From jkiernan <@t> uwo.ca Tue Feb 1 00:11:44 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] aqueous mounting medium References: <1864.132.236.104.213.1107189504.squirrel@132.236.104.213> Message-ID: <41FF1DA0.FC6CA60E@uwo.ca> The product of the indigogenic reaction for beta-galactosidase is a pigment. It doesn't dissolve in the solvents used for microscopy and should be OK in any mounting medium, resinous or aqueous. I've used 3 or 4 different resinous media with products of indigogenic reactions, and all have been OK. Amanda MacFarlane wrote: > > I have stained whole tissues for beta-galactosidase (X-gal) and am now > cryosectioning the tissues. I have been fixing the sections in 4% PFA > overnight at 4 degrees Celsius and cover slipping with 70% glycerol. > Coverslipping with glycerol is a time-consuming pain in the butt so I've > checked out glycerol vinyl alcohol aqueous (GVA) mounting solution (Zymed) > and Supermount aqueous mounting medium (Biogenex) as alternatives. It > appears that these products have not been tested with beta-gal staining, > so I'm wondering if people use these products and/or what people > recommend. Thanks. > > Amanda MacFarlane, PhD > Division of Nutritional Sciences > Cornell University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kim.Osullivan <@t> med.monash.edu.au Tue Feb 1 00:41:06 2005 From: Kim.Osullivan <@t> med.monash.edu.au (Kim O'Sullivan) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Excessive shrinkage Message-ID: <130.194.114.210.1107239881.79442@my.monash.edu.au> Hi all, Can anyone tell me what may cause bouins fixed tissue (in this case mice kidneys's) to shrink after tissue processing (6 hour).This normally does not happen. ta Kim From Kemlo.Rogerson <@t> elht.nhs.uk Tue Feb 1 02:11:01 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Excessive shrinkage[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EEF2@bhrv-nt-11.bhrv.nwest.nhs.uk> If it normally doesn't happen then what makes you think it was the Bouins and is there anything else you changed. Bouins is alcoholic Picric Acid isn't it? I assume it's an additive coagulant fixative and if you leave tissue in it too long it goes rock hard. Dimly in my memory we used to wash out the picric acid but I don't understand why we put it in, in the first place. Has your processing changed? Are you leaving the tissue in Bouins too long? -----Original Message----- From: Kim O'Sullivan [mailto:Kim.Osullivan@med.monash.edu.au] Sent: 01 February 2005 06:41 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Excessive shrinkage[Scanned] Hi all, Can anyone tell me what may cause bouins fixed tissue (in this case mice kidneys's) to shrink after tissue processing (6 hour).This normally does not happen. ta Kim _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Tue Feb 1 02:16:39 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] yeast and bacteria on slides[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EEF3@bhrv-nt-11.bhrv.nwest.nhs.uk> Do you have piped water into the stainer? Is this contaminated? The source water maybe being contaminated within the staining machine. Any of your Staff going rotten? View the slides along the process; when unused, after the water bath, after drying, after staining, after mounting. Eat the elephant in small portions, once you have identified the area in the process you can then focus on the culprit; the 'scatter gun' approach rarely works, be logical. -----Original Message----- From: Melissa Jans [mailto:histomjans@yahoo.com] Sent: 01 February 2005 01:40 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] yeast and bacteria on slides[Scanned] I need some help!! We have been experiencing some yeast and bacteria showing up on some of our special stains. It started about 2 weeks ago and seems to be sporadic (one day...no problems, the next day, several stains are affected). The yeast and bacteria (rods) are clumped together all over the slide, on the tissue sections, around the sections and on the slide in areas where there is no tissue. The stains we have seen the contamination are the Gram, PAS/D, PAS and GMS. All of these stains are automated except the Gram, so I do not think the special stainer is the problem. Here is what I have eliminated as problem areas: -Processor solutions and paraffin (we cut and stained a block from another institute and had contamination) -Waterbaths, slide drying boards, water containers (used to fill waterbathes), stainer containers, ice pans (all of the mentioned have been thoroughly bleached) -Tap water and DI water (all water supplies have been tested by sending a sample to micro...there was no bacteria or yeast detected) -Stay-on used as a slide adhesive (smeared some on a slide and stained it) -Bad batch of slides (we have had contamination on two different slide sources (vendors)) Does anybody have any ideas? What am I missing? Any suggestions would be much appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ 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 Kemlo.Rogerson <@t> elht.nhs.uk Tue Feb 1 02:23:35 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] yeast and bacteria on slides[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EEF6@bhrv-nt-11.bhrv.nwest.nhs.uk> My point, I think? -----Original Message----- From: Mildred Fail [mailto:failm@musc.edu] Sent: 01 February 2005 01:51 To: histonet@lists.utsouthwestern.edu; histomjans@yahoo.com Subject: Re: [Histonet] yeast and bacteria on slides[Scanned] Do you have tubing on your faucets? >>> Melissa Jans 01/31/05 20:41 PM >>> I need some help!! We have been experiencing some yeast and bacteria showing up on some of our special stains. It started about 2 weeks ago and seems to be sporadic (one day...no problems, the next day, several stains are affected). The yeast and bacteria (rods) are clumped together all over the slide, on the tissue sections, around the sections and on the slide in areas where there is no tissue. The stains we have seen the contamination are the Gram, PAS/D, PAS and GMS. All of these stains are automated except the Gram, so I do not think the special stainer is the problem. Here is what I have eliminated as problem areas: -Processor solutions and paraffin (we cut and stained a block from another institute and had contamination) -Waterbaths, slide drying boards, water containers (used to fill waterbathes), stainer containers, ice pans (all of the mentioned have been thoroughly bleached) -Tap water and DI water (all water supplies have been tested by sending a sample to micro...there was no bacteria or yeast detected) -Stay-on used as a slide adhesive (smeared some on a slide and stained it) -Bad batch of slides (we have had contamination on two different slide sources (vendors)) Does anybody have any ideas? What am I missing? Any suggestions would be much appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Martina.Urbanek <@t> uibk.ac.at Tue Feb 1 02:50:35 2005 From: Martina.Urbanek <@t> uibk.ac.at (Martina Urbanek) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Fwd: de Olmos staining Message-ID: <1107247835.41ff42db62ff8@web-mail1.uibk.ac.at> Ms. Martina Urbanek Forschungslabor der Klin.Abt. f?r Neonatologie neonatal neuroscience research laboratory Med. University Innsbruck Innrain 66, 4th floor A-6020 Innsbruck Tel. +43 (0)512 504 27755/27765 Fax: +43 (0)512 504 27766 Email: Martina.Urbanek@uibk.ac.at ----- Weitergeleitete Nachricht von Martina Urbanek ----- Datum: Fri, 28 Jan 2005 11:26:38 +0100 Von: Martina Urbanek Antwort an: Martina Urbanek Betreff: de Olmos staining An: Histonet Hello, Everyone, Does anyone have a detailed protocol for de Olmos amino-cupric-silver stain which works on paraffin sections (does it even work on paraffin?)? We work with mice brains which are fixed in 10% formalin, but not perfused and we are looking for a technique to stain degenerated neuronal cells. Your help is greatly appreciated. Martina Urbanek Ms. Martina Urbanek Forschungslabor der Klin.Abt. f?r Neonatologie neonatal neuroscience research laboratory Med. University Innsbruck Innrain 66, 4th floor A-6020 Innsbruck Tel. +43 (0)512 504 27755/27765 Fax: +43 (0)512 504 27766 Email: Martina.Urbanek@uibk.ac.at ----- Ende der weitergeleiteten Nachricht ----- From c.m.vanderloos <@t> amc.uva.nl Tue Feb 1 02:56:35 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] RE: aqueous mounting medium Message-ID: Hi Amanda, From my experiments with X-gal as chromogen for IHC techniques it appeared that the blue-greenish reaction product (with X-gal + ferro/ferricyanide) is very stable and survives any mounting medium. You may dehydrate and mount organically. 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 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From "Amanda MacFarlane" Date Mon, 31 Jan 2005 11:38:24 -0500 (EST) To Histonet@lists.utsouthwestern.edu Subject [Histonet] aqueous mounting medium I have stained whole tissues for beta-galactosidase (X-gal) and am now cryosectioning the tissues. I have been fixing the sections in 4% PFA overnight at 4 degrees Celsius and cover slipping with 70% glycerol. Coverslipping with glycerol is a time-consuming pain in the butt so I've checked out glycerol vinyl alcohol aqueous (GVA) mounting solution (Zymed) and Supermount aqueous mounting medium (Biogenex) as alternatives. It appears that these products have not been tested with beta-gal staining, so I'm wondering if people use these products and/or what people recommend. Thanks. Amanda MacFarlane, PhD Division of Nutritional Sciences Cornell University References 1. mailto:c.m.vanderloos@amc.uva.nl From JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk Tue Feb 1 03:50:54 2005 From: JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk (JOHN PHILLIPS) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] yeast and bacteria on slides Message-ID: <166A1E642B5B644DA694C08FD29D0ADC3E1350@ztroy.new-tr.wales.nhs.uk> May I suggest you consider this for starters. The most likely cause is your water floatation bath. How many are used? Do you meticulously clean each one at the end of each day and fill up with fresh deionised water? Can you identify the water bath or baths(if several were used). Here, operator marks their slides with initials for audit purposes so we know who picked up the section, labelled the slide and so which water bath they were using. Hey, bloody audit coming in handy!! Water baths filled with water and left on for several days prior to use will grow micro-organisms, especially if they have been used and not cleaned. If you can pin point sections from each bath and have spare unstained sections from each bath from the "day of recogning", then i suggest you stain a couple of spares from each with PAS and you may get your answer. It has happened here a couple of times in the past. Hope this helps. John, Wrexham, Wales. -----Original Message----- From: Melissa Jans [mailto:histomjans@yahoo.com] Sent: 01 February 2005 01:40 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] yeast and bacteria on slides I need some help!! We have been experiencing some yeast and bacteria showing up on some of our special stains. It started about 2 weeks ago and seems to be sporadic (one day...no problems, the next day, several stains are affected). The yeast and bacteria (rods) are clumped together all over the slide, on the tissue sections, around the sections and on the slide in areas where there is no tissue. The stains we have seen the contamination are the Gram, PAS/D, PAS and GMS. All of these stains are automated except the Gram, so I do not think the special stainer is the problem. Here is what I have eliminated as problem areas: -Processor solutions and paraffin (we cut and stained a block from another institute and had contamination) -Waterbaths, slide drying boards, water containers (used to fill waterbathes), stainer containers, ice pans (all of the mentioned have been thoroughly bleached) -Tap water and DI water (all water supplies have been tested by sending a sample to micro...there was no bacteria or yeast detected) -Stay-on used as a slide adhesive (smeared some on a slide and stained it) -Bad batch of slides (we have had contamination on two different slide sources (vendors)) Does anybody have any ideas? What am I missing? Any suggestions would be much appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ 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 Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r rheolwr systemau wybod drwy ddefnyddio'r manylion isod. Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain Cymru. Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau???r Ddeddf Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru ar www.newalesnhstrust.org.uk This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager using the details below. The contents of this email represent the views of the individual(s) named above and do not necessarily represent the views of the North East Wales NHS Trust. Please be aware that, under the terms of the Freedom of Information Act 2000, the North East Wales NHS Trust may be required to make public the content of any emails or correspondence received. For futher information on Freedom of Information, please refer to the North East Wales NHS Trust website at www.newalesnhstrust.org.uk For further assistance, please contact system.administrator@new-tr.wales.nhs.uk. From JWEEMS <@t> sjha.org Tue Feb 1 04:12:06 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] yeast and bacteria on slides Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0B8F28@sjhaexc02.sjha.org> I would suggest you try new stain packs on your automated stainer, just to rule that out. AFTER you have changed and bleached your water baths. Do you change them daily? Good luck, j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Melissa Jans Sent: Mon 1/31/2005 8:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] yeast and bacteria on slides I need some help!! We have been experiencing some yeast and bacteria showing up on some of our special stains. It started about 2 weeks ago and seems to be sporadic (one day...no problems, the next day, several stains are affected). The yeast and bacteria (rods) are clumped together all over the slide, on the tissue sections, around the sections and on the slide in areas where there is no tissue. The stains we have seen the contamination are the Gram, PAS/D, PAS and GMS. All of these stains are automated except the Gram, so I do not think the special stainer is the problem. Here is what I have eliminated as problem areas: -Processor solutions and paraffin (we cut and stained a block from another institute and had contamination) -Waterbaths, slide drying boards, water containers (used to fill waterbathes), stainer containers, ice pans (all of the mentioned have been thoroughly bleached) -Tap water and DI water (all water supplies have been tested by sending a sample to micro...there was no bacteria or yeast detected) -Stay-on used as a slide adhesive (smeared some on a slide and stained it) -Bad batch of slides (we have had contamination on two different slide sources (vendors)) Does anybody have any ideas? What am I missing? Any suggestions would be much appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ 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 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 DSpears <@t> agr.state.il.us Tue Feb 1 05:49:44 2005 From: DSpears <@t> agr.state.il.us (DSpears@agr.state.il.us) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Illinois Website is Up Message-ID: Dear Histonet, Just wanted to let everyone know that Illinois now has a website : www.ilhisto.org It is still a work in progress, but we are excited to have it! From Andrew.MacDuff <@t> ed.ac.uk Tue Feb 1 06:12:51 2005 From: Andrew.MacDuff <@t> ed.ac.uk (Andrew MacDuff) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Mouse liver fixation Message-ID: <001201c50857$5a194670$b9b2d781@universi8lynpq> Dear All Thanks for your great advice last time on fixing mouse lungs. You'll be glad to know we've got it working and have some nice H+E slides and the immunos will be staring soon. Can you help again? My boss has decided to look at the mouse's liver as well (poor thing!) and I'd be grateful if you have any tips on fixing liver. Can you just remove the liver and place it in a vial of formalin or does it need perfused etc? Looking forward to hearing from you all again. Regards Andrew Andrew MacDuff Clinical Research Fellow Wilkie Laboratory Medical School Edinburgh University Teviot Place Edinburgh andrew.macduff@ed.ac.uk From ian.montgomery <@t> bio.gla.ac.uk Tue Feb 1 06:51:36 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:33 2005 Subject: Fwd: [Histonet] Mouse liver fixation Message-ID: <6.0.1.1.2.20050201125022.02d25870@udcf.gla.ac.uk> Andrew, For routine Histology I found the best fixative for mouse liver was Susa. Ian. >From: "Andrew MacDuff" >To: >Date: Tue, 1 Feb 2005 12:12:51 -0000 >X-Mailer: Microsoft Outlook Express 6.00.2900.2180 >X-Edinburgh-Scanned: at lawnmarket.ucs.ed.ac.uk > with MIMEDefang 2.33, Sophie 3.04rc1, Sophos Anti-Virus 3.88 >X-Scanned-By: MIMEDefang 2.33 (www . roaringpenguin . com / mimedefang) >X-Scan-Signature: 7175f3aa946646b32d1e6d3ef45b12b6 >X-Content-Filtered-By: Mailman/MimeDel 2.1.5 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >List-Id: For the exchange of information pertaining to histotechnology and > related fields >List-Unsubscribe: >, > > >List-Archive: >List-Post: >List-Help: >List-Subscribe: , > >Sender: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 05628b3a735271e1629bc35bb8abda07 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] Mouse liver fixation >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=5.5 tests=none autolearn=no version=2.63 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >X-GLA-Spam-Scan: R >X-GLA-Spam-Score: 0.0 (/) >X-GLA-Spam-Report: > >Dear All > >Thanks for your great advice last time on fixing mouse lungs. You'll be >glad to know we've got it working and have some nice H+E slides and the >immunos will be staring soon. >Can you help again? My boss has decided to look at the mouse's liver as >well (poor thing!) and I'd be grateful if you have any tips on fixing >liver. Can you just remove the liver and place it in a vial of formalin or >does it need perfused etc? >Looking forward to hearing from you all again. >Regards >Andrew > >Andrew MacDuff >Clinical Research Fellow >Wilkie Laboratory >Medical School >Edinburgh University >Teviot Place >Edinburgh >andrew.macduff@ed.ac.uk >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk From bruyntjes <@t> voeding.tno.nl Tue Feb 1 07:10:49 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Formalin pigment Message-ID: <3B070848E7C2204F9DEB8BCFD767728004A694C9@ntexch1.voeding.tno.nl> Is there anyone who can tell me what other reason could cause the formation of formalin pigment except a too acidic formalin? Joost Bruijntjes TNO Nutrition and Food research Zeist The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From celebrej <@t> HHSC.CA Tue Feb 1 07:13:49 2005 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Millers Elastin Stain HELP Message-ID: <3AADFB88753AD31189C100902786B91C14E6DAE4@hch_nt_exchange.hhsc.ca> Good morning all.... Our lab makes our own Millers solution, and I think it's just for plain old punishment because if anyone has had to make it they understand what I mean... Normally the stain works beautifully, but since we've run out of 'Victoria Blue 4R' and have used 'Victoria Blue B' as a substitute the stain doesn't seem to be working too well. The reason why we substituted because Victoria Blue 4R is no longer availalbe.. What I'm asking all you histology guru's out there... 