PM To: 'Histonet' Subject: [Histonet] Xylene substitutes I use Clear-rite 3 and our pathologists have no problem with it. They all realize how bad Xylene is. I use H-2 Blue beads to help eliminate the water from our high humidity. _______________________________________________ Histonet mailing list Histonet@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 rebecca.riesen <@t> dsilabs.com Mon Jun 20 17:53:03 2005 From: rebecca.riesen <@t> dsilabs.com (Riesen, Rebecca) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] JOB OPPORTUNITIES Message-ID: <05844092236E7749A1BBBCB1077C34A2DEA013@dsi-ex01.gateway.dom> I know, right now it's beautiful and sunny there up north, but before we all know it "old man winter" will roll in and you'll all be saying how cold it is and I gotta get out-a-here. WELL, I have two positions available down here in SUNNY NAPLES FLORIDA!!!!!!! One is a daytime position and the other is an evening position (gives you the daytime to work on that tan). We have an excellent benefits program and tuition reimbursement. If interested please contact our HR department by phone 1-239-561-8200 or send your resume to them at kquick@dsilabs.com or jgood@dsilabs.com . Opportunity is knocking on the door! Don't ignore! THANKS! DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(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 pmarcum <@t> vet.upenn.edu Mon Jun 20 17:53:03 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] RE: Xylene Substitutes[Scanned] In-Reply-To: <566FB0B522443D43AF02D2ADBE35A6F001B0DDCA@UTHEVS3.mail.utho uston.edu> References: <566FB0B522443D43AF02D2ADBE35A6F001B0DDCA@UTHEVS3.mail.uthouston.edu> Message-ID: <6.1.1.1.2.20050620103826.0199a098@mail.vet.upenn.edu> Well, stated and the rules we should live by. Unfortunately many of the people most concerned about the effects of the chemicals only read a warning label not the MSDS for full safety usage of the chemical (in the lab) or cleaner in the home, where only the label is available. We hear stories about how people have had really bad reactions to chemicals without knowing the whole story and then everyone panics. We need to be aware of what we are using and how to use it safely then some of the dangers will be handled by correct practice not fear of contamination. The use of chemicals in histology will not change and some things just work better - I use xylene and will continue to under a hood or in a well ventilated area as we all should. (Remember the old days when no one even had a hood and even the pathologist would wash the paraffin off their hands with xylene. That was before we went too far the other way.) Personally I am allergic to the d-limoene based clearants and can't even be in the room with them. However, I also have a problem with citrus if I over do it in my diet. These two problems are related and yet most people hear I can't stand the orange stuff and assume the problem is the only the chemical in the lab when in fact it is a problem in my daily life too. Don't use any orange or citrus based stuff anywhere due to this so all labels for cleaners are a must read for me. They should be for everyone. Pam Marcum At 10:35 AM 6/20/2005, Rittman, Barry R wrote: >I must in general agree with Chuck. I have not used any clearing agents >that I consider to be as good as xylenes. > >I think that we are now in an era where the pendulum has swung away from >an almost total disregard for the use of chemicals to an era where we >are sometimes overly cautious. This is not just with chemicals but many >aspects in life. >While it is critical to be very careful in the use of all chemicals and >various practices (in the lab and at home)there is a point beyond which >we would never work in a lab in case we absorbed some chemical or even >leave the house in case we might get hit by a falling piece from an >aircraft. >When training in a lab we were taught to be aware of potential dangers >from chemicals, equipment and especially trainees. Safe practices should >not only be confined to the lab but should be a way of life for us all. >After all we are exposed to enough pollution without our adding to it. >If you cut your grass are you careful about not breathing exhaust fumes >or getting gasoline on your hands? It seems at times as if we are trying >to eliminate nature including us). >Does this mean that we should not be using chemicals to clean ovens, to >scrub sinks or to bleach clothes? I suggest that there is an acceptable >limit to all these substances and practices but in order be realistic we >always make ourselves fully aware of possible consequences of use. >I make such decisions every day that I drive in Houston. >Barry > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >Charles.Embrey >Sent: Monday, June 20, 2005 8:24 AM >To: Rogerson Kemlo (ELHT) Pathology >Cc: histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: Xylene Substitutes[Scanned] > >Kemlo, There is an acceptable limit to water ingestion. If you go past >that it becomes toxic and even fatal. Sorry, I'm not going to stop >drinking water just because at some point it becomes toxic. Many of >these "warnings" in the lab boil down to junk science. I just can't >take the "Henny Penny" approach to things and run around screaming "The >sky is falling, the sky is falling". (My apologies to those that don't >understand this analogy). Relax a little, life can be >fun........................ > >Chuck Embrey > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rogerson >Kemlo (ELHT) Pathology >Sent: Monday, June 20, 2005 1:45 AM >To: Barbara Stancel; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: Xylene Substitutes[Scanned] > >Acceptable limit????? Define an acceptable limit! There is no acceptable >limit as far as I know, all exposure brings with it danger, but I >concede there is a limit beneath which that danger is acceptable. But >what are the unacceptable elements of this acceptability? Only 1 in a >1,000 women has a spontaneous abortions and I suppose that may be >acceptable. > >Complete removal and replacement or extraction that results in zero >exposure is acceptable, isn't it? > >-----Original Message----- >From: Barbara Stancel [mailto:stancelb@msn.com] >Sent: 18 June 2005 20:03 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] RE: Xylene Substitutes[Scanned] > >I'm with Charles. > >We have been using xylene for 31 years. We have tried the xylene >substitutes, but we and our pathologist prefer xylene. We recycle all >the >xylene to reduce cost of purchasing and cut hazardous waste. We have >great >ventilation in the lab. Our twice-a-year STEL monitoring has come back >at >way below the acceptable limits. > >If it ain't broke, don't fix it. > > >Barbara Stancel >USDA, FSIS, OPHS, EL, Pathology >Athens, Georgia > > >-----Original Message----- >From: Charles.Embrey >Sent: Friday, June 17, 2005 1:40 PM >To: 'Joyce Cline' >Subject: RE: [Histonet] Xylene substitutes > >Your quote "They all realize how bad Xylene is". Just how bad IS xylene? >I have been using it for 25 years and am still alive and kicking. We >recycle ours so we don't even have an environmental issue to deal with. >I have never used any substitute that worked as well or better than >xylene itself. Many claim to be almost as good as xylene and that is >about as close as any get. As for me, I am sticking with what works. > >Charles Embrey PA(ASCP) > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce >Cline >Sent: Friday, June 17, 2005 12:05 PM >To: 'Histonet' >Subject: [Histonet] Xylene substitutes > >I use Clear-rite 3 and our pathologists have no problem with it. They >all realize how bad Xylene is. I use H-2 Blue beads to help eliminate >the water from our high humidity. > > > >_______________________________________________ >Histonet mailing list >Histonet@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 Thanks, Pam Marcum From ljb <@t> medicine.wisc.edu Mon Jun 20 17:53:03 2005 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Vender help please, Looking for EG2, Message-ID: Good Morning Netters, Those of us involved in allergy or asthma studies are very interested in eosinophils, as are folks in many other lines of inflammatory research. About five years ago I was able to buy my best eo ab called EG2. It came from a Swedish company called Pharmacia-Upjohn. I have another eo marker that's OK, but I'd love to get EG2 back again. I bet I've made a dozen phone calls and, here's what I've been told. Pharmacia-Upjohn was sold at least in part to Monsanto, who then sold it to Amersham Biosciences, who then sold it in some form to GE Healthcare. Along the way the company was fractured into various sub-companies each of whom kept one part or another of the original company and it's products. No person along the way has been able to give me any leads in finding EG2. If any other detective out there has snooped it out I would be ever so grateful!!! Thank You!!!! 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 MSafron <@t> wilresearch.com Mon Jun 20 17:53:03 2005 From: MSafron <@t> wilresearch.com (Mike Safron) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] (no subject) Message-ID: Michael Safron A.S.,HT(ASCP) Manager, Histology WIL Research Laboratories LLC 419-289-8700 ext: 3040 Ashland, Ohio 44805 From rebecca.riesen <@t> dsilabs.com Mon Jun 20 17:53:03 2005 From: rebecca.riesen <@t> dsilabs.com (Riesen, Rebecca) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] OPPORTUNITY FOR EMPLOYMENT Message-ID: <05844092236E7749A1BBBCB1077C34A2DEA014@dsi-ex01.gateway.dom> My previous email had an incorrect address for HR. If you would love to work with a great team here in Naples, Florida you may contact me directly or through HR at karen.quick@dislabs.com her fax is 239-561-8236. Sorry about the first email address. DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(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 tpmorken <@t> labvision.com Mon Jun 20 17:53:03 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:14 2005 Subject: Vendor view on expiration dates RE: Here I go again[Histonet] Ex piration date (Joe Nocito) Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CE73@usca0082k08.labvision.apogent.com> Just a note about expiration dates from the vendor viewpoint. The expiration date is not a date that a vendor expects an antibody to go bad, rather it is the longest date the vendor is willing to GUARANTEE that an antibody will work. If the antibody quits working before that date the customer gets a new antibody lot. After that date the vendor cannot be held responsible for performance. So it's more of a financial equation and a way for the vendor to avoid eternal responsibility for a product than anything else. I agree that the CAP is being too strict about the expiration dates and I have had the same experience as others in using 10-y.o. frozen aliquots that work just as well as the first one. If the antibody is aliquoted and frozen and sucessfully QC'd regularly, there is no reason it cannot be used. I think the problem the CAP is addressing is more of a QC issue than an antibody longevity. I the past a lab could simply do QC testing of each lot and show it worked and keep it in use. Why that was dropped is not clear. Maybe the CAP simply does not trust labortory QC methods. My own feeling about CAP is that they make up rules and see how people react to them, then changes them according to the reaction. So maybe this topic needs a strong reaction. Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Todd Sherman Sent: Sunday, June 19, 2005 1:41 PM To: histonet@lists.utsouthwestern.edu Subject: Re: Here I go again[Histonet] Expiration date (Joe Nocito) Joe, Regarding "If an antibody expires June 30, 2005, does it automatically go dead on July 1?" - according to the corporate legal teams and their liability concerns, of course. ;) But in all seriousness, you bring up a good point. I can understand a manufacturer needing to maintain QC and establish liability limits, not to mention bump potential premature boosts in sales from the dumping of viable product by enduser, but some modification should be considered. You mention a nice protocol of aliquoting, inventorying, and testing that seems entirely reasonable. Consider another lab that dumps lots according to date but that doesn't aliquot properly and lets an undiluted lot freeze and thaw such that it "expires" before the label date would indicate it has deteriorated. Would such a scenario be better? Obviously a more scaled weighting that considers the continual QC testing against positive and negative controls should be incorporated since that is the fundamental concern. If a lab can produce a series of time-dated QC test samples/slides associated with a particular lot/vial/aliquot that proves efficacy, then I should think that the Ab is still "good". Granted, the manufacturers should not be held responsible for extended usage beyond what they deem reasonable/optimal; however, thoughtless disposal of good product should not be a mandate by the governing histological agency either. $0.02 worth of validation. Regards, Todd Todd Sherman President HistoSoft Corporation www.histosoft.com Biology In A New Form (c) On Sun, 19 Jun 2005 12:00:23 -0500, wrote: > > Today's Topics: > > 1. Here I go again[Histonet] Expiration date (Joe Nocito) > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sat, 18 Jun 2005 12:20:47 -0500 > From: "Joe Nocito" > Subject: Here I go again[Histonet] Expiration date > To: "Katia Cristina Catunda" , "histonet" > > Message-ID: <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=response > > Okay, > I've been too quiet for too long. Here I go again. > > I am so tired of CAP and their ridiculous regulations. Every CAP > inspection > I fight this same question. When I was supervising the Immuno lab at > AFIP, > we froze concentrated antibodies at -70 for years. Antibodies that were > frozen in 1080, were still viable in 1990. That means we were running > immunos in 1990 with 1980 prices. What is so wrong about that? > We also ran a known positive control with each batch. If the positive > control worked, guess what? The antibody was still good. If the positive > control did not work, we threw out that lot number and repeated it with > another lot number. When we saw that there was a drop in the staining > intensity, we tossed that lot and started another. My experience with > antibodies is that they just don't go bad over night, they start staining > with less intensity. > I think that the CAP board members have stocks in the biochemical > companies. Please don't get me wrong, I have many close friends that > work in > the biochemical side of this, but why would this drastic change in view? > The > FDA? I doubt it. > Maybe I'm too one sides on this issue, but give me a break! If the > positive control works in any other antibody, doesn't that mean the > antibody > is viable? > I know companies have to put an expiration date on their products, > but > come on. If an antibody expires June 30, 2005, does it automatically go > dead > on July 1? > As managers and supervisors, we are continuously bashed about saving > money. This would be a great place to start, don't you think? > That is all. Thank you > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Jun 20 17:53:03 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo ELHTPathology) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] RE: Xylene Substitutes[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD2FD@elht-exch1.xelht.nhs.uk> I'm relaxed, a bit light headed but totally relaxed!!! -----Original Message----- From: Rittman, Barry R [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: 20 June 2005 15:35 To: histonet Subject: RE: [Histonet] RE: Xylene Substitutes[Scanned] I must in general agree with Chuck. I have not used any clearing agents that I consider to be as good as xylenes. I think that we are now in an era where the pendulum has swung away from an almost total disregard for the use of chemicals to an era where we are sometimes overly cautious. This is not just with chemicals but many aspects in life. While it is critical to be very careful in the use of all chemicals and various practices (in the lab and at home)there is a point beyond which we would never work in a lab in case we absorbed some chemical or even leave the house in case we might get hit by a falling piece from an aircraft. When training in a lab we were taught to be aware of potential dangers from chemicals, equipment and especially trainees. Safe practices should not only be confined to the lab but should be a way of life for us all. After all we are exposed to enough pollution without our adding to it. If you cut your grass are you careful about not breathing exhaust fumes or getting gasoline on your hands? It seems at times as if we are trying to eliminate nature including us). Does this mean that we should not be using chemicals to clean ovens, to scrub sinks or to bleach clothes? I suggest that there is an acceptable limit to all these substances and practices but in order be realistic we always make ourselves fully aware of possible consequences of use. I make such decisions every day that I drive in Houston. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, June 20, 2005 8:24 AM To: Rogerson Kemlo (ELHT) Pathology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Xylene Substitutes[Scanned] Kemlo, There is an acceptable limit to water ingestion. If you go past that it becomes toxic and even fatal. Sorry, I'm not going to stop drinking water just because at some point it becomes toxic. Many of these "warnings" in the lab boil down to junk science. I just can't take the "Henny Penny" approach to things and run around screaming "The sky is falling, the sky is falling". (My apologies to those that don't understand this analogy). Relax a little, life can be fun........................ Chuck Embrey -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rogerson Kemlo (ELHT) Pathology Sent: Monday, June 20, 2005 1:45 AM To: Barbara Stancel; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Xylene Substitutes[Scanned] Acceptable limit????? Define an acceptable limit! There is no acceptable limit as far as I know, all exposure brings with it danger, but I concede there is a limit beneath which that danger is acceptable. But what are the unacceptable elements of this acceptability? Only 1 in a 1,000 women has a spontaneous abortions and I suppose that may be acceptable. Complete removal and replacement or extraction that results in zero exposure is acceptable, isn't it? -----Original Message----- From: Barbara Stancel [mailto:stancelb@msn.com] Sent: 18 June 2005 20:03 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Xylene Substitutes[Scanned] I'm with Charles. We have been using xylene for 31 years. We have tried the xylene substitutes, but we and our pathologist prefer xylene. We recycle all the xylene to reduce cost of purchasing and cut hazardous waste. We have great ventilation in the lab. Our twice-a-year STEL monitoring has come back at way below the acceptable limits. If it ain't broke, don't fix it. Barbara Stancel USDA, FSIS, OPHS, EL, Pathology Athens, Georgia -----Original Message----- From: Charles.Embrey Sent: Friday, June 17, 2005 1:40 PM To: 'Joyce Cline' Subject: RE: [Histonet] Xylene substitutes Your quote "They all realize how bad Xylene is". Just how bad IS xylene? I have been using it for 25 years and am still alive and kicking. We recycle ours so we don't even have an environmental issue to deal with. I have never used any substitute that worked as well or better than xylene itself. Many claim to be almost as good as xylene and that is about as close as any get. As for me, I am sticking with what works. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Cline Sent: Friday, June 17, 2005 12:05 PM To: 'Histonet' Subject: [Histonet] Xylene substitutes I use Clear-rite 3 and our pathologists have no problem with it. They all realize how bad Xylene is. I use H-2 Blue beads to help eliminate the water from our high humidity. _______________________________________________ Histonet mailing list Histonet@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 sjchtascp <@t> yahoo.com Mon Jun 20 17:53:03 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] tissue dye Message-ID: <20050620154744.73767.qmail@web90202.mail.scd.yahoo.com> Good morning all, I have heard of some techs using eosin in an alcohol of the tissue processor to dye tissues. I'd like to avoid this, but would like to dye specific tissues for easier orientation. Would a dilute eosin work if used prior to putting the tissues on the processor? If so, aqueous or alcoholic? What percentage? Steve --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football From koellinr <@t> amgen.com Mon Jun 20 17:53:03 2005 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Vender help please, Looking for EG2, Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CC785@wa-mb4-sea.amgen.com> A PubMed search on EG2 shows a lot of articles including the most recent in 2005 staining eosinophils in sputum of asthmatics with a Mab called EG2. Koh YI is lead author and contact and has an e-mail address (is in Korea) so must know the story of EG2 and how they got it. Ray -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LaCinda Burchell Sent: Monday, June 20, 2005 7:59 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Vender help please, Looking for EG2, Good Morning Netters, Those of us involved in allergy or asthma studies are very interested in eosinophils, as are folks in many other lines of inflammatory research. About five years ago I was able to buy my best eo ab called EG2. It came from a Swedish company called Pharmacia-Upjohn. I have another eo marker that's OK, but I'd love to get EG2 back again. I bet I've made a dozen phone calls and, here's what I've been told. Pharmacia-Upjohn was sold at least in part to Monsanto, who then sold it to Amersham Biosciences, who then sold it in some form to GE Healthcare. Along the way the company was fractured into various sub-companies each of whom kept one part or another of the original company and it's products. No person along the way has been able to give me any leads in finding EG2. If any other detective out there has snooped it out I would be ever so grateful!!! Thank You!!!! 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon Jun 20 17:53:03 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] xylene substitutes Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717547@lsexch.lsmaster.lifespan.org> I have used xylene for close to forty years, and consider it the safest of all commonly available clearing agents. For certain special projects over the years I have also used toluene, carbon tetrachloride, dioxane, tetrahydrofuran, and a few other compounds that require much more stringent safety precautions than xylene does. Certainly xylene does pose some health concerns. What chemical doesn't? You shouldn't wash your hands in it to remove paraffin. But my attitude toward the various recently-offered "xylene substitutes" is this: I would rather use xylene, the potential hazards of which are well documented after many decades of general use, than an unknown mixture of organic solvents which have not been in use long enough to allow accurate assessment of possible longterm health hazards. Most of the "xylene substitutes" on the market are not pure compounds, but mixtures of several (in some cases many) different organic solvents. The manufacturers won't reveal the solvents contained in the mix, and in some cases they have not even identified all the organic compounds present in the product! I don't feel comfortable exposing myself on a daily basis to large volumes of a product of unknown chemical composition and largely untested health effects. That having been said, I recently did test several "xylene substitutes" because of a special project where I needed to paraffin-embed synthetic hollow fibers which were broken down by xylene. First I tested a number of pure compounds, many of which attacked the fibers much more vigorously than xylene did. Some of them completely dissolved the fibers within seconds. I did discover that carbon tetrachloride worked exceptionally well, but was reluctant to use it due to the combination of high toxicity and high price. Then I turned my attention to "xylene substitutes". I tested eight different ones, from various manufacturers, and while most of them caused no apparent damage to the fibers, they differed greatly in their ability to dissolve paraffin, as well as in their odor. I know from past experience that they also differ greatly in their ability to dissolve fat, though that was not an issue in this study. I finally settled on "Safe-Clear" from Fisher Scientific, which gave me results comparable to carbon tetrachloride, and has a mild odor. But is it really safe? I don't know! I handle it with care since I don't really know what I am handling! Of course, the fact that this product produced good results for this specific purpose doesn't necessarily mean that it would be the best choice for general tissue processing. Paul M. From kappeler <@t> patho.unibe.ch Mon Jun 20 17:53:03 2005 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] HIF-1 immunohistochemistry Message-ID: <017701c575b2$55afcbe0$27955c82@patho.unibe.ch> Dear all I have a vet who wants to do immunohistochemistry for HIF-1 alpha (hypoxia inducible factor 1 alpha) on fresh frozen and/or paraffin embedded tissue from CATS. Would anybody know a source for an anti-HIF-1 antibody that would work on cat tissue? The MSRS database revealed quite a few antibodies, but - of course - no one of them was mentioned to be suitable for cats... Many thanks! Andi Kappeler Institute of Pathology, University of Bern, Switzerland From JNocito <@t> Pathreflab.com Mon Jun 20 17:53:03 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:14 2005 Subject: Here I go again[Histonet] Expiration date In-Reply-To: Message-ID: Patti, this year we labeled the reagents "For Student Use Only" Since we use the Ventana XTs, it's hard to by-pass the expiration dates. Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patti Loykasek Sent: Monday, June 20, 2005 9:32 AM To: histonet Subject: Re: Here I go again[Histonet] Expiration date Joe - I totally agree with you. My only question is this: with the new CAP rule in place, have you been cited for using expired reagents? While I would be most concerned with proper QC, I know most inspectors go by the letter of the rule, and aren't open for debate. We got written up for having a box with an expired date on it - we were using it to store some excess non-expired reagent ( I know, exactly a smart thing to do). Patti Loykasek PhenoPath Laboratories Seattle, WA > Okay, > I've been too quiet for too long. Here I go again. > > I am so tired of CAP and their ridiculous regulations. Every CAP inspection > I fight this same question. When I was supervising the Immuno lab at AFIP, > we froze concentrated antibodies at -70 for years. Antibodies that were > frozen in 1080, were still viable in 1990. That means we were running > immunos in 1990 with 1980 prices. What is so wrong about that? > We also ran a known positive control with each batch. If the positive > control worked, guess what? The antibody was still good. If the positive > control did not work, we threw out that lot number and repeated it with > another lot number. When we saw that there was a drop in the staining > intensity, we tossed that lot and started another. My experience with > antibodies is that they just don't go bad over night, they start staining > with less intensity. > I think that the CAP board members have stocks in the biochemical > companies. Please don't get me wrong, I have many close friends that work in > the biochemical side of this, but why would this drastic change in view? The > FDA? I doubt it. > Maybe I'm too one sides on this issue, but give me a break! If the > positive control works in any other antibody, doesn't that mean the antibody > is viable? > I know companies have to put an expiration date on their products, but > come on. If an antibody expires June 30, 2005, does it automatically go dead > on July 1? > As managers and supervisors, we are continuously bashed about saving > money. This would be a great place to start, don't you think? > That is all. Thank you > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > ----- Original Message ----- > From: "Katia Cristina Catunda" > To: "histonet" > Sent: Wednesday, June 15, 2005 8:20 PM > Subject: [Histonet] Expiration date > > >> What should we do with antibodies and reagents after their indicated >> expiration dates? Theorectically they shouldn't be estable after the >> expiration date but we do know that most of them still works perfectly... >> should we just throw them out and not use them anymore, wouldn't this be a >> way for the manufacturers manipulate their sellings? Need some opinions... >> >> Tks >> >> Katia >> >> >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> -- >> No virus found in this incoming message. >> Checked by AVG Anti-Virus. >> Version: 7.0.323 / Virus Database: 267.7.8/22 - Release Date: 6/17/2005 >> >> > > > _______________________________________________ > Histonet mailing list > Histonet@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 Luis.Chiriboga <@t> med.nyu.edu Mon Jun 20 17:53:03 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] HIF-1 immunohistochemistry In-Reply-To: <017701c575b2$55afcbe0$27955c82@patho.unibe.ch> Message-ID: I would recommend you try the antibody from Neomarkers/labvision. works well in human and mouse. They also say works in Sheep, rat and Ferret so perhaps it may work in cat. you may want to visit the website or give them a call to see if the have done any further species testing since I ordered the antibody well over 2 years ago..... LC>>> ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andi Kappeler Sent: Monday, June 20, 2005 12:08 PM To: Histonet Subject: [Histonet] HIF-1 immunohistochemistry Dear all I have a vet who wants to do immunohistochemistry for HIF-1 alpha (hypoxia inducible factor 1 alpha) on fresh frozen and/or paraffin embedded tissue from CATS. Would anybody know a source for an anti-HIF-1 antibody that would work on cat tissue? The MSRS database revealed quite a few antibodies, but - of course - no one of them was mentioned to be suitable for cats... Many thanks! Andi Kappeler Institute of Pathology, University of Bern, Switzerland _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon Jun 20 17:53:03 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Help, slide adhesive for wood Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717548@lsexch.lsmaster.lifespan.org> I have extremely limited experience with sectioning wood (a few times, years ago). But I think that PVA (polyvinyl acetate) would be a good choice. I made some comments about PVA last week, in a thread on sectioning nails, so I won't re-post that information here. PVA is used in making wood glue, so I assume it will bind very well to wood! And I know from experience that it adheres very well to glass. Of course, the first consideration in attaching any section to a slide, regardless of what adhesive you are using, is creating intimate contact between the surface of the slide and the surface of the tissue section. In other words, getting the section to lie flat. No adhesive will fill gaps between the tissue and the slide. So, if the section looks "wavy" on the water bath when you pick it up, it probably won't adhere to the slide during staining, regardless of the adhesive used. Methods for flattening the section include: - longer time on the water bath - higher temperature of the water bath - placing the slide on a "slide warmer" at 80 degrees after draining the slide briefly - holding the section against the slide with physical pressure while drying, as is often done with methyl methacrylate sections Paul M. From ekh9535 <@t> bjc.org Mon Jun 20 17:53:03 2005 From: ekh9535 <@t> bjc.org (Erin Herter) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] QUALITY ASSURANC/CONTROL Message-ID: Would there be anyone out there willing to share their QA procedures? I am mostly interested in QA for cytopathology. Also what kinds of things do you keep track for for CAP "surveilllance" Thanks From cdemarinis <@t> SARATOGACARE.ORG Mon Jun 20 17:53:03 2005 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis, Carolyn) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Cost per test Message-ID: Does ASCP, CAP, or other regulatory agency have guidelines established to calculate cost per test in the clinical laboratory? *************************************************************************************** CONFIDENTIALITY NOTICE: This e-mail communication and any attachmentsmay contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. From ksamayoa <@t> genomichealth.com Mon Jun 20 17:53:03 2005 From: ksamayoa <@t> genomichealth.com (Kimberly Samayoa) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] JOB OPPORTUNITY IN NORTHERN CALIFORNIA Message-ID: <5C9B539E7DD26C4AA41D4E27ACECBB9101F4696C@jaguar.genomichealth.com> Histotechnologist Opening in Redwood City, California (Northern California) Are you interested in working on an exciting, cutting edge technology? Do you have a desire to prolong and enhance the lives of cancer patients? Are you ready to utilize your specialized knowledge of histological testing? Then come join Genomic Health as a Histotechnologist and play a part in realizing an innovative new approach to disease diagnosis and treatment. Click on this link to view the job description: http://www.genomichealth.com/careers/jobs.aspx?JobID=42 Genomic Health offers competitive salaries, an environment that encourages career growth, rewards for accomplishments and many other excellent benefits. Vist www.genomichealth.com for more information. For immediate consideration, send resumes to jobs@genomichealth.com or fax to (650) 569-2464. The contents of this electronic message, including any attachments, are intended only for the use of the individual or entity to which they are addressed and may contain confidential information. If you are not the intended recipient, you are hereby notified that any use, dissemination, distribution, or copying of this message or any attachment is strictly prohibited. If you have received this transmission in error, please send an e-mail to postmaster@genomichealth.com and delete this message, along with any attachments, from your computer. From lfidgen <@t> vt.edu Mon Jun 20 17:53:03 2005 From: lfidgen <@t> vt.edu (Laura Fidgen) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] looking for Debbie Vigil Message-ID: <6.0.0.22.0.20050620135927.026b0600@pop.vt.edu> If anyone has her e-mail, I would appreciate it. Debbie, if you receive this e-mail please contact me. Laura Fidgen Laura L. Fidgen, MScF, BSc, HT(ASCP) Laboratory and Research Practioner VMRCVM, Histopatholgy Blacksburg, VA 24060-0443 From cestanecki <@t> ucdavis.edu Mon Jun 20 17:53:03 2005 From: cestanecki <@t> ucdavis.edu (Catherine Stanecki) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Frozen sections for IF Message-ID: <200506201802.j5KI2Bun014652@lymeon.ucdavis.edu> Hi Histonetters, So, I'm pretty new to histology and I have several questions about frozen sections. I would really appreciate your help! First of all, how long are frozen tissues good for after freezing in tissue tek OTC? Is it better to store at -20 C or -80 C? Also, how long are sections on slides good for before you have stained and how should you store them (i.e. in a slide box in the freezer)? How far in advance can you cut your sections before staining them? Thanks so much for you help! Cathy From tpmorken <@t> labvision.com Mon Jun 20 17:53:03 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Cost per test Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CE7B@usca0082k08.labvision.apogent.com> Carolyn, The Clincal Laboratory Standards Institute (formerly NCCLS) has a guideline for cost accounting that can be downloaded from their website. Unfortunately it costs US $120. But it is a good basic guide and give examples of spreadsheets that can be set up to do cost accounting. It may be that your lab management already has a copy of this, and if they don't, and are members of CLSI, they can get it cheaper ($60). Alternatively, you can sign up for the Cost Accounting course that I and Jan Gardner are giving at the NSH meeting in Ft. Lauderdale this year. It is based both on our combined experience and the guideline mentioned above and we give you an extensive spreadsheet that has been used in the real world of the lab. Worshope #49, Cost Accounting for the Histology Laboratory. GP11-A Basic cost accounting forClinical Services, electronic or paper document http://www.nccls.org/ (looking under the "shop" menu, "General Laboratory Practices", will give you a list of publications). Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Demarinis, Carolyn Sent: Monday, June 20, 2005 10:24 AM To: HISTONET (E-mail) Subject: [Histonet] Cost per test Does ASCP, CAP, or other regulatory agency have guidelines established to calculate cost per test in the clinical laboratory? **************************************************************************** *********** CONFIDENTIALITY NOTICE: This e-mail communication and any attachmentsmay contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon Jun 20 17:53:03 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] formic acid decal Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717549@lsexch.lsmaster.lifespan.org> You might want to consider adding phloroglucinol (1,3,5-Trihydroxybenzene) to your decalcifying solution while increasing the concentration of acid. This compound, for reasons not fully understood, seems to protect the morphology of tissue during extended exposure to strong acids. I have read that tissues can be decalcified in 40% nitric acid when 2 to 4% phloroglucinol is added. I haven't tried this. However, for rapid decalcification I use the following solution, which works well for me: distilled water 750 ml formic acid 150 ml hydrochloric acid 100 ml phloroglucinol 20 gm The phloroglucinol is a dry powder which takes about 10 minutes to dissolve on a stirrer. Paul M. From emerald_lake77 <@t> yahoo.com Mon Jun 20 17:53:03 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Demonstration of antigen on cell surface - question? Message-ID: <20050620194833.18528.qmail@web31713.mail.mud.yahoo.com> Hello, How would one go about determining if your specific antigen (say for IHC) is located on the cell surface? Right now it seems that the antigen is staining everywhere. (I am running a sheep anti-mouse 'antigen' and using a rabbit anti-sheep biotin conjugated with ABC + DAB for detection. (My negative is normal sheep IgG -- is negative). Is there a way through immuno or other that I can specifically say that the antigen is present on the cell surface? Are there general cell surface markers that I can use along with my primary? Thank you all. --------------------------------- Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. From JNocito <@t> Pathreflab.com Mon Jun 20 17:53:03 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:14 2005 Subject: Here I go again[Histonet] Expiration date (Joe Nocito) In-Reply-To: Message-ID: Todd, I do not expect a manufacturer to guarantee any reagent beyond its expiration date, but if the QC is done properly, with the proper documentation to back it up, throwing away good reagents just doesn't make any sense to me. Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Todd Sherman Sent: Sunday, June 19, 2005 3:41 PM To: histonet@lists.utsouthwestern.edu Subject: Re: Here I go again[Histonet] Expiration date (Joe Nocito) Joe, Regarding "If an antibody expires June 30, 2005, does it automatically go dead on July 1?" - according to the corporate legal teams and their liability concerns, of course. ;) But in all seriousness, you bring up a good point. I can understand a manufacturer needing to maintain QC and establish liability limits, not to mention bump potential premature boosts in sales from the dumping of viable product by enduser, but some modification should be considered. You mention a nice protocol of aliquoting, inventorying, and testing that seems entirely reasonable. Consider another lab that dumps lots according to date but that doesn't aliquot properly and lets an undiluted lot freeze and thaw such that it "expires" before the label date would indicate it has deteriorated. Would such a scenario be better? Obviously a more scaled weighting that considers the continual QC testing against positive and negative controls should be incorporated since that is the fundamental concern. If a lab can produce a series of time-dated QC test samples/slides associated with a particular lot/vial/aliquot that proves efficacy, then I should think that the Ab is still "good". Granted, the manufacturers should not be held responsible for extended usage beyond what they deem reasonable/optimal; however, thoughtless disposal of good product should not be a mandate by the governing histological agency either. $0.02 worth of validation. Regards, Todd Todd Sherman President HistoSoft Corporation www.histosoft.com Biology In A New Form (c) On Sun, 19 Jun 2005 12:00:23 -0500, wrote: > > Today's Topics: > > 1. Here I go again[Histonet] Expiration date (Joe Nocito) > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sat, 18 Jun 2005 12:20:47 -0500 > From: "Joe Nocito" > Subject: Here I go again[Histonet] Expiration date > To: "Katia Cristina Catunda" , "histonet" > > Message-ID: <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=response > > Okay, > I've been too quiet for too long. Here I go again. > > I am so tired of CAP and their ridiculous regulations. Every CAP > inspection > I fight this same question. When I was supervising the Immuno lab at > AFIP, > we froze concentrated antibodies at -70 for years. Antibodies that were > frozen in 1080, were still viable in 1990. That means we were running > immunos in 1990 with 1980 prices. What is so wrong about that? > We also ran a known positive control with each batch. If the positive > control worked, guess what? The antibody was still good. If the positive > control did not work, we threw out that lot number and repeated it with > another lot number. When we saw that there was a drop in the staining > intensity, we tossed that lot and started another. My experience with > antibodies is that they just don't go bad over night, they start staining > with less intensity. > I think that the CAP board members have stocks in the biochemical > companies. Please don't get me wrong, I have many close friends that > work in > the biochemical side of this, but why would this drastic change in view? > The > FDA? I doubt it. > Maybe I'm too one sides on this issue, but give me a break! If the > positive control works in any other antibody, doesn't that mean the > antibody > is viable? > I know companies have to put an expiration date on their products, > but > come on. If an antibody expires June 30, 2005, does it automatically go > dead > on July 1? > As managers and supervisors, we are continuously bashed about saving > money. This would be a great place to start, don't you think? > That is all. Thank you > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Mon Jun 20 17:53:03 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Frozen sections for IF In-Reply-To: <200506201802.j5KI2Bun014652@lymeon.ucdavis.edu> Message-ID: Cathy, welcome to histology. My experience with tissue frozen in OCT is that it can last for about a year, if the tissue doesn't go through a freeze-thaw cycle, like in those no-frost freezers. Slides always should be stained as soon as possible after cutting. The most we ever let slides sit for IF is over a 3 day weekend. -80 is always better than -20. We cut control slides for IF two months ago and the tissue started loosing its antigenicity 6 weeks after at -20. Good luck 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 Catherine Stanecki Sent: Monday, June 20, 2005 1:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozen sections for IF Hi Histonetters, So, I'm pretty new to histology and I have several questions about frozen sections. I would really appreciate your help! First of all, how long are frozen tissues good for after freezing in tissue tek OTC? Is it better to store at -20 C or -80 C? Also, how long are sections on slides good for before you have stained and how should you store them (i.e. in a slide box in the freezer)? How far in advance can you cut your sections before staining them? Thanks so much for you help! Cathy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Luis.Chiriboga <@t> med.nyu.edu Mon Jun 20 17:53:03 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Demonstration of antigen on cell surface - question? In-Reply-To: <20050620194833.18528.qmail@web31713.mail.mud.yahoo.com> Message-ID: I think a bit more info is needed but here's a few suggestions.... If your working in frozen or formalin fixed tissue, serially dilute both primary and secondary antibodies independently to arrive at your best signal to noise. If you are performing antigen retrieval (formalin fixed), run a time series of different HEIR times or various concentrations of enzyme. This should help you improve the S/N to an extent that allows easier interpretation. If your working with a novel protein... If you have access to cell culture, perform cell fractionation and check localization via western blot. Have you checked in the literature? for similar class or family of protein and it's localization? If you have/know the target protein/antigen check for protein localization signal (NCBI-protein database & genbank), or protein sequence homology to other known proteins (NCBI-Blast) LC>>> ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GT Hebert Sent: Monday, June 20, 2005 3:49 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Demonstration of antigen on cell surface - question? Hello, How would one go about determining if your specific antigen (say for IHC) is located on the cell surface? Right now it seems that the antigen is staining everywhere. (I am running a sheep anti-mouse 'antigen' and using a rabbit anti-sheep biotin conjugated with ABC + DAB for detection. (My negative is normal sheep IgG -- is negative). Is there a way through immuno or other that I can specifically say that the antigen is present on the cell surface? Are there general cell surface markers that I can use along with my primary? Thank you all. --------------------------------- Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ftulenko06 <@t> jcu.edu Mon Jun 20 17:53:03 2005 From: ftulenko06 <@t> jcu.edu (ftulenko06@jcu.edu) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] generating 3D models from serial sections Message-ID: <7eddd1d1.f8d4871.825ca00@mirapoint.jcu.edu> I have been generating 3-D models from serially sectioned turtle embryos using Winsurf software. I have imported these models (DFX files) into Maya 6.0 (software) to smooth them out. Has anyone done this? I am able to manipulate objects generated within Maya, but am having a lot of difficulty working on imported files from WINSURF. The points in the model do not appear to be grouped together. I would greatly appreciate any help or advice. Thanks for your time and attention, Frank From anh2006 <@t> med.cornell.edu Mon Jun 20 17:57:50 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Demonstration of antigen on cell surface - question? In-Reply-To: References: Message-ID: Good points .... Also, in addition to background issues which are often the culprit, in my experience membrane antigens when highly expressed - such as say EGFR in certain cancers or in skin - are often detected in both cytoplasmic and membrane staining patterns mainly due to two reasons I can think of: (1) The antigen truly is in the cytoplasm on it's way to the cell membrane therefore the staining you detect is "real" (2) The antibody staining reaction is so strong you are getting a fuzzy deposition of DAB (this is especially the case in high amplification kits when used on overexpressed antigens) (finally I just want to add that sometimes what is membrane can appear cytoplasmic depending on the shape of the cell and the cell type - for example when staining VEGF receptors on endothelial cells it is often VERY hard to tell where on the cell they are located using IHC). Perhaps with a little more info about the cell type and antibody you are using more info can be provided in this specific case. Generally speaking though I would suggest that you not use IHC/DAB etc for determining where in a cell a specific protein is expressed. It is more useful perhaps to use immunofluorescence and confocal microscopy or at minimum epifluorescence. Then you will most certainly get the answer you need. In addition, you can take a non-staining approach and do either of the two following: (1) Make differential cell lysates of membrane and cytosolic fractions and do westerns (2) Make a GFP fusion protein and express it in your cell and view where it is located via confocal Good luck and I hope this makes sense! Andrea At 4:34 PM -0400 6/20/05, Luis Chiriboga wrote: >I think a bit more info is needed but here's a few suggestions.... >If your working in frozen or formalin fixed tissue, serially dilute both >primary and secondary antibodies independently to arrive at your best signal >to noise. >If you are performing antigen retrieval (formalin fixed), run a time series >of different HEIR times or various concentrations of enzyme. >This should help you improve the S/N to an extent that allows easier >interpretation. >If your working with a novel protein... >If you have access to cell culture, perform cell fractionation and check >localization via western blot. >Have you checked in the literature? for similar class or family of protein >and it's localization? >If you have/know the target protein/antigen check for protein localization >signal (NCBI-protein database & genbank), or protein sequence homology to >other known proteins (NCBI-Blast) >LC>>> > > > > > > > > > > >____________________________________ >Luis Chiriboga Ph.D. >NYU Cancer Institute and >Bellevue Hospital Center >New York University School Of Medicine >Department Of Pathology 4W27 >462 First Avenue >New York, N.Y. 10016 >W(212) 562-4667. >F(212) 263-2041 > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GT Hebert >Sent: Monday, June 20, 2005 3:49 PM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Demonstration of antigen on cell surface - question? > > >Hello, > >How would one go about determining if your specific antigen (say for IHC) is >located on the cell surface? Right now it seems that the antigen is >staining everywhere. (I am running a sheep anti-mouse 'antigen' and using a >rabbit anti-sheep biotin conjugated with ABC + DAB for detection. (My >negative is normal sheep IgG -- is negative). Is there a way through immuno >or other that I can specifically say that the antigen is present on the cell >surface? Are there general cell surface markers that I can use along with >my primary? Thank you all. > > > -- From emerald_lake77 <@t> yahoo.com Mon Jun 20 18:22:40 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Demonstration of antigen on cell surface - question? In-Reply-To: Message-ID: <20050620232240.95560.qmail@web31714.mail.mud.yahoo.com> Sharon, I know a group that uses confocal but do not know it very well myself. Can you give me a more detailed explanation of why confocal would be useful ... and suggestion for cell surface markers that can be used? Thank you very much. Sharon Cooperman wrote: You could do confocal microscopy - that would unequivocally demonstrate thqt the antigen is on the cell surface, particularly if you co-stain with a cell surface marker. Sharon >Hello, > >How would one go about determining if your specific antigen (say for >IHC) is located on the cell surface? Right now it seems that the >antigen is staining everywhere. (I am running a sheep anti-mouse >'antigen' and using a rabbit anti-sheep biotin conjugated with ABC + >DAB for detection. (My negative is normal sheep IgG -- is >negative). Is there a way through immuno or other that I can >specifically say that the antigen is present on the cell surface? >Are there general cell surface markers that I can use along with my >primary? Thank you all. > > > > >--------------------------------- >Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From anh2006 <@t> med.cornell.edu Mon Jun 20 18:40:55 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Frozen sections for IF In-Reply-To: References: Message-ID: I have stored tissue (and even slides) for years at -80 deg C in proper containers with no deleterious effects on cutting quality or morphology. In addition, I have stained cut sections stored at -80 deg C for a year in proper containers with no deleterious effects on the antigens I was staining for (CD31 human and mouse, EGFR, B220 and other similar antigens). It all really depends on how the tissue was stored and the antigen of interest. But as Joe pointed out, fresher is always better and preferred. Sincerely, Andrea At 3:29 PM -0500 6/20/05, Joe Nocito wrote: >Cathy, >welcome to histology. My experience with tissue frozen in OCT is that it can >last for about a year, if the tissue doesn't go through a freeze-thaw cycle, >like in those no-frost freezers. >Slides always should be stained as soon as possible after cutting. The most >we ever let slides sit for IF is over a 3 day weekend. -80 is always better >than -20. We cut control slides for IF two months ago and the tissue started >loosing its antigenicity 6 weeks after at -20. Good luck > >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 Catherine >Stanecki >Sent: Monday, June 20, 2005 1:02 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Frozen sections for IF > > > >Hi Histonetters, > > So, I'm pretty new to histology and I have several questions about frozen >sections. I would really appreciate your help! First of all, how long are >frozen tissues good for after freezing in tissue tek OTC? Is it better to >store at -20 C or -80 C? Also, how long are sections on slides good for >before you have stained and how should you store them (i.e. in a slide box >in the freezer)? How far in advance can you cut your sections before >staining them? > > Thanks so much for you help! > >Cathy > >_ -- From Emily.Wiesner <@t> medecine.unige.ch Tue Jun 21 02:07:07 2005 From: Emily.Wiesner <@t> medecine.unige.ch (Emily Jane Wiesner-Camm) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Storing antibodies Message-ID: <42B7BC9B.9010903@medecine.unige.ch> Hi All, I have been talking with colleagues recently and have received mixed responses about whether there is any concern about storing antibodies at -20 degrees if the spec sheets specify 2-8 degrees? I would appreciate your comments. Thanks in advance. Emily From Kemlo.Rogerson <@t> elht.nhs.uk Tue Jun 21 02:53:42 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] tissue dye[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD452@elht-exch1.xelht.nhs.uk> I've used alcoholic eosin to colour core biopsies of breast and pancreas so that you can see them better at embedding. I think it was 1% but be careful; if you are 'messy' as I am you get everything pinky red. The colour did stay in and it did help greatly, the only problem is how to do it without getting carry over. Don't immerse in a common bath of dye, I know that sounds obvious, but I've seen it done. I used a pipette and the top of the biopsy lid; put the biopsies in the lid, then gently drop on the dye and leave a few secs. When you put the lid back on the pot you get dye everywhere!!!! -----Original Message----- From: Steven Coakley [mailto:sjchtascp@yahoo.com] Sent: 20 June 2005 16:48 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] tissue dye[Scanned] Good morning all, I have heard of some techs using eosin in an alcohol of the tissue processor to dye tissues. I'd like to avoid this, but would like to dye specific tissues for easier orientation. Would a dilute eosin work if used prior to putting the tissues on the processor? If so, aqueous or alcoholic? What percentage? Steve --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Tue Jun 21 07:03:29 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:14 2005 Subject: Here I go again[Histonet] Expiration date (Joe Nocito) Message-ID: I believe even manufacturers of antibodies will change the expiration date of a product after it has been re-tested and found to be still potent. They don't want to throw away a ton of good material, either. But, they'll change the lot number. "Joe Nocito" Sent by: histonet-bounces@lists.utsouthwestern.edu 06/20/2005 03:21 PM To: "Todd Sherman" , cc: Subject: RE: Here I go again[Histonet] Expiration date (Joe Nocito) Todd, I do not expect a manufacturer to guarantee any reagent beyond its expiration date, but if the QC is done properly, with the proper documentation to back it up, throwing away good reagents just doesn't make any sense to me. Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Todd Sherman Sent: Sunday, June 19, 2005 3:41 PM To: histonet@lists.utsouthwestern.edu Subject: Re: Here I go again[Histonet] Expiration date (Joe Nocito) Joe, Regarding "If an antibody expires June 30, 2005, does it automatically go dead on July 1?" - according to the corporate legal teams and their liability concerns, of course. ;) But in all seriousness, you bring up a good point. I can understand a manufacturer needing to maintain QC and establish liability limits, not to mention bump potential premature boosts in sales from the dumping of viable product by enduser, but some modification should be considered. You mention a nice protocol of aliquoting, inventorying, and testing that seems entirely reasonable. Consider another lab that dumps lots according to date but that doesn't aliquot properly and lets an undiluted lot freeze and thaw such that it "expires" before the label date would indicate it has deteriorated. Would such a scenario be better? Obviously a more scaled weighting that considers the continual QC testing against positive and negative controls should be incorporated since that is the fundamental concern. If a lab can produce a series of time-dated QC test samples/slides associated with a particular lot/vial/aliquot that proves efficacy, then I should think that the Ab is still "good". Granted, the manufacturers should not be held responsible for extended usage beyond what they deem reasonable/optimal; however, thoughtless disposal of good product should not be a mandate by the governing histological agency either. $0.02 worth of validation. Regards, Todd Todd Sherman President HistoSoft Corporation www.histosoft.com Biology In A New Form (c) On Sun, 19 Jun 2005 12:00:23 -0500, wrote: > > Today's Topics: > > 1. Here I go again[Histonet] Expiration date (Joe Nocito) > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sat, 18 Jun 2005 12:20:47 -0500 > From: "Joe Nocito" > Subject: Here I go again[Histonet] Expiration date > To: "Katia Cristina Catunda" , "histonet" > > Message-ID: <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=response > > Okay, > I've been too quiet for too long. Here I go again. > > I am so tired of CAP and their ridiculous regulations. Every CAP > inspection > I fight this same question. When I was supervising the Immuno lab at > AFIP, > we froze concentrated antibodies at -70 for years. Antibodies that were > frozen in 1080, were still viable in 1990. That means we were running > immunos in 1990 with 1980 prices. What is so wrong about that? > We also ran a known positive control with each batch. If the positive > control worked, guess what? The antibody was still good. If the positive > control did not work, we threw out that lot number and repeated it with > another lot number. When we saw that there was a drop in the staining > intensity, we tossed that lot and started another. My experience with > antibodies is that they just don't go bad over night, they start staining > with less intensity. > I think that the CAP board members have stocks in the biochemical > companies. Please don't get me wrong, I have many close friends that > work in > the biochemical side of this, but why would this drastic change in view? > The > FDA? I doubt it. > Maybe I'm too one sides on this issue, but give me a break! If the > positive control works in any other antibody, doesn't that mean the > antibody > is viable? > I know companies have to put an expiration date on their products, > but > come on. If an antibody expires June 30, 2005, does it automatically go > dead > on July 1? > As managers and supervisors, we are continuously bashed about saving > money. This would be a great place to start, don't you think? > That is all. Thank you > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Tue Jun 21 08:32:59 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:25:14 2005 Subject: Here I go again[Histonet] Expiration date (Joe Nocito) Message-ID: <062120051332.11517.42B8170B00017BFA00002CFD2205886014CECE030E9D0C9A03@comcast.net> Jackie, You weren't suppose to tell everyone. Generally when it is re-tested by the manufacturer it is given a new lot number and the records are combined or noted according GMP or other regulating body so it is legal. This way we can't tell what they did based on our current lot numbers that are expiring and may even buy the same lot with a new number and expiration withnot realizing it. They are generally careful to re-test and comfirm the validity of the product be it antibody or other reagents before doing this. Pam Marcum -------------- Original message -------------- > I believe even manufacturers of antibodies will change the expiration date > of a product after it has been re-tested and found to be still potent. > They don't want to throw away a ton of good material, either. But, > they'll change the lot number. > > > > > > > "Joe Nocito" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 06/20/2005 03:21 PM > > > To: "Todd Sherman" , > > cc: > Subject: RE: Here I go again[Histonet] Expiration date (Joe > Nocito) > > > Todd, > I do not expect a manufacturer to guarantee any reagent beyond its > expiration date, but if the QC is done properly, with the proper > documentation to back it up, throwing away good reagents just doesn't make > any sense to me. > > Joe > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Todd > Sherman > Sent: Sunday, June 19, 2005 3:41 PM > To: histonet@lists.utsouthwestern.edu > Subject: Re: Here I go again[Histonet] Expiration date (Joe Nocito) > > > Joe, > > Regarding "If an antibody expires June 30, 2005, does it automatically go > dead on July 1?" - according to the corporate legal teams and their > liability concerns, of course. ;) > > But in all seriousness, you bring up a good point. I can understand a > manufacturer needing to maintain QC and establish liability limits, not to > mention bump potential premature boosts in sales from the dumping of > viable product by enduser, but some modification should be considered. You > mention a nice protocol of aliquoting, inventorying, and testing that > seems entirely reasonable. Consider another lab that dumps lots according > to date but that doesn't aliquot properly and lets an undiluted lot freeze > and thaw such that it "expires" before the label date would indicate it > has deteriorated. Would such a scenario be better? Obviously a more scaled > weighting that considers the continual QC testing against positive and > negative controls should be incorporated since that is the fundamental > concern. > > If a lab can produce a series of time-dated QC test samples/slides > associated with a particular lot/vial/aliquot that proves efficacy, then I > should think that the Ab is still "good". Granted, the manufacturers > should not be held responsible for extended usage beyond what they deem > reasonable/optimal; however, thoughtless disposal of good product should > not be a mandate by the governing histological agency either. > > $0.02 worth of validation. > > Regards, > Todd > > > Todd Sherman > President > HistoSoft Corporation > > www.histosoft.com > Biology In A New Form (c) > > > On Sun, 19 Jun 2005 12:00:23 -0500, > wrote: > > > > Today's Topics: > > > > 1. Here I go again[Histonet] Expiration date (Joe Nocito) > > > > ---------------------------------------------------------------------- > > > > Message: 1 > > Date: Sat, 18 Jun 2005 12:20:47 -0500 > > From: "Joe Nocito" > > Subject: Here I go again[Histonet] Expiration date > > To: "Katia Cristina Catunda" , "histonet" > > > > Message-ID: <009701c5742a$118674c0$b4bd0b43@yourxhtr8hvc4p> > > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > > reply-type=response > > > > Okay, > > I've been too quiet for too long. Here I go again. > > > > I am so tired of CAP and their ridiculous regulations. Every CAP > > inspection > > I fight this same question. When I was supervising the Immuno lab at > > AFIP, > > we froze concentrated antibodies at -70 for years. Antibodies that were > > frozen in 1080, were still viable in 1990. That means we were running > > immunos in 1990 with 1980 prices. What is so wrong about that? > > We also ran a known positive control with each batch. If the > positive > > control worked, guess what? The antibody was still good. If the positive > > control did not work, we threw out that lot number and repeated it with > > another lot number. When we saw that there was a drop in the staining > > intensity, we tossed that lot and started another. My experience with > > antibodies is that they just don't go bad over night, they start > staining > > with less intensity. > > I think that the CAP board members have stocks in the biochemical > > companies. Please don't get me wrong, I have many close friends that > > work in > > the biochemical side of this, but why would this drastic change in view? > > The > > FDA? I doubt it. > > Maybe I'm too one sides on this issue, but give me a break! If the > > positive control works in any other antibody, doesn't that mean the > > antibody > > is viable? > > I know companies have to put an expiration date on their products, > > but > > come on. If an antibody expires June 30, 2005, does it automatically go > > dead > > on July 1? > > As managers and supervisors, we are continuously bashed about saving > > money. This would be a great place to start, don't you think? > > That is all. Thank you > > > > Joe Nocito BS, HT(ASCP)QIHC > > Histology Manager > > Pathology Reference Lab > > San Antonio, TX > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JEllin <@t> yumaregional.org Tue Jun 21 08:44:59 2005 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] HPV Antibody Message-ID: We are currently looking at doing HPV as a immuno instread of a probe. The company we are looking at is abcam. Has anyone out there tried this product? If so with specifically with the Ventanna immunos stainers?/ Jesus Ellin Yuma Regional Medical Center From sobrien <@t> bthosp.com Tue Jun 21 09:25:34 2005 From: sobrien <@t> bthosp.com (O'Brien, Sue) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Job Opening - New Jersey Message-ID: <4D95C24EC0E4C84787A6919F1165B25E24B15B@btmhems01.BTHOSP.INT> I have a full time (HT or HTL) position available in our Histology department. Anyone interested in the particulars can contact the hospitals Human Resources department at 609-463-2170. Thanks, Susan O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May Court House, NJ 08210 From rebecca.riesen <@t> dsilabs.com Tue Jun 21 09:29:52 2005 From: rebecca.riesen <@t> dsilabs.com (Riesen, Rebecca) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] RE: Histonet Digest, Vol 19, Issue 30 Message-ID: <05844092236E7749A1BBBCB1077C34A2DEA017@dsi-ex01.gateway.dom> We use eosin in the processor and find that if it is in anything earlier than the second absolute it washes out too much. So, I would guess that eosin prior to putting the tissues on the processor wouldn't work. There are many other dyes that are made for the purpose you are proposing. We use Bradley products, but there are others out there. Message: 9 Date: Mon, 20 Jun 2005 08:47:44 -0700 (PDT) From: Steven Coakley Subject: [Histonet] tissue dye To: histonet@lists.utsouthwestern.edu Message-ID: <20050620154744.73767.qmail@web90202.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Good morning all, I have heard of some techs using eosin in an alcohol of the tissue processor to dye tissues. I'd like to avoid this, but would like to dye specific tissues for easier orientation. Would a dilute eosin work if used prior to putting the tissues on the processor? If so, aqueous or alcoholic? What percentage? Steve --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(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 SMAPAISAIC <@t> aol.com Tue Jun 21 09:42:12 2005 From: SMAPAISAIC <@t> aol.com (SMAPAISAIC@aol.com) Date: Fri Sep 16 15:25:14 2005 Subject: [Histonet] Sledge microtome needed Message-ID: <8a.29528524.2fe98144@aol.com> We need a sledge microtome with a cryosledge attachment to do frozens. Anyone know of any new or used? Sharon CRL/Pathology Associates Cary, North Carolina From ljones <@t> pathology.umsmed.edu Tue Jun 21 10:26:17 2005 From: ljones <@t> pathology.umsmed.edu (Linda Jones) Date: Fri Sep 16 15:25:14 2005 Subject: Fwd: [Histonet] Required CEU's Message-ID: Yes, is that true? >>> "Dianne Holmes" 6/16/2005 11:04:04 AM >>> I have heard that there is a required number of CEU's for HT's to get within each year to maintain their certification. Is this so and if it is - how many? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Linda Harper-Jones BS.,HT/ HTL(ASCP) University of Mississippi Medical Center Department of Pathology Chief Histotechnologist Supervisor (601) 984-1576 (601) 984-4968 Fax ljones@pathology.umsmed.edu From relia1 <@t> earthlink.net Tue Jun 21 11:28:19 2005 From: relia1 <@t> earthlink.net (Pam Barker) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Relia Job Alert - Opportunities in the Sunshine State. Message-ID: Hello Histonetters, Enjoying your summer? Keep your tan all year long. I have several opportunities in Southern Florida. I am currently recruiting for 2 clients in Southern Florida. Both of these positions are full time permanent Monday-Friday dayshift opportunities Here is the info: 1. Histotechnologist/Lead Tech in a private lab. They are a growing company with a brand new lab facility. My client offers the opportunity to work the full range of histologic duties. You won't be spending your whole day cutting tissue!! You will be doing grossing and IHC as well. In addition they offer a very competitive salary, benefits and relocation assistance. If you have your Florida technologist's license great!! If not don't worry if you are ASCP certified the Florida license is easier to get than you think. And I can help you with that. 2. Histotechnologist in a hospital lab. They are located in a beautiful area of Florida and offer a great opportunity for a well rounded tech that can handle all areas of histologic duties. They offer a excellent salary benefits and relocation assistance. Again If you have your Florida technologist's license great!! If not don't worry if you are ASCP certified the Florida license is easier to get than you think. And I can help you with that. If you or anyone you know might be interested in hearing more about either or both of these jobs I can be reached at relia1@earthlink.net or toll free at 866-60RELIA (866-607-3542. Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net From david.kinsley <@t> spcorp.com Tue Jun 21 11:28:54 2005 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Required CEU's Message-ID: Is this true for everyone, or just those certified after a certain date? Dave -----Original Message----- From: Linda Jones [mailto:ljones@pathology.umsmed.edu] Sent: Tuesday, June 21, 2005 11:26 AM To: histonet@lists.utsouthwestern.edu Subject: Fwd: [Histonet] Required CEU's Yes, is that true? >>> "Dianne Holmes" 6/16/2005 11:04:04 AM >>> I have heard that there is a required number of CEU's for HT's to get within each year to maintain their certification. Is this so and if it is - how many? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Linda Harper-Jones BS.,HT/ HTL(ASCP) University of Mississippi Medical Center Department of Pathology Chief Histotechnologist Supervisor (601) 984-1576 (601) 984-4968 Fax ljones@pathology.umsmed.edu _______________________________________________ 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 plucas <@t> biopath.org Tue Jun 21 11:58:43 2005 From: plucas <@t> biopath.org (Paula Lucas) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Reagents Message-ID: <001301c57682$7c00f230$7c01a8c0@biopath.org> I appreciate any feedback on this subject: In efforts to reduce our costs, the company I work for is looking into the pricing of our supplies with the various vendors. We currently use Richard-Allan products, and I was asked to seek alternative stains and reagents for our processing and H&E staining. Can you tell me which H&E manufacturer you use on an automatic slide stainer? How about the alcohol, clearing and decolorizing agent? Thank you, Paula Lucas Bio-Path Medical Group Fountain Valley, CA From jcline <@t> wchsys.org Tue Jun 21 12:12:12 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] tissue dye Message-ID: <000101c57684$5df0ccf0$1d2a14ac@wchsys.org> I use Toluidine Blue 0.5% alcoholic to dye all our endoscopy, colonoscopy, prostate and cervical biopsies. The dye affords easier orientation on the cervical biopsies. It lasts through processing and does not affect the staining. The recipe is in the Ann Preece Histology book. ***** 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 emerald_lake77 <@t> yahoo.com Tue Jun 21 13:50:37 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Immunostainers -- in your opinion what's best and why? Message-ID: <20050621185037.72626.qmail@web31712.mail.mud.yahoo.com> Hi everyone, It's that time again when we need to make a list for big budget items and I am in need of a good automated immunostainer. I perform IHC (85%) and ISH (15%) all by hand and at this time it will help me a lot to get some of that work to become automated. So in your opion and based on your experiences, what is the best one out there and why (for both IHC and ISH, or compromise)? I am only familar with the Biogenex system. I learned a lot about the Biocare Nemesis system -- does anyone have any experience with this machine? Biocare claims its machine to be a nemesis ... but the price seems to also rival all others -- $77,000 YIKES. I have been to the NSH convention and discovered there are many others -- but without some experience there is no way I can actually know how good it works, how easy it is to use, how useful it will be and if it breaks down after 3 months. Thank you all for your responses. Sincerely, Gustave T. Hebert Scientist II Wyeth Research Cambridge MA --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football From Lori.Auth <@t> chsli.org Tue Jun 21 14:19:08 2005 From: Lori.Auth <@t> chsli.org (Auth, Lori) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] quality assurance Message-ID: please I need help!!!!!!!!! I am looking for a new quality assurance indicator for Histology. I currently am using specimen/requisition deficiencies. We have been compliant for over 2 years and I need to identify high risk, high volume error prone issue. I can't come up with something new to monitor. What does everyone else use??? Lori Auth, Lead Technician Histology Mercy Medical Center Rockville Centre, N.Y. From failm <@t> musc.edu Tue Jun 21 14:01:53 2005 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] MFH-2 antibody Message-ID: Hello All Has anyone heard of this antibody? Thanks in advance for your help Rena Fail From jcline <@t> wchsys.org Tue Jun 21 14:44:31 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Steve Coakley/toluidine blue Message-ID: <000401c57699$a4e45b30$1d2a14ac@wchsys.org> Toluidine Blue 0.5% Alcoholic Toluidine Blue 0.5 gm 20% reagent alcohol 100.0mL Combine and filter. We put one to two drops on aggregate of specimen. ***** 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 PMonfils <@t> Lifespan.org Tue Jun 21 14:44:34 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] MFH-2 antibody Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717555@lsexch.lsmaster.lifespan.org> I'm not familiar with the specific antibody, but MFH usually refers to malignant fibrous histiocytoma. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Mildred Fail > Sent: Tuesday, June 21, 2005 12:01 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] MFH-2 antibody > > Hello All > Has anyone heard of this antibody? > Thanks in advance for your help > Rena Fail > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From denise.woodward <@t> uconn.edu Tue Jun 21 15:22:05 2005 From: denise.woodward <@t> uconn.edu (Woodward, Denise) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] FIP, Neospora and Canine parvo Antibodies Message-ID: Hello everyone, Can anyone doing veterinary IHC for FIP, Neospora and Canine Parvo get in touch with me? I have some questions about these antibodies. Thanks and have a great day, Dwoodward University of Connecticut From Connie.Sims <@t> dchstx.org Tue Jun 21 15:23:18 2005 From: Connie.Sims <@t> dchstx.org (Connie Sims) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] quality assurance Message-ID: <11906C81B7B03E47A6F10CE5F4784449018A9C65@dchexch01.driscoll.dch> We used a chart check this year to find out what percentage of frozen section reports were being charted...we found out it was 0%! We use Copath and our frozen reports print directly to the OR. They are supposed to be charted. We'll be following this one for a while I think. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Auth, Lori Sent: Tuesday, June 21, 2005 2:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] quality assurance please I need help!!!!!!!!! I am looking for a new quality assurance indicator for Histology. I currently am using specimen/requisition deficiencies. We have been compliant for over 2 years and I need to identify high risk, high volume error prone issue. I can't come up with something new to monitor. What does everyone else use??? Lori Auth, Lead Technician Histology Mercy Medical Center Rockville Centre, N.Y. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ======================================================================================================== Driscoll Children's Hospital Disclaimer: This e-mail message and all information enclosed, 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 of the information enclosed, in its entirety or in part, is strictly prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and immediately destroy all copies of the message. For further assistance or questions please contact the email administrator at Driscoll Children's Hospital at (361) 694-5000. ========================================================================================================= From Charlene.Henry <@t> STJUDE.ORG Tue Jun 21 15:25:37 2005 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] quality assurance. . Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2099A1942@SJMEMXMB02.stjude.sjcrh.local> A few suggestions. 1. Frozen Section Turn Around Time (20 minute TAT) 2. Number of SS Repeated Tests (ours is a target of < 5% of total SS tests) 3. Number of IHC Repeated Tests (ours is a target of < 5% of total IHC tests) 3. Number of surgical cases completed after ____ AM (our target is < 5% after 9:00 AM) Hope these help. Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Auth, Lori Sent: Tuesday, June 21, 2005 2:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] quality assurance. . please I need help!!!!!!!!! I am looking for a new quality assurance indicator for Histology. I currently am using specimen/requisition deficiencies. We have been compliant for over 2 years and I need to identify high risk, high volume error prone issue. I can't come up with something new to monitor. What does everyone else use??? Lori Auth, Lead Technician Histology Mercy Medical Center Rockville Centre, N.Y. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ddittus787 <@t> aol.com Tue Jun 21 15:28:17 2005 From: ddittus787 <@t> aol.com (ddittus787@aol.com) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] quality assurance In-Reply-To: References: Message-ID: <8C744B6E480AD01-D74-134EA@mblk-r25.sysops.aol.com> Lori; did you try demographic failures by location, body part mis id (ie: rt hand versus left hand) req says one thing, bottle another? How about accessioning error rates? dana -----Original Message----- From: Auth, Lori To: histonet@lists.utsouthwestern.edu Sent: Tue, 21 Jun 2005 15:19:08 -0400 Subject: [Histonet] quality assurance please I need help!!!!!!!!! I am looking for a new quality assurance indicator for Histology. I currently am using specimen/requisition deficiencies. We have been compliant for over 2 years and I need to identify high risk, high volume error prone issue. I can't come up with something new to monitor. What does everyone else use??? Lori Auth, Lead Technician Histology Mercy Medical Center Rockville Centre, N.Y. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kbroomal <@t> NEMOURS.ORG Tue Jun 21 16:09:43 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Wrinkles in cartilage sections Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824356@wlmmsx02.nemours.org> I need some "O wise one" suggestions! I'm sectioning sheep trachea & the cartilage layer is coming out full of wrinkles. The epithelium layer is nice, flat & happy (it is, I asked it), as is the muscle. The tissue is FFPE and I'm cutting at 5-6 um. The sections look good in the ribbon, but as soon as they hit the water bath, the cartilage wrinkles up. I've tried: *ethanol in the water bath *ammonia in the water bath *tween in the water bath *soaking the paraffin blocks in ammonia water before cutting *increased water bath temperature Suggestions??? Kristen Broomall, HT (ASCP) From algranth <@t> u.arizona.edu Tue Jun 21 16:21:04 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Oil Red O Message-ID: <4.3.2.7.2.20050621141017.0252eed8@algranth.inbox.email.arizona.edu> I know that this stain has been discussed many times on histonet but I have a question that may not have been asked before: The tissue I am working with is mouse liver from mice who are fed diets very high in fat or alcohol. This makes for very, very fatty livers. So much so that when stained with Oil Red O that the sections just ooze red. If we look at the slides and do quantitative analysis and/or photographs right away there is little or no problem but if the investigator waits a few days before looking at the slides they are a mess. Control slides are OK because they contain a normal amount of fatty tissue. We have also found that doing the hematoxylin counterstain reduces the amount of Oil Red Staining so we do a set counterstained and one without for quantitative analysis. Is there a way to extend the life of this stain when they can't be viewed immediately? 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 shive003 <@t> umn.edu Tue Jun 21 16:50:42 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] IHC for Clostridium perfringens Message-ID: <00e301c576ab$459eb000$41065486@auxs.umn.edu> Does anyone out there do IHC for Clostridium perfringens? If so, would you please point me in the direction of where I could locate an antibody? Thanks in advance. Jan Shivers U of MN Vet Diag Lab From Eric <@t> ategra.com Thu Jun 16 17:40:31 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] HistoTech Manager Job Opening Message-ID: Histonetters - My name is Eric Dye and I am the new Histo Tech recruiter here at Ategra. I took over all Histo Tech recruiting since Pam Barker retired. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job opening is in Central Ohio seeking a HistoTech Manager. I also have bench histo tech jobs available in: Arizona, Pennsylvania, Illinois, New York, Missouri, Georgia, Iowa, Rhode Island, and Florida.. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- 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 Eric <@t> ategra.com Thu Jun 16 17:40:31 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] HistoTech Manager Job Opening Message-ID: Histonetters - My name is Eric Dye and I am the new Histo Tech recruiter here at Ategra. I took over all Histo Tech recruiting since Pam Barker retired. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job opening is in Central Ohio seeking a HistoTech Manager. I also have bench histo tech jobs available in: Arizona, Pennsylvania, Illinois, New York, Missouri, Georgia, Iowa, Rhode Island, and Florida.. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- 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 Eric <@t> ategra.com Thu Jun 16 17:40:31 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] HistoTech Manager Job Opening Message-ID: Histonetters - My name is Eric Dye and I am the new Histo Tech recruiter here at Ategra. I took over all Histo Tech recruiting since Pam Barker retired. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job opening is in Central Ohio seeking a HistoTech Manager. I also have bench histo tech jobs available in: Arizona, Pennsylvania, Illinois, New York, Missouri, Georgia, Iowa, Rhode Island, and Florida.. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- 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 louise.renton <@t> gmail.com Wed Jun 22 01:58:50 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Wrinkles in cartilage sections In-Reply-To: <6E41111281623B4B8A9AB8F9A7EA3437824356@wlmmsx02.nemours.org> References: <6E41111281623B4B8A9AB8F9A7EA3437824356@wlmmsx02.nemours.org> Message-ID: Kristen, try placing your slide briefly on a hot plate after picking the section up from the waterbath before allowing it to drain. This used to work on lungs with bronchus, so i gues it might do for you. The only problem you might encounter is if the tissue is poorly processes in which case the other stuff might "blow-up" Hope this is some help On 6/21/05, Kristen Broomall wrote: > I need some "O wise one" suggestions! > > I'm sectioning sheep trachea & the cartilage layer is coming out full of > wrinkles. The epithelium layer is nice, flat & happy (it is, I asked it), as > is the muscle. The tissue is FFPE and I'm cutting at 5-6 um. The sections > look good in the ribbon, but as soon as they hit the water bath, the > cartilage wrinkles up. > > I've tried: > > *ethanol in the water bath > *ammonia in the water bath > *tween in the water bath > *soaking the paraffin blocks in ammonia water before cutting > *increased water bath temperature > > Suggestions??? > > Kristen Broomall, HT (ASCP) > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Jun 22 02:53:50 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] RE: dyeing specific tissues for processing Message-ID: A method we use when processing tiny biopsies is to wrap them in a small piece of "speci-wrap" tissue like a parcel, then place parcel into a biopsy basket to hold it, place a couple of drops of 1% methylene blue onto the parcel and leave for a few seconds. Remove the parcel and, holding onto it carefully, shwish (a technical term) in a beaker of water for a few secs to wash. Place back in the basket, put lid on, place in cassette etc and process. The dye is enough to lightly stain the tissue so when you unwrap it, you can see it to embed. Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From bruyntjes <@t> voeding.tno.nl Wed Jun 22 05:02:22 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] (no subject) Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079DD9@ntexch1.voeding.tno.nl> Hi all I was wondering if anyone of you is used to cut non-fixed cryoslides of brain tissue of about 30-40 micron thick. I've tried a few peaces on a rather high temperature (approximately 12?C), but it did not make me smile? J.P. Bruijntjes TNO Quality of Life Zeist Holland This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From abright <@t> brightinstruments.com Wed Jun 22 05:35:11 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] (no subject) Message-ID: Dear Mr Bruijntjes, I am assuming your brain tissue and microtome chamber are at -12?C not 12?c, If you lowered only the chamber temperature to -20?C to lower the knife and anti-roll plate it will return your smile. 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: Bruijntjes, J.P. [mailto:bruyntjes@voeding.tno.nl] Sent: 22 June 2005 11:02 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi all I was wondering if anyone of you is used to cut non-fixed cryoslides of brain tissue of about 30-40 micron thick. I've tried a few peaces on a rather high temperature (approximately 12?C), but it did not make me smile? J.P. Bruijntjes TNO Quality of Life Zeist Holland This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Jun 22 07:07:02 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] RE: here I go again Message-ID: In reply to Joe Nocito's message re- use of expired antisera, and using antisera frozen in 1080 - is this BC or AD, and were they frozen in the arctic or Antarctic! I agree wholeheartedly about using (with proper monitoring) expired antisera, but I bet even the top firms would balk at putting an viability period of 910 years1 Only pulling your leg, Joe! I once made a similar typo and felt such a plank after I'd e-mailed it to our institute message forum! Jacqui Lancaster DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From kmerriam2003 <@t> yahoo.com Wed Jun 22 07:23:48 2005 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Immunostainers -- in your opinion what's best and why? In-Reply-To: <20050621185037.72626.qmail@web31712.mail.mud.yahoo.com> Message-ID: <20050622122348.80418.qmail@web50304.mail.yahoo.com> Hello, I purchased a Biocare Nemesis stainer a few months ago, and I absolutely love it. Being in research, it is important to have a stainer that is open, meaning I can use any reagent that I want to. This machine allows me to do just that, I can use any antibody or detection system with this stainer. You can bar code the reagents or the slides, or both. The pricing is pretty competitive, especially considering it has a big 84-slide per run capacity. Also - the support I get from this company is phenomenal! Kim Merriam Novartis Cambridge, MA GT Hebert wrote: Hi everyone, It's that time again when we need to make a list for big budget items and I am in need of a good automated immunostainer. I perform IHC (85%) and ISH (15%) all by hand and at this time it will help me a lot to get some of that work to become automated. So in your opion and based on your experiences, what is the best one out there and why (for both IHC and ISH, or compromise)? I am only familar with the Biogenex system. I learned a lot about the Biocare Nemesis system -- does anyone have any experience with this machine? Biocare claims its machine to be a nemesis ... but the price seems to also rival all others -- $77,000 YIKES. I have been to the NSH convention and discovered there are many others -- but without some experience there is no way I can actually know how good it works, how easy it is to use, how useful it will be and if it breaks down after 3 months. Thank you all for your responses. Sincerely, Gustave T. Hebert Scientist II Wyeth Research Cambridge MA --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football From pathrm35 <@t> adelphia.net Wed Jun 22 08:10:22 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] hair shaft and hair bulb procedures Message-ID: <27634996.1119445822779.JavaMail.root@web5.mail.adelphia.net> Fellow Techs, Does anyone have procedures for microscopic hair shaft evaluation and microscopic hair bulb evaluation they are willing to share? Also, what do you use for controls? Thanks, Ron Martin fax 561-721-1249 From ajennings <@t> unmc.edu Wed Jun 22 08:42:16 2005 From: ajennings <@t> unmc.edu (Anita Jennings) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Oil Red O mountant Message-ID: Andi I had the same problem. initially with my fat stain.....Poly Scientific kit K043 contains the glycerine jelly mounting medium specific for Oil Red coverslipping, havent had a problem since I started ordering this kit 800-645-5825 From Kemlo.Rogerson <@t> elht.nhs.uk Wed Jun 22 09:15:21 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] RE: here I go again[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD318@elht-exch1.xelht.nhs.uk> In the Private Lab that I worked in a few years back they used 'old' reagents on their Analysers; they had to fool the bar code reader and software, but their claim was that if the controls were OK, then what is the problem? They failed to tell CPA that this was standard practice. -----Original Message----- From: Malam Jacqueline (Morecambe Bay Hospitals NHS Trust) Sent: 22 June 2005 13:07 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: here I go again[Scanned] In reply to Joe Nocito's message re- use of expired antisera, and using antisera frozen in 1080 - is this BC or AD, and were they frozen in the arctic or Antarctic! I agree wholeheartedly about using (with proper monitoring) expired antisera, but I bet even the top firms would balk at putting an viability period of 910 years1 Only pulling your leg, Joe! I once made a similar typo and felt such a plank after I'd e-mailed it to our institute message forum! Jacqui Lancaster DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Wed Jun 22 09:23:48 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] hair shaft and hair bulb procedures In-Reply-To: <27634996.1119445822779.JavaMail.root@web5.mail.adelphia.net> Message-ID: Ron, funny you should ask. I've been pulling my hair out (no pun intended) trying to get these things oriented. My dermpath was impressed with our last attempt. We use Histogel by Richard Allen to orient the hairs and process as usual. Since the orientation is performed at the grossing table, all the embedders have to do is put the hair enbedded in Histogel right into a paraffin block. We cut three levels and perforn a PAS/Fungus on each level. Hope this helps. I have hair growing back now. 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 pathrm35@adelphia.net Sent: Wednesday, June 22, 2005 8:10 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] hair shaft and hair bulb procedures Fellow Techs, Does anyone have procedures for microscopic hair shaft evaluation and microscopic hair bulb evaluation they are willing to share? Also, what do you use for controls? Thanks, Ron Martin fax 561-721-1249 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From koellinr <@t> amgen.com Wed Jun 22 09:40:51 2005 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Immunostainers -- in your opinion what's best and why? Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CC792@wa-mb4-sea.amgen.com> Gustave, If you will be at Nationals in Ft. Lauderdale in Sept. , our lab will have 2 posters describing and detailing an automated immunohistochemistry platform by TECAN. Can run 144 IHC slides at one time. Although we haven't used it for such, it is set up and capable to also do that many ISH slides. Completely open. Ray Raymond Koelling Research Scientist Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of GT Hebert Sent: Tuesday, June 21, 2005 11:51 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Immunostainers -- in your opinion what's best and why? Hi everyone, It's that time again when we need to make a list for big budget items and I am in need of a good automated immunostainer. I perform IHC (85%) and ISH (15%) all by hand and at this time it will help me a lot to get some of that work to become automated. So in your opion and based on your experiences, what is the best one out there and why (for both IHC and ISH, or compromise)? I am only familar with the Biogenex system. I learned a lot about the Biocare Nemesis system -- does anyone have any experience with this machine? Biocare claims its machine to be a nemesis ... but the price seems to also rival all others -- $77,000 YIKES. I have been to the NSH convention and discovered there are many others -- but without some experience there is no way I can actually know how good it works, how easy it is to use, how useful it will be and if it breaks down after 3 months. Thank you all for your responses. Sincerely, Gustave T. Hebert Scientist II Wyeth Research Cambridge MA --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emerald_lake77 <@t> yahoo.com Wed Jun 22 10:14:08 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Immunostainers -- in your opinion what's best and why? In-Reply-To: <16834C6DFFA6004C88DE4507FB8AE544018CC792@wa-mb4-sea.amgen.com> Message-ID: <20050622151408.16519.qmail@web31704.mail.mud.yahoo.com> That's amazing ... do you know how much that system costs?? .. right now I am just handing in info regarding the amount of money I need, so the cost is somewhat crucial though I am not sure what system I am interested in yet.... once I get approved for the money, I will look into the machines and eventually choose one. Right now though, I just don't want to say I need an immunostainer and say it costs around 70,000 when in fact, the one I want is 120,000. So, yes, I will be in FL and will check out that poster. Thank you for your reply. Gustave "Koelling, Ray" wrote: Gustave, If you will be at Nationals in Ft. Lauderdale in Sept. , our lab will have 2 posters describing and detailing an automated immunohistochemistry platform by TECAN. Can run 144 IHC slides at one time. Although we haven't used it for such, it is set up and capable to also do that many ISH slides. Completely open. Ray Raymond Koelling Research Scientist Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of GT Hebert Sent: Tuesday, June 21, 2005 11:51 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Immunostainers -- in your opinion what's best and why? Hi everyone, It's that time again when we need to make a list for big budget items and I am in need of a good automated immunostainer. I perform IHC (85%) and ISH (15%) all by hand and at this time it will help me a lot to get some of that work to become automated. So in your opion and based on your experiences, what is the best one out there and why (for both IHC and ISH, or compromise)? I am only familar with the Biogenex system. I learned a lot about the Biocare Nemesis system -- does anyone have any experience with this machine? Biocare claims its machine to be a nemesis ... but the price seems to also rival all others -- $77,000 YIKES. I have been to the NSH convention and discovered there are many others -- but without some experience there is no way I can actually know how good it works, how easy it is to use, how useful it will be and if it breaks down after 3 months. Thank you all for your responses. Sincerely, Gustave T. Hebert Scientist II Wyeth Research Cambridge MA --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Jun 22 10:20:38 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Cytology fibrin block preps Message-ID: Hi Someone recently mentioned a method of preparing blocks from cytology preps using fibrin. Please would they mention the method again - I can't just locate it. Many thanks Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From dmccaig <@t> ckha.on.ca Wed Jun 22 10:25:36 2005 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Cassette labellers Message-ID: <3E5A3F039F0BD8118B4700C00D0020240435E6@CKHA9> We are having problems with the secureline markers fading considerably during processing. I am trying to present a business case for a cassette labeller. Does anyone have any opinions they would like to share. Do you use your stand alone or can they be interfaced into a pathology program (tamtron, etc) Did you notice a significant difference in pricing of the cassettes from bulk to sleeves? I would appreciate any information that could be provided. Diana McCaig, MLT From josephnerk <@t> hotmail.com Wed Jun 22 10:42:31 2005 From: josephnerk <@t> hotmail.com (Joseph Nerk) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Zues-Michel fixative Message-ID: Hello all Is anyone out there is in Histonet can shed any like on Zues- Michel fixative? I have a renal biopsy for immunofluorescence, and this fixative was recommended. Thank you all Erskine _________________________________________________________________ Express yourself instantly with MSN Messenger! [1]MSN Messenger Download today it's FREE! References 1. http://g.msn.com/8HMBEN/2734??PS=47575 From djohnson14 <@t> hotmail.com Wed Jun 22 10:44:31 2005 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Liquid Particle Count In-Reply-To: <3E5A3F039F0BD8118B4700C00D0020240435E6@CKHA9> Message-ID: Does anyone know what LPC (liquid particulate count) is acceptable for plastics used in cytology cell collection. And specifically the max quantity for a particular size particle. Thanks Dave From TJJ <@t> Stowers-Institute.org Wed Jun 22 10:53:54 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Re: Immunostainers -- in your opinion what's best Message-ID: Gustave, Contact the vendors (or just wait, because no doubt they'll be contacting you!) and find out the list price on their instrumentation and make arrangements for demos. Try before you buy and you will find a unit you absolutely cannot live without. All of the commercially available immunostainers are good at what they do, some just have different bells and whistles. You will have to decide yourself which one works best for your application. Many companies are amenable to knocking some $$$s off the purchase price with reagent purchase agreements. Otherwise, I would budget for roughly $100k, and likely you'll be able to bring the bid in for much less. Better to go that way than to beg for more money later. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From krat18 <@t> aol.com Wed Jun 22 11:22:09 2005 From: krat18 <@t> aol.com (krat18@aol.com) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] RE: dyeing specific tissues for processing In-Reply-To: References: Message-ID: <8C7455DAC39580D-84-17638@mblk-r28.sysops.aol.com> Don't think I've seen this mentioned yet, but our pathologists just drop hematoxylin on the tiny biopsies when they're grossing, put them on sponges or wrap in paper and drop them into the formalin. No rinsing. Works great. Karen_Raterman@ssmhc.com krat18@aol.com -----Original Message----- From: Malam Jacqueline To: Histonet submissions (histonet@lists.utsouthwestern.edu) Sent: Wed, 22 Jun 2005 08:53:50 +0100 Subject: [Histonet] RE: dyeing specific tissues for processing A method we use when processing tiny biopsies is to wrap them in a small piece of "speci-wrap" tissue like a parcel, then place parcel into a biopsy basket to hold it, place a couple of drops of 1% methylene blue onto the parcel and leave for a few seconds. Remove the parcel and, holding onto it carefully, shwish (a technical term) in a beaker of water for a few secs to wash. Place back in the basket, put lid on, place in cassette etc and process. The dye is enough to lightly stain the tissue so when you unwrap it, you can see it to embed. Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lleach <@t> uic.edu Wed Jun 22 11:30:40 2005 From: lleach <@t> uic.edu (Lu Leach) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Fekete's soution Message-ID: <42B99230.7010701@uic.edu> Dear Netters, Does anyone have information on Fekete's Solution? I have exhausted by library trying to find it's composition and use. Thank you, Lu Leach University of Illinois at Chicago From juan.gutierrez <@t> christushealth.org Wed Jun 22 11:38:23 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Fekete's soution Message-ID: It's a fixative containing: acetic acid, formaldehyde and alcohol. 37-40% Formaldehyde...................10ml Glacial acetic acid....................5ml 70% Alcohol..........................100ml Hope this helps. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lu Leach Sent: Wednesday, June 22, 2005 11:31 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fekete's soution Dear Netters, Does anyone have information on Fekete's Solution? I have exhausted by library trying to find it's composition and use. Thank you, Lu Leach University of Illinois at Chicago _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Wed Jun 22 11:40:48 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Fekete's soution Message-ID: I forgot to mention the uses. It's supposed to be excellent for glycogen preservation and good cytoplasmic fixative when working with RNA. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lu Leach Sent: Wednesday, June 22, 2005 11:31 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fekete's soution Dear Netters, Does anyone have information on Fekete's Solution? I have exhausted by library trying to find it's composition and use. Thank you, Lu Leach University of Illinois at Chicago _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bhewlett <@t> cogeco.ca Wed Jun 22 11:43:15 2005 From: bhewlett <@t> cogeco.ca (bhewlett@cogeco.ca) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Fekete's soution Message-ID: <42b99523.95.2f05.28353@cogeco.ca> > Lu, You will find the composition of Fekete's fixative in Lillie's textbook. An approximation is Alcohol 70 mL Formalin(37% formaldehyde) 10mL Acetic acid 5 mL Water to make 100 mL Regards, Bryan > Dear Netters, > > Does anyone have information on Fekete's Solution? I have exhausted by > library trying to find it's composition and use. > > Thank you, > > Lu Leach > University of Illinois at Chicago > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tmmrosla <@t> healtheast.org Wed Jun 22 12:06:40 2005 From: tmmrosla <@t> healtheast.org (Mrosla, Tina M) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] xylene vs. xylene substitutes Message-ID: <22E6C4C32AFB3E44B7056C7EC3E1F20452AFB6@HECLUSTER.HEALTHEAST.LOC> I was wondering what kind of experience people have had with xylene substitutes. I work in a medium-volume hospital laboratory that would like to change to a xylene substitute wherever we can without compromising quality. It probably would have no effect on the tissue processors, but how about the automatic stainer (I should mention that we have a Sakura tape coverslipper)? Hemo-D is not an option because some employees have hypersensitivity to it, but how about Americlear or any others? Tina Mrosla tmmrosla@healtheast.org The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. From ploykasek <@t> phenopath.com Wed Jun 22 12:20:47 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Zues-Michel fixative In-Reply-To: Message-ID: Hi Joseph. Here is our formula for IF transport media ? otherwise known as Zeus or Michel?s or... I just copied & pasted from our reagent prep. Note that this reagent does have a shelf life. I?d be happy to answer any further questions on this reagent. Patti Loykasek PhenoPath Laboratories PURPOSE Transport media is used as a holding media for tissue biopsies. Transport Media prevents degradation of tissue and immune complexes by inhibiting tissue enzymes. Tissue can be held in transport media up to 2 weeks without harm to the tissue. The various components of the buffer and media have the following effects Ammonium sulfate precipitates macromolecules including Antigen-Antibody complexes without loss of antigenicity, while proteolytic and other depolymerizing enzymes would be precipitated and inhibited. A role for such enzymes in dermal-epidermal separation has been postulated. Citrate buffer is used for washing because it is least likely to facilitate dermal-epidermal separation. Magnesium sulfate neutralizes the chelating capacity of the citrate. n-Ethylmaleimide minimizes proteolytic activity. .REAGENTS 1. Ammonium Sulfate, granular (NH4)2SO4 (VWR, Catalog No. MK351206 or equivalent). HMIS: 2-0-0 2. Deionized Water, NCCLS type I 3. 1.0M Potassium Hydroxide HMIS:3-0-2 4. Transport Buffer HMIS: 2-0-0 PROCEDURE 1. Transfer 250mL of transport buffer to a large container. 2. Weigh 137.5g Ammonium Sulfate and transfer to 250mL transport buffer 3. Mix the solution well and adjust the pH to 7.2 to 7.5 with 1M Potassium Hydroxide (1M KOH). Note: The higher pH will extend the shelf life. Transport media with a pH below 7.0 and above 7.5 will give unsatisfactory results 4. Label secondary container with the following information: Name, pH, Date Prepared, Storage Conditions, Expiration Date (6 months), HMIS 2-0-0, and initials. 5. Transport Media is stored at room temperature with a shelf life of 6 months. REFERENCE DOCUMENTS 1. HazCom Training Handbook 2. Material Safety Data Sheet (MSDS) 3. RP057, 1.0M Potassium Hydroxide Stock Solution 4. RP041, Transport Buffer from Stock Solutions 5. 1973 Journal of Investigative Dermatology Vol59. No. 6 ?Preservation of tissue fixed immunoglobins in Skin Biopies of patients with Lupus Erythemalosis and bullous diseases. Prelim report Authors (Micheal Miller and Davied 6. Immunohistochemistopathology, A Practical Approach to Diagnosis. Jules M. Elias, 1990, ASCP Press 7. Chemical SOP SAFETY CAUTION: Ammonium sulfate can be an allergen and an irritant. Transport buffer is toxic and can be an irritant. Potassium Hydroxide is caustic and should be stored in a base cabinet. PROCEDURE 1. Transfer 250mL of transport buffer to a large container. 2. Weigh 137.5g Ammonium Sulfate and transfer to 250mL transport buffer 3. Mix the solution well and adjust the pH to 7.2 to 7.5 with 1M Potassium Hydroxide (1M KOH). Note: The higher pH will extend the shelf life. Transport media with a pH below 7.0 and above 7.5 will give unsatisfactory results 4. Label secondary container with the following information: Name, pH, Date Prepared, Storage Conditions, Expiration Date (6 months), HMIS 2-0-0, and initials. 5. Transport Media is stored at room temperature with a shelf life of 6 months. > Hello all > > Is anyone out there is in Histonet can shed any like on Zues- Michel > fixative? > > I have a renal biopsy for immunofluorescence, and this fixative was > recommended. > > Thank you all > > Erskine > _________________________________________________________________ > > Express yourself instantly with MSN Messenger! [1]MSN Messenger > Download today it's FREE! > > References > > 1. http://g.msn.com/8HMBEN/2734??PS=47575 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Wed Jun 22 12:26:23 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Re: Immunostainers -- in your opinion what's best In-Reply-To: Message-ID: > Gustave, I was wondering how many slides you are planning on running in a day? I think this information is important when considering a stainer. Perhaps all the bells & whistles are not necessary if you are not running a large amount of slides & are on a restricted budget. I have extensively used the Dako Autostainer with good results, however it does not do ISH. I am anxiously awaiting the release of the new Dako stainer ?Eridan?. For ISH we have a couple of different techniques depending on the volume. When we are doing small numbers of ISH we use a manual system that is the Microprobe. It uses capillary gap action (so it is best to use special capillary gap slides) and can do 10 pairs of slides at a time. It uses very little reagent. I?d be happy to answer more questions. > Patti Loykasek PhenoPath Laboratories > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Wed Jun 22 12:48:06 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Zues-Michel fixative In-Reply-To: Message-ID: <000001c57752$8c99cb00$1d2a14ac@wchsys.org> You can order this fixative from Wampole Lab,Inc, order #0102, 12 bottles per box. Pricing is good also. ************************************************************************ * Hello all Is anyone out there is in Histonet can shed any like on Zues- Michel fixative? I have a renal biopsy for immunofluorescence, and this fixative was recommended. Thank you all Erskine _________________________________________________________________ Express yourself instantly with MSN Messenger! [1]MSN Messenger Download today it's FREE! References 1. http://g.msn.com/8HMBEN/2734??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** 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 GDawson <@t> dynacaremilwaukee.com Wed Jun 22 12:46:02 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Zues-Michel fixative Message-ID: Erskine, As I understand it, these two "fixatives" are more like transport mediums. I use them to get renal bx's to me from outside hospitals so that I can perform IF testing on them. They don't "fix" the tissues (this is why they are good for IF procedures since they are geared for unfixed tissue), but they preserve them for transport or maybe overnight in a fridge. I don't like to keep the bx's in these transport mediums for longer than I have to. Just remember to rinse out the bx's with a tris buffer thoroughly before you cryosection them. Good Luck, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: Joseph Nerk [mailto:josephnerk@hotmail.com] Sent: Wednesday, June 22, 2005 9:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Zues-Michel fixative Hello all Is anyone out there is in Histonet can shed any like on Zues- Michel fixative? I have a renal biopsy for immunofluorescence, and this fixative was recommended. Thank you all Erskine _________________________________________________________________ Express yourself instantly with MSN Messenger! [1]MSN Messenger Download today it's FREE! References 1. http://g.msn.com/8HMBEN/2734??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hymclab <@t> hyhc.com Wed Jun 22 13:05:43 2005 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] xylene vs. xylene substitutes Message-ID: We have been using Thermo's Histosolve for approx. 12 years now and love it. I have another Xylene related question to pose: What is everyone doing with their Xylene waste if you don't have a recycler? We don't have much but what we do have we use a 5 gal. catch can with a saftey lid. When that is full we have housekeeping take it and dump it in a 55 gal. drum. When the drum is full we have a waste hauler come and take it away. Risk Management is questioning our decision to have housekeeping transfer it into the drum. I would appreciate your comments on this. Thanks, Dawn -----Original Message----- From: Mrosla, Tina M [mailto:tmmrosla@healtheast.org] Sent: Wednesday, June 22, 2005 12:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] xylene vs. xylene substitutes I was wondering what kind of experience people have had with xylene substitutes. I work in a medium-volume hospital laboratory that would like to change to a xylene substitute wherever we can without compromising quality. It probably would have no effect on the tissue processors, but how about the automatic stainer (I should mention that we have a Sakura tape coverslipper)? Hemo-D is not an option because some employees have hypersensitivity to it, but how about Americlear or any others? Tina Mrosla tmmrosla@healtheast.org The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emry <@t> u.washington.edu Wed Jun 22 13:05:41 2005 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] bone clearing Message-ID: Do any of you find one clearing product works better on bone than others? Thanks, Trisha U of WA Seattle From Jackie.O'Connor <@t> abbott.com Wed Jun 22 13:26:31 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] xylene vs. xylene substitutes Message-ID: I would think that if housekeeping personnel are handling waste xylene, they would be subject to STEL monitoring - I'll betcha this hasn't been done. Also, one would hope that they are trained in the use of anti-static devices when decanting such a volatile solution into a 55 gal drum. One static spark is all it takes. boom. hymclab Sent by: histonet-bounces@lists.utsouthwestern.edu 06/22/2005 01:05 PM To: "'Mrosla, Tina M'" , histonet@lists.utsouthwestern.edu cc: Subject: RE: [Histonet] xylene vs. xylene substitutes We have been using Thermo's Histosolve for approx. 12 years now and love it. I have another Xylene related question to pose: What is everyone doing with their Xylene waste if you don't have a recycler? We don't have much but what we do have we use a 5 gal. catch can with a saftey lid. When that is full we have housekeeping take it and dump it in a 55 gal. drum. When the drum is full we have a waste hauler come and take it away. Risk Management is questioning our decision to have housekeeping transfer it into the drum. I would appreciate your comments on this. Thanks, Dawn -----Original Message----- From: Mrosla, Tina M [mailto:tmmrosla@healtheast.org] Sent: Wednesday, June 22, 2005 12:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] xylene vs. xylene substitutes I was wondering what kind of experience people have had with xylene substitutes. I work in a medium-volume hospital laboratory that would like to change to a xylene substitute wherever we can without compromising quality. It probably would have no effect on the tissue processors, but how about the automatic stainer (I should mention that we have a Sakura tape coverslipper)? Hemo-D is not an option because some employees have hypersensitivity to it, but how about Americlear or any others? Tina Mrosla tmmrosla@healtheast.org The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eca9 <@t> georgetown.edu Wed Jun 22 13:30:14 2005 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Her2 antibody Message-ID: <42B9AE36.9010409@georgetown.edu> Hello, I just ordered mouse anti-Her2 cat.no. 18-7107 from Zymed. There is however no indication given as to which dilution to use. I want to use it for Immunohistochemistry on paraffin embedded sections. It also doesn't say anything about antigen retrieval. Is anyone using this antibody and if so could you tell me what you do? Thanks, Eva Andersson Georgetown University From sharon.osborn <@t> dnax.org Wed Jun 22 13:33:21 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Frozen Tissue Bank tissues Message-ID: <29B25753F6B1D51196110002A589D4440239804E@PALMSG30.us.schp.com> Histonetters, Question: What methods or technics do you use when retrieving a small piece of tissue from a large piece in a -80 freezer? There is only one large piece (quarter sized) smished into a 100 ml centrifuge tube and stored in the -80. Once it is dislodged from the tube, how do you proceed to cut off a small piece (3-4 cm diameter) in a safe contained way? One technic is to wrap the piece in foil, have a small cutting board, a hammer or rubber mallet and a small, sharp chisel. What methods would be used to keep tissue pieces from flying about contaminating the environment? Wrapping the piece in foil does contain a lot of the debris. Of course, the tech is using standard safety procedures with gloves, glasses or face shield, lab coat, etc. Thanks for your input. sharon osborn dnax schering plough biopharma palo alto, ca ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From lleach <@t> uic.edu Wed Jun 22 13:36:16 2005 From: lleach <@t> uic.edu (Lu Leach) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] thank you Message-ID: <42B9AFA0.9080409@uic.edu> Many thanks to all for the help with Fekete's solution. Regards, Lu Leach From Bauer.Karen <@t> mayo.edu Wed Jun 22 13:57:21 2005 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] CDX2 Message-ID: Does anyone in histoland have a working protocol for CDX2? I have a pre-dilute from BioGenex that I'm trying to use on our BenchMark, but nothing seems to work. I've called Ventana and BioGenex and have tried their suggestions, but with no luck. Anybody willing to share their protocol? I'd love to give it a try. Thanks much!! Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From liz <@t> premierlab.com Wed Jun 22 14:01:32 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] bone clearing In-Reply-To: Message-ID: <000901c5775c$cdd21140$a7d48a80@AMY> Trisha We process a lot of bone (rat, guinea pig, dog, sheep joints, etc) and still use xylene as a clearing agent. Our processor has three xylene stages on it. I have used Hemo De in the past and it worked well if I am remembering correctly. To be honest we have not tried many xylene subsititutes, we did once and it was a disaster. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Trisha Emry Sent: Wednesday, June 22, 2005 11:06 AM To: histo Subject: [Histonet] bone clearing Do any of you find one clearing product works better on bone than others? Thanks, Trisha U of WA Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Mara.Fairman <@t> RoperSaintFrancis.com Wed Jun 22 14:03:36 2005 From: Mara.Fairman <@t> RoperSaintFrancis.com (Fairman Mara) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Looking for a copy of the 1st edition Message-ID: One of our pathologists is searching for a copy of the 1st edition of the Quality Improvement Manual in Anatomic Pathology put out by the CAP (College of American Pathologists) and edited by Raouf E. Nakhleh, M.D. and Patrick L. Fitzgibbons, M.D. We have the 2nd edition and the new one they came out with this year, but he is looking for the 1st to research some issue. If you have one that you are wiling to part with, let me know and include your phone number. Thanks! Mara Fairman Anatomic Pathology Supervisor Roper Hospital 316 Calhoun Street Charleston, S.C. 29401 843-724-2264 Fax- 843-724-2356 Beeper 764-1489 mara.fairman@ropersaintfrancis.com Upgrade Your Email - Click here! Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain confidential and/or legally privileged information. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Any unauthorized review, use, disclosure, or distribution is prohibited. Thank you. From settembr <@t> umdnj.edu Wed Jun 22 14:07:45 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] CDX2 Message-ID: Karen, I use BioGenex' pre-diluted Ab also. My control is a colon cancer, my pretreatment is DakoCytomation's Target Retrieval Soln 40 minutes in a steamer, Dako's LSAB2 detection kit, and their DAB kit. It looks great. Dana Settembre University Hospital - UMDNJ Newark, NJ >>> "Bauer, Karen" 6/22/2005 2:57:21 PM >>> Does anyone in histoland have a working protocol for CDX2? I have a pre-dilute from BioGenex that I'm trying to use on our BenchMark, but nothing seems to work. I've called Ventana and BioGenex and have tried their suggestions, but with no luck. Anybody willing to share their protocol? I'd love to give it a try. Thanks much!! Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ejandrew <@t> med.unc.edu Wed Jun 22 14:18:08 2005 From: ejandrew <@t> med.unc.edu (Liz Andrews) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] histotechnologist looking for work Message-ID: <000101c5775f$1ff4e9f0$0b2e1398@peds.med.unc.edu> Hi, my name is Liz Andrews. I am currently working in a histology lab at UNC-Chapel Hill. I am interested in relocating to St. Louis, MO. I have 3 and a half years of full-time experience over a 5-year span. I am in the middle of getting my HTL certification. I have passed the written part and am currently working on the practical portion. I have some supervisory duties with part-time employees along with training responsibilities. For a copy of my resume, you can contact me at ejandrew@med.unc.edu or 919-360-0910. Thank you and I look forward to moving to St. Louis. Go Rams! Liz Andrews UNC-Chapel Hill CF Center Chapel Hill, NC From vazquezr <@t> ohsu.edu Wed Jun 22 14:20:38 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Zues-Michel fixative Message-ID: Are you talking about Zeus transport (fixative) medium? I use it. It is a great transport medium. I believe that Michels is another brand. My doctor didn't care for it as well as Zeus. Robyn OHSU From krat18 <@t> aol.com Wed Jun 22 14:29:43 2005 From: krat18 <@t> aol.com (krat18@aol.com) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] xylene vs. xylene substitutes In-Reply-To: <22E6C4C32AFB3E44B7056C7EC3E1F20452AFB6@HECLUSTER.HEALTHEAST.LOC> References: <22E6C4C32AFB3E44B7056C7EC3E1F20452AFB6@HECLUSTER.HEALTHEAST.LOC> Message-ID: <8C74577E0191CB1-B30-196F5@FWM-R28.sysops.aol.com> Just a note: the rep from CBG Biotech told me that Formula 83 (which, as I understand, is mainly naphtha) does NOT work for the coverslipping tape, so you'd have to use xylene only on the coverslipper. Otherwise, the Formula 83 works fine. Karen_Raterman@ssmhc.com krat18@aol.com -----Original Message----- From: Mrosla, Tina M To: histonet@lists.utsouthwestern.edu Sent: Wed, 22 Jun 2005 12:06:40 -0500 Subject: [Histonet] xylene vs. xylene substitutes I was wondering what kind of experience people have had with xylene substitutes. I work in a medium-volume hospital laboratory that would like to change to a xylene substitute wherever we can without compromising quality. It probably would have no effect on the tissue processors, but how about the automatic stainer (I should mention that we have a Sakura tape coverslipper)? Hemo-D is not an option because some employees have hypersensitivity to it, but how about Americlear or any others? Tina Mrosla tmmrosla@healtheast.org The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eca9 <@t> georgetown.edu Wed Jun 22 14:27:08 2005 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] ZO-1 Message-ID: <42B9BB8C.4000306@georgetown.edu> Hi, I am looking to order some ZO-1 antibody for FFPE staining. Can anyone give me some suggestions on a company? Eva Andersson Georgetown University From failm <@t> musc.edu Wed Jun 22 14:38:40 2005 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] CDX2 Message-ID: Karen, I use Biogenex concentrate at 1:200, EDTA in the steamer for 20 minutes, cool down for ten minutes. DAKO's Envision + system on the autostainer. Rena Fail Lead Tech IHC/SS MUSC >>> "Bauer, Karen" 06/22/05 02:57PM >>> Does anyone in histoland have a working protocol for CDX2? I have a pre-dilute from BioGenex that I'm trying to use on our BenchMark, but nothing seems to work. I've called Ventana and BioGenex and have tried their suggestions, but with no luck. Anybody willing to share their protocol? I'd love to give it a try. Thanks much!! Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lwhite <@t> lakeridgehealth.on.ca Wed Jun 22 15:02:52 2005 From: lwhite <@t> lakeridgehealth.on.ca (White, Lori) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] magnifying glass for embedding Message-ID: Hi, I need to replace a magnifying glass for an old Tissue Tek embedding station. Does anybody have suggestions other than through the manufacturer? Thanks in advance, Lori From slappycraw <@t> yahoo.com Wed Jun 22 15:14:36 2005 From: slappycraw <@t> yahoo.com (Larry Woody) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Histonet] magnifying glass for embedding Message-ID: <20050622201437.95348.qmail@web52609.mail.yahoo.com> there are lights that swivel with a magnifying glass in the center that work pretty well. __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From ddittus787 <@t> aol.com Wed Jun 22 15:29:03 2005 From: ddittus787 <@t> aol.com (ddittus787@aol.com) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Her2 antibody In-Reply-To: <42B9AE36.9010409@georgetown.edu> References: <42B9AE36.9010409@georgetown.edu> Message-ID: <8C745802A0837E0-F84-175FB@mblk-r39.sysops.aol.com> anti-her 2 cb11 clone works very well with citrate hier dilution(depending on your method is 1:40-1:100) and a 30 minute incubation at 37 degrees. dana -----Original Message----- From: Eva C Andersson To: histonet@lists.utsouthwestern.edu Sent: Wed, 22 Jun 2005 14:30:14 -0400 Subject: [Histonet] Her2 antibody Hello, I just ordered mouse anti-Her2 cat.no. 18-7107 from Zymed. There is however no indication given as to which dilution to use. I want to use it for Immunohistochemistry on paraffin embedded sections. It also doesn't say anything about antigen retrieval. Is anyone using this antibody and if so could you tell me what you do? Thanks, Eva Andersson Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ameeker <@t> mail.jhmi.edu Wed Jun 22 15:52:38 2005 From: ameeker <@t> mail.jhmi.edu (ALAN K MEEKER) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] No staining with monoclonal to Cytokeratins 5+8 Message-ID: <901195c8f902.42b99756@jhmimail.jhmi.edu> We have been trying to get epithelial cell staining in formalin-fixed mouse prostate tissue sections with anti-CK5+8 mouse monoclonal antibody (clone RCK 102) but are getting no signals at whatsoever. So far we've tried antibodies from Fitzgerald and Abcam (same clone). For antigen-retrieval we've tried either 20 minute Citrate steaming or 40 minute steaming in target retrieval solution from DAKO. Primary antibody was diluted in Ventana "Chem Mate" antibody dilution buffer (up to 1:5) and incubated for 60 min. or overnight and detection attempted with DAKO Envision kit which is working well for us with other antibodies on these mouse prostate sections. We are now going to try some protease treatments. Does anyone have any experience with this monoclonal or any other suggestions. Our main goal here is to have a good epithelial stain for the mouse prostate that hits both basal and luminal cells. Thanks in advance, Alan Meeker, PhD Assistant Professor of Pathology and Urology The Johns Hopkins Medical Institutions Department of Pathology Division of Genitourinary Pathology Bunting-Blaustein Cancer Research Building Room 124 1650 Orleans Street Baltimore, MD 21231-1000 ph (410) 502-7880/502-7354 fax (410) 502-9817 e-mail: [1]ameeker@mail.jhmi.edu References 1. mailto:ameeker@mail.jhmi.edu From jbaez <@t> interscopepath.com Wed Jun 22 16:08:46 2005 From: jbaez <@t> interscopepath.com (Baez, Janet) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] quality assurance Message-ID: <9E956D8FEB06C2408B08AC16498325E90139D9@scopemx1.interscope.com> Hi Lori, You can monitor specimen integrity (quality of processing and staining), Turn-around-time, employee competency and much more. Contact me if you need more information. Janet Baez Histology Manager Interscope Pathology Canoga Park, Ca. -----Original Message----- From: ddittus787@aol.com [mailto:ddittus787@aol.com] Sent: Tuesday, June 21, 2005 1:28 PM To: Lori.Auth@chsli.org; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] quality assurance Lori; did you try demographic failures by location, body part mis id (ie: rt hand versus left hand) req says one thing, bottle another? How about accessioning error rates? dana -----Original Message----- From: Auth, Lori To: histonet@lists.utsouthwestern.edu Sent: Tue, 21 Jun 2005 15:19:08 -0400 Subject: [Histonet] quality assurance please I need help!!!!!!!!! I am looking for a new quality assurance indicator for Histology. I currently am using specimen/requisition deficiencies. We have been compliant for over 2 years and I need to identify high risk, high volume error prone issue. I can't come up with something new to monitor. What does everyone else use??? Lori Auth, Lead Technician Histology Mercy Medical Center Rockville Centre, N.Y. _______________________________________________ Histonet mailing list Histonet@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 gisela <@t> vetmed.wsu.edu Wed Jun 22 16:56:31 2005 From: gisela <@t> vetmed.wsu.edu (Bailey, Gisela) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Job Posting Message-ID: <95111571D1C6B5498C64F68DD0769BB54A3BB2@cvm36.vetmed.wsu.edu> WASHINGTON STATE UNIVERSITY COLLEGE OF VETERINARY MEDICINE HISTOLOGY LABORATORY SUPERVISOR This full time permanent staff position is located in the Washington Animal Disease Diagnostic Laboratory - a high volume and service oriented tissue processing (histopathology) laboratory. This position supervises four full time employees and several student hourly employees. Implementation of new histologic techniques, and subsequent training of personnel and quality control will be ongoing managerial responsibilities. MINIMUM QUALS: American Society of Clinical Pathology Certification or eligibility as a histotechnician/histotechnologist is required, and a minimum of two years experience directing the operation of a histology laboratory (or equivalent). DESIRED QUALS: ASCP certification, computer proficiency, supervisory experience, strong communication and interpersonal skills. For additional information, contact Dr. Charles W. Leathers (509) 335-9696. Please apply on line at www.hrs.wsu.edu. Applications must be received in Human Resource Services by July 11, 2005. WSU offers a competitive salary and generous benefit package. WSU is an EO/AA educator and employer. From debbiekeith <@t> cox.net Wed Jun 22 17:50:32 2005 From: debbiekeith <@t> cox.net (Debbie Keith) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] magnifying glass for embedding In-Reply-To: Message-ID: <5.2.0.9.0.20050622154938.02621640@pop.central.cox.net> try this... http://www.staples.com/Catalog/Browse/Sku.asp?PageType=1&Sku=ELE7268 i've always used a bigger one... but this looks great! deb At 04:02 PM 6/22/05 -0400, you wrote: >Hi, >I need to replace a magnifying glass for an old Tissue Tek embedding >station. Does anybody have suggestions other than through the >manufacturer? >Thanks in advance, >Lori > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DDDeltour <@t> mar.med.navy.mil Thu Jun 23 05:05:12 2005 From: DDDeltour <@t> mar.med.navy.mil (Deltour, Douglas D. (HM2)) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Fibrohistocytic marker Message-ID: <080566D001A3D9459FFC0A391A646C91064533CA@marxchg03.mar.med.navy.mil> Can anyone recommend a Fibrohistocytic marker? We currently need one for Histiocytoma's. We are using a FAC XIIIa currently from Cell Marque but it is not giving us good results. Thanks Douglas D Deltour Histology Technician NMCP Portsmouth Va (757) 953-1525/1523 From ree3 <@t> leicester.ac.uk Thu Jun 23 05:31:19 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] zebrafish histology Message-ID: Any hints on zebrafish histology, fixation processing etcetc and whereabouts of any relevant websites all gratefully received Many thanks Richard Edwards MRC TOXICOLOGY UNIT...U.K..... From billingconsultants <@t> yahoo.com Thu Jun 23 05:45:37 2005 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Re: xylene vs xylene substitute Message-ID: <20050623104537.90055.qmail@web54209.mail.yahoo.com> I would recommend using Xylene for processing, but for staining and coverslipping I prefer using MicroClear from Micron (using MicroCover coverslipping media). It has virtually no odor. Hope this helps. Louri Roberts --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football From koellinr <@t> amgen.com Thu Jun 23 07:40:59 2005 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Immunostainers -- in your opinion what's best and why? Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CC79D@wa-mb4-sea.amgen.com> Gustave (and others who inquired off-line), This product can be found at www.tecan.com which is a liquid handling/robotics company. It is certainly not an IHC platform for everyone. You build it the way you want to do the procedures you want with reagents you want. Don't work for TECAN or have any relationship with them other than normal purchases. . But the machine is ideal for us. One thing that I really like- it utilizes 8 simultaneous pipetting tips. So staining 144 slides in a 5.5 hour protocol takes that amount of time and not many more extra hours as a single tip would take to handle 144 individual slides, one at a time. Ray -----Original Message----- From: GT Hebert [mailto:emerald_lake77@yahoo.com] Sent: Wednesday, June 22, 2005 8:14 AM To: Koelling, Ray Cc: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Immunostainers -- in your opinion what's best and why? That's amazing ... do you know how much that system costs?? .. right now I am just handing in info regarding the amount of money I need, so the cost is somewhat crucial though I am not sure what system I am interested in yet.... once I get approved for the money, I will look into the machines and eventually choose one. Right now though, I just don't want to say I need an immunostainer and say it costs around 70,000 when in fact, the one I want is 120,000. So, yes, I will be in FL and will check out that poster. Thank you for your reply. Gustave "Koelling, Ray" wrote: Gustave, If you will be at Nationals in Ft. Lauderdale in Sept. , our lab will have 2 posters describing and detailing an automated immunohistochemistry platform by TECAN. Can run 144 IHC slides at one time. Although we haven't used it for such, it is set up and capable to also do that many ISH slides. Completely open. Ray Raymond Koelling Research Scientist Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of GT Hebert Sent: Tuesday, June 21, 2005 11:51 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Immunostainers -- in your opinion what's best and why? Hi everyone, It's that time again when we need to make a list for big budget items and I am in need of a good automated immunostainer. I perform IHC (85%) and ISH (15%) all by hand and at this time it will help me a lot to get some of that work to become automated. So in your opion and based on your experiences, what is the best one out there and why (for both IHC and ISH, or compromise)? I am only familar with the Biogenex system. I learned a lot about the Biocare Nemesis system -- does anyone have any experience with this machine? Biocare claims its machine to be a nemesis ... but the price seems to also rival all others -- $77,000 YIKES. I have been to the NSH convention and discovered there are many others -- but without some experience there is no way I can actually know how good it works, how easy it is to use, how useful it will be and if it breaks down after 3 months. Thank you all for your responses. Sincerely, Gustave T. Hebert Scientist II Wyeth Research Cambridge MA --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. From josephnerk <@t> hotmail.com Thu Jun 23 08:27:52 2005 From: josephnerk <@t> hotmail.com (Joseph Nerk) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Chromosomal analysis Message-ID: A good day to all you in Histonet land Can anyone provide any information on the best way to ship specimen for chromosomal analysis? Thanks Erskine _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar [1]MSN Toolbar Get it now! References 1. http://g.msn.com/8HMAEN/2755??PS=47575 From 1dpeterson <@t> meriter.com Thu Jun 23 08:36:12 2005 From: 1dpeterson <@t> meriter.com (Peterson, Dan) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Blue glands Message-ID: <328CBAE62F31C642B422970E879DFADC01A7FF98@pcwex01> OK netters, A question that I'm sure has been covered: What can we do to get rid of blue color in the glands of our colon bx's, prostate bx's, etc. When it happens, it usually seemed to be a clearing problem (contaminated xylene), but it's only happening on one machine. All machines get the same recycled xylene. The machine is immaculate. No clogging of the screen visible. WIDE open for suggestions. Daniel R Peterson HT(ASCP) Histopathology Section Head General Medical Laboratories 1dpeterson@meriter.com CONFIDENTIALITY NOTICE: This message (including any attachments) is intended for the sole use of the individual and entity to whom it is addressed. This message may contain information that is confidential and is protected by law. If you are not the intended recipient, you are hereby notified that any disclosure, copying or distribution of this message is strictly prohibited. If you received this message in error, please immediately notify the sender by reply email and then delete the message. Thank you. From NLemke <@t> asterand.com Thu Jun 23 08:59:26 2005 From: NLemke <@t> asterand.com (Nancy Lemke) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] IHC for Alzheimers for frozens Message-ID: <56E63440ABF1FC4897947D5BA4CFA44F0534B1BE@ATL1VEXC005.usdom004.tco.tc> Good Morning Histoland, I would appreciate information about what everyone is using to demonstrate Alzheimer's Disease in frozen sections. Thanks, Nancy Lemke This e-mail message, together with any attachments, contains information belonging to Asterand, Inc. that may be confidential, proprietary, copyrighted and/or legally protected or privileged, and is intended for the sole use of the individual or entity named on this message. If you are not the intended recipient, and have received this e-mail message in error, please immediately return this message to its original sender and permanently delete it from your electronic and paper records. Thank You. From Jackie.O'Connor <@t> abbott.com Thu Jun 23 09:29:57 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Blue glands Message-ID: My guess would be your hematoxylin. What kind do you use? There are some commercial brands/types that will stain mucin - check the specs on yours. There is no reason why the xylene would cause this problem to my knowledge. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist GPRD Cancer Research Abbott Laboratories, Abbott Park, IL Jackie.OConnor@abbott.com "Peterson, Dan" <1dpeterson@meriter.com> Sent by: histonet-bounces@lists.utsouthwestern.edu 06/23/2005 08:36 AM To: cc: Subject: [Histonet] Blue glands OK netters, A question that I'm sure has been covered: What can we do to get rid of blue color in the glands of our colon bx's, prostate bx's, etc. When it happens, it usually seemed to be a clearing problem (contaminated xylene), but it's only happening on one machine. All machines get the same recycled xylene. The machine is immaculate. No clogging of the screen visible. WIDE open for suggestions. Daniel R Peterson HT(ASCP) Histopathology Section Head General Medical Laboratories 1dpeterson@meriter.com CONFIDENTIALITY NOTICE: This message (including any attachments) is intended for the sole use of the individual and entity to whom it is addressed. This message may contain information that is confidential and is protected by law. If you are not the intended recipient, you are hereby notified that any disclosure, copying or distribution of this message is strictly prohibited. If you received this message in error, please immediately notify the sender by reply email and then delete the message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Thu Jun 23 09:46:34 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Blue glands Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42E0@fh2k093.fhmis.net> Are the biopsies pre-dyed with the Toluidine or Methylene Blue? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 6/23/2005 9:36 AM Subject: [Histonet] Blue glands OK netters, A question that I'm sure has been covered: What can we do to get rid of blue color in the glands of our colon bx's, prostate bx's, etc. When it happens, it usually seemed to be a clearing problem (contaminated xylene), but it's only happening on one machine. All machines get the same recycled xylene. The machine is immaculate. No clogging of the screen visible. WIDE open for suggestions. Daniel R Peterson HT(ASCP) Histopathology Section Head General Medical Laboratories 1dpeterson@meriter.com CONFIDENTIALITY NOTICE: This message (including any attachments) is intended for the sole use of the individual and entity to whom it is addressed. This message may contain information that is confidential and is protected by law. If you are not the intended recipient, you are hereby notified that any disclosure, copying or distribution of this message is strictly prohibited. If you received this message in error, please immediately notify the sender by reply email and then delete the message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Thu Jun 23 10:49:37 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] IHC for Clostridium perfringens Message-ID: I use a rabbit polyclonal antibody from ViroStat (Portland, ME) labeled "Clostridium sp." to stain clostridia in formalin-fixed, paraffin-embedded tissue. This antibody is not specific for C. perfringens; it reacts with proteins from common clostridia species. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Jan Shivers" 06/21/05 05:50PM >>> Does anyone out there do IHC for Clostridium perfringens? If so, would you please point me in the direction of where I could locate an antibody? Thanks in advance. Jan Shivers U of MN Vet Diag Lab _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From ddittus787 <@t> aol.com Thu Jun 23 11:24:19 2005 From: ddittus787 <@t> aol.com (ddittus787@aol.com) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Her2 antibody In-Reply-To: <8C745802A0837E0-F84-175FB@mblk-r39.sysops.aol.com> References: <42B9AE36.9010409@georgetown.edu> <8C745802A0837E0-F84-175FB@mblk-r39.sysops.aol.com> Message-ID: <8C7462724236049-8AC-1D7E3@mblk-r40.sysops.aol.com> As a side note zymed also has Tab 250 clone of cerb-2 that only need enzyme and then incubation and gives awesome reuslts as well. dana -----Original Message----- From: ddittus787@aol.com To: eca9@georgetown.edu; histonet@lists.utsouthwestern.edu Sent: Wed, 22 Jun 2005 16:29:03 -0400 Subject: Re: [Histonet] Her2 antibody anti-her 2 cb11 clone works very well with citrate hier dilution(depending on your method is 1:40-1:100) and a 30 minute incubation at 37 degrees. dana -----Original Message----- From: Eva C Andersson To: histonet@lists.utsouthwestern.edu Sent: Wed, 22 Jun 2005 14:30:14 -0400 Subject: [Histonet] Her2 antibody Hello, I just ordered mouse anti-Her2 cat.no. 18-7107 from Zymed. There is however no indication given as to which dilution to use. I want to use it for Immunohistochemistry on paraffin embedded sections. It also doesn't say anything about antigen retrieval. Is anyone using this antibody and if so could you tell me what you do? Thanks, Eva Andersson Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From denise <@t> pclv.net Thu Jun 23 11:45:07 2005 From: denise <@t> pclv.net (Denise) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Employment opportunity Seattle/Everett Area Message-ID: Phoenix Central Laboratory for Veterinarians, Inc. has a position available for a Histology Technician with ASCP certification (or eligibility). Phoenix Central Laboratory, (PCLV), is a veterinary reference laboratory that provides diagnostic services to more than 800 clinics located throughout the Pacific Northwest and Alaska. We are dedicated to providing rapid, quality service. Our laboratory is located in Everett, Washington, just thirty minutes north of downtown Seattle. Visit our web site at www.pclv.net. PCLV offers a competitive salary and benefit package including health and dental coverage, personal time, and a 401K plan. Please submit your resume to Denise DeRouchey, c/o Phoenix Central Laboratory, 11620 Airport Road, Everett, WA 98204. Alternately, you may fax your resume to 425-355-3676. From rocan <@t> mac.com Thu Jun 23 12:21:36 2005 From: rocan <@t> mac.com (Rocan) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] RESEARCH JOB IN SOUTHERN CALIFORNIA Message-ID: <718E9F9A-C222-49BE-96CF-1588EF14EDAA@mac.com> Dear Histoneters, One of my colleagues at the CEDARS SINAI RESEARCH INSTITUTE has asked me to post this position: ________________________________________________________________________ ____________________________________________________________ Research Histotechnology Position There is an opening for a Research Histotechnologist Technologist at the Research Institute of the Cedars-Sinai Medical Center in Los Angeles. The position requires experience in a biomedical research laboratory with particular expertise in general histology, immunohistochemistry and immunofluorescence of mouse and rat tissues. We are seeking a highly motivated individual to work in a challenging and diverse Cardiovascular Biology Research Laboratory. Responsibilities of this position include animal necropsy, wet tissue trimming, routine histology procedures, image processing and analysis, laboratory equipment maintenance, quality control of work and database entries, Requirements Candidate must have 5 years of relevant experience including cryotomy, microtomy, embedding, tissue processing, H&E staining, special staining and immunohistochemistry. The candidate is not required to be a Certified histotechnician, HT (ASCP) or histotechnologist, HTL (ASCP). However, these qualifications are desirable. Experience using confocal microscope, micro dissection and EM techniques are also a plus. Must be able to follow verbal and written instructions, have good organizational and interpersonal skills, and be able to work independently with minimal supervision. Attention to detail is critical. Experience using image analysis and processing software as well as basic computer skills (Word and Excel) are required. Please contact: Dr. Berhrooz Sharifi Principal Investigator and Laboratory Director Davis 1016 Department of Cardiology CSHS 8700 Beverly Blvd. Los Angeles, CA 90048: Behrooz.Sharifi@cshs.org 310-423-7621 ________________________________________________________________________ ____________________________________________________________ Thanks, Dr.Rocio Sierra-Honigmann Director Engineered Wound Repair Laboratory Cedars Sinai Research Institute Davis 1091 310-423-1882 Honigmannr@cshs.org From ohenry <@t> dfw.net Thu Jun 23 12:42:39 2005 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] re: magnifying glass for embedding Message-ID: <000601c5781b$335a4050$b2dd3040@your4f1261a8e5> For anyone interested in a magnifier for embedding or at the grossing table, try an "OptiVISOR", model DA-5. I just got a new one at my local 'hobby shop' (cost 25.00).. My first one is 20 years old and just doesn't look very nice. BUT it works great. The OptiVISOR is worn on the head, it's binocular. The magnifier pulls down to view. But once you get the hang of it, you can set the magnifier part in one place and just tilt your head. They come in different lens. I use a number 5 which magnifies 2.5 times. (A loop accessory is available which adds another 2.5 monocular magnification,if you need it) This can be worn over prescription or safety glasses. http://www.doneganoptical.com/catalog/opti/ You can see it here, but you will have to email them for a price. Like I said, I got mine at the local Hobby Shop. SUSAN OWENS-TX ohenry@dfw.net From anh2006 <@t> med.cornell.edu Thu Jun 23 12:59:49 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Survey: What fixative are you using for your fresh frozens? Message-ID: I want to take a broad survey of what fixative people are using for their fresh frozen cryosections (any species) for immunohistochemistry. If you only do H&E please let me know what you use but make sure to point out it is tested for H&E only. I would love to hear pros and cons of each and if there are any you swear by! I am doing some protocol development and want to compare as many fixations as possible. I will gladly compile data and distribute to those interested. Thanks in advance. Andrea -- From rebecca.riesen <@t> dsilabs.com Thu Jun 23 13:38:08 2005 From: rebecca.riesen <@t> dsilabs.com (Riesen, Rebecca) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] CDX2 Message-ID: <05844092236E7749A1BBBCB1077C34A2DEA01C@dsi-ex01.gateway.dom> We have had no trouble at all with this antibody from BioGenex. I use Benchmark XT. We do Mild pretreatment with CC1 for 30 minutes and incubate the primary at 42 degrees for 16 min, with a standard detection kit. Good Luck! Message: 12 Date: Wed, 22 Jun 2005 13:57:21 -0500 From: "Bauer, Karen" Subject: [Histonet] CDX2 To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Does anyone in histoland have a working protocol for CDX2? I have a pre-dilute from BioGenex that I'm trying to use on our BenchMark, but nothing seems to work. I've called Ventana and BioGenex and have tried their suggestions, but with no luck. Anybody willing to share their protocol? I'd love to give it a try. Thanks much!! Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. ------------------------------ DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(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 pruegg <@t> ihctech.net Thu Jun 23 14:21:42 2005 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] ZO-1 In-Reply-To: <42B9BB8C.4000306@georgetown.edu> Message-ID: <200506231921.j5NJLYsI068310@pro12.abac.com> I just got some from Zymed but have yet to get it to work on ffpe tissue, it is suggested for FITC which to me usually means frozens. Let me know if you get it to work on ffpe tissue. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C Andersson Sent: Wednesday, June 22, 2005 1:27 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] ZO-1 Hi, I am looking to order some ZO-1 antibody for FFPE staining. Can anyone give me some suggestions on a company? Eva Andersson Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lhadley <@t> iupui.edu Thu Jun 23 14:49:39 2005 From: lhadley <@t> iupui.edu (Baldridge, Lee Ann) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] ZO-1 Message-ID: Me too! I've tried just about everything I know. Zymed says it will work in FFPE tissue. Lee Ann IUSM Indpls., IN. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of patsy ruegg Sent: Thursday, June 23, 2005 2:22 PM To: 'Eva C Andersson'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] ZO-1 I just got some from Zymed but have yet to get it to work on ffpe tissue, it is suggested for FITC which to me usually means frozens. Let me know if you get it to work on ffpe tissue. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C Andersson Sent: Wednesday, June 22, 2005 1:27 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] ZO-1 Hi, I am looking to order some ZO-1 antibody for FFPE staining. Can anyone give me some suggestions on a company? Eva Andersson Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SMAPAISAIC <@t> aol.com Thu Jun 23 15:30:51 2005 From: SMAPAISAIC <@t> aol.com (SMAPAISAIC@aol.com) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Positions in Cary/Raleigh NC Message-ID: We are growing into our brand new facility. A histology lab with windows! We are looking for experienced techs at all levels if anyone is interested, feel free to email me! Sharon Ambrose Pathology Associates Charles River Laboratories Cary, NC From kerry.l.crabb <@t> gsk.com Thu Jun 23 16:03:43 2005 From: kerry.l.crabb <@t> gsk.com (kerry.l.crabb@gsk.com) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] NSH Office Staff Retirement Message-ID: Joe and Sylvia Palmer will be retiring from the NSH office this summer with over 18 years of service to NSH. The NSH office staff and the Palmer family will celebrate a retirement lunch in the middle of July to honor their dedication to the Society. Many of you have inquired about sending best wishes to them. We will present them with a wishing well at the lunch, if you would like to include a congratulatory note - please mail to: Joe and Sylvia Palmer C/O Carrie Diamond 314 Montpelier Ct. Westminster, MD 21157 We would be happing to include you in this special part of their lives. Thank you, Carrie Diamond Executive Director National Society for Histotechnology From dimension0007 <@t> hotmail.com Thu Jun 23 16:54:04 2005 From: dimension0007 <@t> hotmail.com (Sarah Dickerson) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Rat embryo perfusions Message-ID: Has anyone on the list ever perfused rat embryos, or know anyone who has? If so, any suggestions you have will be greatly appreciated. My IHC's on embryonic brain tissue have a ton of autofluorescence, and I'm hoping to reduce it by perfusing the embryos. Thanks, Sarah Dickerson University of Texas at Austin Division of Pharmacology & Toxicology From MTowers <@t> prdus.jnj.com Thu Jun 23 17:19:43 2005 From: MTowers <@t> prdus.jnj.com (Towers, Meghan [PRDUS]) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] IHC run help Message-ID: Ok, i have called several ihc labs about this problem i am having at my lab and we are all stumped. all of our ihc runs are coming up positive when we put our dab on, even our negatives. here are some factors to consider in this problem that has us stumped. 1. This procedure has worked for years and we have not changed a single reagent at all throughout the entire procedure. 2. it's only happening with polyclonal antibodies (goat and rabbit) and not monoclonal antibodies (mouse). 3. since our mouse is coming out fine and our polyclonals are not, that means any reagents in common with the two clonalities (peroxide block, streptavidin, dab, etc.) are not the problem or else we would see the same reaction in the monoclonal runs, therefore, the problem must lie in the only different steps between the two (blocking serum, primary, or secondary). 4. since the negatives are coming out the same as the positives, the primary must no be the issue (plus, what are the chances of all our polyclonals going bad at the same time), yes, we've tried several different polyclonal ab.s 5. we have tried different tissue types cut from different sources at different times, as well as repeating exact runs we've done in the past and they all turned brown, meaning it's not tissue or time specific. 6. when we thought it was our blocking or secondary, we remade them both from an unopened, unexpired kit and still the same problem (even though it is the same kit we always use). 7. we do our runs manually, not automatically, so mechanics being contaminated is not the problem. 8. Please help us! keep in mind while possibly pondering the problem, it's always happening on the polyclonals, not the monoclonals, so if you think of another reagent that is in common with the two, it's not or else it would happen on the monoclonal slides as well. 9. the labeling we see, by the way, is completely nonspecific, like painting the slide brown, even the glass area around the tissue is turning slightly brown. if anyone has ever had this happen before or has any idea what it might be, please let me know. thanks so much. Meghan A. Towers Johnson and Johnson Research Associate-Target Validation P.O. Box 776 Welsh and McKean Rds. Spring House, PA 19477 Phone: 215-628-5291 Fax: 215-628-5966 From dusko.trajkovic <@t> pfizer.com Thu Jun 23 17:21:36 2005 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] CD31 IF protocol Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B2D894CD@lajamrexm01.amer.pfizer.com> Good day my Histo colleagues, Can anyone share their protocol for CD31 Immunofluorescence staining on FFPE tissue? Thank you Dusko Trajkovic 858-638-6202 LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From cfranci <@t> rigel.com Thu Jun 23 18:14:18 2005 From: cfranci <@t> rigel.com (Christian Franci) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Zeus media Message-ID: Can one use this transport media prior to formalin and paraffin embedding? or, is it not recommended? some times, due to scheduling issues, we harvest tissues (mainly xenografts ) at different times and they end up being fixed for different amounts of time. I'm always afraid that some are over-fixed so I'm always on the look out for tips for avoiding this. If there is a simple way to store the tissues til they're all ready for fixing and embedding, it would be great. any pointers? Thanks in advance for any help. Chris From katri <@t> cogeco.ca Thu Jun 23 21:34:54 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] IHC run help References: Message-ID: <000601c57865$4e2b0780$6a9a9618@Katri> Hi Meghan, It sounds to me as if you have a blocking serum raised in an animal that is not compatible with the rest of the system. Make sure your blocking serum is not raised in the same species as either of your polyclonal primary antibodies (rabbit or goat). Your blocking serum should be raised in the same species as your respective biotinylated secondary antibodies. For instance: when you are using rabbit polyclonal antibody, your secondary is most likely biotinylated goat-anti-rabbit, so the blocking serum should be normal goat serum. When you are using goat polyclonal antibody, the secondary would possibly be biotinylated rabbit-anti-goat and your blocking serum normal rabbit serum. If you were to switch the two blocking serums by mistake, you would get the reaction you are talking about. If you are using kits, some of them don't tell you what the blocking serum is, so your best bet is to talk to the supplier, in case there has been a packaging error. Ask for another lot number... Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Towers, Meghan [PRDUS]" To: Sent: Thursday, June 23, 2005 6:19 PM Subject: [Histonet] IHC run help > Ok, i have called several ihc labs about this problem i am having at my > lab > and we are all stumped. > all of our ihc runs are coming up positive when we put our dab on, even > our > negatives. here are some factors to consider in this problem that has us > stumped. > 1. This procedure has worked for years and we have not changed a single > reagent at all throughout the entire procedure. > 2. it's only happening with polyclonal antibodies (goat and rabbit) and > not > monoclonal antibodies (mouse). > 3. since our mouse is coming out fine and our polyclonals are not, that > means any reagents in common with the two clonalities (peroxide block, > streptavidin, dab, etc.) are not the problem or else we would see the same > reaction in the monoclonal runs, therefore, the problem must lie in the > only > different steps between the two (blocking serum, primary, or secondary). > 4. since the negatives are coming out the same as the positives, the > primary must no be the issue (plus, what are the chances of all our > polyclonals going bad at the same time), yes, we've tried several > different > polyclonal ab.s > 5. we have tried different tissue types cut from different sources at > different times, as well as repeating exact runs we've done in the past > and > they all turned brown, meaning it's not tissue or time specific. > 6. when we thought it was our blocking or secondary, we remade them both > from an unopened, unexpired kit and still the same problem (even though it > is the same kit we always use). > 7. we do our runs manually, not automatically, so mechanics being > contaminated is not the problem. > 8. Please help us! keep in mind while possibly pondering the problem, > it's > always happening on the polyclonals, not the monoclonals, so if you think > of > another reagent that is in common with the two, it's not or else it would > happen on the monoclonal slides as well. > 9. the labeling we see, by the way, is completely nonspecific, like > painting the slide brown, even the glass area around the tissue is turning > slightly brown. > > if anyone has ever had this happen before or has any idea what it might > be, > please let me know. > thanks so much. > > Meghan A. Towers > Johnson and Johnson > Research Associate-Target Validation > P.O. Box 776 > Welsh and McKean Rds. > Spring House, PA 19477 > Phone: 215-628-5291 > Fax: 215-628-5966 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JGordon <@t> cellmarque.com Thu Jun 23 21:34:44 2005 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] IHC run help Message-ID: Meghan, according to your description, the only difference would be the negative serum, unless you are using separate mouse and rabbit detections rather than a universal detection, in which case that could also be a possible cause. There may be a contaminant in your rabbit or goat negative serum, such as some Fc receptor proteins that the secondary is reacting with. Have you tried using a different manufacturer's negative serum to see if that eliminated the problem? I have heard of that happening before, and a lab had to switch negative serums to eliminate the negative staining. If the negative serum doesn't end up being the issue, then you can try to put the step-by-step components of your detection on your negatives to see what it is that is reacting. For instance, if you just put the chromogen on the negative and it lights up, then you know that you have endogenous peroxidase (or endogenous alkaline phosphatase) in your tissue that needs to be blocked, since the chromogen molecules bind to those enzymes. If there is no staining with just the chromogen, then put the label AND chromogen on the negative tissue. If you see staining then, you can attribute that to endogenous avidin or biotin (since the streptavidin label will bind to any biotin molecules in the tissue), so you need to put an A/B block on it. If you put the label and chromogen on the tissue, and it doesn't stain, then you can put the link, label and chromogen on it and see if you get staining. If you do, then you have charged sites that are mimicing the heavy chain of the primary antibody (also known as the Fc receptor site) that the link is reacting with, so you would need to put a background block on it to eliminate the charged sites (but be judicious with this blocking reagent because it can possibly diminish your primary stain). I hope that this helps. Jeff Gordon Cell Marque Corp. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [ mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Towers, Meghan [PRDUS] Sent: Thursday, June 23, 2005 5:20 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] IHC run help Ok, i have called several ihc labs about this problem i am having at my lab and we are all stumped. all of our ihc runs are coming up positive when we put our dab on, even our negatives. here are some factors to consider in this problem that has us stumped. 1. This procedure has worked for years and we have not changed a single reagent at all throughout the entire procedure. 2. it's only happening with polyclonal antibodies (goat and rabbit) and not monoclonal antibodies (mouse). 3. since our mouse is coming out fine and our polyclonals are not, that means any reagents in common with the two clonalities (peroxide block, streptavidin, dab, etc.) are not the problem or else we would see the same reaction in the monoclonal runs, therefore, the problem must lie in the only different steps between the two (blocking serum, primary, or secondary). 4. since the negatives are coming out the same as the positives, the primary must no be the issue (plus, what are the chances of all our polyclonals going bad at the same time), yes, we've tried several different polyclonal ab.s 5. we have tried different tissue types cut from different sources at different times, as well as repeating exact runs we've done in the past and they all turned brown, meaning it's not tissue or time specific. 6. when we thought it was our blocking or secondary, we remade them both from an unopened, unexpired kit and still the same problem (even though it is the same kit we always use). 7. we do our runs manually, not automatically, so mechanics being contaminated is not the problem. 8. Please help us! keep in mind while possibly pondering the problem, it's always happening on the polyclonals, not the monoclonals, so if you think of another reagent that is in common with the two, it's not or else it would happen on the monoclonal slides as well. 9. the labeling we see, by the way, is completely nonspecific, like painting the slide brown, even the glass area around the tissue is turning slightly brown. if anyone has ever had this happen before or has any idea what it might be, please let me know. thanks so much. Meghan A. Towers Johnson and Johnson Research Associate-Target Validation P.O. Box 776 Welsh and McKean Rds. Spring House, PA 19477 Phone: 215-628-5291 Fax: 215-628-5966 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Eric <@t> ategra.com Tue Jun 21 11:37:09 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] HistoTech Job Openings Message-ID: Histonetters - My name is Eric Dye and I am the new Histo Tech recruiter here at Ategra. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job openings are in Central Ohio and Arizona seeking a HistoTech Manager/supervisor. I also have bench histo tech jobs available in: Arizona, Pennsylvania, Illinois, New York, Missouri, Georgia, Iowa, Rhode Island,Oregon,California and Florida.. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- 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 Eric <@t> ategra.com Tue Jun 21 11:40:19 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] HistoTech Job Openings Message-ID: Histonetters - My name is Eric Dye and I am the new Histo Tech recruiter here at Ategra. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job opening is in Central Ohio,Colorado and Arizona seeking a HistoTech Manager/Supervisor. I also have bench histo tech jobs available in: Arizona, Pennsylvania, Illinois, New York, Missouri, Georgia, Iowa, Rhode Island,California,Colorado,Oregon and Florida.. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- 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 abright <@t> brightinstruments.com Fri Jun 24 03:57:47 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:15 2005 Subject: [Histonet] Brain sectioning problems Message-ID: I am pleased to report back to those who are interested that the procedure I recommended for brain sectioning problems solved Joost's problems. 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: Bruijntjes, J.P. [mailto:bruyntjes@voeding.tno.nl] Sent: 23 June 2005 09:55 To: Alan Bright Subject: RE: [Histonet] (no subject) Hi Alan You are right, my smile is back. Thanks for your remarks. Joost Bruijntjes TNO Quality of Life Zeist Holland -----Original Message----- From: Alan Bright [mailto:abright@brightinstruments.com] Sent: woensdag 22 juni 2005 12:35 To: Bruijntjes, J.P.; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] (no subject) Dear Mr Bruijntjes, I am assuming your brain tissue and microtome chamber are at -12?C not 12?c, If you lowered only the chamber temperature to -20?C to lower the knife and anti-roll plate it will return your smile. 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: Bruijntjes, J.P. [mailto:bruyntjes@voeding.tno.nl] Sent: 22 June 2005 11:02 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi all I was wondering if anyone of you is used to cut non-fixed cryoslides of brain tissue of about 30-40 micron thick. I've tried a few peaces on a rather high temperature (approximately 12?C), but it did not make me smile? J.P. Bruijntjes TNO Quality of Life Zeist Holland This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From 1dpeterson <@t> meriter.com Fri Jun 24 07:45:29 2005 From: 1dpeterson <@t> meriter.com (Peterson, Dan) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Blue Glands II Message-ID: <328CBAE62F31C642B422970E879DFADC01A7FF99@pcwex01> Thanks for all the responses to my Blue Gland problem. However the problem is not a staining problem, it's a processing problem. I should have specified that one processor was the culprit out of four. So to recap: One processor out of four produces bx's with bluing in the glands. All slides run through the same stainer, that's how we traced it to the one processor Sorry for the confusion, but still looking for clues. Daniel R Peterson HT(ASCP) Histopathology Section Head General Medical Laboratories 1dpeterson@meriter.com CONFIDENTIALITY NOTICE: This message (including any attachments) is intended for the sole use of the individual and entity to whom it is addressed. This message may contain information that is confidential and is protected by law. If you are not the intended recipient, you are hereby notified that any disclosure, copying or distribution of this message is strictly prohibited. If you received this message in error, please immediately notify the sender by reply email and then delete the message. Thank you. From DonnaWillis <@t> texashealth.org Fri Jun 24 08:57:32 2005 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Blue Glands II Message-ID: <4483B6EE0FA82A4EAED2B1E161E5E41147C899@ftwex02.txhealth.org> Dan, What Fixative are you using. We had issues many years ago with biopsies that were fixed in Hollande. Had to add iodine to our stainers to get out the blue. 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 Peterson, Dan Sent: Friday, June 24, 2005 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Blue Glands II Thanks for all the responses to my Blue Gland problem. However the problem is not a staining problem, it's a processing problem. I should have specified that one processor was the culprit out of four. So to recap: One processor out of four produces bx's with bluing in the glands. All slides run through the same stainer, that's how we traced it to the one processor Sorry for the confusion, but still looking for clues. Daniel R Peterson HT(ASCP) Histopathology Section Head General Medical Laboratories 1dpeterson@meriter.com CONFIDENTIALITY NOTICE: This message (including any attachments) is intended for the sole use of the individual and entity to whom it is addressed. This message may contain information that is confidential and is protected by law. If you are not the intended recipient, you are hereby notified that any disclosure, copying or distribution of this message is strictly prohibited. If you received this message in error, please immediately notify the sender by reply email and then delete the message. Thank you. _______________________________________________ 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 sjchtascp <@t> yahoo.com Fri Jun 24 11:02:11 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] plus slides question Message-ID: <20050624160211.33353.qmail@web90209.mail.scd.yahoo.com> When using plus slides shouldn't the sections almost attract the slide when picked up on the water bath and not easily float off again. I'm using VWR, and Surgipath plus slides that act just like regular un-charged slides. I'll pick up sections, let them remain on the slide for several seconds then float the sections right off the slide. Is that odd for plus slides. Some I've used in the past certainly attracted the section w/o floating off again. I'm asking because I've been trying to correct a minor problem I'm having with sections detatching during routine staining. Steve --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football From hymclab <@t> hyhc.com Fri Jun 24 11:23:42 2005 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] plus slides question Message-ID: We use Surgipath plus slides and our sections are attracted extremely well. In fact if we mess up picking them up we have to wipe the slide really good or make a new slide they attract so well. We have had no problems what so ever witht the Surgipath plus slides keeping tissues attached to them for routine H&E's, routine specials or immunos. We use plain distilled water in our baths. Hope you solve your problem. Dawn -----Original Message----- From: Steven Coakley [mailto:sjchtascp@yahoo.com] Sent: Friday, June 24, 2005 11:02 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] plus slides question When using plus slides shouldn't the sections almost attract the slide when picked up on the water bath and not easily float off again. I'm using VWR, and Surgipath plus slides that act just like regular un-charged slides. I'll pick up sections, let them remain on the slide for several seconds then float the sections right off the slide. Is that odd for plus slides. Some I've used in the past certainly attracted the section w/o floating off again. I'm asking because I've been trying to correct a minor problem I'm having with sections detatching during routine staining. Steve --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Fri Jun 24 11:29:01 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] tau-8 antibody Message-ID: Hello and happy weekend, Our neuropathologist would like us to switch to tau-8 from the tau-5 we're currently using. We're having a bit of trouble finding a tau-8 antibody. Is there anyone out there using one? 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 mlgiebel <@t> mail2.vcu.edu Fri Jun 24 10:49:52 2005 From: mlgiebel <@t> mail2.vcu.edu (Mary Lee Giebel) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] plus slides question In-Reply-To: <20050624160211.33353.qmail@web90209.mail.scd.yahoo.com> Message-ID: <200506241549.LAA23284@arrakis.vcu.edu> Steve, It sounds like your slides have loss their charge. The charge only last around 18 months. You can also loose the charge by handling the slide incorrectly, but your problem sounds like more than just the occasional slide. I use Plus slides from VWR and Fisher. I usually call the company to get the manufacture date and then mark each box and rotate my stock. For paraffin sections I use Platinum StarFrost Adhesive slides from Mercedes Medical. I have never had a slide shed water as well as the StarFrost Adhesive. I have not had a problem with them rejecting tissue, but they don't stay in the lab long. I stain by hand, so I am not sure about using them in machines. I hope this helps. Regards, Mary Lee Mary Lee Giebel, Manager & HTL Massey Cancer Center Research Histopathology Core Virginia Commonwealth University 1101 East Marshall Street Sanger Hall Room 4-036 P.O. Box 980662 Richmond, VA 23298 USA phone (804)828-5092 fax (804)828-9749 voice (804)278-1513 From lfidgen <@t> vt.edu Fri Jun 24 12:50:18 2005 From: lfidgen <@t> vt.edu (Laura Fidgen) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] S100 Message-ID: <6.0.0.22.0.20050624134204.0271bec0@pop.vt.edu> We are a Veterinary Hospital evaluating a Ventana Benchmark LT. I am having a lot of problems staining for S100. It is very dark with background. The people at Ventana think the secondary antibody is cross-reacting and suggest using a different secondary (our secondary currently is goat/rabbit). Can anyone in veterinary medicine help me out with this? So far my experience with this machine has not been favorable. Any advice would be appreciated. Laura L. Fidgen, MScF, BSc, HT(ASCP) Laboratory and Research Practioner VMRCVM, Histopatholgy Blacksburg, VA 24060-0443 From JonesLy <@t> mir.wustl.edu Fri Jun 24 13:13:12 2005 From: JonesLy <@t> mir.wustl.edu (JonesLy@mir.wustl.edu) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] PAR staining - Japanese source? In-Reply-To: <20050624170848.578334193@expurgate1.wustl.edu> Message-ID: Hello - The person who started this project is currently in China, but my PI asked if I could query the list anyway - apologies if my question is vague. I collect the tissues we stain, but have little involvement in their subsequent staining and am NOT a histologist. We need to stain formalin-fixed parafin embedded tissue (from streptozotocin-treated rats) for PAR, Poly-ADP-ribose - not PARP, poly-ADP-ribose polymerase. When we started this project, we had an antibody that gave beautiful results; unfortunately subsequent lot numbers show poor staining of our control block. (The vendor has acknowleded that the problem is on their end, but that doesn't help our research.) While at a meeting in DC, the PI spoke with someone who was also working with PAR and has excellent results using a stain (monoclonal antibody?) from a vendor in Japan. It is apparently expensive, but gives very reproducable results. The person working with this stain was supposed to e-mail our PI with the details, but he hasn't heard from her. Even if no-one has worked with the stain, a list of Japanese suppliers of antibodies would help. (Again, we need to stain for PAR, not PARP.) Thanks in advance, Lynne Jones Senior Research Technician Dept. of Radiological Sciences Washington University Medical School St. Louis, MO From BRIAN.CHELACK <@t> usask.ca Sat Jun 25 12:20:32 2005 From: BRIAN.CHELACK <@t> usask.ca (Brian Chelack) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Re: S100 Staining References: <20050625170118.E6164B85243@spamf3.usask.ca> Message-ID: <42BD9260.BF1870FB@sask.usask.ca> Hello Laura; Regarding the Ventana secondaries in veterinary species, If I assume that you are trying this staining on canine tissues, our experience has been similar. The Ventana secondaries do have a tendancy to crossreact with tissue components (immunoglobulin)from some small animal species. It does limit the usefulness of this platform for small animal tumour markers, you should see how much fun it is for B-cell markers... regards, Brian Chelack Prairie Diagnostic Services > Date: Fri, 24 Jun 2005 13:50:18 -0400 > From: Laura Fidgen > Subject: [Histonet] S100 > To: histonet@lists.utsouthwestern.edu > Message-ID: <6.0.0.22.0.20050624134204.0271bec0@pop.vt.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > We are a Veterinary Hospital evaluating a Ventana Benchmark LT. I am > having a lot of problems staining for S100. It is very dark with > background. The people at Ventana think the secondary antibody is > cross-reacting and suggest using a different secondary (our secondary > currently is goat/rabbit). Can anyone in veterinary medicine help me out > with this? So far my experience with this machine has not been > favorable. Any advice would be appreciated. > > Laura L. Fidgen, MScF, BSc, HT(ASCP) > Laboratory and Research Practioner > VMRCVM, Histopatholgy > Blacksburg, VA 24060-0443 > *** From CrochiereSteve <@t> aol.com Sat Jun 25 22:10:07 2005 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] immunostainers Message-ID: <1e9.3ec0135d.2fef768f@aol.com> I wholeheartedly recommend the Nemesis Immunostainer from Biocare Medical. I have been using this instrument for almost a year and have had no technical problems with this machine. The increased capacity compared to other available instruments has been a great help in handling the workload in my laboratory. The instrument can stain up to 84 slides per run and has 40 reagent dispensers available. The slide layout can be altered to a 72 slide mode allowing for up to 80 reagent dispensers, however I have not need to do so. I prefer more slides to more reagents. The system is open and staining protocols can be altered to meet your specific needs. I have been getting acceptable results with the staining protocols provided with the antibodies purchased from Biocare and have also used antibodies from other companies with good results. The antibodies and detection kits from Biocare have proven to be reliable and are less expensive compared to other commercial vendors resulting in a savings this past year on budgeted expenses set aside for immunohistochemistry testing. Another nice feature of this instrument is the ability to set up a delayed start run so that slides can be stained overnight and be ready the next morning. Although counterstaining can be done on the Nemesis, I prefer to do my counterstain off- line in my routine H&E stainer. This keeps the Nemesis cleaner and allows me to use the spaces that would be taken up by the hematoxylin and bluing reagents for antibody dispensers. I use Biocare?s hematoxylin and bluing for a counterstain. Here is a short list of the reasons that I purchased the instrument: Greater slide and reagent capacity Less expensive to use Open and flexible programming Overnight staining Good experience with Biocare?s antibodies in the past Reliable results I hope this will help you in considering this instrument for use in your facility. If I can be of any further assistance, feel free to call or e-mail. Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 From pathrm35 <@t> adelphia.net Sun Jun 26 07:13:10 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] immunostainers Message-ID: <21812371.1119787990993.JavaMail.root@web8.mail.adelphia.net> We are in the process of purchasing the Biogenex i6000 immunostainer. I was not interested at first because of bad experiences in the past with Biogenex. I (personally) think Biogenex has made a complete turn around with their quality of customer service, the stainer itself and the antibodies. I have been getting great results with the stains and the Biogenex rep (Allen Younger) has been a pleasure to work with. The price of the Biogenex stainer was also more in our range compared to other stainers. Just my personal and professional opinion. Ron Martin ---- CrochiereSteve@aol.com wrote: > I wholeheartedly recommend the Nemesis Immunostainer from Biocare Medical. > I have been using this instrument for almost a year and have had no technical > problems with this machine. The increased capacity compared to other > available instruments has been a great help in handling the workload in my laboratory. > The instrument can stain up to 84 slides per run and has 40 reagent > dispensers available. The slide layout can be altered to a 72 slide mode allowing for > up to 80 reagent dispensers, however I have not need to do so. I prefer more > slides to more reagents. > The system is open and staining protocols can be altered to meet > your specific needs. I have been getting acceptable results with the staining > protocols provided with the antibodies purchased from Biocare and have also > used antibodies from other companies with good results. The antibodies and > detection kits from Biocare have proven to be reliable and are less expensive > compared to other commercial vendors resulting in a savings this past year on > budgeted expenses set aside for immunohistochemistry testing. > Another nice feature of this instrument is the ability to set up > a delayed start run so that slides can be stained overnight and be ready the > next morning. Although counterstaining can be done on the Nemesis, I prefer to > do my counterstain off- line in my routine H&E stainer. This keeps the Nemesis > cleaner and allows me to use the spaces that would be taken up by the > hematoxylin and bluing reagents for antibody dispensers. I use Biocare?s hematoxylin > and bluing for a counterstain. > Here is a short list of the reasons that I purchased the > instrument: > > Greater slide and reagent capacity > Less expensive to use > Open and flexible programming > Overnight staining > Good experience with Biocare?s antibodies in the past > Reliable results > > I hope this will help you in considering this instrument for use in your > facility. If I can be of any further assistance, feel free to call or e-mail. > > > > Steven M. Crochiere, HT(ASCP) > Histology Supervisor > LifePath Partners @ Mercy Medical Center > Springfield, MA 01104 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From chesarato <@t> hotmail.com Mon Jun 27 00:03:14 2005 From: chesarato <@t> hotmail.com (Cesar Francisco Romero) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Freezing Antobodies ! Message-ID: Dear People from Histonet I would like to know if you freeze your antibodies in Kitchen like Freezers ( - 18 or - 20 ? C ). I can not afford a - 80 ? C Freezer. Thank You in advance Cesar Romero Buenos Aires Argentina _________________________________________________________________ Nuevo MSN Messenger [1]Una forma r?pida y divertida de enviar mensajes References 1. http://g.msn.com/8HMAESAR/2734??PS=47575 From Jacqueline.Malam <@t> rli.mbht.nhs.uk Mon Jun 27 02:44:38 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] RE: S100 staining Message-ID: Hi Laura We have a Benchmark XT and it works fine for us - can you let me know your ab source, protocol and dilution - see if I can shed some light. Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From louise.renton <@t> gmail.com Mon Jun 27 02:51:30 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:16 2005 Subject: OT:: [Histonet] immunostainers In-Reply-To: <1e9.3ec0135d.2fef768f@aol.com> References: <1e9.3ec0135d.2fef768f@aol.com> Message-ID: On 6/26/05, CrochiereSteve@aol.com wrote: > I wholeheartedly recommend the Nemesis Immunostainer from Biocare Medical. > I have been using this instrument for almost a year and have had no technical > problems with this machine. Does anyone think about the meaning of words when they name these machines? NEMISIS: A source of harm or ruin: Retributive justice in its execution or outcome: An opponent that cannot be beaten or overcome. One that inflicts retribution or vengeance. Nemesis Greek Mythology. The goddess of retributive justice or vengeance. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Mon Jun 27 04:30:50 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] RE: pos control for copper Message-ID: Many thanks to everyone who responded. I am now the proud possessor of a positive control block - my world is complete ! Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From cloukas <@t> hotmail.com Mon Jun 27 07:08:00 2005 From: cloukas <@t> hotmail.com (C. Loukas) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] prognostic markers and IHC staining Message-ID: Hello all, Although I'm new in this list, it seems that is viewed by loads of Histologists, so I would be grateful if someone could recommend me a couple of review articles/textbook focusing on the assessment of the various prognostic markers currently in use (e.g. ER in breast cancer, MIB-1 in oesophageal cancers), through immunohistochemical staining of tumor tissue sections. In particular I'm after a list of: established prognostic markers (or stains) vs tumor type, if such a think exists. Thanks in advance! cl From katri <@t> cogeco.ca Mon Jun 27 08:01:26 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Freezing Antobodies ! References: Message-ID: <001001c57b18$5471c1a0$6a9a9618@Katri> Cesar, If your freezer is a self defrosting kind, it is not a good idea to keep antibodies or frozen tissue in it. There is too much temperature fluctuation as it defrosts itself. Chest freezers keep the temperature more stable and may be acceptable. Keep a thermometer in it to record the temperature daily. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Cesar Francisco Romero" To: Sent: Monday, June 27, 2005 1:03 AM Subject: [Histonet] Freezing Antobodies ! > > Dear People from Histonet > > I would like to know if you freeze your antibodies in Kitchen like > Freezers ( - 18 or - 20 ? C ). > > I can not afford a - 80 ? C Freezer. > > Thank You in advance > > Cesar Romero > > Buenos Aires > > Argentina > _________________________________________________________________ > > Nuevo MSN Messenger [1]Una forma r?pida y divertida de enviar mensajes > > References > > 1. http://g.msn.com/8HMAESAR/2734??PS=47575 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sobrien <@t> bthosp.com Mon Jun 27 08:38:47 2005 From: sobrien <@t> bthosp.com (O'Brien, Sue) Date: Fri Sep 16 15:25:16 2005 Subject: : [Histonet] immunostainers Message-ID: <4D95C24EC0E4C84787A6919F1165B25E24B16A@btmhems01.BTHOSP.INT> Steve, Sounds like the company is focusing on the "An opponent that cannot be beaten or overcome" part of the definition. I also have a Nemesis (since Feb) and have been VERY pleased with its performance. I would highly recommend it to anyone considering an automated Immunostainer. Susan O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May court House, NJ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Monday, June 27, 2005 3:52 AM To: CrochiereSteve@aol.com; Histonet@lists.utsouthwestern.edu Subject: OT:: [Histonet] immunostainers On 6/26/05, CrochiereSteve@aol.com wrote: > I wholeheartedly recommend the Nemesis Immunostainer from Biocare Medical. > I have been using this instrument for almost a year and have had no technical > problems with this machine. Does anyone think about the meaning of words when they name these machines? NEMISIS: A source of harm or ruin: Retributive justice in its execution or outcome: An opponent that cannot be beaten or overcome. One that inflicts retribution or vengeance. Nemesis Greek Mythology. The goddess of retributive justice or vengeance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Mon Jun 27 08:55:20 2005 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] S/P Pro-Texx* Mounting Medium Message-ID: <6.0.1.1.0.20050627084308.01b1a090@mailhost.vetmed.auburn.edu> Good morning All, do any of you have a source for S/P Pro-Texx* Mounting Medium, Cat. M7635-1? If so will you please share it with me ASAP? If it's no longer on the market will someone please provide ordering info for a comparable mounting medium? Your prompt reply will be much appreciated. Atoska From Rcartun <@t> harthosp.org Mon Jun 27 09:58:13 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] prognostic markers and IHC staining Message-ID: A good article dealing with breast cancer is, "Prognostic and Predictive Factors in Breast Cancer by Immunohistochemical Analysis" by D. Craig Allred et al. in Modern Pathology 1998:11(2):155-168. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "C. Loukas" 06/27/05 08:08AM >>> Hello all, Although I'm new in this list, it seems that is viewed by loads of Histologists, so I would be grateful if someone could recommend me a couple of review articles/textbook focusing on the assessment of the various prognostic markers currently in use (e.g. ER in breast cancer, MIB-1 in oesophageal cancers), through immunohistochemical staining of tumor tissue sections. In particular I'm after a list of: established prognostic markers (or stains) vs tumor type, if such a think exists. Thanks in advance! cl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From pzeitlow <@t> bbpllab.com Mon Jun 27 10:29:39 2005 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] MUM1/IRF4 and CD138 Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A47D652@bbplsrv1.bbpl> Is anyone using MUM1/IRF4 and/or CD138 that would be willing to share vendors, clones or protocols for use with routine FFPE IHC? Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. From TMcNemar <@t> lmhealth.org Mon Jun 27 11:00:24 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] S/P Pro-Texx* Mounting Medium Message-ID: <6CD94D97ED7D924BA5C2B588FA9528213967D5@mail.lmhealth.org> We used Pro-tex for many years and got from Scientific Products. Two maybe three years ago they (SP) substituted Cytoseal 60 (made by Richard Alan). Works just as well so that's what we are using to this day. Pro-tex is no longer available throgh Scientific Products. The order numbe for Cytoseal 60 M7630-2A and the phone number for Scientific Products is (800) 964-5227. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Atoska S. Gentry [mailto:gentras@vetmed.auburn.edu] Sent: Monday, June 27, 2005 9:55 AM To: Histonet Subject: [Histonet] S/P Pro-Texx* Mounting Medium Good morning All, do any of you have a source for S/P Pro-Texx* Mounting Medium, Cat. M7635-1? If so will you please share it with me ASAP? If it's no longer on the market will someone please provide ordering info for a comparable mounting medium? Your prompt reply will be much appreciated. Atoska _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Mon Jun 27 13:05:02 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Freezing Antobodies ! Message-ID: I have been storing my antibodies at -20 with no problems. I only have a six months supply at most. Robyn OHSU From jluis.palazon <@t> icman.csic.es Mon Jun 27 13:09:03 2005 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Bowies method Message-ID: <20050627180903.9267281A6C8@perceval.uca.es> Dear List-fellows Could any of you send me the protocol of Bowie?s method for juxtaglomerular cells in kidney. thanks in advance Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From SMcMurtrey <@t> UniPathLLC.com Mon Jun 27 16:34:30 2005 From: SMcMurtrey <@t> UniPathLLC.com (Samantha McMurtrey) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] MMP-1 Message-ID: Hello all, I am having a lot of trouble getting any staining for MMP-1. I am using Santa Cruz, catalog # sc-21731, lot # A1405. It's a mouse monoclonal antibody. I have tried dilutions from 1:200 down to 1:25. I incubated the primary antibody for 30 minutes and the secondary for 30 minutes. I have also tried using proteinase K, Target, EDTA, and a pH 9 retrieval. I am working it up on formalin fixed paraffin embedded tissue. I am mostly using breast tissue that has atypical ductal hyperplasia (per the pathologist's request). I haven't had any luck at all. I am getting no staining. Please help! Thank you, Samantha McMurtrey, BS, QIHC UniPath, LLC 2180 South Leyden Street Denver, CO 80222 (303) 512-2220 From Secholsb <@t> aol.com Mon Jun 27 19:06:13 2005 From: Secholsb <@t> aol.com (Secholsb@aol.com) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] ROTAVIRUS Message-ID: <1fe.4776b11.2ff1ee75@aol.com> Would anybody have an unstained paraffin section or block containing human rotavirus-infected intestine? Thanks Sara State Vet Lab Auburn, Alabama From bills <@t> icpmr.wsahs.nsw.gov.au Mon Jun 27 20:16:52 2005 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Problem with CC1 on Benchmark. Message-ID: <000001c57b7f$11f8aca0$0ecd080a@wsahs.nsw.gov.au> Dear All, Particularly those using Ventana IHC Stainers. We have a Ventana Benchmark and recently (from the middle of May) we have a problem with antibodies which have had a pre-treatment in CC1. 1 After CC1, whether mild or standard we are getting non-specific biotin type staining. However, when you look at the slides they look specific in their staining although for the wrong antibody. The negative controls are displaying this phenomenon as well. 2. The staining can be blocked with the Avidin-Biotin block, but this does not always remove the staining completely. We are using Batch #'s 48091 & 48127 of CC1. Any advice would be greatly appreciated. Thanks Jane Milliken, IHC Benchmark Driver Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 __________________________________________________________________ This electronic message and any attachments may be confidential. 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From CrochiereSteve <@t> aol.com Mon Jun 27 21:18:24 2005 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] nemesis Message-ID: <1ea.3f9c052b.2ff20d70@aol.com> It's just a little star trek reference. From cloukas <@t> hotmail.com Tue Jun 28 01:32:02 2005 From: cloukas <@t> hotmail.com (C. Loukas) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] prognostic markers and IHC staining Message-ID: Thanks Richard, Do you or anyone else know where I can get more info on IHC markers currently in use for deriving prognostic information from other tumour types?? Thanks again, I really appreciate your help, cl > > > >Message: 9 >Date: Mon, 27 Jun 2005 10:58:13 -0400 >From: "Richard Cartun" >Subject: Re: [Histonet] prognostic markers and IHC staining >To: , >Message-ID: >Content-Type: text/plain; charset=US-ASCII > >A good article dealing with breast cancer is, "Prognostic and Predictive >Factors in Breast Cancer by Immunohistochemical Analysis" by D. Craig >Allred et al. in Modern Pathology 1998:11(2):155-168. > >Richard > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 >(860) 545-1596 >(860) 545-0174 Fax > > >>> "C. Loukas" 06/27/05 08:08AM >>> >Hello all, > >Although I'm new in this list, it seems that is viewed by loads of >Histologists, so I would be grateful if someone could recommend me a >couple >of review articles/textbook focusing on the assessment of the various >prognostic markers currently in use (e.g. ER in breast cancer, MIB-1 in >oesophageal cancers), through immunohistochemical staining of tumor tissue >sections. > >In particular I'm after a list of: established prognostic markers (or >stains) vs tumor type, if such a think exists. > >Thanks in advance! > >cl From arrrid702 <@t> skmc.gov.ae Tue Jun 28 02:23:23 2005 From: arrrid702 <@t> skmc.gov.ae (Ridhwaan Arries) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] RE: Histonet Digest, Vol 19, Issue 39 Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02A7CF42@SKMCEMAIL.skmc.gov.ae> TO PAT ZEITLOW- CD138 ANTIBODY PROTOCOL DAKO CD138 PRODUCT CODE:M7228 CLONE MI15 HIER DAKO S1699 OR S1700 DILUTION 1:25 PRIMARY INCUBATION 1HOUR DETECTION BY DAKO LSAB+/HRP OR ENVISION+/HRP as per packet insert. I hope this helps. -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Monday, June 27, 2005 9:03 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 19, Issue 39 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. Freezing Antobodies ! (Cesar Francisco Romero) 2. RE: S100 staining (Malam Jacqueline) 3. OT:: [Histonet] immunostainers (louise renton) 4. RE: pos control for copper (Malam Jacqueline) 5. prognostic markers and IHC staining (C. Loukas) 6. Re: Freezing Antobodies ! (Katri Tuomala) 7. RE: : [Histonet] immunostainers (O'Brien, Sue) 8. S/P Pro-Texx* Mounting Medium (Atoska S. Gentry) 9. Re: prognostic markers and IHC staining (Richard Cartun) 10. MUM1/IRF4 and CD138 (Pat Zeitlow) 11. RE: S/P Pro-Texx* Mounting Medium (Tom McNemar) ---------------------------------------------------------------------- Message: 1 Date: Mon, 27 Jun 2005 02:03:14 -0300 From: "Cesar Francisco Romero" Subject: [Histonet] Freezing Antobodies ! To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1"; format="flowed" Dear People from Histonet I would like to know if you freeze your antibodies in Kitchen like Freezers ( - 18 or - 20 ? C ). I can not afford a - 80 ? C Freezer. Thank You in advance Cesar Romero Buenos Aires Argentina _________________________________________________________________ Nuevo MSN Messenger [1]Una forma r?pida y divertida de enviar mensajes References 1. http://g.msn.com/8HMAESAR/2734??PS=47575 ------------------------------ Message: 2 Date: Mon, 27 Jun 2005 08:44:38 +0100 From: Malam Jacqueline Subject: [Histonet] RE: S100 staining To: "Histonet submissions (histonet@lists.utsouthwestern.edu)" Message-ID: Content-Type: text/plain Hi Laura We have a Benchmark XT and it works fine for us - can you let me know your ab source, protocol and dilution - see if I can shed some light. Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 3 Date: Mon, 27 Jun 2005 09:51:30 +0200 From: louise renton Subject: OT:: [Histonet] immunostainers To: CrochiereSteve@aol.com, Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 On 6/26/05, CrochiereSteve@aol.com wrote: > I wholeheartedly recommend the Nemesis Immunostainer from Biocare Medical. > I have been using this instrument for almost a year and have had no technical > problems with this machine. Does anyone think about the meaning of words when they name these machines? NEMISIS: A source of harm or ruin: Retributive justice in its execution or outcome: An opponent that cannot be beaten or overcome. One that inflicts retribution or vengeance. Nemesis Greek Mythology. The goddess of retributive justice or vengeance. ------------------------------ Message: 4 Date: Mon, 27 Jun 2005 10:30:50 +0100 From: Malam Jacqueline Subject: [Histonet] RE: pos control for copper To: "Histonet submissions (histonet@lists.utsouthwestern.edu)" Message-ID: Content-Type: text/plain Many thanks to everyone who responded. I am now the proud possessor of a positive control block - my world is complete ! Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 5 Date: Mon, 27 Jun 2005 12:08:00 +0000 From: "C. Loukas" Subject: [Histonet] prognostic markers and IHC staining To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hello all, Although I'm new in this list, it seems that is viewed by loads of Histologists, so I would be grateful if someone could recommend me a couple of review articles/textbook focusing on the assessment of the various prognostic markers currently in use (e.g. ER in breast cancer, MIB-1 in oesophageal cancers), through immunohistochemical staining of tumor tissue sections. In particular I'm after a list of: established prognostic markers (or stains) vs tumor type, if such a think exists. Thanks in advance! cl ------------------------------ Message: 6 Date: Mon, 27 Jun 2005 09:01:26 -0400 From: "Katri Tuomala" Subject: Re: [Histonet] Freezing Antobodies ! To: "Cesar Francisco Romero" , Message-ID: <001001c57b18$5471c1a0$6a9a9618@Katri> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=response Cesar, If your freezer is a self defrosting kind, it is not a good idea to keep antibodies or frozen tissue in it. There is too much temperature fluctuation as it defrosts itself. Chest freezers keep the temperature more stable and may be acceptable. Keep a thermometer in it to record the temperature daily. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Cesar Francisco Romero" To: Sent: Monday, June 27, 2005 1:03 AM Subject: [Histonet] Freezing Antobodies ! > > Dear People from Histonet > > I would like to know if you freeze your antibodies in Kitchen like > Freezers ( - 18 or - 20 ? C ). > > I can not afford a - 80 ? C Freezer. > > Thank You in advance > > Cesar Romero > > Buenos Aires > > Argentina > _________________________________________________________________ > > Nuevo MSN Messenger [1]Una forma r?pida y divertida de enviar mensajes > > References > > 1. http://g.msn.com/8HMAESAR/2734??PS=47575 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Mon, 27 Jun 2005 09:38:47 -0400 From: "O'Brien, Sue" Subject: RE: : [Histonet] immunostainers To: "louise renton" , , Message-ID: <4D95C24EC0E4C84787A6919F1165B25E24B16A@btmhems01.BTHOSP.INT> Content-Type: text/plain; charset="us-ascii" Steve, Sounds like the company is focusing on the "An opponent that cannot be beaten or overcome" part of the definition. I also have a Nemesis (since Feb) and have been VERY pleased with its performance. I would highly recommend it to anyone considering an automated Immunostainer. Susan O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May court House, NJ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Monday, June 27, 2005 3:52 AM To: CrochiereSteve@aol.com; Histonet@lists.utsouthwestern.edu Subject: OT:: [Histonet] immunostainers On 6/26/05, CrochiereSteve@aol.com wrote: > I wholeheartedly recommend the Nemesis Immunostainer from Biocare Medical. > I have been using this instrument for almost a year and have had no technical > problems with this machine. Does anyone think about the meaning of words when they name these machines? NEMISIS: A source of harm or ruin: Retributive justice in its execution or outcome: An opponent that cannot be beaten or overcome. One that inflicts retribution or vengeance. Nemesis Greek Mythology. The goddess of retributive justice or vengeance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Mon, 27 Jun 2005 08:55:20 -0500 From: "Atoska S. Gentry" Subject: [Histonet] S/P Pro-Texx* Mounting Medium To: Histonet Message-ID: <6.0.1.1.0.20050627084308.01b1a090@mailhost.vetmed.auburn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Good morning All, do any of you have a source for S/P Pro-Texx* Mounting Medium, Cat. M7635-1? If so will you please share it with me ASAP? If it's no longer on the market will someone please provide ordering info for a comparable mounting medium? Your prompt reply will be much appreciated. Atoska ------------------------------ Message: 9 Date: Mon, 27 Jun 2005 10:58:13 -0400 From: "Richard Cartun" Subject: Re: [Histonet] prognostic markers and IHC staining To: , Message-ID: Content-Type: text/plain; charset=US-ASCII A good article dealing with breast cancer is, "Prognostic and Predictive Factors in Breast Cancer by Immunohistochemical Analysis" by D. Craig Allred et al. in Modern Pathology 1998:11(2):155-168. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "C. Loukas" 06/27/05 08:08AM >>> Hello all, Although I'm new in this list, it seems that is viewed by loads of Histologists, so I would be grateful if someone could recommend me a couple of review articles/textbook focusing on the assessment of the various prognostic markers currently in use (e.g. ER in breast cancer, MIB-1 in oesophageal cancers), through immunohistochemical staining of tumor tissue sections. In particular I'm after a list of: established prognostic markers (or stains) vs tumor type, if such a think exists. Thanks in advance! cl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 10 Date: Mon, 27 Jun 2005 10:29:39 -0500 From: "Pat Zeitlow" Subject: [Histonet] MUM1/IRF4 and CD138 To: "Histonet" Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A47D652@bbplsrv1.bbpl> Content-Type: text/plain; charset="US-ASCII" Is anyone using MUM1/IRF4 and/or CD138 that would be willing to share vendors, clones or protocols for use with routine FFPE IHC? Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. ------------------------------ Message: 11 Date: Mon, 27 Jun 2005 12:00:24 -0400 From: Tom McNemar Subject: RE: [Histonet] S/P Pro-Texx* Mounting Medium To: "'Atoska S. Gentry'" , Histonet Message-ID: <6CD94D97ED7D924BA5C2B588FA9528213967D5@mail.lmhealth.org> Content-Type: text/plain; charset="iso-8859-1" We used Pro-tex for many years and got from Scientific Products. Two maybe three years ago they (SP) substituted Cytoseal 60 (made by Richard Alan). Works just as well so that's what we are using to this day. Pro-tex is no longer available throgh Scientific Products. The order numbe for Cytoseal 60 M7630-2A and the phone number for Scientific Products is (800) 964-5227. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Atoska S. Gentry [mailto:gentras@vetmed.auburn.edu] Sent: Monday, June 27, 2005 9:55 AM To: Histonet Subject: [Histonet] S/P Pro-Texx* Mounting Medium Good morning All, do any of you have a source for S/P Pro-Texx* Mounting Medium, Cat. M7635-1? If so will you please share it with me ASAP? If it's no longer on the market will someone please provide ordering info for a comparable mounting medium? Your prompt reply will be much appreciated. Atoska _______________________________________________ Histonet mailing list Histonet@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 19, Issue 39 **************************************** From mpaula <@t> rpa.fmrp.usp.br Mon Jun 27 15:26:32 2005 From: mpaula <@t> rpa.fmrp.usp.br (mpaula) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Oil Red O mountant References: Message-ID: <000001c57bd8$9a439580$24cc6b8f@fmrp.usp.br> Hello Andi, I have monting my fat slide with a monting medium specific for Oil Red O Protocol is: Apathy's Mouting Media 50 g sucrose PA 50 g Acacia (gum arabic) PA 150 ml Distilled Water 0.1 g Thymol Mix Acacia and distilled water in a Becker. Place the Becker in a pan of boiling water until Aacia is dissolved. Than mix sucrose until dissolved and put on a Thymol. Refrigarete to remove air bubbles. Carffull with bubbles. MariaPaula Montiani Scandar Department of Pathology Labory of Neuropathology School of Medicine of Ribeir?o Preto University of S?o Paulo Brazil ----- Original Message ----- From: "Anita Jennings" Cc: Sent: Wednesday, June 22, 2005 10:42 AM Subject: [Histonet] Oil Red O mountant > > > > > Andi > I had the same problem. initially with my fat stain.....Poly Scientific kit > K043 contains the glycerine jelly mounting medium specific for Oil Red > coverslipping, havent had a problem since I started ordering this kit > 800-645-5825 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.7.11/26 - Release Date: 22/06/05 > > From fillsigh <@t> hotmail.com Tue Jun 28 07:26:18 2005 From: fillsigh <@t> hotmail.com (phil tsai) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] post-fixing quick frozen tissues for IHC Message-ID: Hello all, I was wondering if anyone has had any problems with tissues that were quick frozen before being being fixed and cryoprotected? We usually obtain our tissues fresh, fix in some sort of paraformaldehyde solution (Zamboni's, Lanas), cryoprotect in sucrose solution for around 1 week, and then quick freeze. However, for our positive controls, we are only able to obtain quick frozen tissues that haven't yet been fixed. I suppose that this might affect the achitecture a bit, but does anyone have any knowledge of whether immunohistochemistry might be affected? We are looking for nerve innervation, and will be obtaining muscle and skin for these purposes.... -Phil From andrew.macduff <@t> ed.ac.uk Tue Jun 28 09:02:33 2005 From: andrew.macduff <@t> ed.ac.uk (andrew.macduff@ed.ac.uk) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Adrenomedullin Ab for use in Mouse tissue Message-ID: <1119967353.42c15879c6f97@staffmail.ed.ac.uk> Dear All Does anyone have experience in staining for Adrenomedullin in mouse tissue? Who supplied the antibody and at what concentration did you use it? Thanks for any help Andy Andrew MacDuff Clinical Research Fellow MRC Centre for Inflammation Research Medical School Edinburgh University From alightbown <@t> cellsignal.com Tue Jun 28 07:23:53 2005 From: alightbown <@t> cellsignal.com (Lightbown, Angela) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Re: Histonet Digest, Vol 19, Issue 37 Message-ID: <7DCC6AD7-E7CF-11D9-8F63-00039391CC7C@cellsignal.com> On Tuesday, June 28, 2005, at 08:21 AM, histonet-request@lists.utsouthwestern.edu wrote: > 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. S100 (Laura Fidgen) > 2. PAR staining - Japanese source? (JonesLy@mir.wustl.edu) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 24 Jun 2005 13:50:18 -0400 > From: Laura Fidgen > Subject: [Histonet] S100 > To: histonet@lists.utsouthwestern.edu > Message-ID: <6.0.0.22.0.20050624134204.0271bec0@pop.vt.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > We are a Veterinary Hospital evaluating a Ventana Benchmark LT. I am > having a lot of problems staining for S100. It is very dark with > background. The people at Ventana think the secondary antibody is > cross-reacting and suggest using a different secondary (our secondary > currently is goat/rabbit). Can anyone in veterinary medicine help me > out > with this? So far my experience with this machine has not been > favorable. Any advice would be appreciated. > > Laura L. Fidgen, MScF, BSc, HT(ASCP) > Laboratory and Research Practioner > VMRCVM, Histopatholgy > Blacksburg, VA 24060-0443 > > > > > ------------------------------ > > Message: 2 > Date: Fri, 24 Jun 2005 13:13:12 -0500 > From: JonesLy@mir.wustl.edu > Subject: [Histonet] PAR staining - Japanese source? > To: histonet@lists.utsouthwestern.edu > Message-ID: > 8625702A.00641606@msnotes.wustl.edu> > > Content-Type: text/plain; charset=US-ASCII > > > Hello - > The person who started this project is currently in China, but my PI > asked > if I could query the list anyway - apologies if my question is vague. > I > collect the tissues we stain, but have little involvement in their > subsequent staining and am NOT a histologist. > > We need to stain formalin-fixed parafin embedded tissue (from > streptozotocin-treated rats) for PAR, Poly-ADP-ribose - not PARP, > poly-ADP-ribose polymerase. When we started this project, we had an > antibody that gave beautiful results; unfortunately subsequent lot > numbers > show poor staining of our control block. (The vendor has acknowleded > that > the problem is on their end, but that doesn't help our research.) > While at > a meeting in DC, the PI spoke with someone who was also working with > PAR > and has excellent results using a stain (monoclonal antibody?) from a > vendor in Japan. It is apparently expensive, but gives very > reproducable > results. The person working with this stain was supposed to e-mail > our PI > with the details, but he hasn't heard from her. > > Even if no-one has worked with the stain, a list of Japanese suppliers > of > antibodies would help. (Again, we need to stain for PAR, not PARP.) > Thanks in advance, > Lynne Jones > > Senior Research Technician > Dept. of Radiological Sciences > Washington University Medical School > St. Louis, MO > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 19, Issue 37 > **************************************** > > Angela Lightbown Clinical Applications/IHC Cell Signaling Technology 166B Cummings Center Beverly, MA 01915 phone: (978) 867-2440 email: alightbown@cellsignal.com From lu_ze <@t> sbcglobal.net Tue Jun 28 10:01:05 2005 From: lu_ze <@t> sbcglobal.net (Ze Lu) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] question on Nikon DIAPHOT-TMD microscope and camera Message-ID: <001501c57bf2$358be5b0$5b02a8c0@OPTIMUM2> Hello, histonet friend, We have a Nikon DIAPHOT-TMD microscope and camera in lab. We want to have a digital camera connected. I have some questions as I have not much microscopy experience. Does this model microscope has a C-mount? Also we are using the microscope for fluorescence study, and thinking to buy a ProgRes C-10 plus camera. I am wodering whether this camera that will meet regular requirement. However if you have experience with other camera, we will also appreciate your input. We are planning to buy a camera that is not very expensive as this microscope is old. Thank you. Ze Lu, Ph.D. Optimum Therapeutics, LLC From pruegg <@t> ihctech.net Tue Jun 28 10:11:53 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] post-fixing quick frozen tissues for IHC In-Reply-To: Message-ID: <200506281511.j5SFBikM012810@chip.viawest.net> I have the same issues, I get tissue quick frozen without any OCT or surcrose cryoprotectant from the molecular biologists all the time, sometimes the tissue is not even quick frozen it is just taken fresh and stuck in a tube in the -80dc freezer and the morphology is terrible. There was a technique a year ago in Histologic about thawing and refreezing poorly prepared samples, I have tried this with some improvement but it is never as good as doing it right in the first place. I would be interested in others approach to this issue as I don't see an end to getting samples not properly prepared for frozen sectioning. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of phil tsai Sent: Tuesday, June 28, 2005 5:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] post-fixing quick frozen tissues for IHC Hello all, I was wondering if anyone has had any problems with tissues that were quick frozen before being being fixed and cryoprotected? We usually obtain our tissues fresh, fix in some sort of paraformaldehyde solution (Zamboni's, Lanas), cryoprotect in sucrose solution for around 1 week, and then quick freeze. However, for our positive controls, we are only able to obtain quick frozen tissues that haven't yet been fixed. I suppose that this might affect the achitecture a bit, but does anyone have any knowledge of whether immunohistochemistry might be affected? We are looking for nerve innervation, and will be obtaining muscle and skin for these purposes.... -Phil _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bruyntjes <@t> voeding.tno.nl Tue Jun 28 10:28:00 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] GFAP Message-ID: <3B070848E7C2204F9DEB8BCFD7677280055CE4F3@ntexch1.voeding.tno.nl> Hi all Is anyone of you familiar with the staining of GFAP on muscular structures of blood vessels and bronchi/bronchioli in na?ve rats. The staining which appeared in my sections was not slight or weak, but really strong. As a 'positive control' I used slides of the brain (rat tissue). In these brain slides I found a rather big artery and that vessel was 100% negative? The antibody I have used is a polyclonal antibody from Dako (code Z0334). Does anyone of you know why those vessels in the lungs are positive for GFAP? Joost Bruijntjes TNO Quality of Life Zeist Holland This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From mpaula <@t> rpa.fmrp.usp.br Tue Jun 28 12:09:19 2005 From: mpaula <@t> rpa.fmrp.usp.br (mpaula) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] OIL RED O - Mounting Media Message-ID: <003301c57c04$20fb9840$24cc6b8f@fmrp.usp.br> I mounted my fat slide with a mounting medium specific for Oil Red O The Protocol is: Apathy's Mouting Media 50 g sucrose PA 50 g Acacia (gum arabic) PA 150 ml Distilled Water 0.1 g Thymol Mix Acacia and distilled water in a Becker. Place the Becker in a pan of boiling water until the Acacia is dissolved. Than mix sucrose until dissolved and put in a Thymol. Refrigerate to remove air bubbles. Carefful with bubbles. Thanks MariaPaula Montiani Scandar Department of Pathology Labory of Neuropathology School of Medicine of Ribeir?o Preto University of S?o Paulo Brazil ----- Original Message ----- From: "Anita Jennings" Cc: Sent: Wednesday, June 22, 2005 10:42 AM Subject: [Histonet] Oil Red O mountant > > > > > Andi > I had the same problem. initially with my fat stain.....Poly Scientific kit > K043 contains the glycerine jelly mounting medium specific for Oil Red > coverslipping, havent had a problem since I started ordering this kit > 800-645-5825 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.7.11/26 - Release Date: 22/06/05 > > From mcauliff <@t> umdnj.edu Tue Jun 28 12:13:19 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] GFAP In-Reply-To: <3B070848E7C2204F9DEB8BCFD7677280055CE4F3@ntexch1.voeding.tno.nl> References: <3B070848E7C2204F9DEB8BCFD7677280055CE4F3@ntexch1.voeding.tno.nl> Message-ID: <42C1852F.2070300@umdnj.edu> So what does the staining look like? Smooth muscle cells, fibroblasts, endothelial cells or ?? Exactly where in the artery (intima, media, adventitia)? Did you run a negative control, one without primary antibody? If not you could be picking up endogenous peroxidase activity. Not all arteries are the same so variations may not mean much. Geoff Bruijntjes, J.P. wrote: >Hi all > >Is anyone of you familiar with the staining of GFAP on muscular structures of blood vessels and bronchi/bronchioli in na?ve rats. The staining which appeared in my sections was not slight or weak, but really strong. > >As a 'positive control' I used slides of the brain (rat tissue). In these brain slides I found a rather big artery and that vessel was 100% negative? > >The antibody I have used is a polyclonal antibody from Dako (code Z0334). > >Does anyone of you know why those vessels in the lungs are positive for GFAP? > >Joost Bruijntjes >TNO Quality of Life >Zeist >Holland > > This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From int09018 <@t> alphahunt.com Tue Jun 28 12:14:37 2005 From: int09018 <@t> alphahunt.com (HCS) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] looking for used shandon embedding center , working or not Message-ID: <0b6401c57c04$ddb7c990$6601a8c0@hp> Hi, my heating unit on the storage side of the embedder has gone out. Does anyone have a Shandon embedding center they would like to sell of perhaps a parts machine. This is the mostly white embedder with the black front. Thanks, LeRoy Brown HT(ASCP) HTL Histology Consultation Services Everson, WA 98247 1-360-966-7300 www.histocs.com rhbrown@histocs.com From dmccaig <@t> ckha.on.ca Tue Jun 28 12:19:18 2005 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] uneven haematoxylin/eosin staining Message-ID: <3E5A3F039F0BD8118B4700C00D0020240435FB@CKHA9> We have been experiencing uneven staining recently. Even when all sections are cut on the same microtome and put in the same rack on an autostainer, their macroscopic appearance show incredible variations. It is so random we can not identify the problem. The Harris hemotoxylin is filtered daily. The tissue type does not make any difference. We can have 4 prostate slides and 2 will be good and 2 will show random staining intensities. Recuts may show variation in a different area which makes me think it is the staining and not the cutting. Any suggestions would be truly appreciated. Diana McCaig, MLT From eca9 <@t> georgetown.edu Tue Jun 28 12:24:38 2005 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] anti-HER2 ab Message-ID: <42C187D6.3090900@georgetown.edu> Hello Everyone, I ordered mouse anti-HER2, clone CB11 (cat.no. 18-7107) from Zymed. The spec sheet does however not say anything about a concentration. I want to use it for FFPE tissue. Has anyone out there used this antibody and if so are you willing to share yours conditions? Thanks, Eva Andersson Georgetown University From MDiCarlo <@t> KaleidaHealth.Org Tue Jun 28 12:32:48 2005 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] sharpening tungsten carbide knives Message-ID: <139141F8BAF4A642A945ECC528511AF001F5ECE6@kalmb02.kaleidahealth.org> Hello histonetters, I was told I can not sharpen tungsten carbide blades using Shandon's Autosharp 5 to cut large decalcified paraffin blocks on a Polycut E. I just purchased a knife holder, two plates and pastes and now I won't be able to use them. What do you use or where do you send your tungsten carbide knives to be resharpened? I appreciate your responses. Thank you. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From lwhite <@t> lakeridgehealth.on.ca Tue Jun 28 12:31:44 2005 From: lwhite <@t> lakeridgehealth.on.ca (White, Lori) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Problem with CC1 on Benchmark Message-ID: Bill, We have also been experiencing non-specific staining and do not use CC1 but retrieve offline. The problem appears to be with specific lot # of detection kits. Ventana is aware of this problem and should be working with you to resolve it. Good luck! Lori Message: 5 Date: Tue, 28 Jun 2005 11:16:52 +1000 From: "Bill Sinai" Subject: [Histonet] Problem with CC1 on Benchmark. To: "histonet \(E-mail\)" Message-ID: <000001c57b7f$11f8aca0$0ecd080a@wsahs.nsw.gov.au> Content-Type: text/plain; charset="iso-8859-1" Dear All, Particularly those using Ventana IHC Stainers. We have a Ventana Benchmark and recently (from the middle of May) we have a problem with antibodies which have had a pre-treatment in CC1. 1 After CC1, whether mild or standard we are getting non-specific biotin type staining. However, when you look at the slides they look specific in their staining although for the wrong antibody. The negative controls are displaying this phenomenon as well. 2. The staining can be blocked with the Avidin-Biotin block, but this does not always remove the staining completely. We are using Batch #'s 48091 & 48127 of CC1. Any advice would be greatly appreciated. Thanks Jane Milliken, IHC Benchmark Driver Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 Lori White, B.Sc., MLT Charge Technologist, Histopathology Lakeridge Health Corporation Tel 905 576-8711 ext. 4358 Fax 905 905 721-4757 From Julie.Sanders <@t> med.va.gov Tue Jun 28 12:42:57 2005 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] RE: Histonet Digest, Vol 19, Issue 40 Message-ID: <457381D92B01BD44B21CF37CC02EBDFD02927690@vhacinexc2.v10.med.va.gov> Jane, Check the pH of your buffer solution. We had trouble with non specific staining and turned out to be high pH of the buffer. Of course it happened on more than the CC1 treated slides, but its worth a try. Julie Julie Sanders, Supervisor, Anatomic Pathology VAMC Cincinnati, Ohio "Dear All, Particularly those using Ventana IHC Stainers. We have a Ventana Benchmark and recently (from the middle of May) we have a problem with antibodies which have had a pre-treatment in CC1. 1 After CC1, whether mild or standard we are getting non-specific biotin type staining. However, when you look at the slides they look specific in their staining although for the wrong antibody. The negative controls are displaying this phenomenon as well. 2. The staining can be blocked with the Avidin-Biotin block, but this does not always remove the staining completely. We are using Batch #'s 48091 & 48127 of CC1. Any advice would be greatly appreciated. Thanks Jane Milliken," From Melissa.Gonzalez <@t> cellgenesys.com Tue Jun 28 12:44:31 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] frozen cryoprotected tissue Message-ID: Dear Phil and Patsy, Unfortunately, I do not have an answer for your question, and would be curious myself to see if anyone has a solution to this problem. While I have you guys on the fixed, sucrose cryoprotection and then freezing method of processing tissues, I have some questions regarding the process. 1. First, how do you freeze the tissue when it is ready (after sucrose)? I have known people to freeze it directly in OCT in the cryostat or -20C, or also a quick freeze as you would fresh frozen tissue in a dry-ice isopentane bath. I myself prefer the slower freeze, as the tissues always seem to cut much better, and have never seen any compromise in morphology/staining. When the tissues are already fixed and cryoprotected, does the temp really matter? 2. I have however, noticed that regardless of the freezing temp, the H&E's appear strange. The cells are usually separated and the nuclear detail is very poor. Even though the H&E process is the same we would use for fresh frozens. So the fresh frozen slides actually turn out better than the fixed, cryoprotected samples. (Our processing procedure is as follows: after resection, post-fix tissue in 4% NB paraformaldehyde 3 hrs RT, followed by 30% sucrose immersion overnight at +4C or until tissue sinks, followed by (slow/quick) freezing in OCT. Any comments? Thanks Melissa --------------------------------------------------------------------- Hello all, I was wondering if anyone has had any problems with tissues that were quick frozen before being being fixed and cryoprotected? We usually obtain our tissues fresh, fix in some sort of paraformaldehyde solution (Zamboni's, Lanas), cryoprotect in sucrose solution for around 1 week, and then quick freeze. However, for our positive controls, we are only able to obtain quick frozen tissues that haven't yet been fixed. I suppose that this might affect the achitecture a bit, but does anyone have any knowledge of whether immunohistochemistry might be affected? We are looking for nerve innervation, and will be obtaining muscle and skin for these purposes.... -Phil Message: 14 Date: Tue, 28 Jun 2005 09:11:53 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] post-fixing quick frozen tissues for IHC To: "'phil tsai'" , Message-ID: <200506281511.j5SFBikM012810@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" I have the same issues, I get tissue quick frozen without any OCT or surcrose cryoprotectant from the molecular biologists all the time, sometimes the tissue is not even quick frozen it is just taken fresh and stuck in a tube in the -80dc freezer and the morphology is terrible. There was a technique a year ago in Histologic about thawing and refreezing poorly prepared samples, I have tried this with some improvement but it is never as good as doing it right in the first place. I would be interested in others approach to this issue as I don't see an end to getting samples not properly prepared for frozen sectioning. Patsy From Evelyn.Flynn <@t> childrens.harvard.edu Tue Jun 28 12:50:11 2005 From: Evelyn.Flynn <@t> childrens.harvard.edu (Flynn, Evelyn) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] sharpening tungsten carbide knives References: <139141F8BAF4A642A945ECC528511AF001F5ECE6@kalmb02.kaleidahealth.org> Message-ID: Dear Peggy, I have received excellent service from Delaware Diamond Knives in Wilmington, DE (www.ddk.com). Their turn-around time is about one week. The cost is approximately $215 to resharpen a 16 cm tungsten carbide knife. Evelyn Flynn Dept. Orthopedic Research Children's Hospital, Boston ---Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of DiCarlo, Margaret Sent: Tue 6/28/2005 1:32 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] sharpening tungsten carbide knives Hello histonetters, I was told I can not sharpen tungsten carbide blades using Shandon's Autosharp 5 to cut large decalcified paraffin blocks on a Polycut E. I just purchased a knife holder, two plates and pastes and now I won't be able to use them. What do you use or where do you send your tungsten carbide knives to be resharpened? I appreciate your responses. Thank you. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Tue Jun 28 13:27:08 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] uneven haematoxylin/eosin staining In-Reply-To: <3E5A3F039F0BD8118B4700C00D0020240435FB@CKHA9> References: <3E5A3F039F0BD8118B4700C00D0020240435FB@CKHA9> Message-ID: <42C1967C.7060609@umdnj.edu> In my experince incomplete removal of wax is a very common cause of uneven staining. I suggest fresh xylene (or whatever you use) and more changes, longer times, agitation or all of these. Geoff Diana McCaig wrote: >We have been experiencing uneven staining recently. Even when all sections >are cut on the same microtome and put in the same rack on an autostainer, >their macroscopic appearance show incredible variations. It is so random we >can not identify the problem. The Harris hemotoxylin is filtered daily. >The tissue type does not make any difference. We can have 4 prostate slides >and 2 will be good and 2 will show random staining intensities. Recuts may >show variation in a different area which makes me think it is the staining >and not the cutting. > >Any suggestions would be truly appreciated. >Diana McCaig, MLT > > >_______________________________________________ >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 41dmb41 <@t> gmail.com Tue Jun 28 13:30:32 2005 From: 41dmb41 <@t> gmail.com (Drew Meyer) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] uneven haematoxylin/eosin staining In-Reply-To: <3E5A3F039F0BD8118B4700C00D0020240435FB@CKHA9> Message-ID: <42c19748.100c8b23.5c76.54fc@mx.gmail.com> Are you making your own Hematoxylin and Eosin, or are you buying it? If you're buying it, what are you using? Drew -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: Tuesday, June 28, 2005 1:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] uneven haematoxylin/eosin staining We have been experiencing uneven staining recently. Even when all sections are cut on the same microtome and put in the same rack on an autostainer, their macroscopic appearance show incredible variations. It is so random we can not identify the problem. The Harris hemotoxylin is filtered daily. The tissue type does not make any difference. We can have 4 prostate slides and 2 will be good and 2 will show random staining intensities. Recuts may show variation in a different area which makes me think it is the staining and not the cutting. Any suggestions would be truly appreciated. Diana McCaig, MLT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From 41dmb41 <@t> gmail.com Tue Jun 28 13:34:00 2005 From: 41dmb41 <@t> gmail.com (Drew Meyer) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] AE1/AE3 (Cytokeratin) Procedure Message-ID: <42c19816.57df51c8.62b2.59b7@mx.gmail.com> I was wondering if anyone out there using the Dako Artisan Stainer could post their procedure for the AE1/AE3. I'm having problems with the intensity of the Ig staining. The Ig is staining to light and I want to compare our procedure with others that I know work well. Thanks, Drew From ddittus787 <@t> aol.com Tue Jun 28 14:38:54 2005 From: ddittus787 <@t> aol.com (ddittus787@aol.com) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] anti-HER2 ab In-Reply-To: <42C187D6.3090900@georgetown.edu> References: <42C187D6.3090900@georgetown.edu> Message-ID: <8C74A30275B11D6-CD4-8D88@mblk-r33.sysops.aol.com> Eva: I have used this antibody with great results, if you are automated staining try 1:40 to 1:80,and this is probably a good starting point for manual as well. The antibody likes HIER with citrate and incubation at 37 degrees, it only needs about 30 minutes,go longer (60 minutes) for manual room temp staining. Dana -----Original Message----- From: Eva C Andersson To: histonet@lists.utsouthwestern.edu Sent: Tue, 28 Jun 2005 13:24:38 -0400 Subject: [Histonet] anti-HER2 ab Hello Everyone, I ordered mouse anti-HER2, clone CB11 (cat.no. 18-7107) from Zymed. The spec sheet does however not say anything about a concentration. I want to use it for FFPE tissue. Has anyone out there used this antibody and if so are you willing to share yours conditions? Thanks, Eva Andersson Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From azdudley <@t> hotmail.com Tue Jun 28 14:40:30 2005 From: azdudley <@t> hotmail.com (anita dudley) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Her2 antibody In-Reply-To: <8C7462724236049-8AC-1D7E3@mblk-r40.sysops.aol.com> Message-ID: I also use the tab 250 her-2 from zymed, with proteinase, and get great results. anita dudley providence hosp. mobile alabama >From: ddittus787@aol.com >To: ddittus787@aol.com, eca9@georgetown.edu, >histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Her2 antibody >Date: Thu, 23 Jun 2005 12:24:19 -0400 > >As a side note zymed also has Tab 250 clone of cerb-2 that only need enzyme >and then incubation and gives awesome reuslts as well. > dana > >-----Original Message----- >From: ddittus787@aol.com >To: eca9@georgetown.edu; histonet@lists.utsouthwestern.edu >Sent: Wed, 22 Jun 2005 16:29:03 -0400 >Subject: Re: [Histonet] Her2 antibody > > >anti-her 2 cb11 clone works very well with citrate hier dilution(depending >on >your method is 1:40-1:100) >and a 30 minute incubation at 37 degrees. > >dana > > > > > > > > > > > > > >-----Original Message----- >From: Eva C Andersson >To: histonet@lists.utsouthwestern.edu >Sent: Wed, 22 Jun 2005 14:30:14 -0400 >Subject: [Histonet] Her2 antibody > > >Hello, >I just ordered mouse anti-Her2 cat.no. 18-7107 from Zymed. There is however >no >indication given as to which dilution to use. I want to use it for >Immunohistochemistry on paraffin embedded sections. It also doesn't say >anything >about antigen retrieval. >Is anyone using this antibody and if so could you tell me what you do? >Thanks, >Eva Andersson >Georgetown University > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcresor <@t> lcpath.com Tue Jun 28 15:59:27 2005 From: jcresor <@t> lcpath.com (Jennifer N. Cresor) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Phenol green gram stain Message-ID: <200506282059.j5SKxTY06072@fauna.host4u.net> Hello, Does anyone have a procedure for a gram stain sounding similar to phenol green gram stain? I understand it has a green background. I am trying to locate this. Any information would be helpful. Thank you, Jennifer Lower Columbia Pathologists jcresor@lcpath.com (360)575-9419 From histology.bc <@t> shaw.ca Tue Jun 28 18:23:27 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] uneven haematoxylin/eosin staining References: <3E5A3F039F0BD8118B4700C00D0020240435FB@CKHA9> Message-ID: <42C1DBEF.6090008@shaw.ca> Sounds like a classical case of incomplete removal of wax before staining begins. Check the de-waxing reagents and times. Paul Bradbury Kamloops, BC Canada Diana McCaig wrote: >We have been experiencing uneven staining recently. Even when all sections >are cut on the same microtome and put in the same rack on an autostainer, >their macroscopic appearance show incredible variations. It is so random we >can not identify the problem. The Harris hemotoxylin is filtered daily. >The tissue type does not make any difference. We can have 4 prostate slides >and 2 will be good and 2 will show random staining intensities. Recuts may >show variation in a different area which makes me think it is the staining >and not the cutting. > >Any suggestions would be truly appreciated. >Diana McCaig, MLT > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From pengbw <@t> sjtu.edu.cn Tue Jun 28 19:35:04 2005 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Blocking serum Message-ID: <20050629003504.7EDCD11104FD@sjtu.edu.cn> Hello, all I\'m doing IHC on mouse frozen sections. The first antibody in my hand is Rabbit polyclonal antibody,the second antibody is Goat anti Rabbit IgG(H+L). I\'m wondering what kind of serum I should use to block non specific staining. Thanks in advance! Baowei Peng 1954 Huashan Road Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China Ph:86-21-62932108 E-mail:pengbw@sjtu.edu.cn From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Jun 29 02:39:34 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] RE: Histonet Digest, Vol 19, Issue 39 - CD 138 Message-ID: We have tried both the Vector clone 5F7 and the Lab Vision clone B-B4. On our Ventana Benchmark XT, the B-B4 worked really well, but it didn't like Vector's and only occasional cells stained. Jacqui Malam Royal Infirmary Lancaster England -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 27 June 2005 18:02 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 19, Issue 39 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. Freezing Antobodies ! (Cesar Francisco Romero) 2. RE: S100 staining (Malam Jacqueline) 3. OT:: [Histonet] immunostainers (louise renton) 4. RE: pos control for copper (Malam Jacqueline) 5. prognostic markers and IHC staining (C. Loukas) 6. Re: Freezing Antobodies ! (Katri Tuomala) 7. RE: : [Histonet] immunostainers (O'Brien, Sue) 8. S/P Pro-Texx* Mounting Medium (Atoska S. Gentry) 9. Re: prognostic markers and IHC staining (Richard Cartun) 10. MUM1/IRF4 and CD138 (Pat Zeitlow) 11. RE: S/P Pro-Texx* Mounting Medium (Tom McNemar) ---------------------------------------------------------------------- Message: 1 Date: Mon, 27 Jun 2005 02:03:14 -0300 From: "Cesar Francisco Romero" Subject: [Histonet] Freezing Antobodies ! To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1"; format="flowed" Dear People from Histonet I would like to know if you freeze your antibodies in Kitchen like Freezers ( - 18 or - 20 ? C ). I can not afford a - 80 ? C Freezer. Thank You in advance Cesar Romero Buenos Aires Argentina _________________________________________________________________ Nuevo MSN Messenger [1]Una forma r?pida y divertida de enviar mensajes References 1. http://g.msn.com/8HMAESAR/2734??PS=47575 ------------------------------ Message: 2 Date: Mon, 27 Jun 2005 08:44:38 +0100 From: Malam Jacqueline Subject: [Histonet] RE: S100 staining To: "Histonet submissions (histonet@lists.utsouthwestern.edu)" Message-ID: Content-Type: text/plain Hi Laura We have a Benchmark XT and it works fine for us - can you let me know your ab source, protocol and dilution - see if I can shed some light. Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 3 Date: Mon, 27 Jun 2005 09:51:30 +0200 From: louise renton Subject: OT:: [Histonet] immunostainers To: CrochiereSteve@aol.com, Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 On 6/26/05, CrochiereSteve@aol.com wrote: > I wholeheartedly recommend the Nemesis Immunostainer from Biocare Medical. > I have been using this instrument for almost a year and have had no > technical problems with this machine. Does anyone think about the meaning of words when they name these machines? NEMISIS: A source of harm or ruin: Retributive justice in its execution or outcome: An opponent that cannot be beaten or overcome. One that inflicts retribution or vengeance. Nemesis Greek Mythology. The goddess of retributive justice or vengeance. ------------------------------ Message: 4 Date: Mon, 27 Jun 2005 10:30:50 +0100 From: Malam Jacqueline Subject: [Histonet] RE: pos control for copper To: "Histonet submissions (histonet@lists.utsouthwestern.edu)" Message-ID: Content-Type: text/plain Many thanks to everyone who responded. I am now the proud possessor of a positive control block - my world is complete ! Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 5 Date: Mon, 27 Jun 2005 12:08:00 +0000 From: "C. Loukas" Subject: [Histonet] prognostic markers and IHC staining To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hello all, Although I'm new in this list, it seems that is viewed by loads of Histologists, so I would be grateful if someone could recommend me a couple of review articles/textbook focusing on the assessment of the various prognostic markers currently in use (e.g. ER in breast cancer, MIB-1 in oesophageal cancers), through immunohistochemical staining of tumor tissue sections. In particular I'm after a list of: established prognostic markers (or stains) vs tumor type, if such a think exists. Thanks in advance! cl ------------------------------ Message: 6 Date: Mon, 27 Jun 2005 09:01:26 -0400 From: "Katri Tuomala" Subject: Re: [Histonet] Freezing Antobodies ! To: "Cesar Francisco Romero" , Message-ID: <001001c57b18$5471c1a0$6a9a9618@Katri> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=response Cesar, If your freezer is a self defrosting kind, it is not a good idea to keep antibodies or frozen tissue in it. There is too much temperature fluctuation as it defrosts itself. Chest freezers keep the temperature more stable and may be acceptable. Keep a thermometer in it to record the temperature daily. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Cesar Francisco Romero" To: Sent: Monday, June 27, 2005 1:03 AM Subject: [Histonet] Freezing Antobodies ! > > Dear People from Histonet > > I would like to know if you freeze your antibodies in Kitchen like > Freezers ( - 18 or - 20 ? C ). > > I can not afford a - 80 ? C Freezer. > > Thank You in advance > > Cesar Romero > > Buenos Aires > > Argentina > _________________________________________________________________ > > Nuevo MSN Messenger [1]Una forma r?pida y divertida de enviar mensajes > > References > > 1. http://g.msn.com/8HMAESAR/2734??PS=47575 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Mon, 27 Jun 2005 09:38:47 -0400 From: "O'Brien, Sue" Subject: RE: : [Histonet] immunostainers To: "louise renton" , , Message-ID: <4D95C24EC0E4C84787A6919F1165B25E24B16A@btmhems01.BTHOSP.INT> Content-Type: text/plain; charset="us-ascii" Steve, Sounds like the company is focusing on the "An opponent that cannot be beaten or overcome" part of the definition. I also have a Nemesis (since Feb) and have been VERY pleased with its performance. I would highly recommend it to anyone considering an automated Immunostainer. Susan O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May court House, NJ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Monday, June 27, 2005 3:52 AM To: CrochiereSteve@aol.com; Histonet@lists.utsouthwestern.edu Subject: OT:: [Histonet] immunostainers On 6/26/05, CrochiereSteve@aol.com wrote: > I wholeheartedly recommend the Nemesis Immunostainer from Biocare Medical. > I have been using this instrument for almost a year and have had no technical > problems with this machine. Does anyone think about the meaning of words when they name these machines? NEMISIS: A source of harm or ruin: Retributive justice in its execution or outcome: An opponent that cannot be beaten or overcome. One that inflicts retribution or vengeance. Nemesis Greek Mythology. The goddess of retributive justice or vengeance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Mon, 27 Jun 2005 08:55:20 -0500 From: "Atoska S. Gentry" Subject: [Histonet] S/P Pro-Texx* Mounting Medium To: Histonet Message-ID: <6.0.1.1.0.20050627084308.01b1a090@mailhost.vetmed.auburn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Good morning All, do any of you have a source for S/P Pro-Texx* Mounting Medium, Cat. M7635-1? If so will you please share it with me ASAP? If it's no longer on the market will someone please provide ordering info for a comparable mounting medium? Your prompt reply will be much appreciated. Atoska ------------------------------ Message: 9 Date: Mon, 27 Jun 2005 10:58:13 -0400 From: "Richard Cartun" Subject: Re: [Histonet] prognostic markers and IHC staining To: , Message-ID: Content-Type: text/plain; charset=US-ASCII A good article dealing with breast cancer is, "Prognostic and Predictive Factors in Breast Cancer by Immunohistochemical Analysis" by D. Craig Allred et al. in Modern Pathology 1998:11(2):155-168. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "C. Loukas" 06/27/05 08:08AM >>> Hello all, Although I'm new in this list, it seems that is viewed by loads of Histologists, so I would be grateful if someone could recommend me a couple of review articles/textbook focusing on the assessment of the various prognostic markers currently in use (e.g. ER in breast cancer, MIB-1 in oesophageal cancers), through immunohistochemical staining of tumor tissue sections. In particular I'm after a list of: established prognostic markers (or stains) vs tumor type, if such a think exists. Thanks in advance! cl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 10 Date: Mon, 27 Jun 2005 10:29:39 -0500 From: "Pat Zeitlow" Subject: [Histonet] MUM1/IRF4 and CD138 To: "Histonet" Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A47D652@bbplsrv1.bbpl> Content-Type: text/plain; charset="US-ASCII" Is anyone using MUM1/IRF4 and/or CD138 that would be willing to share vendors, clones or protocols for use with routine FFPE IHC? Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. ------------------------------ Message: 11 Date: Mon, 27 Jun 2005 12:00:24 -0400 From: Tom McNemar Subject: RE: [Histonet] S/P Pro-Texx* Mounting Medium To: "'Atoska S. Gentry'" , Histonet Message-ID: <6CD94D97ED7D924BA5C2B588FA9528213967D5@mail.lmhealth.org> Content-Type: text/plain; charset="iso-8859-1" We used Pro-tex for many years and got from Scientific Products. Two maybe three years ago they (SP) substituted Cytoseal 60 (made by Richard Alan). Works just as well so that's what we are using to this day. Pro-tex is no longer available throgh Scientific Products. The order numbe for Cytoseal 60 M7630-2A and the phone number for Scientific Products is (800) 964-5227. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Atoska S. Gentry [mailto:gentras@vetmed.auburn.edu] Sent: Monday, June 27, 2005 9:55 AM To: Histonet Subject: [Histonet] S/P Pro-Texx* Mounting Medium Good morning All, do any of you have a source for S/P Pro-Texx* Mounting Medium, Cat. M7635-1? If so will you please share it with me ASAP? If it's no longer on the market will someone please provide ordering info for a comparable mounting medium? Your prompt reply will be much appreciated. Atoska _______________________________________________ Histonet mailing list Histonet@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 19, Issue 39 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From c.m.vanderloos <@t> amc.uva.nl Wed Jun 29 03:26:32 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] RE: MMP-1 Message-ID: <100f1da100b813.100b813100f1da@amc.uva.nl> Dear Samantha, You have done already a lot to make this MMP-1 antibody work. There are only a few things left that you may try from here on: * A new titration series (1:20 - 1:60 - 1:180) with an overnight primary antibody incubation at 4C. Tissue pretreatment: no treatment, HIER with EDTA pH9.0, HIER with citrate pH 6.0 and pronase * Test some different anti-MMP1 antibodies from different sources! To my opinion Santa Cruz antibodies are not the best guarantee for optimal staining results. With respect to other MMP antibodies (3 and 9) we have good staining results with LabVision reagents. Perhaps one of their anti-MMP-1 antibodies will work for you. * In case of negative staining, you need perhaps a more sensitive/efficient detection system. Tyramide amplification techniques can be helpful in this respect (Perkin&Elmer TSA NEL-700 kit or DakoCytomation CSA II kit). Good luck, Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Mon, 27 Jun 2005 15:34:30 -0600 From: "Samantha McMurtrey" Subject: [Histonet] MMP-1 To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hello all, I am having a lot of trouble getting any staining for MMP-1. I am using Santa Cruz, catalog # sc-21731, lot # A1405. It's a mouse monoclonal antibody. I have tried dilutions from 1:200 down to 1:25. I incubated the primary antibody for 30 minutes and the secondary for 30 minutes. I have also tried using proteinase K, Target, EDTA, and a pH 9 retrieval. I am working it up on formalin fixed paraffin embedded tissue. I am mostly using breast tissue that has atypical ductal hyperplasia (per the pathologist's request). I haven't had any luck at all. I am getting no staining. Please help! Thank you, Samantha McMurt! rey, BS, From BMolinari <@t> heart.thi.tmc.edu Wed Jun 29 06:00:37 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] sharpening tungsten carbide knives Message-ID: Symantec Mail Security replaced Message Body with this text message. The original text contained prohibited content and was quarantined. ID:THIMAIL::SYQd2caf72e From KPercival <@t> wyeth.com Wed Jun 29 06:43:26 2005 From: KPercival <@t> wyeth.com (Karen Percival) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] uneven haematoxylin/eosin staining Message-ID: Hi Diana, It sounds like a processing problem problem to me. You are not getting good infiltration. I would check the tissue processor for proper functioning and change all reagents as well if the processor functions look as if they are working properly. Check all reagents on the processor, too, for quality. Start with fresh everything for the next run. I have found that it's usually a reagent problem (usually technician error) if everything has been looking great and then all of a sudden something is wrong. Karen Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com >>> Diana McCaig 6/28/2005 1:19:18 PM >>> We have been experiencing uneven staining recently. Even when all sections are cut on the same microtome and put in the same rack on an autostainer, their macroscopic appearance show incredible variations. It is so random we can not identify the problem. The Harris hemotoxylin is filtered daily. The tissue type does not make any difference. We can have 4 prostate slides and 2 will be good and 2 will show random staining intensities. Recuts may show variation in a different area which makes me think it is the staining and not the cutting. Any suggestions would be truly appreciated. Diana McCaig, MLT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Wed Jun 29 07:03:24 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] tungsten knives Message-ID: Peggy, I used to sharpen my TC knives on the Autosharp but due to time constraints I now send them to Delaware Diamond Kinves (1-800-222-5143) Their turn around time is very good. I cut GMA samples and I have 2 knives so I always have one in the lab while the other is being sharpened. Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston, TX 77030 832-355-6524 From mhorne <@t> upei.ca Wed Jun 29 08:44:01 2005 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] evaporating Xylene Message-ID: <42C26D60.21398.28C3D1@localhost> I am a research tech and so don't do a run of slides every week. I am about to get a full set-up for deparafining and staining. What do other people do to reduce the xylenes from evaporating out from under the loose fitting glass lids? Or is there a staining station with tight lids out there? Any ideas appreciated, Margaret ps. Do the glass racks break often? They are cheaper than the metal racks. Any tips on the pro's and con's? 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 Histopatty <@t> aol.com Wed Jun 29 07:49:17 2005 From: Histopatty <@t> aol.com (Histopatty@aol.com) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Slide labelers Message-ID: <158.538904aa.2ff3f2cd@aol.com> Dose anyone have a slide labeler that is interfaced with their meditech histology data section? Patty Eneff Histopatty@aol.com Patricia.Eneff@hcahealthcare.com From jluis.palazon <@t> icman.csic.es Wed Jun 29 08:00:41 2005 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Bowies method Message-ID: <20050629130041.C67348238AD@perceval.uca.es> Dear List-fellows Could any of you send me the protocol of Bowie?s method for juxtaglomerular cells in kidney. thanks in advance Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From twheelock <@t> mclean.harvard.edu Wed Jun 29 08:42:27 2005 From: twheelock <@t> mclean.harvard.edu (Tim Wheelock) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] XYLENE EVAPORATION AND GLASS RACKS Message-ID: <42C2A543.8010400@mclean.harvard.edu> Hi Margaret: Perhaps the way to stop xylene evaporation is to put something on top of the containers like a cork board, or heavy cookie sheet, anything heavy enough that will help seal the xylene in the containers. Speaking of glass racks, they do seem to be somewhat fragile. I tried them out in an attempt to batch my silver stains years ago but they did not hold enough slides. I was wondering if anyone out there knew of glass racks that held 60 slides, not back-to-back, but just one slide per slot. I am still trying to find a way to batch the Bielschowsky silver stains, instead of dealing with 14 different coplan jars. Tim Wheelock Harvard Brain Bank McLean Hospital From jmahoney <@t> alegent.org Wed Jun 29 08:57:14 2005 From: jmahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] uneven haematoxylin/eosin staining Message-ID: I have experienced this problem at several labs where I have worked. Finally figured out it happens more in the summer when the humidity is high. It was due to incomplete removal of paraffin due to moisture in the xylene. Try changing it more often or filtering it. The filter paper will labsorbe the water. Jan Omaha NE >>> Paul Bradbury 06/28/2005 6:23:27 PM >>> Sounds like a classical case of incomplete removal of wax before staining begins. Check the de-waxing reagents and times. Paul Bradbury Kamloops, BC Canada Diana McCaig wrote: >We have been experiencing uneven staining recently. Even when all sections >are cut on the same microtome and put in the same rack on an autostainer, >their macroscopic appearance show incredible variations. It is so random we >can not identify the problem. The Harris hemotoxylin is filtered daily. >The tissue type does not make any difference. We can have 4 prostate slides >and 2 will be good and 2 will show random staining intensities. Recuts may >show variation in a different area which makes me think it is the staining >and not the cutting. > >Any suggestions would be truly appreciated. >Diana McCaig, MLT > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eca9 <@t> georgetown.edu Wed Jun 29 08:56:36 2005 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Silly question about deparaffinization Message-ID: <42C2A894.7070708@georgetown.edu> Good morning, I am probably asking a very silly question but I recently spoke to someone about deparaffinization and hydration. I found that the times that they use and the times that we use are very different. So my question to you is this: What lenghts of time do you use for Xylenes, 100% alcohol, 95% alcohol and 70% alcohol? It is about deparaffinising charged (+) slides. So nothing fancy. Just curious. Maybe I can cut back some on the timed that we use which are: 30min Xylenes, 10min 100%, 10min 95% and 6min 70%. Thanks for your responses. Eva Andersson Georgetown University From la.sebree <@t> hosp.wisc.edu Wed Jun 29 09:13:44 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Silly question about deparaffinization Message-ID: We deparaffinize in 3 changes of xylene, 2 changes of 100% ETOH, 1 95% ETOH and 1 70% ETOH, all for 3 minutes each. We never skimp on the xylene times but have been known to shorten the ETOH times if we manually agitate the slides and are in a real hurry. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C Andersson Sent: Wednesday, June 29, 2005 8:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly question about deparaffinization Good morning, I am probably asking a very silly question but I recently spoke to someone about deparaffinization and hydration. I found that the times that they use and the times that we use are very different. So my question to you is this: What lenghts of time do you use for Xylenes, 100% alcohol, 95% alcohol and 70% alcohol? It is about deparaffinising charged (+) slides. So nothing fancy. Just curious. Maybe I can cut back some on the timed that we use which are: 30min Xylenes, 10min 100%, 10min 95% and 6min 70%. Thanks for your responses. Eva Andersson Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Wed Jun 29 09:21:59 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Silly question about deparaffinization In-Reply-To: <42C2A894.7070708@georgetown.edu> References: <42C2A894.7070708@georgetown.edu> Message-ID: <6.1.1.1.2.20050629101442.019659d8@mail.vet.upenn.edu> Eva, Depending on the thickness of the sections my times can vary. Sections of 4 to 6 micron are deparaffinized in our laboratory at 2 changes at 5 minutes each with alcohols at 3 minutes each per station to water. Sections thicker than 6 microns may go as long 15 minutes X 2 in xylenes and 5 to 10 minutes in each alcohol. I am very paranoid about getting all of the paraffin (or MMA where I use one hour per change for xylenes) so I may go over board. I also change my reagents more often in humid weather. I do not use an automated stainer so agitation only takes place at the beginning of the changes to assure the last reagent is rinsed off. Hope this helps. At 09:56 AM 6/29/2005, Eva C Andersson wrote: >Good morning, >I am probably asking a very silly question but I recently spoke to someone >about deparaffinization and hydration. I found that the times that they >use and the times that we use are very different. So my question to you is >this: What lenghts of time do you use for Xylenes, 100% alcohol, 95% >alcohol and 70% alcohol? >It is about deparaffinising charged (+) slides. So nothing fancy. >Just curious. Maybe I can cut back some on the timed that we use which are: >30min Xylenes, 10min 100%, 10min 95% and 6min 70%. >Thanks for your responses. >Eva Andersson >Georgetown University > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From c.m.vanderloos <@t> amc.uva.nl Wed Jun 29 09:29:34 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] RE: Blocking serum Message-ID: <10b1c3d10b6fff.10b6fff10b1c3d@amc.uva.nl> Dear Baowei Peng, Since you have a goat anti-rabbit IgG as second step reagent you should use normal goat serum for pre-blocking step. Otherwise use Protein Block, serum-free (DakoCyto X0909). Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Wed, 29 Jun 2005 08:35:04 +0800 (BEIST) From: Baowei Peng Subject: [Histonet] Blocking serum To: histonet@lists.utsouthwestern.edu Message-ID: <20050629003504.7EDCD11104FD@sjtu.edu.cn> Content-Type: text/plain; charset="gb2312" Hello, all I\'m doing IHC on mouse frozen sections. The first antibody in my hand is Rabbit polyclonal antibody,the second antibody is Goat anti Rabbit IgG(H+L). I\'m wondering what kind of serum I should use to block non specific staining. Thanks in advance! Baowei Peng 1954 Huashan Road Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China Ph:86-21-62932108 E-mail:pengbw@sjtu.edu.cn From Barry.R.Rittman <@t> uth.tmc.edu Wed Jun 29 09:31:25 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Silly question about deparaffinization Message-ID: Eva I would suggest that you do a trial and error run. Deparaffinization time with xylene will depend on several factors including: thickness of sections, type of paraffin, number of slides, agitation, temperature of solution, volume of fluid, number of times that xylene bath was used, quality of the xylenes solution and so on. I have always erred on the conservative side and given a longer time as you have done in the past. However, for many labs, as Linda has indicated, 3 minutes in each bath is fine. Suggest that you try a set of slides and deparaffinize them for 3, 5 7 and 10 minutes and compare your results. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sebree Linda A. Sent: Wednesday, June 29, 2005 9:14 AM To: Eva C Andersson; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Silly question about deparaffinization We deparaffinize in 3 changes of xylene, 2 changes of 100% ETOH, 1 95% ETOH and 1 70% ETOH, all for 3 minutes each. We never skimp on the xylene times but have been known to shorten the ETOH times if we manually agitate the slides and are in a real hurry. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C Andersson Sent: Wednesday, June 29, 2005 8:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly question about deparaffinization Good morning, I am probably asking a very silly question but I recently spoke to someone about deparaffinization and hydration. I found that the times that they use and the times that we use are very different. So my question to you is this: What lenghts of time do you use for Xylenes, 100% alcohol, 95% alcohol and 70% alcohol? It is about deparaffinising charged (+) slides. So nothing fancy. Just curious. Maybe I can cut back some on the timed that we use which are: 30min Xylenes, 10min 100%, 10min 95% and 6min 70%. Thanks for your responses. Eva Andersson Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Wed Jun 29 09:32:07 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Phenol green gram stain Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171755E@lsexch.lsmaster.lifespan.org> I'm not sure what "phenol green" is? But the Gram-Twort technique is a gram stain with a green background, due to Fast Green FCF added to the red stain. Perhaps that's what you are looking for. You can find that technique here: http://www.scienceboard.net/resources/protocols.asp?action=article&protocol_ id=310&criteria= From nancy.troiano <@t> yale.edu Wed Jun 29 09:55:20 2005 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] tungsten carbide knife resharpening Message-ID: <5.2.1.1.2.20050629105118.00c05630@email.med.yale.edu> We have our tungsten carbide knives sharpened by Dorn/Hart Microedge, Inc. 135 Home Avenue Villa Park, IL 60181. Telephone number is 630-832-3843. They are great - quality work at a reasonable price, quick turnaround time. I have sent my tungsten carbide knives to them for the last 25 years for resharpening! Nancy Troiano, M.S., Associate in Research III Yale Core Center for Musculoskeletal Disorders Yale University P.O. Box 208071 New Haven, CT 06520 (203)785-5136 From nancy.troiano <@t> yale.edu Wed Jun 29 09:57:31 2005 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] tungsten carbide resharpening Message-ID: <5.2.1.1.2.20050629105626.00c11870@email.med.yale.edu> Forgot to mention that the price to resharpen a knife from Dorn/Hart is $150.00 for a 16 cm tungsten carbide knife. Turnaround time is approximately one week. From dmarsha3 <@t> utmem.edu Wed Jun 29 10:24:23 2005 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] mouse ankles for IHC Message-ID: <000a01c57cbe$a0d08de0$f623c084@DanaM> Hi everyone, I am gearing up for mouse ankle-joint histology. (I may do vertebral column eventually as well if anyone has done that.) I have a Cryojane tape transfer system and have done a bit of sectioning of the undecalcified bones and they look adequate. I haven't done any fancy staining yet. My question is a fairly broad one in that I wanted to ask those who might be doing this for a brief outline of their procedure from beginning to end. So far I am fixing in 10% formalin (in H2O) for a couple of days. I embed the joint with the heel towards the bottom of the block (I thought it might be more stable to cut up from the heel but perhaps not?) Ultimately I will do IHC staining and, possibly, some in situ hybridization. I have some previous info in regards to knives and angles of cutting etc and would appreciate any additional insights into that as well. I am having trouble stringing together the information that will allow me to make wise choices early on to maximize the use of my samples (and my mice). Any advice would be GREATLY appreciated. Thanks again. Dana (who dabbles in histology every 10 years whether I need to or not:-) From sbreeden <@t> nmda.nmsu.edu Wed Jun 29 10:29:01 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] New Mexico Society for Histology Message-ID: The New Mexico Society for Histology, after having been rather inactive for the past couple years, is preparing to go "back on track"! If you are a histologist (or have an interest therein) and live in New Mexico or one of our adjoining states, please consider joining us. If you have been a member and would like to reactivate, we'd love to have you back. There will be a general business meeting and short presentation on Saturday, August 6th, beginning at 10:00 a.m.; the meeting will be held at Tricore Reference Laboratories, 1001 Woodward Place NE, Albuquerque. If interested in either the meeting, or membership, please contact me at this email address, or at NMHISTO@AOL.COM. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From LewisS <@t> pediatrics.ohio-state.edu Tue Jun 28 15:03:42 2005 From: LewisS <@t> pediatrics.ohio-state.edu (Lewis, Sarah) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] OCT Message-ID: <714B9F12B4E18C4C843B66E8E190F2AD447526@res2k3ms01.CRII.ORG> Hello! I use 7% gum Tragacanth that I make up myself using 100ml of distilled water, 7gms of gum Tragacanth, and 5 grains of Thymol. Dissolve by manually stirring at rm temp.(may take several hours) Store half refrigerated(4c) and half frozen (-20c). I work with only muscle and nerve biopsy tissue but this works great for me. I am sure this is a bit "old school", but I have never had any trouble with cracking or stability of the tissue. It also seems to be great for long term storage in -80 freezer. Good luck!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net -----Original Message----- From: ssq5977@yahoo.com.cn [mailto:ssq5977@yahoo.com.cn] Sent: Thursday, June 16, 2005 9:30 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Is there anything better than OCT? Hi all: We are currently using OCT to embed renal biopsy tissue in cryostat mold. First of all ,we put a drop of OCT on the mold ,then we put the tissue on the top of the OCT and dip the mold into liquid nitrogen for 10 seconds. then the tissue is sent to cryosection. Does anyone know if there is anything better than OCT to embed the tissue? Thanks! Shuqiong Shen(ssq5977@yahoo.com.cn) Research Institute of Nephrology, Jinling Hospital 305 East Zhongshan Road, Nanjing Jiangsu Province,P.R of China Zip code: 210002 Tel: +86 25 85912177 Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net Upgrade Your Email - Click here! From PMonfils <@t> Lifespan.org Wed Jun 29 10:35:51 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] detachment of tissue from OCT compound Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171755F@lsexch.lsmaster.lifespan.org> Someone has sent me 60 rat brains frozen in OCT compound (whole brain except for the cerebellum and the anterior 1 cm). He wants multiple 30-micron sections from each brain. They appear to be well infiltrated with sucrose, and they cut smoothly on the cryostat at -20C. However, the sections of tissue fall out of the surrounding OCT, which makes it very difficult to pick them up on slides without a lot of wrinkles and bubbles. Any tricks to deal with this situation? Paul M. From JosefaNava <@t> texashealth.org Wed Jun 29 10:56:47 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Full time Histotech Opening Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944B6@phdex03.txhealth.org> We are looking for a full time Histotech or ASCP eligible position at Presbyterian Hospital of Dallas, Texas. Interested candidates may contact Maureen Hoops at 214-345-7795 for details and benefit information. 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 biswas.16 <@t> osu.edu Wed Jun 29 11:33:50 2005 From: biswas.16 <@t> osu.edu (SABYASACHI BISWAS) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Pig Skin immunohistochemistry-losing sections during antigen retrieval Message-ID: <23a1309239d9f7.239d9f723a1309@osu.edu> Hi All We are facing a problem with our pig skin biopsy samples. We are involved in wound healing research and therefore have to do histological staining of pig skin. Our biopsies are taken with 6mm biopsy punches around a 3mm biopsy punch taken previously (the wound). The samples are fixed in formalin and embedded in paraffin. We were losing sections even during H&E staining but managed to solve that by baking the slides at 60degrees C for 1hour instead of 37 degrees overnight; we also use Superfrost++ slides. However during antigen retrieval using the microwave method we are losing sections wholly or partially. Does anyone have experience with antigen retrieval in pig skin samples. We appreciate any help/advice that you can give us. Thanks Sabya Biswas The Ohio State University From petepath <@t> yahoo.com Wed Jun 29 12:15:55 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] detachment of tissue from OCT compound Message-ID: <20050629171555.69033.qmail@web30414.mail.mud.yahoo.com> Paul, I do not work with pre-frozen samples but my guess is your OCT has had some drying and coming away from the tissue. You can try a techniqe I call plastering after you have trimmed to where you will take the section. It will get some OCT into the space left by drying. I would use cold OCT to plaster so your tissue does not warm too much by the OCT and resuly in freeze artifact. The OCT will not penetrate very deep so you may have to repeat the procedure if you are cutting through a lot ofr tissue to get your section. See the bottom of the page at this link below to see this technique applied to a variety of situations. http://pathologyinnovations.com/new_page_2.htm Stephen From Rcartun <@t> harthosp.org Wed Jun 29 12:21:56 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] Guidelines for submission of tonsils Message-ID: "Thank you" to all of you who responded to my question of guidelines for submission of tonsils to pathology. There was no consensus. Interestingly, our ENT surgeons at our Children's Hospital have decided to stop sending tonsils to our Department unless there is a "clinical" suspicion of disease. And, from what we can tell from the medical literature, there is no reason to submit "routine" tonsils to Pathology for examination (gross/microscopic), regardless of the patient's age. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From schnegelsberg <@t> xgene.com Wed Jun 29 10:53:30 2005 From: schnegelsberg <@t> xgene.com (Birthe Schnegelsberg) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] preservation of the stratum corneum in skin cryosections Message-ID: Hello. Does anybody have a method to keep the stratum corneum attached to the epithelium using in vitro skin cryosections? I am freezing in vitro skin without fixation in isopentane chilled in liquid nitrogen. I don' t see any stratum corneum in my 5-7um HandE stained samples. Thanks, Birthe From Stephen.J.Scholz <@t> osfhealthcare.org Wed Jun 29 12:59:16 2005 From: Stephen.J.Scholz <@t> osfhealthcare.org (Scholz, Stephen J.) Date: Fri Sep 16 15:25:16 2005 Subject: [Histonet] fixation question Message-ID: <7F1312711CA7474A89B3DF8BA0BA54D01333A4@pmc-rfd-mx01.intranet.osfnet.org> Hello all; I have a question about some fixatives. We have a student whom is using our lab in conjunction with an on-line course to prepare for the exam. She was asked to fix tissue in three different fixatives and compare the results on H&E. She has chosen Bouins, formalin/alcohol and 10% BNF. We use the BNF as routine in our lab. My question is after she has fixed the tissue in the other two fixatives can she put the specimen back into the 10% BNF for routine processing or does she need to wait until the processor goes passed its formalin changes in order to add her specimens? I have had no experience with using a fixative other than 10% BNF as a primary so I don't know what to tell her. Thank you for your help, Steve Scholz From carl.hobbs <@t> kcl.ac.uk Wed Jun 29 13:06:42 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Re:xylene times Message-ID: <000f01c57cd5$4e42dc10$0201a8c0@lynne> Manual: 2x xylene 10mins each 4x IMS 74OP ( Industrial methylated spirit) - 2mins each Rinse well in tap water. Proceed to technique ----- Original Message ----- From: To: Sent: Wednesday, June 29, 2005 6:02 PM Subject: Histonet Digest, Vol 19, 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. Bowies method (Jose Luis Palazon Fernandez) > 2. XYLENE EVAPORATION AND GLASS RACKS (Tim Wheelock) > 3. Re: uneven haematoxylin/eosin staining (Janice A Mahoney) > 4. Silly question about deparaffinization (Eva C Andersson) > 5. RE: Silly question about deparaffinization (Sebree Linda A.) > 6. Re: Silly question about deparaffinization (Pamela Marcum) > 7. RE: Blocking serum (C.M. van der Loos) > 8. RE: Silly question about deparaffinization (Rittman, Barry R) > 9. RE: Phenol green gram stain (Monfils, Paul) > 10. tungsten carbide knife resharpening (Nancy W. Troiano) > 11. tungsten carbide resharpening (Nancy W. Troiano) > 12. mouse ankles for IHC (Dana Marshall) > 13. New Mexico Society for Histology (Breeden, Sara) > 14. OCT (Lewis, Sarah) > 15. detachment of tissue from OCT compound (Monfils, Paul) > 16. Full time Histotech Opening (Nava, Josefa) > 17. Pig Skin immunohistochemistry-losing sections during antigen > retrieval (SABYASACHI BISWAS) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Wed, 29 Jun 2005 15:00:41 +0200 (CEST) > From: Jose Luis Palazon Fernandez > Subject: [Histonet] Bowies method > To: histonet@lists.utsouthwestern.edu > Message-ID: <20050629130041.C67348238AD@perceval.uca.es> > Content-Type: text/plain; charset="iso-8859-1" > > Dear List-fellows > > Could any of you send me the protocol of Bowie?s method for > juxtaglomerular cells in kidney. > > thanks in advance > > Jos? Luis > > > Universidad de Oriente-Isla Margarita-Venezuela > actualmente en: Instituto de Ciencias Marinas de Andalucia > Puerto Real, C?diz, Espa?a. > email: jluis.palazon@icman.csic.es > > > > > > ------------------------------ > > Message: 2 > Date: Wed, 29 Jun 2005 09:42:27 -0400 > From: Tim Wheelock > Subject: [Histonet] XYLENE EVAPORATION AND GLASS RACKS > To: histonet@lists.utsouthwestern.edu > Message-ID: <42C2A543.8010400@mclean.harvard.edu> > Content-Type: text/plain; charset=ISO-8859-1; format=flowed > > Hi Margaret: > > Perhaps the way to stop xylene evaporation is to put something on top > of the containers like a cork board, or heavy cookie sheet, anything > heavy enough that will help seal the xylene in the containers. > > Speaking of glass racks, they do seem to be somewhat fragile. I tried > them out in an attempt to batch my silver stains years ago but they did > not hold enough slides. > > I was wondering if anyone out there knew of glass racks that held 60 > slides, not back-to-back, but just one slide per slot. > I am still trying to find a way to batch the Bielschowsky silver stains, > instead of dealing with 14 different coplan jars. > > Tim Wheelock > Harvard Brain Bank > McLean Hospital > > > > ------------------------------ > > Message: 3 > Date: Wed, 29 Jun 2005 08:57:14 -0500 > From: "Janice A Mahoney" > Subject: Re: [Histonet] uneven haematoxylin/eosin staining > To: > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > I have experienced this problem at several labs where I have worked. > Finally figured out it happens more in the summer when the humidity is > high. It was due to incomplete removal of paraffin due to moisture in the > xylene. Try changing it more often or filtering it. The filter paper > will labsorbe the water. > Jan > Omaha NE > >>>> Paul Bradbury 06/28/2005 6:23:27 PM >>> > Sounds like a classical case of incomplete removal of wax before > staining begins. Check the de-waxing reagents and times. > > Paul Bradbury > Kamloops, BC > Canada > > > Diana McCaig wrote: > >>We have been experiencing uneven staining recently. Even when all >>sections >>are cut on the same microtome and put in the same rack on an autostainer, >>their macroscopic appearance show incredible variations. It is so random >>we >>can not identify the problem. The Harris hemotoxylin is filtered daily. >>The tissue type does not make any difference. We can have 4 prostate >>slides >>and 2 will be good and 2 will show random staining intensities. Recuts >>may >>show variation in a different area which makes me think it is the staining >>and not the cutting. >> >>Any suggestions would be truly appreciated. >>Diana McCaig, MLT >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@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: 4 > Date: Wed, 29 Jun 2005 09:56:36 -0400 > From: Eva C Andersson > Subject: [Histonet] Silly question about deparaffinization > To: histonet@lists.utsouthwestern.edu > Message-ID: <42C2A894.7070708@georgetown.edu> > Content-Type: text/plain; charset=us-ascii; format=flowed > > Good morning, > I am probably asking a very silly question but I recently spoke to > someone about deparaffinization and hydration. I found that the times > that they use and the times that we use are very different. So my > question to you is this: What lenghts of time do you use for Xylenes, > 100% alcohol, 95% alcohol and 70% alcohol? > It is about deparaffinising charged (+) slides. So nothing fancy. > Just curious. Maybe I can cut back some on the timed that we use which > are: > 30min Xylenes, 10min 100%, 10min 95% and 6min 70%. > Thanks for your responses. > Eva Andersson > Georgetown University > > > > ------------------------------ > > Message: 5 > Date: Wed, 29 Jun 2005 09:13:44 -0500 > From: "Sebree Linda A." > Subject: RE: [Histonet] Silly question about deparaffinization > To: "Eva C Andersson" , > > Message-ID: > > Content-Type: text/plain; charset="us-ascii" > > We deparaffinize in 3 changes of xylene, 2 changes of 100% ETOH, 1 95% > ETOH and 1 70% ETOH, all for 3 minutes each. We never skimp on the > xylene times but have been known to shorten the ETOH times if we > manually agitate the slides and are in a real hurry. > > Linda A. Sebree > University of Wisconsin Hospital & Clinics > IHC/ISH Clinical & Research Laboratory > DM223-VA > 600 Highland Ave. > Madison, WI 53792 > (608)265-6596 > FAX: (608)262-7174 > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C > Andersson > Sent: Wednesday, June 29, 2005 8:57 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silly question about deparaffinization > > > Good morning, > I am probably asking a very silly question but I recently spoke to > someone about deparaffinization and hydration. I found that the times > that they use and the times that we use are very different. So my > question to you is this: What lenghts of time do you use for Xylenes, > 100% alcohol, 95% alcohol and 70% alcohol? > It is about deparaffinising charged (+) slides. So nothing fancy. Just > curious. Maybe I can cut back some on the timed that we use which are: > 30min Xylenes, 10min 100%, 10min 95% and 6min 70%. Thanks for your > responses. Eva Andersson Georgetown University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 6 > Date: Wed, 29 Jun 2005 10:21:59 -0400 > From: Pamela Marcum > Subject: Re: [Histonet] Silly question about deparaffinization > To: Eva C Andersson , > histonet@lists.utsouthwestern.edu > Message-ID: <6.1.1.1.2.20050629101442.019659d8@mail.vet.upenn.