1. Does anybody know of a company that sells pre-made Millers? 2. Does anybody know where we can purchase Victoria Blue 4R? 3. Does anybody know if/why Victoria Blue B can be used as a substitute? Thanks.. Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca 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 c.m.vanderloos <@t> amc.uva.nl Tue Feb 1 07:30:23 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] RE: endogenous peroxidase blues Message-ID: Hi all, On Patty's request here is a little list of antibodies that were once tested with the Dual Endogenous Enzyme Block reagent (Dako S2003) and the staining result. GFAP (RbxHu) (Dako) rat duodenum no problem OX6 (MsxRt) (Serotec) rat duodenum negative OX17 (MsxRt) (Serotec) rat duodenum weak OX42 (MsxRt) (Serotec) rat duodenum negative &nbs! p; CD These antibodies were not selected or something. Just, at some point this blocking kit was interesting to evaluate. As you see staining with 6/9 antibodies resulted into staining problems. Don't get too scared of this basically good product, but you guys are warned now for eventual failures! 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 e-mail: [1]c.m.vanderloos@amc.uva.nl -----Original Message----- Chris can you share with us which antigens didn't survive this enzyme block in your hands. Thanks, Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [[2]mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of C.M. van der Loos Sent: Monday, January 31, 2005 12:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: endogenous peroxidase blues Hi all, Before everybody is trying to solve their problem of endogenous peroxidase activity with Dual Enzyme Blocking Reagent as Veronique wrote us, I would like to make a comment. This stuff works great indeed in many cases, that's true. But I also found some! antigens References 1. mailto:c.m.vanderloos@amc.uva.nl 2. javascript:main.compose('new', 't=histonet-bounces@lists.utsouthwestern.edu') From sladd <@t> hsc.usf.edu Tue Feb 1 07:39:00 2005 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Millers Elastin Stain HELP In-Reply-To: <3AADFB88753AD31189C100902786B91C14E6DAE4@hch_nt_exchange.hhsc.ca> Message-ID: Julia, There's a great article on elastic staining in Sakura's Histologic Vol. XXXII No. 1, May 2000, page 12 entitled "Elastic Tissue Staining in Human Skin." That's where I found out that you can buy Miller's from Rowley Biochemical, 10 Electronics Ave. Danvers, MA 01923 978-739-4883 or www.rowleybio.com. A couple years ago it cost $55 for a pint. As for your questions 2 and 3, I have no idea. Sharron -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Celebre Julia Sent: Tuesday, February 01, 2005 8:14 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Millers Elastin Stain HELP Good morning all.... Our lab makes our own Millers solution, and I think it's just for plain old punishment because if anyone has had to make it they understand what I mean... Normally the stain works beautifully, but since we've run out of 'Victoria Blue 4R' and have used 'Victoria Blue B' as a substitute the stain doesn't seem to be working too well. The reason why we substituted because Victoria Blue 4R is no longer availalbe.. What I'm asking all you histology guru's out there... 1. Does anybody know of a company that sells pre-made Millers? 2. Does anybody know where we can purchase Victoria Blue 4R? 3. Does anybody know if/why Victoria Blue B can be used as a substitute? Thanks.. Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca 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 am458 <@t> cornell.edu Tue Feb 1 09:20:38 2005 From: am458 <@t> cornell.edu (Amanda MacFarlane) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] RE: aqueous mounting medium In-Reply-To: References: Message-ID: <1935.128.253.96.73.1107271238.squirrel@128.253.96.73> This is good to know. I was under the impression from what I've read that the chromogenic product of beta-gal was soluble in xylene, thus the need for aqueous mounting medium. Thanks, Amanda > > Hi Amanda, > From my experiments with X-gal as chromogen for IHC techniques it appeared > that the blue-greenish reaction product (with X-gal + ferro/ferricyanide) > is very stable and survives any mounting medium. You may dehydrate and > mount organically. > 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 > e-mail: c.m.vanderloos@amc.uva.nl > > ----- Original Message ----- From "Amanda MacFarlane" Date Mon, 31 > Jan 2005 11:38:24 -0500 (EST) To Histonet@lists.utsouthwestern.edu > Subject [Histonet] aqueous mounting medium > I have stained whole tissues for beta-galactosidase (X-gal) and am now > cryosectioning the tissues. I have been fixing the sections in 4% PFA > overnight at 4 degrees Celsius and cover slipping with 70% glycerol. > Coverslipping with glycerol is a time-consuming pain in the butt so I've > checked out glycerol vinyl alcohol aqueous (GVA) mounting solution (Zymed) > and Supermount aqueous mounting medium (Biogenex) as alternatives. It > appears that these products have not been tested with beta-gal staining, > so I'm wondering if people use these products and/or what people > recommend. Thanks. > > > Amanda MacFarlane, PhD > Division of Nutritional Sciences > Cornell University > Amanda MacFarlane Division of Nutritional Sciences Cornell University From gcallis <@t> montana.edu Tue Feb 1 09:20:47 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Excessive shrinkage[Scanned] In-Reply-To: <1030B679AD69D6119C3F00080210DD9D05A3EEF2@bhrv-nt-11.bhrv.n west.nhs.uk> References: <1030B679AD69D6119C3F00080210DD9D05A3EEF2@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: <6.0.0.22.1.20050201081406.01b4a6f8@gemini.msu.montana.edu> Kemlo, Bouins uses: saturated picric acid, aqueous formaldehyde acetic acid and you are correct about removing picric acid, some use 70% alcohol rinses or 70% alcohol containing lithium carbonate. 8 hours is a common fixation time, with no longer than 72 hours recommended. I recall it made our tissues crunchier. Also, test the temperature of your paraffin, has it changed to higher? All processing creates some hardness, with approx 25% shrinkage in tissues due to water removal and heat of paraffin, and is unavoidable. Have an interesting publication where the group tested the shrinkage of paraffin processing and plastic processing. At 01:11 AM 2/1/2005, you wrote: >If it normally doesn't happen then what makes you think it was the Bouins >and is there anything else you changed. Bouins is alcoholic Picric Acid >isn't it? I assume it's an additive coagulant fixative and if you leave >tissue in it too long it goes rock hard. Dimly in my memory we used to wash >out the picric acid but I don't understand why we put it in, in the first >place. Has your processing changed? Are you leaving the tissue in Bouins too >long? > >-----Original Message----- >From: Kim O'Sullivan [mailto:Kim.Osullivan@med.monash.edu.au] >Sent: 01 February 2005 06:41 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Excessive shrinkage[Scanned] > >Hi all, > >Can anyone tell me what may cause bouins fixed tissue (in this case mice >kidneys's) to shrink after tissue processing (6 hour).This normally does not >happen. > >ta > >Kim > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue Feb 1 09:32:38 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Re: Mouse liver fixation In-Reply-To: <001201c50857$5a194670$b9b2d781@universi8lynpq> References: <001201c50857$5a194670$b9b2d781@universi8lynpq> Message-ID: <6.0.0.22.1.20050201082321.01b5fe20@gemini.msu.montana.edu> Andrew, We had terrible problems with mouse liver when whole livers were immersed in only 50 ml neutral buffered formalin and left to sit around. The tissue brought in a week later was NOT fixed internally, it was still red. If you can perfuse the animal, dissect out liver and immerse into formalin 1:20 volume. Or fix for a day, take liver out, slice it and return to fresh formalin. Liver is very homogenous, dense and surprisingly takes a longer time to fix than mouse lung. If liver is dissected out, bread slicing it for better fixation is advisable. Slices are embedded cut sides down unless you want to embed a whole liver - a rather large chunk of tissue for such a little critter. If you plan on a whole liver, be sure to change the fixative after a day or so to replenish the formalin and this is advisable with sliced liver too. Whatever you do, make sure fixative can reach internal parts of liver or you get some autolyzed tissue. At 05:12 AM 2/1/2005, you wrote: >Dear All > >Thanks for your great advice last time on fixing mouse lungs. You'll be >glad to know we've got it working and have some nice H+E slides and the >immunos will be staring soon. >Can you help again? My boss has decided to look at the mouse's liver as >well (poor thing!) and I'd be grateful if you have any tips on fixing >liver. Can you just remove the liver and place it in a vial of formalin or >does it need perfused etc? >Looking forward to hearing from you all again. >Regards >Andrew > >Andrew MacDuff >Clinical Research Fellow >Wilkie Laboratory >Medical School >Edinburgh University >Teviot Place >Edinburgh >andrew.macduff@ed.ac.uk >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Rcartun <@t> harthosp.org Tue Feb 1 09:33:12 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] p34 Message-ID: Is anyone doing IHC for p34 on formalin-fixed, paraffin-embedded tissue? If so, whose antibody are you using? Thank you! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From Andrew.Prior <@t> Smith-Nephew.com Tue Feb 1 09:53:32 2005 From: Andrew.Prior <@t> Smith-Nephew.com (Prior, Andrew) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Professional Qualifications in UK Message-ID: Does anyone out there know of any professional qualifications in the UK for Histologists? I can find ones for Histo-paths, but I don't do any pathology work. >From what I gather, there are two levels of certification in the US for histologists and pathologists. Is the same true here in Ol' Blighty? Is it possible to do distance-learning for the American one? Thanks in advance Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York UK Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. From Andrew.Prior <@t> Smith-Nephew.com Tue Feb 1 10:11:44 2005 From: Andrew.Prior <@t> Smith-Nephew.com (Prior, Andrew) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Re: Millers Elastin Stain HELP Message-ID: Julia , You can find Miller Elastin Stain on the UK Version of the Raymond Lamb website ( http://www.ralamb.net/ ), but for some reason it's not on the US version of the site. You may be able to get it shipped across or just hassle R A Lamb in the States to sell you some. Hope this helps Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York UK >Message: 20 >Date: Tue, 1 Feb 2005 08:13:49 -0500 >From: Celebre Julia >Subject: [Histonet] Millers Elastin Stain HELP >To: "'histonet@lists.utsouthwestern.edu'" >Message-ID: <3AADFB88753AD31189C100902786B91C14E6DAE4@hch_nt_exchange.hhsc.ca> >Content-Type: text/plain; charset="iso-8859-1" >Good morning all.... >Our lab makes our own Millers solution, and I think it's just for plain old >punishment because if anyone has had to make it they understand what I >mean... Normally the stain works beautifully, but since we've run out of >'Victoria Blue 4R' and have used 'Victoria Blue B' as a substitute the stain >doesn't seem to be working too well. The reason why we substituted because >Victoria Blue 4R is no longer availalbe.. >What I'm asking all you histology guru's out there... >1. Does anybody know of a company that sells pre-made Millers? >2. Does anybody know where we can purchase Victoria Blue 4R? >3. Does anybody know if/why Victoria Blue B can be used as a substitute? >Thanks.. >Julia Celebre MLT >Anatomic Pathology >Hamilton General Hospital >905-527-0271 ext46145 >email: celebrej@hhsc.ca Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. From Janet.Bonner <@t> FLHOSP.ORG Tue Feb 1 10:22:18 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] yeast and bacteria on slides Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB418B@fh2k093.fhmis.net> Melissa, We are a 24-7 operation and had the same problem when our shifts exceeded eight hours and some of the waterbathes were not changed out. Changing the waterbath out when each shift was done totally eliminated the problem. -Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Melissa Jans Sent: Monday, January 31, 2005 8:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] yeast and bacteria on slides I need some help!! We have been experiencing some yeast and bacteria showing up on some of our special stains. It started about 2 weeks ago and seems to be sporadic (one day...no problems, the next day, several stains are affected). The yeast and bacteria (rods) are clumped together all over the slide, on the tissue sections, around the sections and on the slide in areas where there is no tissue. The stains we have seen the contamination are the Gram, PAS/D, PAS and GMS. All of these stains are automated except the Gram, so I do not think the special stainer is the problem. Here is what I have eliminated as problem areas: -Processor solutions and paraffin (we cut and stained a block from another institute and had contamination) -Waterbaths, slide drying boards, water containers (used to fill waterbathes), stainer containers, ice pans (all of the mentioned have been thoroughly bleached) -Tap water and DI water (all water supplies have been tested by sending a sample to micro...there was no bacteria or yeast detected) -Stay-on used as a slide adhesive (smeared some on a slide and stained it) -Bad batch of slides (we have had contamination on two different slide sources (vendors)) Does anybody have any ideas? What am I missing? Any suggestions would be much appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ 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 The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From jkiernan <@t> uwo.ca Tue Feb 1 10:51:43 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Formalin pigment References: <3B070848E7C2204F9DEB8BCFD767728004A694C9@ntexch1.voeding.tno.nl> Message-ID: <41FFB39F.954A3FB@uwo.ca> Formalin pigment is formed by the action of acid (pH <5.6) or alkali (pH >8) on haemoglobin. According to Lillie's big book (p. 488-489) similar pigment occurs around sites of haemorrhage in the gastric mucosa; it's then called HCl pigment. Malaria pigment is also closely similar. Lillie cites his own work in the field, which was published as 3 papers in 1947. -- ------------------------------- 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 _______________________________ "Bruijntjes, J.P." wrote: > > Is there anyone who can tell me what other reason could cause the > formation of formalin pigment except a too acidic formalin? > > > > Joost Bruijntjes > > TNO Nutrition and Food research > > Zeist > > The Netherlands > > > > 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 subratab <@t> bdonline.com Tue Feb 1 11:22:22 2005 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] another background / non-specific staining problem Message-ID: <200502011727.j11HRLYb031576@mailout.proshikanet.com> Dear All I am doing immunohistochemistry for 8-OHdG (a DNA marker for oxidative stress) in rat kidney sections (not perfused) fixed in methacarn. Along with positive nuclear staining, I am getting some background staining in some glomerulus and also in some interstitial area. This non-specific background is mainly in the areas where red cells are abundant (the tissue is not perfused). The same background is also present in my negative control slides (absence of primary Ab). My protocol is 1. Deparaffinization 2. Microwave exposure for 5 min in citrate buffer (10 mM, pH 6) 3. Horse serum blocking (5%) in PBS for 1 hr at room temp 4. Primary Ab (mouse monoclonal) against 8-OHdG (N45.1) overnight at 4 C (diluent 1% BSA and 0.3% triton X 100 in PBS) 5. Horse anti-mouse secondary Ab 1 hr at room temp 6. 3% H2O2 in methanol for 10 min. 7. ABC 8. DAB 9. Counter stain with hematoxilin 10. Dehydration and mounting. I did some experiments to identify if this non-specific staining is due to endogenous peroxidase/biotin. I followed above protocol omitting both the primary and secondary Ab. I did not get any background stain, my slides were clean. So I interpreted that the background is not due to endogenous peroxidase/biotin. Most probably it is due to non-specific binding of secondary Ab with red cell (or other tissue) components. Then I tried to block non-specific binding of secondary Ab in different ways (5% horse serum, 20% horse serum, 1%, and 5% non-fat milk, 1% BSA plus 1% non-fat milk). But I could not able to reduce the background. Lastly, I am writing you all for any suggestion. Thank you in advance Subrata Biswas University of Campinas SP, Brazil From subratab <@t> bdonline.com Tue Feb 1 11:33:43 2005 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] RE: HRP-conjugated secondary and biotinylated-secondary Message-ID: <200502011738.j11HcfYb031689@mailout.proshikanet.com> Dear van der Loos Thank you very much for your reply with historical overview. Thank you everybody who replyed to my question. Subrata. --------- Mensagem Original -------- From: "C.M. van der Loos" To: "histonet@lists.utsouthwestern.edu" Cc: subratab@bdonline.com Subject: RE: HRP-conjugated secondary and biotinylated-secondary Date: 31/01/05 14:29 Hi, The existence of both biotinylated- and HRP-conjuated secondary antibodies reflects the development of IHC techniques into more staining efficiency/sensitivity. In the old days, a 2-step staining method using a HRP-conjugated secondary anti-mouse or anti-rabbit was common practice. Mid 80-ies the 3-step staining method using a biotinylated 2nd step reagent and streptavidin/HRP conjugate or streptavidin-biotinylated HRP complex became very popular because of its superior staining efficiency/sensitivity. The disadvantage here is the occurrence of endogenous biotin in many tissues. Endogenous biotin is not present in untreated formalin-fixed and paraffin-embedded tissue sections, but will be retrieved especially when using Tris-EDTA pH8-9. Only recently we have gone to a 2-step staining procedure again, now based on both secondary antibody and HRP attached to a polymer backbone. The efficiency/sensitivity of these polymers is comparable with the 3-step streptavidin/HRP procedure, but avoids the problem of endogenous biotin. I hope this small historical overview helps. 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 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From Subratab Date Sun, 30 Jan 2005 0:18:56 +0600 To Subject [Histonet] HRP-conjugated secondary and biotinylated-secondary Hi, Is it a common practice to use HRP-conjugated secondary antibody instead of biotinylated secondary antibody for immunohistochemistry (with DAB developing system)? I am confused about the advantage/ disadvantage of using HRP-conjugated secondary antibody for immunos. Subrata Biswas UNICAMP, SP, Brazil. References 1. mailto:c.m.vanderloos@amc.uva.nl From lpwenk <@t> sbcglobal.net Tue Feb 1 12:40:55 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Formalin pigment References: <3B070848E7C2204F9DEB8BCFD767728004A694C9@ntexch1.voeding.tno.nl> Message-ID: <004f01c5088d$90d953e0$e32ed445@domainnotset.invalid> What tissue? Spleen, I have found, gets formalin pigment quite often, even when the formalin is newly made up and at the correct pH. Don't know why. Just does, it seems. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Bruijntjes, J.P." To: Sent: Tuesday, February 01, 2005 8:10 AM Subject: [Histonet] Formalin pigment Is there anyone who can tell me what other reason could cause the formation of formalin pigment except a too acidic formalin? Joost Bruijntjes TNO Nutrition and Food research Zeist The Netherlands 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 carolb <@t> mail.phys.mcw.edu Tue Feb 1 12:42:41 2005 From: carolb <@t> mail.phys.mcw.edu (Carol Bobrowitz) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] microscope objective Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A962B@thor.phys.mcw.edu> I am looking for a used Olympus 60X, achromat, plan, high dry, 160mm tube length objective. Due to new technology they are not made any longer. If anyone has one they are willing to sell, please contact me. Thank you, Carol Ann Bobrowitz Histology Laboratory Department of Physiology - Room 541 Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, Wisconsin 53226 414-456-8179 FAX 414-456-6546 cbobrowi@mcw.edu From juan.gutierrez <@t> christushealth.org Tue Feb 1 12:47:56 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] another background / non-specific staining problem Message-ID: Have you tried an Fc receptor block. Innovex has it along with a Background Buster. I use the Fc receptor block for all my bloody specimens and blood related markers. The Background Buster I have not tried yet. Good luck. Innovex Biosciences 1-800-622-7808 www.innvx.com innvx@ix.netcom.com 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 Subratab Sent: Tuesday, February 01, 2005 11:22 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] another background / non-specific staining problem Dear All I am doing immunohistochemistry for 8-OHdG (a DNA marker for oxidative stress) in rat kidney sections (not perfused) fixed in methacarn. Along with positive nuclear staining, I am getting some background staining in some glomerulus and also in some interstitial area. This non-specific background is mainly in the areas where red cells are abundant (the tissue is not perfused). The same background is also present in my negative control slides (absence of primary Ab). My protocol is 1. Deparaffinization 2. Microwave exposure for 5 min in citrate buffer (10 mM, pH 6) 3. Horse serum blocking (5%) in PBS for 1 hr at room temp 4. Primary Ab (mouse monoclonal) against 8-OHdG (N45.1) overnight at 4 C (diluent 1% BSA and 0.3% triton X 100 in PBS) 5. Horse anti-mouse secondary Ab 1 hr at room temp 6. 3% H2O2 in methanol for 10 min. 7. ABC 8. DAB 9. Counter stain with hematoxilin 10. Dehydration and mounting. I did some experiments to identify if this non-specific staining is due to endogenous peroxidase/biotin. I followed above protocol omitting both the primary and secondary Ab. I did not get any background stain, my slides were clean. So I interpreted that the background is not due to endogenous peroxidase/biotin. Most probably it is due to non-specific binding of secondary Ab with red cell (or other tissue) components. Then I tried to block non-specific binding of secondary Ab in different ways (5% horse serum, 20% horse serum, 1%, and 5% non-fat milk, 1% BSA plus 1% non-fat milk). But I could not able to reduce the background. Lastly, I am writing you all for any suggestion. Thank you in advance Subrata Biswas University of Campinas SP, Brazil _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lwhite <@t> lakeridgehealth.on.ca Tue Feb 1 12:56:24 2005 From: lwhite <@t> lakeridgehealth.on.ca (White, Lori) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] RE: Millers Elastin Stain HELP Message-ID: -----Original Message----- Julia, Inter-medico in Canada has just released a kit stain for Millers. Lori W From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, February 01, 2005 1:04 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 15, Issue 2 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: aqueous mounting medium (Amanda MacFarlane) 2. RE: Excessive shrinkage[Scanned] (Gayle Callis) 3. Re: Mouse liver fixation (Gayle Callis) 4. p34 (Richard Cartun) 5. Professional Qualifications in UK (Prior, Andrew) 6. Re: Millers Elastin Stain HELP (Prior, Andrew) 7. RE: yeast and bacteria on slides (Bonner, Janet) 8. Re: Formalin pigment (John Kiernan) 9. another background / non-specific staining problem (Subratab) 10. RE: HRP-conjugated secondary and biotinylated-secondary (Subratab) ---------------------------------------------------------------------- Message: 1 Date: Tue, 1 Feb 2005 10:20:38 -0500 (EST) From: "Amanda MacFarlane" Subject: [Histonet] RE: aqueous mounting medium To: "C.M. van der Loos" Cc: histonet@lists.utsouthwestern.edu Message-ID: <1935.128.253.96.73.1107271238.squirrel@128.253.96.73> Content-Type: text/plain;charset=iso-8859-1 This is good to know. I was under the impression from what I've read that the chromogenic product of beta-gal was soluble in xylene, thus the need for aqueous mounting medium. Thanks, Amanda > > Hi Amanda, > From my experiments with X-gal as chromogen for IHC techniques it appeared > that the blue-greenish reaction product (with X-gal + ferro/ferricyanide) > is very stable and survives any mounting medium. You may dehydrate and > mount organically. > 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 > e-mail: c.m.vanderloos@amc.uva.nl > > ----- Original Message ----- From "Amanda MacFarlane" Date Mon, 31 > Jan 2005 11:38:24 -0500 (EST) To Histonet@lists.utsouthwestern.edu > Subject [Histonet] aqueous mounting medium > I have stained whole tissues for beta-galactosidase (X-gal) and am now > cryosectioning the tissues. I have been fixing the sections in 4% PFA > overnight at 4 degrees Celsius and cover slipping with 70% glycerol. > Coverslipping with glycerol is a time-consuming pain in the butt so I've > checked out glycerol vinyl alcohol aqueous (GVA) mounting solution (Zymed) > and Supermount aqueous mounting medium (Biogenex) as alternatives. It > appears that these products have not been tested with beta-gal staining, > so I'm wondering if people use these products and/or what people > recommend. Thanks. > > > Amanda MacFarlane, PhD > Division of Nutritional Sciences > Cornell University > Amanda MacFarlane Division of Nutritional Sciences Cornell University ------------------------------ Message: 2 Date: Tue, 01 Feb 2005 08:20:47 -0700 From: Gayle Callis Subject: RE: [Histonet] Excessive shrinkage[Scanned] To: Kemlo Rogerson , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050201081406.01b4a6f8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Kemlo, Bouins uses: saturated picric acid, aqueous formaldehyde acetic acid and you are correct about removing picric acid, some use 70% alcohol rinses or 70% alcohol containing lithium carbonate. 8 hours is a common fixation time, with no longer than 72 hours recommended. I recall it made our tissues crunchier. Also, test the temperature of your paraffin, has it changed to higher? All processing creates some hardness, with approx 25% shrinkage in tissues due to water removal and heat of paraffin, and is unavoidable. Have an interesting publication where the group tested the shrinkage of paraffin processing and plastic processing. At 01:11 AM 2/1/2005, you wrote: >If it normally doesn't happen then what makes you think it was the Bouins >and is there anything else you changed. Bouins is alcoholic Picric Acid >isn't it? I assume it's an additive coagulant fixative and if you leave >tissue in it too long it goes rock hard. Dimly in my memory we used to wash >out the picric acid but I don't understand why we put it in, in the first >place. Has your processing changed? Are you leaving the tissue in Bouins too >long? > >-----Original Message----- >From: Kim O'Sullivan [mailto:Kim.Osullivan@med.monash.edu.au] >Sent: 01 February 2005 06:41 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Excessive shrinkage[Scanned] > >Hi all, > >Can anyone tell me what may cause bouins fixed tissue (in this case mice >kidneys's) to shrink after tissue processing (6 hour).This normally does not >happen. > >ta > >Kim > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 3 Date: Tue, 01 Feb 2005 08:32:38 -0700 From: Gayle Callis Subject: [Histonet] Re: Mouse liver fixation To: "Andrew MacDuff" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050201082321.01b5fe20@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Andrew, We had terrible problems with mouse liver when whole livers were immersed in only 50 ml neutral buffered formalin and left to sit around. The tissue brought in a week later was NOT fixed internally, it was still red. If you can perfuse the animal, dissect out liver and immerse into formalin 1:20 volume. Or fix for a day, take liver out, slice it and return to fresh formalin. Liver is very homogenous, dense and surprisingly takes a longer time to fix than mouse lung. If liver is dissected out, bread slicing it for better fixation is advisable. Slices are embedded cut sides down unless you want to embed a whole liver - a rather large chunk of tissue for such a little critter. If you plan on a whole liver, be sure to change the fixative after a day or so to replenish the formalin and this is advisable with sliced liver too. Whatever you do, make sure fixative can reach internal parts of liver or you get some autolyzed tissue. At 05:12 AM 2/1/2005, you wrote: >Dear All > >Thanks for your great advice last time on fixing mouse lungs. You'll be >glad to know we've got it working and have some nice H+E slides and the >immunos will be staring soon. >Can you help again? My boss has decided to look at the mouse's liver as >well (poor thing!) and I'd be grateful if you have any tips on fixing >liver. Can you just remove the liver and place it in a vial of formalin or >does it need perfused etc? >Looking forward to hearing from you all again. >Regards >Andrew > >Andrew MacDuff >Clinical Research Fellow >Wilkie Laboratory >Medical School >Edinburgh University >Teviot Place >Edinburgh >andrew.macduff@ed.ac.uk >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 4 Date: Tue, 01 Feb 2005 10:33:12 -0500 From: "Richard Cartun" Subject: [Histonet] p34 To: Message-ID: Content-Type: text/plain; charset=US-ASCII Is anyone doing IHC for p34 on formalin-fixed, paraffin-embedded tissue? If so, whose antibody are you using? 