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Eva, > > Depending on the thickness of the sections my times can vary. Sections of > 4 > to 6 micron are deparaffinized in our laboratory at 2 changes at 5 minutes > each with alcohols at 3 minutes each per station to water. Sections > thicker than 6 microns may go as long 15 minutes X 2 in xylenes and 5 to > 10 > minutes in each alcohol. I am very paranoid about getting all of the > paraffin (or MMA where I use one hour per change for xylenes) so I may go > over board. I also change my reagents more often in humid weather. I do > not use an automated stainer so agitation only takes place at the > beginning > of the changes to assure the last reagent is rinsed off. > > Hope this helps. > > At 09:56 AM 6/29/2005, Eva C Andersson wrote: >>Good morning, >>I am probably asking a very silly question but I recently spoke to someone >>about deparaffinization and hydration. I found that the times that they >>use and the times that we use are very different. So my question to you is >>this: What lenghts of time do you use for Xylenes, 100% alcohol, 95% >>alcohol and 70% alcohol? >>It is about deparaffinising charged (+) slides. So nothing fancy. >>Just curious. Maybe I can cut back some on the timed that we use which >>are: >>30min Xylenes, 10min 100%, 10min 95% and 6min 70%. >>Thanks for your responses. >>Eva Andersson >>Georgetown University >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Best Regards, > > Pamela A Marcum > Manager, Histology Special Procedures > University of Pennsylvania > School of Veterinary Medicine > R.S. Reynolds Jr. CORL > New Bolton Center > 382 West Street Road > Kennett Square, PA 19348 > > Phone - 610-925-6278 > Fax - 610-925-8120 > E-mail - pmarcum@vet.upenn.edu > > > > > > ------------------------------ > > Message: 7 > Date: Wed, 29 Jun 2005 16:29:34 +0200 > From: "C.M. van der Loos" > Subject: [Histonet] RE: Blocking serum > To: histonet@lists.utsouthwestern.edu > Message-ID: <10b1c3d10b6fff.10b6fff10b1c3d@amc.uva.nl> > Content-Type: text/plain; charset="us-ascii" > > > Dear Baowei Peng, > > Since you have a goat anti-rabbit IgG as second step reagent you > should use normal goat serum for pre-blocking step. Otherwise use > Protein Block, serum-free (DakoCyto X0909). > > Chris van der Loos, PhD > Dept. of Pathology > Academical Medical Center M2-230 > Meibergdreef 9 > NL-1105 AZ Amsterdam > The Netherlands > > phone: +31 20 5665631 > fax: +31 20 6960389 > > Date: Wed, 29 Jun 2005 08:35:04 +0800 (BEIST) > From: Baowei Peng > Subject: [Histonet] Blocking serum > To: histonet@lists.utsouthwestern.edu > Message-ID: <20050629003504.7EDCD11104FD@sjtu.edu.cn> > Content-Type: text/plain; charset="gb2312" > Hello, all > I\'m doing IHC on mouse frozen sections. The first antibody in my hand > is Rabbit polyclonal antibody,the second antibody is Goat anti Rabbit > IgG(H+L). I\'m wondering what kind of serum I should use to block non > specific staining. > Thanks in advance! > Baowei Peng > 1954 Huashan Road > Pharmacy School > Shanghai Jiaotong University > Shanghai, 200030 > China > Ph:86-21-62932108 > E-mail:pengbw@sjtu.edu.cn > > > ------------------------------ > > Message: 8 > Date: Wed, 29 Jun 2005 09:31:25 -0500 > From: "Rittman, Barry R" > Subject: RE: [Histonet] Silly question about deparaffinization > To: "histonet" > Message-ID: > > Content-Type: text/plain; charset="us-ascii" > > Eva > I would suggest that you do a trial and error run. > Deparaffinization time with xylene will depend on several factors > including: thickness of sections, type of paraffin, number of slides, > agitation, temperature of solution, volume of fluid, number of times > that xylene bath was used, quality of the xylenes solution and so on. > I have always erred on the conservative side and given a longer time as > you have done in the past. However, for many labs, as Linda has > indicated, 3 minutes in each bath is fine. > Suggest that you try a set of slides and deparaffinize them for 3, 5 7 > and 10 minutes and compare your results. > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sebree > Linda A. > Sent: Wednesday, June 29, 2005 9:14 AM > To: Eva C Andersson; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Silly question about deparaffinization > > We deparaffinize in 3 changes of xylene, 2 changes of 100% ETOH, 1 95% > ETOH and 1 70% ETOH, all for 3 minutes each. We never skimp on the > xylene times but have been known to shorten the ETOH times if we > manually agitate the slides and are in a real hurry. > > Linda A. Sebree > University of Wisconsin Hospital & Clinics > IHC/ISH Clinical & Research Laboratory > DM223-VA > 600 Highland Ave. > Madison, WI 53792 > (608)265-6596 > FAX: (608)262-7174 > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C > Andersson > Sent: Wednesday, June 29, 2005 8:57 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silly question about deparaffinization > > > Good morning, > I am probably asking a very silly question but I recently spoke to > someone about deparaffinization and hydration. I found that the times > that they use and the times that we use are very different. So my > question to you is this: What lenghts of time do you use for Xylenes, > 100% alcohol, 95% alcohol and 70% alcohol? > It is about deparaffinising charged (+) slides. So nothing fancy. Just > curious. Maybe I can cut back some on the timed that we use which are: > 30min Xylenes, 10min 100%, 10min 95% and 6min 70%. Thanks for your > responses. Eva Andersson Georgetown University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 9 > Date: Wed, 29 Jun 2005 10:32:07 -0400 > From: "Monfils, Paul" > Subject: RE: [Histonet] Phenol green gram stain > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <09C945920A6B654199F7A58A1D7D1FDE0171755E@lsexch.lsmaster.lifespan.org> > > Content-Type: text/plain; charset="ISO-8859-1" > > I'm not sure what "phenol green" is? But the Gram-Twort technique is a > gram > stain with a green background, due to Fast Green FCF added to the red > stain. > Perhaps that's what you are looking for. You can find that technique here: > > http://www.scienceboard.net/resources/protocols.asp?action=article&protocol_ > id=310&criteria= > > > > ------------------------------ > > Message: 10 > Date: Wed, 29 Jun 2005 10:55:20 -0400 > From: "Nancy W. Troiano" > Subject: [Histonet] tungsten carbide knife resharpening > To: histonet@lists.utsouthwestern.edu > Message-ID: <5.2.1.1.2.20050629105118.00c05630@email.med.yale.edu> > Content-Type: text/plain; format=flowed; charset=us-ascii > > We have our tungsten carbide knives sharpened by Dorn/Hart Microedge, Inc. > 135 Home Avenue Villa Park, IL 60181. Telephone number is > 630-832-3843. They are great - quality work at a reasonable price, quick > turnaround time. I have sent my tungsten carbide knives to them for the > last 25 years for resharpening! > > Nancy Troiano, M.S., Associate in Research III > Yale Core Center for Musculoskeletal Disorders > Yale University > P.O. Box 208071 > New Haven, CT 06520 > (203)785-5136 > > > > > ------------------------------ > > Message: 11 > Date: Wed, 29 Jun 2005 10:57:31 -0400 > From: "Nancy W. Troiano" > Subject: [Histonet] tungsten carbide resharpening > To: histonet@lists.utsouthwestern.edu > Message-ID: <5.2.1.1.2.20050629105626.00c11870@email.med.yale.edu> > Content-Type: text/plain; format=flowed; charset=us-ascii > > Forgot to mention that the price to resharpen a knife from Dorn/Hart is > $150.00 for a 16 cm tungsten carbide knife. Turnaround time is > approximately one week. > > > > > ------------------------------ > > Message: 12 > Date: Wed, 29 Jun 2005 10:24:23 -0500 > From: "Dana Marshall" > Subject: [Histonet] mouse ankles for IHC > To: > Message-ID: <000a01c57cbe$a0d08de0$f623c084@DanaM> > Content-Type: text/plain; charset="iso-8859-1" > > Hi everyone, > > I am gearing up for mouse ankle-joint histology. (I may do vertebral > column eventually as well if anyone has done that.) I have a Cryojane > tape transfer system and have done a bit of sectioning of the > undecalcified bones and they look adequate. I haven't done any fancy > staining yet. My question is a fairly broad one in that I wanted to ask > those who might be doing this for a brief outline of their procedure from > beginning to end. So far I am fixing in 10% formalin (in H2O) for a > couple of days. I embed the joint with the heel towards the bottom of the > block (I thought it might be more stable to cut up from the heel but > perhaps not?) Ultimately I will do IHC staining and, possibly, some in > situ hybridization. I have some previous info in regards to knives and > angles of cutting etc and would appreciate any additional insights into > that as well. > > I am having trouble stringing together the information that will allow me > to make wise choices early on to maximize the use of my samples (and my > mice). Any advice would be GREATLY appreciated. > > Thanks again. > > Dana (who dabbles in histology every 10 years whether I need to or not:-) > > > > ------------------------------ > > Message: 13 > Date: Wed, 29 Jun 2005 09:29:01 -0600 > From: "Breeden, Sara" > Subject: [Histonet] New Mexico Society for Histology > To: > Message-ID: > > Content-Type: text/plain; charset="us-ascii" > > The New Mexico Society for Histology, after having been rather inactive > for the past couple years, is preparing to go "back on track"! If you > are a histologist (or have an interest therein) and live in New Mexico > or one of our adjoining states, please consider joining us. If you have > been a member and would like to reactivate, we'd love to have you back. > There will be a general business meeting and short presentation on > Saturday, August 6th, beginning at 10:00 a.m.; the meeting will be held > at Tricore Reference Laboratories, 1001 Woodward Place NE, Albuquerque. > If interested in either the meeting, or membership, please contact me at > this email address, or at NMHISTO@AOL.COM. > > Sally Breeden, HT(ASCP) > New Mexico Department of Agriculture > Veterinary Diagnostic Services > POBox 4700 > Albuquerque, NM 87196 > (505)841-2576 > (505)841-2518 FAX > > > > ------------------------------ > > Message: 14 > Date: Tue, 28 Jun 2005 16:03:42 -0400 > From: "Lewis, Sarah" > Subject: [Histonet] OCT > To: > Message-ID: > <714B9F12B4E18C4C843B66E8E190F2AD447526@res2k3ms01.CRII.ORG> > Content-Type: text/plain; charset="iso-8859-1" > > Hello! > > I use 7% gum Tragacanth that I make up myself using 100ml of distilled > water, 7gms of gum Tragacanth, and 5 grains of Thymol. Dissolve by > manually stirring at rm temp.(may take several hours) Store half > refrigerated(4c) and half frozen > (-20c). I work with only muscle and nerve biopsy tissue but this works > great for me. I am sure this is a bit "old school", but I have never had > any trouble with cracking or stability of the tissue. It also seems to be > great for long term storage in -80 freezer. Good luck!! > > > Sarah E. Lewis > Histotechnician > Childrens Research > Center for Gene Therapy > 700 Childrens Dr Rm WA3112 > Columbus OH 43205 > (614)-722-2204 > LewisS@ccri.net > > > > -----Original Message----- > From: ssq5977@yahoo.com.cn [mailto:ssq5977@yahoo.com.cn] > Sent: Thursday, June 16, 2005 9:30 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Is there anything better than OCT? > > > > Hi all: > > We are currently using OCT to embed renal biopsy tissue in cryostat mold. > First of all ,we put a drop of OCT on the mold ,then we put the tissue on > the top of the OCT and dip the mold into liquid nitrogen for 10 seconds. > then the tissue is sent to cryosection. Does anyone know if there is > anything better than OCT to embed the tissue? > > Thanks! > > Shuqiong Shen(ssq5977@yahoo.com.cn) > Research Institute of Nephrology, Jinling Hospital > 305 East Zhongshan Road, Nanjing > Jiangsu Province,P.R of China > > Zip code: 210002 > Tel: +86 25 85912177 > > > > > Sarah E. Lewis > Histotechnician > Childrens Research > Center for Gene Therapy > 700 Childrens Dr Rm WA3112 > Columbus OH 43205 > (614)-722-2204 > LewisS@ccri.net > > > > > Upgrade Your Email - Click here! > > > > ------------------------------ > > Message: 15 > Date: Wed, 29 Jun 2005 11:35:51 -0400 > From: "Monfils, Paul" > Subject: [Histonet] detachment of tissue from OCT compound > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <09C945920A6B654199F7A58A1D7D1FDE0171755F@lsexch.lsmaster.lifespan.org> > > Content-Type: text/plain; charset="ISO-8859-1" > > Someone has sent me 60 rat brains frozen in OCT compound (whole brain > except > for the cerebellum and the anterior 1 cm). He wants multiple 30-micron > sections from each brain. They appear to be well infiltrated with sucrose, > and they cut smoothly on the cryostat at -20C. However, the sections of > tissue fall out of the surrounding OCT, which makes it very difficult to > pick them up on slides without a lot of wrinkles and bubbles. > > Any tricks to deal with this situation? > > Paul M. > > > > ------------------------------ > > Message: 16 > Date: Wed, 29 Jun 2005 10:56:47 -0500 > From: "Nava, Josefa" > Subject: [Histonet] Full time Histotech Opening > To: > Message-ID: > <2C515C1049EAF5459EFD8C9B929078A41944B6@phdex03.txhealth.org> > Content-Type: text/plain; charset="us-ascii" > > We are looking for a full time Histotech or ASCP eligible position at > Presbyterian Hospital of Dallas, Texas. Interested candidates may > contact Maureen Hoops at 214-345-7795 for details and benefit > information. > > > > > > > > > > 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. > > ------------------------------ > > Message: 17 > Date: Wed, 29 Jun 2005 12:33:50 -0400 > From: SABYASACHI BISWAS > Subject: [Histonet] Pig Skin immunohistochemistry-losing sections > during antigen retrieval > To: histonet@lists.utsouthwestern.edu > Message-ID: <23a1309239d9f7.239d9f723a1309@osu.edu> > Content-Type: text/plain; charset=us-ascii > > Hi All > We are facing a problem with our pig skin biopsy samples. We are involved > in wound healing research and therefore have to do histological staining > of pig skin. Our biopsies are taken with 6mm biopsy punches around a 3mm > biopsy punch taken previously (the wound). The samples are fixed in > formalin and embedded in paraffin. We were losing sections even during H&E > staining but managed to solve that by baking the slides at 60degrees C for > 1hour instead of 37 degrees overnight; we also use Superfrost++ slides. > However during antigen retrieval using the microwave method we are losing > sections wholly or partially. Does anyone have experience with antigen > retrieval in pig skin samples. We appreciate any help/advice that you can > give us. > > Thanks > > Sabya Biswas > The Ohio State University > > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 19, Issue 42 > **************************************** > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.8.6/33 - Release Date: 28/06/2005 > > -- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.323 / Virus Database: 267.8.6/33 - Release Date: 28/06/2005 From llewllew <@t> shaw.ca Wed Jun 29 13:22:41 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] fixation question References: <7F1312711CA7474A89B3DF8BA0BA54D01333A4@pmc-rfd-mx01.intranet.osfnet.org> Message-ID: <001601c57cd7$895ddcd0$7e034246@yourlk4rlmsu> If she is comparing the effects of three different fixatives, then she should go into the post-fixative part of the schedule. Otherwise she is comparing double fixation of two of them. That is another, seperate, question. Bryan Llewellyn ----- Original Message ----- From: "Scholz, Stephen J." To: Sent: Wednesday, June 29, 2005 10:59 AM Subject: [Histonet] fixation question > Hello all; > > I have a question about some fixatives. We have a student whom is using > our lab in conjunction with an on-line course to prepare for the exam. > She was asked to fix tissue in three different fixatives and compare the > results on H&E. She has chosen Bouins, formalin/alcohol and 10% BNF. We > use the BNF as routine in our lab. My question is after she has fixed the > tissue in the other two fixatives can she put the specimen back into the > 10% BNF for routine processing or does she need to wait until the > processor goes passed its formalin changes in order to add her specimens? > I have had no experience with using a fixative other than 10% BNF as a > primary so I don't know what to tell her. > > Thank you for your help, > > Steve Scholz > -------------------------------------------------------------------------------- > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From JEllin <@t> yumaregional.org Wed Jun 29 13:23:29 2005 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Tissue proccesor Message-ID: We are looming at buying a proccesor this budget year, but currently our proccessor is starting to have problem. This pruchase has now been fast tracked, we want to demo a proccesor that does routine proccesor as well as stat, GI biopsies, Non Gyn fluids, a short cycle. Can anyone give me some information on what is being used. Jesus Ellin Yuma Regional Medical Center From JosefaNava <@t> texashealth.org Wed Jun 29 13:39:08 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] MSH 2 and MSH 1 antibodies for colon CA Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944B9@phdex03.txhealth.org> Can someone tell me where they order these antibodies that will work very with the Ventana IHC Machines. I thank you so much. Josie Nava Presbyterian Hospital of Dallas 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 ddittus787 <@t> aol.com Wed Jun 29 14:19:24 2005 From: ddittus787 <@t> aol.com (ddittus787@aol.com) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] fixation question In-Reply-To: <001601c57cd7$895ddcd0$7e034246@yourlk4rlmsu> References: <7F1312711CA7474A89B3DF8BA0BA54D01333A4@pmc-rfd-mx01.intranet.osfnet.org> <001601c57cd7$895ddcd0$7e034246@yourlk4rlmsu> Message-ID: <8C74AF69817B603-634-7920@mblk-r24.sysops.aol.com> Once tissue has been properly fixed ,NBF should have no huge effect on H&E, sometimes over-fixing (2 or more fixatives) can affect special stains, IHC,IN-situ. If I were a student and had 3 fixatives I would wait to get past the formalin step to be able to purely judge fixation issues of each independent fixative. You know what we learn and what happens in the real world is always different. dana -----Original Message----- From: Bryan Llewellyn To: Histonet Sent: Wed, 29 Jun 2005 11:22:41 -0700 Subject: Re: [Histonet] fixation question If she is comparing the effects of three different fixatives, then she should go into the post-fixative part of the schedule. Otherwise she is comparing double fixation of two of them. That is another, seperate, question. Bryan Llewellyn ----- Original Message ----- From: "Scholz, Stephen J." To: Sent: Wednesday, June 29, 2005 10:59 AM Subject: [Histonet] fixation question > Hello all; > > I have a question about some fixatives. We have a student whom is using > our lab in conjunction with an on-line course to prepare for the exam. > She was asked to fix tissue in three different fixatives and compare the > results on H&E. She has chosen Bouins, formalin/alcohol and 10% BNF. We > use the BNF as routine in our lab. My question is after she has fixed the > tissue in the other two fixatives can she put the specimen back into the > 10% BNF for routine processing or does she need to wait until the > processor goes passed its formalin changes in order to add her specimens? > I have had no experience with using a fixative other than 10% BNF as a > primary so I don't know what to tell her. > > Thank you for your help, > > Steve Scholz > -------------------------------------------------------------------------------- > _______________________________________________ > Histonet mailing list > Histonet@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 Janet.Bonner <@t> FLHOSP.ORG Wed Jun 29 15:31:38 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] fixation question Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB42FB@fh2k093.fhmis.net> In practice, do we use NBF after Bouin's fixation?, after alcohol/formalin? after BNF? She would probably be better off doing what is done in practice and not on Mars for a realistic study on comparisons of the fixatives. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: llewllew@shaw.ca; histonet@lists.utsouthwestern.edu Sent: 6/29/2005 3:19 PM Subject: Re: [Histonet] fixation question Once tissue has been properly fixed ,NBF should have no huge effect on H&E, sometimes over-fixing (2 or more fixatives) can affect special stains, IHC,IN-situ. If I were a student and had 3 fixatives I would wait to get past the formalin step to be able to purely judge fixation issues of each independent fixative. You know what we learn and what happens in the real world is always different. dana -----Original Message----- From: Bryan Llewellyn To: Histonet Sent: Wed, 29 Jun 2005 11:22:41 -0700 Subject: Re: [Histonet] fixation question If she is comparing the effects of three different fixatives, then she should go into the post-fixative part of the schedule. Otherwise she is comparing double fixation of two of them. That is another, seperate, question. Bryan Llewellyn ----- Original Message ----- From: "Scholz, Stephen J." To: Sent: Wednesday, June 29, 2005 10:59 AM Subject: [Histonet] fixation question > Hello all; > > I have a question about some fixatives. We have a student whom is using > our lab in conjunction with an on-line course to prepare for the exam. > She was asked to fix tissue in three different fixatives and compare the > results on H&E. She has chosen Bouins, formalin/alcohol and 10% BNF. We > use the BNF as routine in our lab. My question is after she has fixed the > tissue in the other two fixatives can she put the specimen back into the > 10% BNF for routine processing or does she need to wait until the > processor goes passed its formalin changes in order to add her specimens? > I have had no experience with using a fixative other than 10% BNF as a > primary so I don't know what to tell her. > > Thank you for your help, > > Steve Scholz > ------------------------------------------------------------------------ -------- > _______________________________________________ > Histonet mailing list > Histonet@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 euan.mcnaughtone <@t> agresearch.co.nz Wed Jun 29 17:43:32 2005 From: euan.mcnaughtone <@t> agresearch.co.nz (McNaughton, Euan) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Silly question about deparaffinization Message-ID: Wow... Those times all seem huge! We've always just used 2 x 2 min Xylene, 2 x 1 min 100% EtOH, 1 x 1 min 90% EtOH, 1 x 1 min 70% EtOH then to water. No problems so long as the solutions are changed when they start to get dirty. Euan McNaughton -----Original Message----- From: Sebree Linda A. [mailto:la.sebree@hosp.wisc.edu] Sent: Thursday, 30 June 2005 2:14 a.m. To: Eva C Andersson; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Silly question about deparaffinization We deparaffinize in 3 changes of xylene, 2 changes of 100% ETOH, 1 95% ETOH and 1 70% ETOH, all for 3 minutes each. We never skimp on the xylene times but have been known to shorten the ETOH times if we manually agitate the slides and are in a real hurry. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C Andersson Sent: Wednesday, June 29, 2005 8:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly question about deparaffinization Good morning, I am probably asking a very silly question but I recently spoke to someone about deparaffinization and hydration. I found that the times that they use and the times that we use are very different. So my question to you is this: What lenghts of time do you use for Xylenes, 100% alcohol, 95% alcohol and 70% alcohol? It is about deparaffinising charged (+) slides. So nothing fancy. Just curious. Maybe I can cut back some on the timed that we use which are: 30min Xylenes, 10min 100%, 10min 95% and 6min 70%. Thanks for your responses. Eva Andersson Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ======================================================================= Attention: The information contained in this message and/or attachments from AgResearch Limited is intended only for the persons or entities to which it is addressed and may contain confidential and/or privileged material. Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon, this information by persons or entities other than the intended recipients is prohibited by AgResearch Limited. If you have received this message in error, please notify the sender immediately. ======================================================================= From fladanielski <@t> yahoo.com.br Wed Jun 29 18:25:33 2005 From: fladanielski <@t> yahoo.com.br (Flavia Danielski) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] help for brazilian student Message-ID: <20050629232533.90287.qmail@web30613.mail.mud.yahoo.com> Hello... I would be really glad if someone could help me with XGal staining for tibia... the protocols that I have work well for soft tissues, but not for bone. How should I process the sample? I need to obtain sections of the bone ... should I do it before XGal staining or after??? If after, how should I process the slices to reveal beta gal? Does the parafine interfire with the process? Thank you very much and greetings from Brazil sorry for my bad english Flavia Fl?via Helena da Silva Lab. de Imunogen?tica, Depto. de Gen?tica, UFRGS Av. Bento Gon?alves, 9500 pr?dio 43323/lab 212C Bairro Agronomia. POA-RS-BRAZIL phone: 0055-(51)33166737 _______________________________________________________ Yahoo! Acesso Gr?tis - Internet r?pida e gr?tis. Instale o discador agora! http://br.acesso.yahoo.com/ From portera203 <@t> yahoo.com Wed Jun 29 18:52:18 2005 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Tissue proccesor In-Reply-To: Message-ID: <20050629235218.65425.qmail@web40907.mail.yahoo.com> We have a ThermoElectron Excelsior in our laboratory and we love it. Very user friendly, saves on reagents, and exposure to chemicals through a self contained rotation system. Jesus Ellin wrote:We are looming at buying a proccesor this budget year, but currently our proccessor is starting to have problem. This pruchase has now been fast tracked, we want to demo a proccesor that does routine proccesor as well as stat, GI biopsies, Non Gyn fluids, a short cycle. Can anyone give me some information on what is being used. Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Amy S.Porter, HT(ASCP), QIHC Michigan State University - Department of Physiology Division of Human Pathology Investigative Histopathology Laboratory 2201 Biomedical Physical Sciences Room 2133 East Lansing, MI 48824-3320 Ph: 517-355-6475 ext 1480/1481 Fax: 517-432-1368 portera@msu.edu --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football From histology.bc <@t> shaw.ca Wed Jun 29 21:38:04 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Silly question about deparaffinization References: <42C2A894.7070708@georgetown.edu> Message-ID: <42C35B0C.9080904@shaw.ca> Hi Eva, Assuming that you are dealing with routine 4 micron sections of paraffin wax, a room temperature of 18-21 C (65-70 F for you guys South of the 49th), reagents that are changed on a regular basis, adequate volumes of each reagent, and some form of agitation, you can afford to make some major cuts in your timing. Assuming that all the above criteria are in place, I would suggest the following: Xylene 5 minutes Xylene 5 minutes Absolute alcohol 2 minutes Absolute alcohol 2 minutes 95% alcohol 2 minutes 70% alcohol can be skipped without any harm to the tissues. Water 1 minute Carry on with staining procedure of choice. For routine staining, the above times will work just fine. While it is essential that all traces of wax are removed, 30 minutes in xylene is grossly excessive, Once the xylene has removed the wax, the time spent in alcohol does not need to be very long. All you are trying to do is replace one fluid with another ... and that happens pretty quickly ... 2 minutes in each alcohol is ample. The most contaminated reagents should be replaced each day or on a regular basis, or if the reagents begin to look contaminated, discard them. There are references in the literature to methods that require prolonged treatment with solvents prior to staining. The original procedure for the Lendrum's MSB fibrin stain specified 24 hours treatment with trichlorethylene to "degrease" the sections, but I don't know anybody who still does this! Paul Bradbury Kamloops, BC Canada PS ... What are you going to do with all this spare time ? ********************************************************* Eva C Andersson wrote: > Good morning, > I am probably asking a very silly question but I recently spoke to > someone about deparaffinization and hydration. I found that the times > that they use and the times that we use are very different. So my > question to you is this: What lenghts of time do you use for Xylenes, > 100% alcohol, 95% alcohol and 70% alcohol? > It is about deparaffinising charged (+) slides. So nothing fancy. > Just curious. Maybe I can cut back some on the timed that we use which > are: > 30min Xylenes, 10min 100%, 10min 95% and 6min 70%. > Thanks for your responses. > Eva Andersson > Georgetown University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From lpwenk <@t> sbcglobal.net Thu Jun 30 05:26:07 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Bowies method In-Reply-To: <20050629130041.C67348238AD@perceval.uca.es> Message-ID: Just wondering if doing an immunohistochemical stain for renin or cathepsin D would be easier, and more reliable, than doing the Bowie. Also, JG granules will stain positive with PAS (periodic acid-Schiff) and TFT (thioflavin T), though neither of these stains are specific for JG granules. I'll cover the basics of the Bowie stain (taken from the Sheehan book), and you can let me know if you want still want me to send you the procedure. Fix for 48 hours in Helly fixative (mercuric chloride, potassium dichromate, formaldehyde)(Yes, it must be Helly. Bowie will not work with formalin fixed tissue) Wash tissue in running water overnight, then process and embed. Cut sections at 4 um. Deparafinize slide to water. Lugolize (de-Zenkerize) with iodine and sodium thiosulfate Mordant in 2.5% potassium dichromate at 40 degrees C overnight Rinse in d. water Stain in Bowie solution overnight(mixture of Biebrich scarlet, ethyl violet water, filtered, filter paper with dye allowed to dry overnight, dissolving dry dye in alcohol) Blot slides Dip in acetone Differentiate in xylene:clove oil mixture (hint from me - clove oil makes the lab smell great, but is very expensive) Xylene Why the stain isn't used any more: - Fixation and staining takes several days (4 by my count). - Tissue must be fixed in Helly. - Mercury and potassium dichromate waste must be correctly disposed - Expensive clove oil must be bought(100 mL = $60+ US) Let me know if you still would like the Bowie procedure. Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jose Luis Palazon Fernandez Sent: Wednesday, June 29, 2005 9:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bowies method Dear List-fellows Could any of you send me the protocol of Bowie?s method for juxtaglomerular cells in kidney. thanks in advance Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mauger <@t> email.chop.edu Thu Jun 30 07:41:28 2005 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] RE:Renin by IHC Message-ID: Hi Lee and Peggy, Noting your reference to renin staining, I have been working up 2 renin abs,and my problem is the need for a good positive control. Does anyone know where I can purchase one? Or does anyone have a renin producing tumor that they are willing to share? Thanks, Jo Mauger >>> "Lee & Peggy Wenk" 06/30/05 6:26 AM >>> Just wondering if doing an immunohistochemical stain for renin or cathepsin D would be easier, and more reliable, than doing the Bowie. Also, JG granules will stain positive with PAS (periodic acid-Schiff) and TFT (thioflavin T), though neither of these stains are specific for JG granules. I'll cover the basics of the Bowie stain (taken from the Sheehan book), and you can let me know if you want still want me to send you the procedure. Fix for 48 hours in Helly fixative (mercuric chloride, potassium dichromate, formaldehyde)(Yes, it must be Helly. Bowie will not work with formalin fixed tissue) Wash tissue in running water overnight, then process and embed. Cut sections at 4 um. Deparafinize slide to water. Lugolize (de-Zenkerize) with iodine and sodium thiosulfate Mordant in 2.5% potassium dichromate at 40 degrees C overnight Rinse in d. water Stain in Bowie solution overnight(mixture of Biebrich scarlet, ethyl violet water, filtered, filter paper with dye allowed to dry overnight, dissolving dry dye in alcohol) Blot slides Dip in acetone Differentiate in xylene:clove oil mixture (hint from me - clove oil makes the lab smell great, but is very expensive) Xylene Why the stain isn't used any more: - Fixation and staining takes several days (4 by my count). - Tissue must be fixed in Helly. - Mercury and potassium dichromate waste must be correctly disposed - Expensive clove oil must be bought(100 mL = $60+ US) Let me know if you still would like the Bowie procedure. Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jose Luis Palazon Fernandez Sent: Wednesday, June 29, 2005 9:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bowies method Dear List-fellows Could any of you send me the protocol of Bowie s method for juxtaglomerular cells in kidney. thanks in advance Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cmconway <@t> usgs.gov Thu Jun 30 09:34:30 2005 From: cmconway <@t> usgs.gov (Carla M Conway) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Dako EnVision G2 or Vector ImmPRESS comments wanted Message-ID: Hello all, I would like any comments (pro and con) regarding Dako's EnVision G2/AP system or Vector's ImmPRESS kit. We are currently using EnVision + , but are wondering if these new systems could be even better (more sensitive). Thanks for your help, Carla Conway Western Fisheries Research Center 6505 NE 65th St Seattle, WA 98115 ph: 206-526-6282 ext. 242 fax: 206-526-6654 cmconway@usgs.gov From kbroomal <@t> NEMOURS.ORG Thu Jun 30 11:02:48 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] TUNEL Resources Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824362@wlmmsx02.nemours.org> Does anyone know of any good TUNEL for Dummies resources? I don't know much about it and would like to learn. Thanks! Kristen Broomall, HT (ASCP) A.I. DuPont Hospital for Children 1600 Rockland Road ARB 264 Wilmington, DE 19899 kbroomal@nemours.org > NOTICE...This electronic transmission is intended only for the person(s) > named. It may contain information that is (i) proprietary to the sender, > and/or (ii) privileged, confidential and/or otherwise exempt from > disclosure under applicable State and Federal law, including, but not > limited to, privacy standards imposed pursuant to the federal Health > Insurance Portability and Accountability Act of 1996 (HIPAA). Receipt by > anyone other than the named recipient(s) is not a waiver of any applicable > privilege. If you received this confidential communication in error, > please notify the sender immediately by reply e-mail message and > permanently delete the original message from your system. > > > From Charles.Embrey <@t> carle.com Thu Jun 30 11:36:29 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Looking for Sandra Garvin Message-ID: Sorry to bother the list but I am looking for a Cytotech friend, Sandra Garvin, I have lost track of. Any help will be appreciated. Thanks, Charles Embrey From shawnster73 <@t> aol.com Thu Jun 30 12:24:00 2005 From: shawnster73 <@t> aol.com (shawnster73@aol.com) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] TUNEL Resources In-Reply-To: <6E41111281623B4B8A9AB8F9A7EA3437824362@wlmmsx02.nemours.org> References: <6E41111281623B4B8A9AB8F9A7EA3437824362@wlmmsx02.nemours.org> Message-ID: <8C74BAFA3DB513C-B38-2D9B@FWM-D29.sysops.aol.com> These are the sources that were given to me when I was curious. 1: Duan WR, Garner DS, Williams SD, Funckes-Shippy CL, Spath IS, Blomme EA. Comparison of immunohistochemistry for activated caspase-3 and cleaved cytokeratin 18 with the TUNEL method for quantification of apoptosis in histological sections of PC-3 subcutaneous xenografts. J Pathol. 2003 Feb;199(2):221-8. PMID: 12533835 [PubMed - indexed for MEDLINE] 2: Austgulen R, Chedwick L, Vogt Isaksen C, Vatten L, Craven C. Trophoblast apoptosis in human placenta at term as detected by expression of a cytokeratin 18 degradation product of caspase. Arch Pathol Lab Med. 2002 Dec;126(12):1480-6. PMID: 12456208 [PubMed - indexed for MEDLINE] 3: Walker JA, Quirke P. Viewing apoptosis through a 'TUNEL'. J Pathol. 2001 Oct;195(3):275-6. PMID: 11673822 [PubMed - indexed for MEDLINE] 4: Barrett KL, Willingham JM, Garvin AJ, Willingham MC. Advances in cytochemical methods for detection of apoptosis. J Histochem Cytochem. 2001 Jul;49(7):821-32. PMID: 11410607 [PubMed - indexed for MEDLINE] 5: Valavanis C, Naber S, Schwartz LM. In situ detection of dying cells in normal and pathological tissues. Methods Cell Biol. 2001;66:393-415. No abstract available. PMID: 11396013 [PubMed - indexed for MEDLINE] 6: Kadyrov M, Kaufmann P, Huppertz B. Expression of a cytokeratin 18 neo-epitope is a specific marker for trophoblast apoptosis in human placenta. Placenta. 2001 Jan;22(1):44-8. PMID: 11162351 [PubMed - indexed for MEDLINE] -----Original Message----- From: Kristen Broomall To: Histonet@lists.utsouthwestern.edu Sent: Thu, 30 Jun 2005 12:02:48 -0400 Subject: [Histonet] TUNEL Resources Does anyone know of any good TUNEL for Dummies resources? I don't know much about it and would like to learn. Thanks! Kristen Broomall, HT (ASCP) A.I. DuPont Hospital for Children 1600 Rockland Road ARB 264 Wilmington, DE 19899 kbroomal@nemours.org > NOTICE...This electronic transmission is intended only for the person(s) > named. It may contain information that is (i) proprietary to the sender, > and/or (ii) privileged, confidential and/or otherwise exempt from > disclosure under applicable State and Federal law, including, but not > limited to, privacy standards imposed pursuant to the federal Health > Insurance Portability and Accountability Act of 1996 (HIPAA). Receipt by > anyone other than the named recipient(s) is not a waiver of any applicable > privilege. If you received this confidential communication in error, > please notify the sender immediately by reply e-mail message and > permanently delete the original message from your system. > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Thu Jun 30 12:53:45 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] pap pen Message-ID: <20050630175345.29853.qmail@web90201.mail.scd.yahoo.com> Any good progress in Pap pens for Immunofluoresence. Steve --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football From fladanielski <@t> yahoo.com.br Thu Jun 30 14:24:37 2005 From: fladanielski <@t> yahoo.com.br (Flavia Danielski) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] immunocytochemistry anti-gfp protocols Message-ID: <20050630192437.12465.qmail@web30603.mail.mud.yahoo.com> I would be really glad if you could help me with immunocytochemistry anti-gfp... I have no idea what kind of fixative should I use in order to avoid "problems" with gfp detection... we have mnAB (Molecular Probes) and our intention is to detect GFP from our cell cultures (monolayers). Thank you very much and once again sorry for my bad english greetings from Brazil Flavia Fl?via Helena da Silva Lab. de Imunogen?tica, Depto. de Gen?tica, UFRGS Av. Bento Gon?alves, 9500 pr?dio 43323/lab 212C Bairro Agronomia. POA-RS-BRAZIL phone: 0055-(51)33166737 _______________________________________________________ Yahoo! Acesso Gr?tis - Internet r?pida e gr?tis. Instale o discador agora! http://br.acesso.yahoo.com/ From eshields <@t> bhset.org Thu Jun 30 15:13:36 2005 From: eshields <@t> bhset.org (E Sharon Shields) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Pathology dictation system Message-ID: Histonetters, Our lab is in the market for a server driven pathology dictation system. What is out there in Histoland? Anyone willing to share their views on what you have? Vendor replies welcome. Sharon Shields Baptist Hospital of East Tennessee Knoxville, TN 865 549-4351 Fax 865 632-5316 The information in this e-mail message, including any attachments, may contain confidential and privileged information that is protected by law. It is intended for the sole use of the recipient named above. If you are not the intended recipient or the agent responsible for delivering it to the intended recipient, you are hereby notified that any unauthorized review, use, dissemination or copying is strictly prohibited. If you have received this electronic mail transmission in error please notify us immediately at bellis@bhset.org and delete any copies from your system. <<<>>> From zumbor <@t> email.cs.nsw.gov.au Thu Jun 30 17:37:50 2005 From: zumbor <@t> email.cs.nsw.gov.au (Rosalba) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Leica SM2500 Message-ID: <000001c57dd2$bbada250$1e7b4c98@dofm.cs.nsw.gov.au> Hi all, Does anyone have or has used a Leica SM2500 slege microtome. Our Reichert - Jung tetrander has finally packed it in after 40 years and we are looking for a replacement. Unfortunately there are none in Australia at the present and we cannot trial one. Thanks Rosalba Zumbo Laboratory Manager Histology Dept. Department of Forensic Medicine 42-50 Parramatta rd Glebe, 2037 Australia Ph: 61 2 85847842 Fax: 61 2 95664573 email: zumbor@email.cs.nsw.gov.au "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 Sydney South West Area Health Service." From haldana <@t> unimoron.edu.ar Thu Jun 30 20:09:51 2005 From: haldana <@t> unimoron.edu.ar (Hernan Aldana) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] buckground nuclear stain in Immunohistochemistry In-Reply-To: <200506301329562.SM01380@swlx162.swmed.edu> Message-ID: <001e01c57dd9$959d0cc0$9d25e818@b3w6zzmtb6juvbs> Dear all I made immunohistochemistry for androgen receptor in testicular tissue with Hydrochloric Acid Epitope Retrieval Method. I have a background stain remarkably in chromosomes in spermatocytes. Some advises to stop this reaction? Thanks Dr. Hern?n J. Aldana Marcos Secretario Acad?mico Facultad de Medicina. Universidad de Mor?n Machado 914. B1708JPD. Buenos Aires. Argentina e-mail alternativo hernanjavier@yahoo.com web:http://www.ht.org.ar/histologia/NUEVAS%20UNIDADES/home.htm From bills <@t> icpmr.wsahs.nsw.gov.au Thu Jun 30 20:40:30 2005 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Cytospin chambers Message-ID: <000101c57ddd$ddb4b860$0ecd080a@wsahs.nsw.gov.au> Dear All, I have just been informed that ThermoElectron are no longer supplying the re-usable chambers and SS clips for the Cytospin. Does anyone know of an alternate supplier for these items? Many thanks Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From int09018 <@t> alphahunt.com Thu Jun 30 21:15:50 2005 From: int09018 <@t> alphahunt.com (HCS) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] looking for used stainer Message-ID: <000601c57de2$cd9cbf90$6601a8c0@hp> My auto slide stainer is beyond repair. Anyone have a unit sitting around for parts for a IL code-on slide stainer or would anyone have an extra stainer they might be willing to sell cheap. We are in western Washington. I am also looking for a replacement embedding center or parts for a Shandon embedding center II (the white one with black front). Thanks LeRoy Brown HT(ASCP) HTL HCS 360-966-7300 Everson, WA 98247