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 ------------------------------ Message: 5 Date: Tue, 1 Feb 2005 15:53:32 -0000 From: "Prior, Andrew" Subject: [Histonet] Professional Qualifications in UK To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Does anyone out there know of any professional qualifications in the UK for Histologists? I can find ones for Histo-paths, but I don't do any pathology work. >From what I gather, there are two levels of certification in the US for histologists and pathologists. Is the same true here in Ol' Blighty? Is it possible to do distance-learning for the American one? Thanks in advance Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York UK Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. ------------------------------ Message: 6 Date: Tue, 1 Feb 2005 16:11:44 -0000 From: "Prior, Andrew" Subject: [Histonet] Re: Millers Elastin Stain HELP To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Julia , You can find Miller Elastin Stain on the UK Version of the Raymond Lamb website ( http://www.ralamb.net/ ), but for some reason it's not on the US version of the site. You may be able to get it shipped across or just hassle R A Lamb in the States to sell you some. Hope this helps Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York UK >Message: 20 >Date: Tue, 1 Feb 2005 08:13:49 -0500 >From: Celebre Julia >Subject: [Histonet] Millers Elastin Stain HELP >To: "'histonet@lists.utsouthwestern.edu'" >Message-ID: <3AADFB88753AD31189C100902786B91C14E6DAE4@hch_nt_exchange.hhsc.ca> >Content-Type: text/plain; charset="iso-8859-1" >Good morning all.... >Our lab makes our own Millers solution, and I think it's just for plain old >punishment because if anyone has had to make it they understand what I >mean... Normally the stain works beautifully, but since we've run out of >'Victoria Blue 4R' and have used 'Victoria Blue B' as a substitute the stain >doesn't seem to be working too well. The reason why we substituted because >Victoria Blue 4R is no longer availalbe.. >What I'm asking all you histology guru's out there... >1. Does anybody know of a company that sells pre-made Millers? >2. Does anybody know where we can purchase Victoria Blue 4R? >3. Does anybody know if/why Victoria Blue B can be used as a substitute? >Thanks.. >Julia Celebre MLT >Anatomic Pathology >Hamilton General Hospital >905-527-0271 ext46145 >email: celebrej@hhsc.ca Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. ------------------------------ Message: 7 Date: Tue, 1 Feb 2005 11:22:18 -0500 From: "Bonner, Janet" Subject: RE: [Histonet] yeast and bacteria on slides To: "'Melissa Jans'" , histonet@lists.utsouthwestern.edu Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB418B@fh2k093.fhmis.net> Content-Type: text/plain; charset="iso-8859-1" Melissa, We are a 24-7 operation and had the same problem when our shifts exceeded eight hours and some of the waterbathes were not changed out. Changing the waterbath out when each shift was done totally eliminated the problem. -Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Melissa Jans Sent: Monday, January 31, 2005 8:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] yeast and bacteria on slides I need some help!! We have been experiencing some yeast and bacteria showing up on some of our special stains. It started about 2 weeks ago and seems to be sporadic (one day...no problems, the next day, several stains are affected). The yeast and bacteria (rods) are clumped together all over the slide, on the tissue sections, around the sections and on the slide in areas where there is no tissue. The stains we have seen the contamination are the Gram, PAS/D, PAS and GMS. All of these stains are automated except the Gram, so I do not think the special stainer is the problem. Here is what I have eliminated as problem areas: -Processor solutions and paraffin (we cut and stained a block from another institute and had contamination) -Waterbaths, slide drying boards, water containers (used to fill waterbathes), stainer containers, ice pans (all of the mentioned have been thoroughly bleached) -Tap water and DI water (all water supplies have been tested by sending a sample to micro...there was no bacteria or yeast detected) -Stay-on used as a slide adhesive (smeared some on a slide and stained it) -Bad batch of slides (we have had contamination on two different slide sources (vendors)) Does anybody have any ideas? What am I missing? Any suggestions would be much appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ 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 The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. ------------------------------ Message: 8 Date: Tue, 01 Feb 2005 11:51:43 -0500 From: John Kiernan Subject: Re: [Histonet] Formalin pigment To: "Bruijntjes, J.P." , histonet@lists.utsouthwestern.edu Message-ID: <41FFB39F.954A3FB@uwo.ca> Content-Type: text/plain; charset=us-ascii Formalin pigment is formed by the action of acid (pH <5.6) or alkali (pH >8) on haemoglobin. According to Lillie's big book (p. 488-489) similar pigment occurs around sites of haemorrhage in the gastric mucosa; it's then called HCl pigment. Malaria pigment is also closely similar. Lillie cites his own work in the field, which was published as 3 papers in 1947. -- ------------------------------- 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 _______________________________ "Bruijntjes, J.P." wrote: > > Is there anyone who can tell me what other reason could cause the > formation of formalin pigment except a too acidic formalin? > > > > Joost Bruijntjes > > TNO Nutrition and Food research > > Zeist > > The Netherlands > > > > 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 ------------------------------ Message: 9 Date: Tue, 1 Feb 2005 23:22:22 +0600 From: Subratab Subject: [Histonet] another background / non-specific staining problem To: Message-ID: <200502011727.j11HRLYb031576@mailout.proshikanet.com> Content-Type: text/plain; charset="iso-8859-1"; Dear All I am doing immunohistochemistry for 8-OHdG (a DNA marker for oxidative stress) in rat kidney sections (not perfused) fixed in methacarn. Along with positive nuclear staining, I am getting some background staining in some glomerulus and also in some interstitial area. This non-specific background is mainly in the areas where red cells are abundant (the tissue is not perfused). The same background is also present in my negative control slides (absence of primary Ab). My protocol is 1. Deparaffinization 2. Microwave exposure for 5 min in citrate buffer (10 mM, pH 6) 3. Horse serum blocking (5%) in PBS for 1 hr at room temp 4. Primary Ab (mouse monoclonal) against 8-OHdG (N45.1) overnight at 4 C (diluent 1% BSA and 0.3% triton X 100 in PBS) 5. Horse anti-mouse secondary Ab 1 hr at room temp 6. 3% H2O2 in methanol for 10 min. 7. ABC 8. DAB 9. Counter stain with hematoxilin 10. Dehydration and mounting. I did some experiments to identify if this non-specific staining is due to endogenous peroxidase/biotin. I followed above protocol omitting both the primary and secondary Ab. I did not get any background stain, my slides were clean. So I interpreted that the background is not due to endogenous peroxidase/biotin. Most probably it is due to non-specific binding of secondary Ab with red cell (or other tissue) components. Then I tried to block non-specific binding of secondary Ab in different ways (5% horse serum, 20% horse serum, 1%, and 5% non-fat milk, 1% BSA plus 1% non-fat milk). But I could not able to reduce the background. Lastly, I am writing you all for any suggestion. Thank you in advance Subrata Biswas University of Campinas SP, Brazil ------------------------------ Message: 10 Date: Tue, 1 Feb 2005 23:33:43 +0600 From: Subratab Subject: [Histonet] RE: HRP-conjugated secondary and biotinylated-secondary To: "C.M. van der Loos" Cc: histonet@lists.utsouthwestern.edu Message-ID: <200502011738.j11HcfYb031689@mailout.proshikanet.com> Content-Type: text/plain; charset="iso-8859-1"; Dear van der Loos Thank you very much for your reply with historical overview. Thank you everybody who replyed to my question. Subrata. --------- Mensagem Original -------- From: "C.M. van der Loos" To: "histonet@lists.utsouthwestern.edu" Cc: subratab@bdonline.com Subject: RE: HRP-conjugated secondary and biotinylated-secondary Date: 31/01/05 14:29 Hi, The existence of both biotinylated- and HRP-conjuated secondary antibodies reflects the development of IHC techniques into more staining efficiency/sensitivity. In the old days, a 2-step staining method using a HRP-conjugated secondary anti-mouse or anti-rabbit was common practice. Mid 80-ies the 3-step staining method using a biotinylated 2nd step reagent and streptavidin/HRP conjugate or streptavidin-biotinylated HRP complex became very popular because of its superior staining efficiency/sensitivity. The disadvantage here is the occurrence of endogenous biotin in many tissues. Endogenous biotin is not present in untreated formalin-fixed and paraffin-embedded tissue sections, but will be retrieved especially when using Tris-EDTA pH8-9. Only recently we have gone to a 2-step staining procedure again, now based on both secondary antibody and HRP attached to a polymer backbone. The efficiency/sensitivity of these polymers is comparable with the 3-step streptavidin/HRP procedure, but avoids the problem of endogenous biotin. I hope this small historical overview helps. 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 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From Subratab Date Sun, 30 Jan 2005 0:18:56 +0600 To Subject [Histonet] HRP-conjugated secondary and biotinylated-secondary Hi, Is it a common practice to use HRP-conjugated secondary antibody instead of biotinylated secondary antibody for immunohistochemistry (with DAB developing system)? I am confused about the advantage/ disadvantage of using HRP-conjugated secondary antibody for immunos. Subrata Biswas UNICAMP, SP, Brazil. References 1. mailto:c.m.vanderloos@amc.uva.nl ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 15, Issue 2 *************************************** From lchung <@t> ppmh.org Tue Feb 1 13:04:05 2005 From: lchung <@t> ppmh.org (Chung, Luong) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Problem with mucicarmine stain Message-ID: All, When we do mucicarmine (Richar/Allen pre-made) according to our protocol with dilution of one to four. We counterstain with tretrazine. We use appendix as a control and the stain turn out to be very pale. Is there any remedy for this? It used to work very well, but not lately. Anne From sbanwait <@t> buckinstitute.org Tue Feb 1 13:45:04 2005 From: sbanwait <@t> buckinstitute.org (Surita Banwait) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] intranasal delivery dye tracer Message-ID: <4AA34A707932424EBE2D973764D226A9026F89D1@inverness.buckcenter.org> Hello We are interested in trying a new method of delivery in mice (intranasal delivery) to avoid dealing with the blood brain barrier. Eventually a fluorescence-tagged marker will be delivered and analyzed. Before we do this however, my PI is interested in delivering a few different solutions (preferably a colored dye) that we can track through the mouse brain. Are there any suggestions as to some solutions we might try? Thanks for your consideration, Surita Surita Banwait Morphology Core Research Associate II Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2221 sbanwait@buckinstitute.org From Gareth <@t> gdavies24.freeserve.co.uk Tue Feb 1 13:37:17 2005 From: Gareth <@t> gdavies24.freeserve.co.uk (Gareth Davies) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] UK Biomedical scientists in Canada? Message-ID: <001001c50895$708c0120$831e4f51@garethda> Are there any UK qualified Biomedical scientists (or MLSO) working in Canada either perment or as locum (agencies)? I ask, to see if anyone has had experience with the International Credential Evaluation Service (ICES) and the CSMLS evaluation. The handbook guide seems straight forward, but it is always good to get advice from anyone who has been through it. Thank you Gareth Davies BMS 1 (Histology) POWH, Bridgend, Wales, UK From dsnider <@t> shrinenet.org Tue Feb 1 14:15:33 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Glass knife makers Message-ID: Hi all again! Has anybody used other knife makers than the LKB? If so what are the brands, and the high points of that particular piece of equipment? Down side? Ease of use? Service? etc. We are currently looking into the purchase of a new knife breaker and my superior wants to find out and hopefully even demo different ones before making the final purchase. So far I have contacted Leica, who now owns the LKB line, Messer, and GPK. Hopefully someone out there will know something about these or others I am unaware of. Thanks in advance, Deanna Snider Shriners Hospital Research Dept. Cincinnati, Oh 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 godsgirlnow <@t> MSN.COM Tue Feb 1 15:11:40 2005 From: godsgirlnow <@t> MSN.COM (Roxanne Soto) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Problem with mucicarmine stain References: Message-ID: A few more questions before we can help..... a.. Are you using tap water or distilled to make the working mucicarmine? b.. How long are you leaving it in the mucicarmine? c.. Is this done at room temp? d.. Do you use hematoxylin? Roxanne ----- Original Message ----- From: Chung, Luong To: Histonet (E-mail) Sent: Tuesday, February 01, 2005 2:04 PM Subject: [Histonet] Problem with mucicarmine stain All, When we do mucicarmine (Richar/Allen pre-made) according to our protocol with dilution of one to four. We counterstain with tretrazine. We use appendix as a control and the stain turn out to be very pale. Is there any remedy for this? It used to work very well, but not lately. Anne _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Feb 1 15:17:21 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Re: Glass knife makers In-Reply-To: References: Message-ID: <6.0.0.22.1.20050201135126.01b68130@gemini.msu.montana.edu> We have the older version of GKM triangular glass knife breaker, formerly under Sorvall (original manufacturer) name. 1/4" knives were made for cutting EM resin thicker sections, 1/4, 1/2 and 3/4" knives for glycol methacrylate embedded tissues. The joy of the knife breaker - it has been in our dept for over 25 years after heavy usage and needed parts, cleaning, basically reconditioning. It has been a workhorse, simple to operate, easy to use, and totally reliabled. Best of all - it has NOT been discontinued so service, repair and replacement parts are still available (our bills was minimal (less than $400) considering the units age and heavy use. The service from Energy Beam Sciences was excellent, prompt and Cindy Gaspari was very helpful. One can also purchase a little kit to replace the glass scoring wheels in house. You can go to their website for pricing, view the unit, etc. Although new units are pricey, and if I had to buy one based on company service, ease of use, but mainly unit longevity, it would be the now GKM aka Sorvall unit. It has been a excellent investment without downsides. Hmm does this sound like a sale pitch! Well, EBS can enjoy it from a happy customer. At 01:15 PM 2/1/2005, you wrote: >Hi all again! > >Has anybody used other knife makers than the LKB? If so what are the >brands, and the high points of that particular piece of equipment? Down >side? Ease of use? Service? etc. We are currently looking into the >purchase of a new knife breaker and my superior wants to find out and >hopefully even demo different ones before making the final purchase. So far >I have contacted Leica, who now owns the LKB line, Messer, and GPK. >Hopefully someone out there will know something about these or others I am >unaware of. > >Thanks in advance, >Deanna Snider >Shriners Hospital >Research Dept. >Cincinnati, Oh > >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. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From mcauliff <@t> umdnj.edu Tue Feb 1 18:19:09 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] intranasal delivery dye tracer In-Reply-To: <4AA34A707932424EBE2D973764D226A9026F89D1@inverness.buckcenter.org> References: <4AA34A707932424EBE2D973764D226A9026F89D1@inverness.buckcenter.org> Message-ID: <42001C7D.1040200@umdnj.edu> Hmmm. If the "practice" solutions work, does that mean that the fluor-tagged marker will work? I think not. I would try something of the same size, molecule wise, otherwise the practice may be for naught. If you want to get substance X into the CNS, I don't understand practicing with substance Y. Also, it sounds like your boss is assuming that the tracer will be picked up by olfactory neurons and transported to a particular target in the CNS? If the marker gets into the circulatory system (via the olfactory mucosa), the blood brain barrier will still be in the way. Geoff Surita Banwait wrote: >Hello > >We are interested in trying a new method of delivery in mice (intranasal >delivery) to avoid dealing with the blood brain barrier. Eventually a >fluorescence-tagged marker will be delivered and analyzed. Before we do >this however, my PI is interested in delivering a few different >solutions (preferably a colored dye) that we can track through the mouse >brain. Are there any suggestions as to some solutions we might try? > > > > > >Thanks for your consideration, > >Surita > > > >Surita Banwait > >Morphology Core > >Research Associate II > >Buck Institute for Age Research > >8001 Redwood Blvd. > >Novato, CA 94945 > >415-209-2221 > >sbanwait@buckinstitute.org > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From RCHIOVETTI <@t> aol.com Tue Feb 1 15:22:44 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Glass knife makers Message-ID: <1ea.35771305.2f314d24@aol.com> In a message dated 2/1/2005 1:25:55 PM US Mountain Standard Time, dsnider@shrinenet.org writes: > I have contacted Leica, who now owns the LKB line, Messer, and GPK. > Hopefully someone out there will know something about these or others I am > unaware of. > Deanna, You can also try RMC Products, now a part of Boeckeler Instruments, in Tucson, Arizona. They can be reached at: Tel. 520-745-0001 Fax 520-745-0004 www.rmcproducts.com Good luck, hope this helps. Bob Robert (Bob) Chiovetti, Ph.D. Prime Focus Consulting Group Advertising, Sales and Marketing Strategies Specializing in The Scientific, Biomedical and Biotechnology Sectors Tucson, Arizona USA Tel./Fax 520.546.4986 Member, Arizona Small Business Association - ASBA From jkiernan <@t> uwo.ca Tue Feb 1 16:13:20 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] de Olmos staining References: <1106907998.41fa135ee5ed0@web-mail1.uibk.ac.at> Message-ID: <41FFFF00.D3E4BD0@uwo.ca> The reference is: De Olmos JS, Beltramino CA, De Lorenzo SDO (1994) Use of an amino-cupric-silver technique for the detection of early and semiacute neuronal degeneration caused by neurotoxicants, hypoxia, and physical trauma. Neurotoxicology and Teratology 16: 545-561. For a complicated silver method like this one, it's important to read the original, not just a list of instructions. You can get the Abstract of the De Olmos paper on line at http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?holding=npg&cmd=Retrieve&db=PubMed&list_uids=7532272&dopt=Abstract but this doesn't say how to do the method. For that there's no alternative to a trip to the library. -- ------------------------------- 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 _______________________________ Martina Urbanek wrote: > > Hello, Everyone, > > Does anyone have a detailed protocol for de Olmos amino-cupric-silver stain > which works on paraffin sections (does it even work on paraffin?)? We work with > mice brains which are fixed in 10% formalin, but not perfused and we are > looking for a technique to stain degenerated neuronal cells. > Your help is greatly appreciated. > > Martina Urbanek > > Ms. Martina Urbanek > Forschungslabor der > Klin.Abt. f?r Neonatologie > neonatal neuroscience research laboratory > Med. University Innsbruck > Innrain 66, 4th floor > A-6020 Innsbruck > Tel. +43 (0)512 504 27755/27765 > Fax: +43 (0)512 504 27766 > Email: Martina.Urbanek@uibk.ac.at > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Tue Feb 1 16:22:19 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Problem with mucicarmine stain Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB418E@fh2k093.fhmis.net> Your control may be old - sometimes, depending on where you are and the conditions it's stored at, the reactivity diminishes. Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet (E-mail); Chung, Luong Sent: 2/1/2005 4:11 PM Subject: Re: [Histonet] Problem with mucicarmine stain A few more questions before we can help..... a.. Are you using tap water or distilled to make the working mucicarmine? b.. How long are you leaving it in the mucicarmine? c.. Is this done at room temp? d.. Do you use hematoxylin? Roxanne ----- Original Message ----- From: Chung, Luong To: Histonet (E-mail) Sent: Tuesday, February 01, 2005 2:04 PM Subject: [Histonet] Problem with mucicarmine stain All, When we do mucicarmine (Richar/Allen pre-made) according to our protocol with dilution of one to four. We counterstain with tretrazine. We use appendix as a control and the stain turn out to be very pale. Is there any remedy for this? It used to work very well, but not lately. Anne _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From cbass <@t> bidmc.harvard.edu Tue Feb 1 17:16:06 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] lacZ staining in the liver Message-ID: <3FB089D8-74A7-11D9-9415-000A95EC3B78@bidmc.harvard.edu> Hello Everyone, I was wondering if I could get some opinions (or protocols) on the best way to see lacZ expression in the liver. I am injecting the liver directly with a virus that should express lacZ. I would like to see the efficiency of expression with this virus. This is not a histology lab, nor am I very experienced so I have a number of constraints about how I can best process this tissue. I have easy access to a sliding microtome with a dry ice reservoir that will section efficiently in the 20-40 micron range. Due to the large number of animals, I would prefer to collect the tissue fresh and fix by immersion in formalin. I would collect floating sections, and probably stain the sections by floating them in the staining solution (containing NP40 and SDS), mount them onto superfrost plus slides, and counterstain with eosin, then coverslip with mount-quick. Does this sound like it should work? Is there anything I should look out for? For example, am I better off using some sort of subbed slide? Is eosin the best counterstain? If there are any other suggestions I would greatly appreciate it. I used this methodology for lacZ staining in the brain and it seems to work for me. However, I know very little about liver histology. Thanks, Caroline From robint <@t> mediom.qc.ca Tue Feb 1 20:17:24 2005 From: robint <@t> mediom.qc.ca (Robin Turcotte) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] toluidine bleu Message-ID: <003c01c508cd$5b46c650$be8ceccf@client> Dear All I am doing a microtomy on eye sections embedded in GMA. I am looking for a staining recipe with toluidine Blue .Are there special precautions to take for the staining or a special mounting medium to use with sections in GMA. Thank you Robin From cuttingedgehistology <@t> yahoo.com Tue Feb 1 20:26:18 2005 From: cuttingedgehistology <@t> yahoo.com (Brandon Stokes) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] CD-20 nuclear staining.... Message-ID: <20050202022618.47222.qmail@web41404.mail.yahoo.com> Hello fellow histo-netters, I was wondering if anyone else has ever experienced nuclear staining occuring when using the CD-20 antibody. The tonsil control will look great....then you look at the patient tissue(almost always skin for us) and some of the nuclei of the keratinocytes in the epidermis and even some nuclei in the dermis might light up bright red(we use AEC). You can actually see the nuclear detail very well. This problem doesn't happen all the time so I am not sure what it could be. Any advice would be greatly appreciated. Brandon Stokes, HT(ASCP) Lab Manager Cutting Edge Histology Services, LLC --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From rockbeki <@t> ufl.edu Tue Feb 1 21:42:22 2005 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] another background / non-specific staining problem Message-ID: <2009284135.1107315742346.JavaMail.osg@osgjas04.cns.ufl.edu> I had a similar problem when staining non-perfused sheep placenta with eNOS and someone (I think it was Gayle) on Histonet suggest putting my slides in 1% NaOH (the other 99 being ethanol) solution. I've been putting my slides in 1% NaOH for 15 min after the peroxide and it has worked well. -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From rockbeki <@t> ufl.edu Tue Feb 1 21:51:08 2005 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] RE: aqueous mounting medium Message-ID: <794885634.1107316268724.JavaMail.osg@osgjas04.cns.ufl.edu> I haven't used X-gal, but I work with AEC quite a bit and I use Crystal Mount with that. I used GVA before but I was getting a lot of bubbles and just found that it was just a pain to work with (not as thick as permount, but not thin enough to be able to just drop it on the slide like Crystal Mount). I do post-mount after the Crystal Mount dries with Permount and a normal coverslip because it makes my slides sharper looking. (I think this might be partially to due to the texture of the tissue I'm working with-placenta.) -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From lpwenk <@t> sbcglobal.net Wed Feb 2 04:29:57 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Problem with mucicarmine stain References: Message-ID: <003301c50912$2577d9c0$8324d445@domainnotset.invalid> 1. CONTROL: Not all GI have lots of mucin in the goblet cells. It can vary from patient to patient. The goblet cells may have "extruded" some of the mucin recently, so there might not be as much, so the goblet cells stain paler. Try a control from a different patient. There may be more mucin in the goblet cells from another patient. Small or large intestine (not autolyzed) also make good controls. 2. COUNTERSTAIN: Counterstaining with metanil yellow or tartrazine can mask minimal deposits, or reduce the contrast between the pink goblet cells and the background. How about counterstaining with just a 30 second aluminum hematoxylin, such as Gill or Mayer? Easier to see minimal deposits. That's how our pathologists like us to do the stain. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Chung, Luong" To: "Histonet (E-mail)" Sent: Tuesday, February 01, 2005 2:04 PM Subject: [Histonet] Problem with mucicarmine stain All, When we do mucicarmine (Richar/Allen pre-made) according to our protocol with dilution of one to four. We counterstain with tretrazine. We use appendix as a control and the stain turn out to be very pale. Is there any remedy for this? It used to work very well, but not lately. Anne _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From subratab <@t> bdonline.com Wed Feb 2 07:11:47 2005 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] another background / non-specific staining problem: Thank you all Message-ID: <200502021316.j12DGuYb025017@mailout.proshikanet.com> Dear all Thank you very much for your prompt reply to my problem. I have received several responses. Now I am trying to follow the suggestions. Thank you all of you for your nice suggestions. Subrata. --------- Mensagem Original -------- From: SMITH,REBEKAH FELICIA To: GUTIERREZ, JUAN , Subratab , histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] another background / non-specific staining problem Date: 02/02/05 09:48 > > I had a similar problem when staining non-perfused sheep placenta > with eNOS and someone (I think it was Gayle) on Histonet suggest > putting my slides in 1% NaOH (the other 99 being ethanol) > solution. I've been putting my slides in 1% NaOH for 15 min after > the peroxide and it has worked well. > -- > SMITH,REBEKAH FELICIA > "You are a child of the universe, no less than the trees and the > stars > You have a right to be here and whether or not it is clear to you, > no doubt the universe is unfolding as it should. Therefore be at > peace with G-d, whatever you conceive Him to be. And whatever your > labors and aspirations,in the noisy confusion of life, keep peace > in your soul.-Max Ehrmann,"Desiderata" > > > > > From TMcNemar <@t> lmhealth.org Wed Feb 2 06:55:07 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Alcian Blue problem Message-ID: <6CD94D97ED7D924BA5C2B588FA952821396780@nt_exchange.lmhealth.org> We have started to see a hazy/cloudiness to our alcian blue PAS slides. We don't see this on any other slide/stain. We use the same slides, reagents, and procedure that we have always used. The stain works fine and the haze is on the entire slide, not just the tissue. We routinely use the Fisherbrand red control slides and have tried other slides and still have the same problem. I use the Poly Scientific reagents (alcian blue ph 2.5) and have switched them all out for new reagents but still have the haze. We have switched out, altered, and tried everything I can think of. I am totally baffled. Hope you folks have some ideas. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 From Rcartun <@t> harthosp.org Wed Feb 2 09:23:02 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] CD-20 nuclear staining.... Message-ID: I have never seen nuclear staining with CD20 (clone L26). Which pretreatment and detection system are you using? 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 >>> Brandon Stokes 02/01/05 09:26PM >>> Hello fellow histo-netters, I was wondering if anyone else has ever experienced nuclear staining occuring when using the CD-20 antibody. The tonsil control will look great....then you look at the patient tissue(almost always skin for us) and some of the nuclei of the keratinocytes in the epidermis and even some nuclei in the dermis might light up bright red(we use AEC). You can actually see the nuclear detail very well. This problem doesn't happen all the time so I am not sure what it could be. Any advice would be greatly appreciated. Brandon Stokes, HT(ASCP) Lab Manager Cutting Edge Histology Services, LLC --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Feb 2 09:28:28 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:34 2005 Subject: Reference to Gayle on RE: [Histonet] another background / non-specific staining problem In-Reply-To: <2009284135.1107315742346.JavaMail.osg@osgjas04.cns.ufl.edu > References: <2009284135.1107315742346.JavaMail.osg@osgjas04.cns.ufl.edu> Message-ID: <6.0.0.22.1.20050202082630.01b47ea8@gemini.msu.montana.edu> Rebekah, Sorry folks but this is not a recommendation from me. I have never done this! At 08:42 PM 2/1/2005, you wrote: >I had a similar problem when staining non-perfused sheep placenta with >eNOS and someone (I think it was Gayle) on Histonet suggest putting my >slides in 1% NaOH (the other 99 being ethanol) solution. I've been putting >my slides in 1% NaOH for 15 min after the peroxide and it has worked well. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From aep10 <@t> cornell.edu Wed Feb 2 09:53:54 2005 From: aep10 <@t> cornell.edu (Anna Elisse Beaudin) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] storage of frozen cryosections (again) Message-ID: <1937.128.253.96.73.1107359634.squirrel@128.253.96.73> Dear Histonet, I have a question regarding the storage and preservation of frozen cryosections. I apologize if I have asked this question before, but I'm still unclear as to the best approach. Currently, when I collect cryosections, I dry them overnight at room temp and than use them immediately the next day. however, I would like to be able to collect sections to use at later timepoints. My concern is that O/N drying at room temperature followed by freezing and storage at -20 or -80 might cause ice crystals or otherwise affect the quality and morphology of my sections. Additionally, even if I were to freeze the sections immediately following sectioning, they are still collected on slides at room temperature, and thus I wonder if I would have the same problem with thaw/freeze. I would greatly appreciate anyone's advice with this... I would really like to be able to preserve more sections for later use! Thanks so much in advance! Anna Beaudin Division of Nutritional Sciences Cornell University From jkiernan <@t> uwo.ca Wed Feb 2 10:29:47 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] storage of frozen cryosections (again) References: <1937.128.253.96.73.1107359634.squirrel@128.253.96.73> Message-ID: <4200FFFB.C3FD8AF7@uwo.ca> If you put the slides in the freezer after the sections have dried, there shouldn't be enough water in the tissue to form visible ice crystals. It's important to wrap the the slides up in something airtight, and with some dessicant (drierite or silica gel). Don't unwrap the packet until its contents have warmed to room temperature, or water will condense on the cold sections - quite damaging if the tissue is unfixed. -- ------------------------------- 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 _______________________________ Anna Elisse Beaudin wrote: > > Dear Histonet, > > I have a question regarding the storage and preservation of frozen > cryosections. I apologize if I have asked this question before, but > I'm still unclear as to the best approach. Currently, when I collect > cryosections, I dry them overnight at room temp and than use them > immediately the next day. however, I would like to be able to collect > sections to use at later timepoints. My concern is that O/N drying at > room temperature followed by freezing and storage at -20 or -80 might > cause ice crystals or otherwise affect the quality and morphology of my > sections. Additionally, even if I were to freeze the sections > immediately following sectioning, they are still collected on slides at > room temperature, and thus I wonder if I would have the same problem > with thaw/freeze. I would greatly appreciate anyone's advice with > this... I would really like to be able to preserve more sections for > later use! > > Thanks so much in advance! > Anna Beaudin > Division of Nutritional Sciences > Cornell University From BoozerKA <@t> pa1.ah.org Wed Feb 2 10:08:51 2005 From: BoozerKA <@t> pa1.ah.org (Kathleen Boozer) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Alcian Blue problem Message-ID: I also have occasional problems with the 2.5 Alcian Blue. Greenish parcipitants on top of the slide. Same solutions...no consistency of trouble... >>> Tom McNemar 2/2/2005 4:55:07 AM >>> We have started to see a hazy/cloudiness to our alcian blue PAS slides. We don't see this on any other slide/stain. We use the same slides, reagents, and procedure that we have always used. The stain works fine and the haze is on the entire slide, not just the tissue. We routinely use the Fisherbrand red control slides and have tried other slides and still have the same problem. I use the Poly Scientific reagents (alcian blue ph 2.5) and have switched them all out for new reagents but still have the haze. We have switched out, altered, and tried everything I can think of. I am totally baffled. Hope you folks have some ideas. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peoshel <@t> wisc.edu Wed Feb 2 10:36:37 2005 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Alcian Blue problem In-Reply-To: References: Message-ID: Do you filter your stain? Phil >I also have occasional problems with the 2.5 Alcian Blue. Greenish >parcipitants on top of the slide. Same solutions...no consistency of >trouble... > >>>> Tom McNemar 2/2/2005 4:55:07 AM >>> >We have started to see a hazy/cloudiness to our alcian blue PAS slides. > We >don't see this on any other slide/stain. We use the same slides, >reagents, >and procedure that we have always used. The stain works fine and the >haze >is on the entire slide, not just the tissue. We routinely use the >Fisherbrand red control slides and have tried other slides and still >have >the same problem. I use the Poly Scientific reagents (alcian blue ph >2.5) >and have switched them all out for new reagents but still have the >haze. >We have switched out, altered, and tried everything I can think of. I >am >totally baffled. Hope you folks have some ideas. > >Tom Mc Nemar HT(ASCP) >Histology Supervisor >Licking Memorial Hospital >Newark, Ohio 43055 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >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) From NMargaryan <@t> childrensmemorial.org Wed Feb 2 10:53:29 2005 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] IL-6 Message-ID: <63B8B599DE283148B92E83C78B32C15D4C8731@cmhexbe2.childrensmemorial.org> Hi everyone, Is anybody have been tried to use IL-6 for the FFPE Breast tissue (normal and carcinoma)? If yes, what protocol did you use? I have a Monoclonal anti-human IL-6 Ab from R&D (cat#: MAB206) and I tried to IHC with DAB staining on the breast tissue. Unfortunately, I didn't get any staining. Is anybody can send me FFPE tonsil tissue as a control? Thanks, Naira Naira V. Margaryan, Ph.D, D.V.M. Research Associate II Children's Memorial Research Center 2300 Children's Plaza, Box 222 Chicago, IL 60614-3394 Tel: 773-880-4000/5-6740 Fax: 773-755-6594 nmargaryan@childrensmemorial.org For Express Mail: CMRC, Room C.473 2430 N. Halsted Street Chicago, IL 60614-4314 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 kspencer <@t> utmem.edu Wed Feb 2 11:08:42 2005 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] storage of frozen cryosections (again) In-Reply-To: <1937.128.253.96.73.1107359634.squirrel@128.253.96.73> References: <1937.128.253.96.73.1107359634.squirrel@128.253.96.73> Message-ID: <0c83378159b2835ad98931fe35e28681@utmem.edu> We pick up our sections and put them in a slide box that is in the cryostat and keep them there until we are finished sectioning. Then we rush them to the -80 freezer. We put a dessicant pack or two in the box with the slides, and we wrap the slide box in foil, then stick that into a plastic bag that we label as to what the slides are. When we are ready to do the next step, we remove the box from the freezer, quickly take out the slides we need & put the box back. We then place a warm finger under the section for half a second, then put the slide into cold fix or 70% alcohol, just depends on what we are using the slides for. It is usually LCM for RNA. but it could work for other things like IHC. The morphology is terrible on fresh frozen tissue sections, so for IHC we always perfuse the animal with PFA and store the tissue in in 20% sucrose, then freeze the tissue for cryosections. Then they can air dry because they are fixed. Make sense? I hope this helps. Kathleen Spencer HT (ASCP) Lab Manager/LCM Supervisor UTHSC On Feb 2, 2005, at 9:53 AM, Anna Elisse Beaudin wrote: > Dear Histonet, > > I have a question regarding the storage and preservation of frozen > cryosections. I apologize if I have asked this question before, but > I'm still unclear as to the best approach. Currently, when I collect > cryosections, I dry them overnight at room temp and than use them > immediately the next day. however, I would like to be able to collect > sections to use at later timepoints. My concern is that O/N drying at > room temperature followed by freezing and storage at -20 or -80 might > cause ice crystals or otherwise affect the quality and morphology of my > sections. Additionally, even if I were to freeze the sections > immediately following sectioning, they are still collected on slides at > room temperature, and thus I wonder if I would have the same problem > with thaw/freeze. I would greatly appreciate anyone's advice with > this... I would really like to be able to preserve more sections for > later use! > > Thanks so much in advance! > Anna Beaudin > Division of Nutritional Sciences > Cornell University > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Wed Feb 2 11:09:51 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] IL-6 In-Reply-To: <63B8B599DE283148B92E83C78B32C15D4C8731@cmhexbe2.childrensmemorial.org> Message-ID: <001001c5094a$0200e490$76d48a80@AMY> Naira I have stained for IL-6 on formalin fixed, formic acid decalcifed rat joints. I use the antibody from Abcam ab6672. I used proteinase K digestion with envision +. This antibody is directed against human IL-6. My dilution was 1:300 for 30 minutes at RT. 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 Margaryan, Naira Sent: Wednesday, February 02, 2005 9:53 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IL-6 Hi everyone, Is anybody have been tried to use IL-6 for the FFPE Breast tissue (normal and carcinoma)? If yes, what protocol did you use? I have a Monoclonal anti-human IL-6 Ab from R&D (cat#: MAB206) and I tried to IHC with DAB staining on the breast tissue. Unfortunately, I didn't get any staining. Is anybody can send me FFPE tonsil tissue as a control? Thanks, Naira Naira V. Margaryan, Ph.D, D.V.M. Research Associate II Children's Memorial Research Center 2300 Children's Plaza, Box 222 Chicago, IL 60614-3394 Tel: 773-880-4000/5-6740 Fax: 773-755-6594 nmargaryan@childrensmemorial.org For Express Mail: CMRC, Room C.473 2430 N. Halsted Street Chicago, IL 60614-4314 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 mprice26 <@t> juno.com Wed Feb 2 11:23:59 2005 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] RE: Zeta Co. Message-ID: <20050202.092435.13815.790@webmail25.nyc.untd.com> Histonetters, Does anyone have any contact information for a company called Zeta Co. I found an antibody (P504S) that someone apparently ordered before I was employed here and I do not find an spec sheet on it. I appreciate your help. Marsha Price From vargasb <@t> nova.edu Wed Feb 2 11:45:10 2005 From: vargasb <@t> nova.edu (Bernardo Vargas-Angel) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Nuclear staining Message-ID: <200502021745.j12HjAUN014364@r2d2.acast.nova.edu> I am working with scleractinian corals and I am experiencing problems with nuclear staining (H&E), particularly one species: Montastraea cavernosa. Initially, we attributed lack of basophilia to overdecalcification, however after decreasing the acid concentration from 5% to 2.5% and being really careful to assess the decal endpoint, we still have very weak nuclear staining (for the aforementioned species). Other species of corals that I have been working with have responded quite favorably to the decreased acid concentration (increased basophilia); however, not M. cavernosa. Has any out there encountered similar problems with other kinds of tissues? Have any of you observed differential tissue sensitivity to acid decal? Any insight would be greatly appreciated. Best and many thanks in advance. Bernardo Vargas-Angel Ph.D. Research Scientist National Coral Reef Institute Nova Southeastern University 8000 N Ocean Drive, Dania Beach, FL, 33004 Voice +(954) 262-3677 Fax +(954) 262-4027 http://www.nova.edu/ocean/vargas-angel/index.html From travers.3 <@t> osu.edu Wed Feb 2 14:51:35 2005 From: travers.3 <@t> osu.edu (Susan Travers) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] fluorescent counterstain Message-ID: Can anyone recommend a fluorescent counterstain for cell bodies in neural tissue (similar to cresyl violet). The tissue is formalin fixed. Thanks Susan Travers -- Susan Travers, Ph.D. Professor, Oral Biology The Ohio State University College of Dentistry 305 W. 12th Avenue Columbus, Ohio 43210-1267 (614)292-6366(V) (614)247-6945(F) From gcallis <@t> montana.edu Wed Feb 2 12:02:02 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:34 2005 Subject: LCM and IHC Re: [Histonet] storage of frozen cryosections (again) In-Reply-To: <0c83378159b2835ad98931fe35e28681@utmem.edu> References: <1937.128.253.96.73.1107359634.squirrel@128.253.96.73> <0c83378159b2835ad98931fe35e28681@utmem.edu> Message-ID: <6.0.0.22.1.20050202104739.01b07b20@gemini.msu.montana.edu> I think the key point here is cryostat storage works for LCM protocols. However, for general IHC without using LCM in the end, we avoid cryostat storage. Sections taken from cryostat temperature get water condensation formation on sections which contributes to morphology and antigen damage. We do not have problems with fresh frozen section morphology but I think that is more dependent on fixation.. Acetone fixation is notorious for poor morphology, but if the antigen can be fixed with another fixative, say acetone/alcohol mixture, one sees a huge improvement in morphology preservation. Obviously, PFA is even better. We know in our lab, as do many others, that PFA fixation for our murine CD4 and CD8 markers leads to no staining of these lymphocytes. I patiently await the day when we could use PFA for murine CD4 and CD8, alas - no clones yet nor a retrieval that works. At 10:08 AM 2/2/2005, you wrote: >We pick up our sections and put them in a slide box that is in the >cryostat and keep them there until we are finished sectioning. Then we >rush them to the -80 freezer. We put a dessicant pack or two in the box >with the slides, and we wrap the slide box in foil, then stick that into a >plastic bag that we label as to what the slides are. When we are ready to >do the next step, we remove the box from the freezer, quickly take out >the slides we need & put the box back. We then place a warm finger under >the section for half a second, then put the slide into cold fix or 70% >alcohol, just depends on what we are using the slides for. It is usually >LCM for RNA. but it could work for other things like IHC. The morphology >is terrible on fresh frozen tissue sections, so for IHC we always perfuse >the animal with PFA and store the tissue in in 20% sucrose, then freeze >the tissue for cryosections. Then they can air dry because they are fixed. >Make sense? I hope this helps. > >Kathleen Spencer HT (ASCP) >Lab Manager/LCM Supervisor >UTHSC > > >On Feb 2, 2005, at 9:53 AM, Anna Elisse Beaudin wrote: > >>Dear Histonet, >> >> I have a question regarding the storage and preservation of frozen >>cryosections. I apologize if I have asked this question before, but >>I'm still unclear as to the best approach. Currently, when I collect >>cryosections, I dry them overnight at room temp and than use them >>immediately the next day. however, I would like to be able to collect >>sections to use at later timepoints. My concern is that O/N drying at >>room temperature followed by freezing and storage at -20 or -80 might >>cause ice crystals or otherwise affect the quality and morphology of my >>sections. Additionally, even if I were to freeze the sections >>immediately following sectioning, they are still collected on slides at >>room temperature, and thus I wonder if I would have the same problem >>with thaw/freeze. I would greatly appreciate anyone's advice with >>this... I would really like to be able to preserve more sections for >>later use! >> >>Thanks so much in advance! >>Anna Beaudin >>Division of Nutritional Sciences >>Cornell 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 Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From vazquezr <@t> ohsu.edu Wed Feb 2 12:22:18 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] storage of frozen cryosections (again) Message-ID: Anna, What we do is cut our sections of frozen tissue, place them in slide boxes and put them in -20 and -80 with the same results of the tissue diagnosis. The tissue is our controls. Robyn OHSU >>> "Anna Elisse Beaudin" 2/2/2005 7:53:54 AM >>> Dear Histonet, I have a question regarding the storage and preservation of frozen cryosections. I apologize if I have asked this question before, but I'm still unclear as to the best approach. Currently, when I collect cryosections, I dry them overnight at room temp and than use them immediately the next day. however, I would like to be able to collect sections to use at later timepoints. My concern is that O/N drying at room temperature followed by freezing and storage at -20 or -80 might cause ice crystals or otherwise affect the quality and morphology of my sections. Additionally, even if I were to freeze the sections immediately following sectioning, they are still collected on slides at room temperature, and thus I wonder if I would have the same problem with thaw/freeze. I would greatly appreciate anyone's advice with this... I would really like to be able to preserve more sections for later use! Thanks so much in advance! Anna Beaudin Division of Nutritional Sciences Cornell University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Wed Feb 2 12:22:40 2005 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] RE: Zeta Co. Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5C1D7@EXCHANGE1.huntingtonhospital.com> Their number is (626) 355-2053. We also use this antibody. Laurie Colbert -----Original Message----- From: mprice26@juno.com [mailto:mprice26@juno.com] Sent: Wednesday, February 02, 2005 9:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Zeta Co. Histonetters, Does anyone have any contact information for a company called Zeta Co. I found an antibody (P504S) that someone apparently ordered before I was employed here and I do not find an spec sheet on it. I appreciate your help. Marsha Price _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Feb 2 14:24:32 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] CD-20 nuclear staining.... In-Reply-To: Message-ID: <200502022024.j12KOSG7020566@chip.viawest.net> Over heating during hier can cause false nuclear staining? Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Wednesday, February 02, 2005 8:23 AM To: Histonet@lists.utsouthwestern.edu; cuttingedgehistology@yahoo.com Subject: Re: [Histonet] CD-20 nuclear staining.... I have never seen nuclear staining with CD20 (clone L26). Which pretreatment and detection system are you using? 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 >>> Brandon Stokes 02/01/05 09:26PM >>> Hello fellow histo-netters, I was wondering if anyone else has ever experienced nuclear staining occuring when using the CD-20 antibody. The tonsil control will look great....then you look at the patient tissue(almost always skin for us) and some of the nuclei of the keratinocytes in the epidermis and even some nuclei in the dermis might light up bright red(we use AEC). You can actually see the nuclear detail very well. This problem doesn't happen all the time so I am not sure what it could be. Any advice would be greatly appreciated. Brandon Stokes, HT(ASCP) Lab Manager Cutting Edge Histology Services, LLC --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bwhitaker <@t> brownpathology.com Wed Feb 2 14:37:24 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] CD-20 nuclear staining.... In-Reply-To: <200502022024.j12KOSG7020566@chip.viawest.net> Message-ID: <000001c50967$008031d0$3601a8c0@brownpathology.net> If the pH of the HIER solution is off (too acid) you can see this artifact, as well. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Wednesday, February 02, 2005 2:25 PM To: 'Richard Cartun'; Histonet@lists.utsouthwestern.edu; cuttingedgehistology@yahoo.com Subject: RE: [Histonet] CD-20 nuclear staining.... Over heating during hier can cause false nuclear staining? Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Wednesday, February 02, 2005 8:23 AM To: Histonet@lists.utsouthwestern.edu; cuttingedgehistology@yahoo.com Subject: Re: [Histonet] CD-20 nuclear staining.... I have never seen nuclear staining with CD20 (clone L26). Which pretreatment and detection system are you using? 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 >>> Brandon Stokes 02/01/05 09:26PM >>> Hello fellow histo-netters, I was wondering if anyone else has ever experienced nuclear staining occuring when using the CD-20 antibody. The tonsil control will look great....then you look at the patient tissue(almost always skin for us) and some of the nuclei of the keratinocytes in the epidermis and even some nuclei in the dermis might light up bright red(we use AEC). You can actually see the nuclear detail very well. This problem doesn't happen all the time so I am not sure what it could be. Any advice would be greatly appreciated. Brandon Stokes, HT(ASCP) Lab Manager Cutting Edge Histology Services, LLC --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@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 nena.dimaano <@t> stryker.com Wed Feb 2 15:07:14 2005 From: nena.dimaano <@t> stryker.com (Dimaano, Nena) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Leitz Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD13F340@HOS2KEXCHCL.howost.strykercorp.com> Hi Everyone, I was wondering if anyone has any idea or uses the special microtome for hard tissue with implants called Leitz 1600. Thanks, Nena Dimaano, MT/HT(ASCP) Advanced Technology Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com From katri <@t> cogeco.ca Wed Feb 2 15:51:10 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] CD-20 nuclear staining.... References: <20050202022618.47222.qmail@web41404.mail.yahoo.com> Message-ID: <006501c50971$4f659920$6a9a9618@Katri> I have seen this occasionally with various antibodies. It seems to be a combination of too short a fixation and HIER. Properly fixed control material does not show this false positivity. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Brandon Stokes" To: Sent: Tuesday, February 01, 2005 9:26 PM Subject: [Histonet] CD-20 nuclear staining.... > Hello fellow histo-netters, > > I was wondering if anyone else has ever experienced nuclear staining > occuring when using the CD-20 antibody. The tonsil control will look > great....then you look at the patient tissue(almost always skin for us) > and some of the nuclei of the keratinocytes in the epidermis and even some > nuclei in the dermis might light up bright red(we use AEC). You can > actually see the nuclear detail very well. This problem doesn't happen > all the time so I am not sure what it could be. > > Any advice would be greatly appreciated. > > > Brandon Stokes, HT(ASCP) > Lab Manager > Cutting Edge Histology Services, LLC > > > > --------------------------------- > Do you Yahoo!? > Yahoo! Search presents - Jib Jab's 'Second Term' > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jkiernan <@t> uwo.ca Wed Feb 2 16:26:17 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] De Olmos silver stain - more information Message-ID: <42015389.4D5EEEAB@uwo.ca> The Neuroscience Associates web site has some information about the technique, with nice photomicrographs. http://www.NSALabs.com/Stains/silver_degen.htm There are also informative, well illustrated links from this page. The method is called "Amino Cupric Silver." The name De Olmos isn't mentioned; possibly they've modified the original and aren't saying how. NA is a firm in Knoxville TN owned by a Dr Robert Switzer. They do staining and other lab work for researchers. -- ------------------------------- 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 _______________________________ From rockbeki <@t> ufl.edu Wed Feb 2 16:25:12 2005 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:34 2005 Subject: Reference to Gayle on RE: [Histonet] another background / non-specific staining problem Message-ID: <1913502526.1107383112033.JavaMail.osg@osgjas04.cns.ufl.edu> Sorry 'bout that, I guess I got people mixed up. On Wed Feb 02 10:28:28 EST 2005, Gayle Callis wrote: > Rebekah, > > Sorry folks but this is not a recommendation from me. > > I have never done this! > > At 08:42 PM 2/1/2005, you wrote: >> I had a similar problem when staining non-perfused sheep >> placenta with eNOS and someone (I think it was Gayle) on >> Histonet suggest putting my slides in 1% NaOH (the other 99 >> being ethanol) solution. I've been putting my slides in 1% NaOH >> for 15 min after the peroxide and it has worked well. > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From naje1972 <@t> yahoo.com Wed Feb 2 16:43:49 2005 From: naje1972 <@t> yahoo.com (cynthia haynes) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] microtomes Message-ID: <20050202224349.65329.qmail@web41723.mail.yahoo.com> Would the sales rep from the Microm company please contact me at 773-484-4133. We are putting a budget together for new equipment and we are in need of a new microtome. We are looking for either a rotary or semi-automatic machine. My name is Cynthia Haynes. We would like to hear from you soon. Thanks in advance. Cynthia Haynes H.T. Laboratory Director __________________________________ Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. http://info.mail.yahoo.com/mail_250 From gcallis <@t> montana.edu Wed Feb 2 16:45:42 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Leitz for implants In-Reply-To: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD13F340@HOS2KEXCHCL.howost. strykercorp.com> References: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD13F340@HOS2KEXCHCL.howost.strykercorp.com> Message-ID: <6.0.0.22.1.20050202154236.01b1d918@gemini.msu.montana.edu> Nena, Metal implant or soft implant? Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From travers.3 <@t> osu.edu Wed Feb 2 19:50:14 2005 From: travers.3 <@t> osu.edu (Susan Travers) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] GABAa, alpha1 subunit Message-ID: Does anyone have a suggestion for an antibody for immunohistochemistry (para-fixed brain sections) for the GABAa receptor, alpha1 subunit. Thanks so much. Susan Travers -- Susan Travers, Ph.D. Professor, Oral Biology The Ohio State University College of Dentistry 305 W. 12th Avenue Columbus, Ohio 43210-1267 (614)292-6366(V) (614)247-6945(F) From AnthonyH <@t> chw.edu.au Wed Feb 2 17:32:06 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Nuclear staining Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E17F@simba.kids> Bernardo, Try treating the cut sections with 1% Periodic acid for 10-15 minutes and then rinse in dilute Lithium Carbonate for 10 minutes prior to haematoxylin staining. You might also double your Hx staining time and decrease the eosin staining time to give you better results. 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: Bernardo Vargas-Angel [mailto:vargasb@nova.edu] Sent: Thursday, 3 February 2005 4:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Nuclear staining I am working with scleractinian corals and I am experiencing problems with nuclear staining (H&E), particularly one species: Montastraea cavernosa. Initially, we attributed lack of basophilia to overdecalcification, however after decreasing the acid concentration from 5% to 2.5% and being really careful to assess the decal endpoint, we still have very weak nuclear staining (for the aforementioned species). Other species of corals that I have been working with have responded quite favorably to the decreased acid concentration (increased basophilia); however, not M. cavernosa. Has any out there encountered similar problems with other kinds of tissues? Have any of you observed differential tissue sensitivity to acid decal? Any insight would be greatly appreciated. Best and many thanks in advance. Bernardo Vargas-Angel Ph.D. Research Scientist National Coral Reef Institute Nova Southeastern University 8000 N Ocean Drive, Dania Beach, FL, 33004 Voice +(954) 262-3677 Fax +(954) 262-4027 http://www.nova.edu/ocean/vargas-angel/index.html _______________________________________________ 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 jkiernan <@t> uwo.ca Wed Feb 2 21:45:25 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] fluorescent counterstain References: Message-ID: <42019E55.B970E9F8@uwo.ca> There are many. My favourite is neutral red (CI 40040; Basic red 5). Stain sections for 5 min with a 0.002% aqueous solution, then dehydrate, clear and cover. Use a non-fluorescent mountant such as DPX. Orange-red fluorescence with a wide range of workable excitation wavelength (325-500 nm). For more information and a picture, see Neuroscience 99: 755-764 (1994). John Kiernan London, Canada --------------------------------------------------- Susan Travers wrote: > > Can anyone recommend a fluorescent counterstain for cell bodies in > neural tissue (similar to cresyl violet). The tissue is formalin > fixed. > > Thanks > Susan Travers > -- > Susan Travers, Ph.D. > Professor, Oral Biology > > The Ohio State University > College of Dentistry > 305 W. 12th Avenue > Columbus, Ohio 43210-1267 > > (614)292-6366(V) > (614)247-6945(F) > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Thu Feb 3 02:01:24 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] RE: storage of frozen cryosections (again) Message-ID: <105120410559fc.10559fc1051204@amc.uva.nl> Hi Anna, I fully agree with John's opinion with respect to drying of cyrosections. I would like to add that we found out that storage at -20C leads to a loss of specific antigens like CD4, CD3, CD25. Storage of the cryosections at -80C gives a much better preservation. Even after many years, those "vulnerable" antigens can be detected without any problem. 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 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From "Anna Elisse Beaudin" Date Wed, 2 Feb 2005 10:53:54 -0500 (EST) To Histonet@lists.utsouthwestern.edu Subject [Histonet] storage of frozen cryosections (again) Dear Histonet, I have a question regarding the storage and preservation of frozen cryosections. I apologize if I have asked this question before, but I'm still unclear as to the best approach. Currently, when I collect cryosections, I dry them overnight at room temp and than use them immediately the next day. however, I would like to be able to collect sections to use at later timepoints. My concern is that O/N drying at room temperature followed by freezing and storage at -20 or -80 might cause ice crystals or otherwise affect the quality and morphology of my sections. Additionally, even if I were to freeze the sections immediately following sectioning, they are still collected on slides at room temperature, and thus I wonder if I would have the same problem with thaw/freeze. I would greatly appreciate anyone's advice with this... I would really like to be ! able to p ----- Original Message ----- From John Kiernan Date Wed, 02 Feb 2005 11:29:47 -0500 To Anna Elisse Beaudin Cc Histonet@lists.utsouthwestern.edu Subject Re: [Histonet] storage of frozen cryosections (again) If you put the slides in the freezer after the sections have dried, there shouldn't be enough water in the tissue to form visible ice crystals. It's important to wrap the the slides up in something airtight, and with some dessicant (drierite or silica gel). Don't unwrap the packet until its contents have warmed to room temperature, or water will condense on the cold sections - quite damaging if the tissue is unfixed. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 References 1. mailto:c.m.vanderloos@amc.uva.nl From Kemlo.Rogerson <@t> elht.nhs.uk Thu Feb 3 02:34:17 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Macropath[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF10@bhrv-nt-11.bhrv.nwest.nhs.uk> We have just received the SurgiPath MACROPATH and find there is no microphone to capture audio evidence. I intend to use a USB microphone to capture voice as a .wav file and then send it to the server so that the Medical Secretaries can gain access to the dictation. Has anyone any experience? From Inga.Hansson <@t> neuro.uu.se Thu Feb 3 07:28:44 2005 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] paraffin problem Message-ID: Hi everyone, Has anyone experienced a difference in the quality of paraffin from different suppliers? Since we use rather small amounts, I found a company selling it in 1 kg packages. I noticed lately that it seems to be more opaque than before and although the melting-point is said to be the same it seems to take longer time to melt. Perhaps I?m imagining it but there has been occasional problems in sectioning as well. Comments? Thanks ever so much! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 From Don.Birgerson <@t> leica-microsystems.com Thu Feb 3 07:58:49 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Leitz Message-ID: Hi Nena, The SP1600 Saw Microtome is used to make slices of hard mixes of material. The original use was found in dental schools where it made 50 to 100 micron slices of samples of teeth with the metal fillings intact. The advantages are its ability to make optically flat slices of material with different harnesses The limitations are the specimen sizes of less than 25 mm diameter and the loss of material between sections. Leica Microsystems still manufactures this instrument. If you have further questions, please feel free to phone . Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Dimaano, Nena" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Leitz western.edu 02/02/2005 03:07 PM Hi Everyone, I was wondering if anyone has any idea or uses the special microtome for hard tissue with implants called Leitz 1600. Thanks, Nena Dimaano, MT/HT(ASCP) Advanced Technology Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From bernaweston <@t> hotmail.com Thu Feb 3 08:24:35 2005 From: bernaweston <@t> hotmail.com (Bernadette Weston) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] paraffin Message-ID: Our batch of paraffin this week seems to be bluer than usual, we haven't had any problems yet. Bernadette Weston HT Barberton Citizens Hopital Barberton, OH From histo <@t> kvl.dk Thu Feb 3 08:36:23 2005 From: histo <@t> kvl.dk (=?ISO-8859-1?Q?F=E6lles=20IVP?=) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] IHC detection of Yersinia pseudotuberculosis in formalin-fixed, paraffin-embedded tissue. Message-ID: Hi... Does anyone know a antibody for Yersinia pseudotuberculosis or a polyclonal antibody for all Yersinia bacteria?? I would like to make an IHC detection of Yersinia pseudotuberculosis in formalin-fixed, paraffin-embedded tissue. Thanks. Lene Jensen From mcauliff <@t> umdnj.edu Thu Feb 3 12:11:25 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Alcian Blue problem In-Reply-To: <6CD94D97ED7D924BA5C2B588FA952821396780@nt_exchange.lmhealth.org> References: <6CD94D97ED7D924BA5C2B588FA952821396780@nt_exchange.lmhealth.org> Message-ID: <4202694D.8080301@umdnj.edu> Many years ago I noticed that Alcian Blue stained/coated the whole slide to a slight extent. I seem to remember a short paper in Stain Technology, the former name of Biotechnique and Histochemistry, that described Alcian Blue as a coating for slides that helped attach sections. Geoff Tom McNemar wrote: >We have started to see a hazy/cloudiness to our alcian blue PAS slides. We >don't see this on any other slide/stain. We use the same slides, reagents, >and procedure that we have always used. The stain works fine and the haze >is on the entire slide, not just the tissue. We routinely use the >Fisherbrand red control slides and have tried other slides and still have >the same problem. I use the Poly Scientific reagents (alcian blue ph 2.5) >and have switched them all out for new reagents but still have the haze. >We have switched out, altered, and tried everything I can think of. I am >totally baffled. Hope you folks have some ideas. > >Tom Mc Nemar HT(ASCP) >Histology Supervisor >Licking Memorial Hospital >Newark, Ohio 43055 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > -- -- ********************************************** 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 PDavala <@t> sjh-nh.org Thu Feb 3 09:25:46 2005 From: PDavala <@t> sjh-nh.org (Davala, Pam) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Dissolving fat around lymphnodes of breast tissue Message-ID: Does some one out there have a specialize solution for dissolving fat from around the lymphnodes of breast or bowel tissue..... Our pathologists seem to have a hard time taking the fat off of the nodes and it makes it harder for the tech to cut due to unfixed fat... anyone's expertise would be appreciate... Is there a fixative out there from a company or is there one that we can make.... thank you Pam Davala HT(ASCP) Supervisor of Histology From JNocito <@t> Pathreflab.com Thu Feb 3 09:37:12 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Receiving Cytology Specimens in Glass Containers Message-ID: Morning Histonetters, we have come across a little problem. We are performing some work from local small hospitals. Our concern is that we are receiving many large volumes of cytology fluids in glass containers. I checked the JCAHO website and could not find anything relating to transporting specimens in glass containers. Does anyone know any regulations concerning transporting specimens in glass containers? Once again, I thank you for your efforts and vast knowledge. Joe Nocito Histology Manager Pathology Reference Lab San Antonio, TX From Kemlo.Rogerson <@t> elht.nhs.uk Thu Feb 3 09:40:59 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Dissolving fat around lymphnodes of breast tissue[ Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF1C@bhrv-nt-11.bhrv.nwest.nhs.uk> Um.... xylene? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Davala, Pam [mailto:PDavala@sjh-nh.org] Sent: 03 February 2005 15:26 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dissolving fat around lymphnodes of breast tissue[Scanned] Does some one out there have a specialize solution for dissolving fat from around the lymphnodes of breast or bowel tissue..... Our pathologists seem to have a hard time taking the fat off of the nodes and it makes it harder for the tech to cut due to unfixed fat... anyone's expertise would be appreciate... Is there a fixative out there from a company or is there one that we can make.... thank you Pam Davala HT(ASCP) Supervisor of Histology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rocan <@t> mac.com Thu Feb 3 09:41:03 2005 From: rocan <@t> mac.com (Rocan) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Dissolving fat around lymphnodes of breast tissue In-Reply-To: References: Message-ID: <3d83f091112a70bc487b02766b8d2124@mac.com> We have used cold acetone (-20oC) for about 1 hour. We did this for small fragments of fat tissue. This could work as you would delipidate the fat and firm the tissue. I am not sure if this is acceptable for a clinical sample. On Feb 3, 2005, at 7:25 AM, Davala, Pam wrote: > Does some one out there have a specialize solution for dissolving fat > from > around the lymphnodes of breast or bowel tissue..... Our pathologists > seem > to have a hard time taking the fat off of the nodes and it makes it > harder > for the tech to cut due to unfixed fat... anyone's expertise would be > appreciate... Is there a fixative out there from a company or is there > one > that we can make.... thank you Pam Davala HT(ASCP) Supervisor of > Histology > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MICHAEL.OWEN <@t> fda.gov Thu Feb 3 09:46:40 2005 From: MICHAEL.OWEN <@t> fda.gov (Owen, Michael P) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] IHC detection of Yersinia pseudotuberculosis in fo rmalin-fixed, paraffin-embedded tissue. Message-ID: Dear Dr. Jensen, The Yersinia expert in my laboratory (and the rest of FDA), Steve Weagent, would probably know the answer to your question. Steve Weagent Research Microbiologist FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021 Phone: 425-483-4873 E-Mail: steve.weagent@fda.gov Steve is one of the authors of the Yersinia chapter in the FDA CFSAN Bacteriological Analytical Manual. FDA CFSAN Bacteriological Analytical Manual (BAM) Online Chapter 8: Yersinia enterocolitica and Yersinia pseudotuberculosis http://www.cfsan.fda.gov/~ebam/bam-toc.html http://www.cfsan.fda.gov/~ebam/bam-8.html Wishing you success in your project, Michael P. Owen From: F?lles IVP [mailto:histo@kvl.dk] Sent: Thursday, February 03, 2005 6:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC detection of Yersinia pseudotuberculosis in formalin-fixed, paraffin-embedded tissue. Hi... Does anyone know a antibody for Yersinia pseudotuberculosis or a polyclonal antibody for all Yersinia bacteria?? I would like to make an IHC detection of Yersinia pseudotuberculosis in formalin-fixed, paraffin-embedded tissue. Thanks. Lene Jensen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Michael P. Owen, Regulatory Microbiologist FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From ian.montgomery <@t> bio.gla.ac.uk Thu Feb 3 11:12:50 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Paraffin wax. Message-ID: <6.2.1.2.2.20050203171013.0300ca00@udcf.gla.ac.uk> After several years my stock of paraffin wax needs replacing. Currently, in the UK, who sells good reliable wax. Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk From emerald_lake77 <@t> yahoo.com Thu Feb 3 11:20:50 2005 From: emerald_lake77 <@t> yahoo.com (- -) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Rubber cement (for in situ) Message-ID: <20050203172050.41652.qmail@web42301.mail.yahoo.com> Hello, I am starting to work up a protocol for in situ hybridization and discovered I need to seal my slides because of the high temps and long incubation. Can anyone help me with the following questions: a. I noticed the rubber cement is quite fluid. Using the brush that it comes with the can (or another method), how does one go about applying it after your sample is coverslipped? b. I've heard about fumes ... what concerns should I have; safety precautions should I take? (my normal range of temperature will be between 43 and 55 degrees Celcius within a Boekel Slide Moat or all purpose incubator) c. Are there better methods to use when sealing slides? d. What is the best way to remove the rubber cement/coverslip after performing hybridization? THANK YOU ALL VERY MUCH!!!!! --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. From TJJ <@t> Stowers-Institute.org Thu Feb 3 11:37:47 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Nomenclature: HT equivalent in France Message-ID: For those histologists working in France (and other European countries), what is the nomenclature used for your position? In the UK we are biomedical scientists? And what are the education requirements for such? Any info (or links to helpful websites) is appreciated. Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From brett_connolly <@t> merck.com Thu Feb 3 11:46:24 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Rubber cement (for in situ) Message-ID: Don't use the brush. Use 22 mm wide coverslips instead of 24 mm. With a plastic transfer pipet lay a thin line of rubber cement at the junction of the coverslip and the slide. After hybridization the solidified rubber cement easily peels away. Actually, your temps are relatively low for ISH, we've hybridized at >65C. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of - - Sent: Thursday, February 03, 2005 12:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Rubber cement (for in situ) Hello, I am starting to work up a protocol for in situ hybridization and discovered I need to seal my slides because of the high temps and long incubation. Can anyone help me with the following questions: a. I noticed the rubber cement is quite fluid. Using the brush that it comes with the can (or another method), how does one go about applying it after your sample is coverslipped? b. I've heard about fumes ... what concerns should I have; safety precautions should I take? (my normal range of temperature will be between 43 and 55 degrees Celcius within a Boekel Slide Moat or all purpose incubator) c. Are there better methods to use when sealing slides? d. What is the best way to remove the rubber cement/coverslip after performing hybridization? THANK YOU ALL VERY MUCH!!!!! --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. _______________________________________________ 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 liz <@t> premierlab.com Thu Feb 3 11:52:09 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Rubber cement (for in situ) In-Reply-To: <20050203172050.41652.qmail@web42301.mail.yahoo.com> Message-ID: <000001c50a19$16180420$76d48a80@AMY> We use Frame-Seal from MJ Research, Inc. www.mjr.com (888)735-8437 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 - - Sent: Thursday, February 03, 2005 10:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Rubber cement (for in situ) Hello, I am starting to work up a protocol for in situ hybridization and discovered I need to seal my slides because of the high temps and long incubation. Can anyone help me with the following questions: a. I noticed the rubber cement is quite fluid. Using the brush that it comes with the can (or another method), how does one go about applying it after your sample is coverslipped? b. I've heard about fumes ... what concerns should I have; safety precautions should I take? (my normal range of temperature will be between 43 and 55 degrees Celcius within a Boekel Slide Moat or all purpose incubator) c. Are there better methods to use when sealing slides? d. What is the best way to remove the rubber cement/coverslip after performing hybridization? THANK YOU ALL VERY MUCH!!!!! --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MElliott <@t> mrl.ubc.ca Thu Feb 3 11:59:45 2005 From: MElliott <@t> mrl.ubc.ca (Mark Elliott) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] CX3-CR1 antibody Message-ID: Does anyone have any experience with the polyclonal antibody to CX3-CR1 from Abcam (ab7200) on formalin-fixed paraffin-embedded sections? The data sheet says it can be used for this type of tissue but doesn't say if retrieval of some sort is required. Is anybody using a different antibody? Any input is greatly appreciated. W. Mark Elliott, PhD James Hogg iCAPTURE Centre Vancouver BC Canada From cthouron <@t> ochsner.org Thu Feb 3 12:18:35 2005 From: cthouron <@t> ochsner.org (Carol Thouron) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Cryostat Recomendations Message-ID: Our lab is in the market for a new cryostat. We are presently looking at the UltraPro 5000 and the Leica CM 1900. Budget allowance is $25,000.00. We are particularly interested in a model that has the handwheel located close to the front of the unit, as the reach on our old cryostat is way too far. Any input will be appreciated. Carol Thouron Ochsner Clinic Foundation New Orleans, LA From jkiernan <@t> uwo.ca Thu Feb 3 12:27:42 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Alcian Blue problem References: <6CD94D97ED7D924BA5C2B588FA952821396780@nt_exchange.lmhealth.org> <4202694D.8080301@umdnj.edu> Message-ID: <42026D1E.DBB3E5A6@uwo.ca> It also stains ceramic sinks pretty well! The dye cations are supposed to stick to the silicate anions of the glass or pot. The slide adhesive paper is: Archimbaud E, Islam A, Preisler HD (1986) Alcian blue method for attaching glycol methacrylate sections to glass slides. Stain Technology 61: 121-123. To obtain a positively charged surface this way you'd have to stain the slides immediately before using, and wash in something a bit acidic. Anything as basic as water removes the pendant cationic groups from bound alcian blue, leaving behind uncharged and completely insoluble copper phthalocyanine. I don't have that issue of Stain Technology to hand, so cannot check up on the method. -- ------------------------------- 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 _______________________________ Geoff McAuliffe wrote: > > Many years ago I noticed that Alcian Blue stained/coated the whole > slide to a slight extent. I seem to remember a short paper in Stain > Technology, the former name of Biotechnique and Histochemistry, that > described Alcian Blue as a coating for slides that helped attach sections. > > Geoff > > Tom McNemar wrote: > > >We have started to see a hazy/cloudiness to our alcian blue PAS slides. We > >don't see this on any other slide/stain. We use the same slides, reagents, > >and procedure that we have always used. The stain works fine and the haze > >is on the entire slide, not just the tissue. We routinely use the > >Fisherbrand red control slides and have tried other slides and still have > >the same problem. I use the Poly Scientific reagents (alcian blue ph 2.5) > >and have switched them all out for new reagents but still have the haze. > >We have switched out, altered, and tried everything I can think of. I am > >totally baffled. Hope you folks have some ideas. > > > >Tom Mc Nemar HT(ASCP) > >Histology Supervisor > >Licking Memorial Hospital > >Newark, Ohio 43055 > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > > -- > -- > ********************************************** > Geoff McAuliffe, Ph.D. > Neuroscience and Cell Biology > Robert Wood Johnson Medical School > 675 Hoes Lane, Piscataway, NJ 08854 > voice: (732)-235-4583; fax: -4029 > mcauliff@umdnj.edu > ********************************************** > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BlazekL <@t> childrensdayton.org Thu Feb 3 12:48:13 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] 37% formaldehyde Message-ID: Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org From swaram <@t> myrealbox.com Thu Feb 3 12:59:32 2005 From: swaram <@t> myrealbox.com (Swaram) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Same Host Ab(fluorescence IHC) HELP...!!! In-Reply-To: References: Message-ID: <42027494.3010308@myrealbox.com> Dear Histonetters, Here is the problem that is giving me a lot of grey hairs even though I am only 26. Its making me crazy and I am in deep despair. I cant go on like this.. Please help..! :'( I have to do Fluorescent based IHC on two proteins on old paraffin fixed skin tissue. The first antibody telomerase is from Mouse. I use goat antimouse Fab2 fragments of Alexa488 for the secondary detection. The second Ab that I have to stain is melanoma cells and I use Pan Melanoma cocktail from Biocare Medicals for this purpose. This is again from Mouse and has several IgG subtypes like IgG1K, IgG2a, IgG2b etc. I use Goat antimouse TRITC as my secondary for this step. Before the application of my second Primary(Pan Melanoma), I use Mouse (H+L) and incubate for 30 minutes, and then add (Mouse Fab(H+L) for blocking any epitopes on the first primary Ab (telomerase) so that my second secondary (TRITC) will not bind to that. However, I have been far from successful. I have run out of ideas. There are no good Ab for using against Melanoma (especially basal melanocytes) from any other hosts that work well. Here is a scheme that I use from Jacksonlab website. [1]http://www.jacksonimmuno.com/technical/examplec.asp Please help... ! Thank you all.. SWARAM References 1. http://www.jacksonimmuno.com/technical/examplec.asp From swaram <@t> myrealbox.com Thu Feb 3 12:59:32 2005 From: swaram <@t> myrealbox.com (Swaram) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Same Host Ab(fluorescence IHC) HELP...!!! In-Reply-To: References: Message-ID: <42027494.3010308@myrealbox.com> Dear Histonetters, Here is the problem that is giving me a lot of grey hairs even though I am only 26. Its making me crazy and I am in deep despair. I cant go on like this.. Please help..! :'( I have to do Fluorescent based IHC on two proteins on old paraffin fixed skin tissue. The first antibody telomerase is from Mouse. I use goat antimouse Fab2 fragments of Alexa488 for the secondary detection. The second Ab that I have to stain is melanoma cells and I use Pan Melanoma cocktail from Biocare Medicals for this purpose. This is again from Mouse and has several IgG subtypes like IgG1K, IgG2a, IgG2b etc. I use Goat antimouse TRITC as my secondary for this step. Before the application of my second Primary(Pan Melanoma), I use Mouse (H+L) and incubate for 30 minutes, and then add (Mouse Fab(H+L) for blocking any epitopes on the first primary Ab (telomerase) so that my second secondary (TRITC) will not bind to that. However, I have been far from successful. I have run out of ideas. There are no good Ab for using against Melanoma (especially basal melanocytes) from any other hosts that work well. Here is a scheme that I use from Jacksonlab website. [1]http://www.jacksonimmuno.com/technical/examplec.asp Please help... ! Thank you all.. SWARAM References 1. http://www.jacksonimmuno.com/technical/examplec.asp From pruegg <@t> ihctech.net Thu Feb 3 13:14:57 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Rubber cement (for in situ) In-Reply-To: <000001c50a19$16180420$76d48a80@AMY> Message-ID: <200502031914.j13JEpG7016996@chip.viawest.net> I have used just a glass coverslip laying on over the hybridizing reagent without sealing it at all, then the glass can be floated off in buffer. I hope I am understanding this inquiry. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Thursday, February 03, 2005 10:52 AM To: '- -'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Rubber cement (for in situ) We use Frame-Seal from MJ Research, Inc. www.mjr.com (888)735-8437 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 - - Sent: Thursday, February 03, 2005 10:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Rubber cement (for in situ) Hello, I am starting to work up a protocol for in situ hybridization and discovered I need to seal my slides because of the high temps and long incubation. Can anyone help me with the following questions: a. I noticed the rubber cement is quite fluid. Using the brush that it comes with the can (or another method), how does one go about applying it after your sample is coverslipped? b. I've heard about fumes ... what concerns should I have; safety precautions should I take? (my normal range of temperature will be between 43 and 55 degrees Celcius within a Boekel Slide Moat or all purpose incubator) c. Are there better methods to use when sealing slides? d. What is the best way to remove the rubber cement/coverslip after performing hybridization? THANK YOU ALL VERY MUCH!!!!! --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. _______________________________________________ Histonet mailing list Histonet@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 scoop <@t> mail.nih.gov Thu Feb 3 13:21:01 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Same Host Ab(fluorescence IHC) HELP...!!! In-Reply-To: <42027494.3010308@myrealbox.com> References: <42027494.3010308@myrealbox.com> Message-ID: Hi, I have had the same problem in the past with blocking with Fab fragment. My solution to the problem was to label the primary antibody and use a secondary antibody directed against that label. Suggestions for labels for the primary antibody are biotin (kit from Vector or Molecular Probes - you could use a fluor conjugated Streptavidin as your secondary) or digoxigenin (kit from Roche). You could also just try to label the primary with a fluor. I did this once with a kit from molecular probes that worked well. Another possibility suggested to me by someone was to incubate the primary and secondary antibodies together in a tube and then put that complex on the tissue. You would need to work out the concentration of each. I never tried this because it was quicker for me to label the primary Abs. Sharon >Dear Histonetters, > Here is the problem that is giving me a lot of grey hairs even though > I am only 26. Its making me crazy and I am in deep despair. I cant go > on like this.. Please help..! :'( > I have to do Fluorescent based IHC on two proteins on old paraffin > fixed skin tissue. The first antibody telomerase is from Mouse. I use > goat antimouse Fab2 fragments of Alexa488 for the secondary detection. > The second Ab that I have to stain is melanoma cells and I use Pan > Melanoma cocktail from Biocare Medicals for this purpose. This is > again from Mouse and has several IgG subtypes like IgG1K, IgG2a, IgG2b > etc. I use Goat antimouse TRITC as my secondary for this step. Before > the application of my second Primary(Pan Melanoma), I use Mouse (H+L) > and incubate for 30 minutes, and then add (Mouse Fab(H+L) for blocking > any epitopes on the first primary Ab (telomerase) so that my second > secondary (TRITC) will not bind to that. However, I have been far from > successful. I have run out of ideas. There are no good Ab for using > against Melanoma (especially basal melanocytes) from any other hosts > that work well. > Here is a scheme that I use from Jacksonlab website. > [1]http://www.jacksonimmuno.com/technical/examplec.asp > Please help... ! > Thank you all.. > SWARAM > >References > > 1. http://www.jacksonimmuno.com/technical/examplec.asp >_______________________________________________ >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 From pruegg <@t> ihctech.net Thu Feb 3 13:24:09 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Receiving Cytology Specimens in Glass Containers In-Reply-To: Message-ID: <200502031924.j13JO4G7020415@chip.viawest.net> There are new rules for transporting biological samples for 2005, this is handled by the Department of Transportation, I am told you can do a web search for dot.gov and find the regs. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, February 03, 2005 8:37 AM To: Histonet Subject: [Histonet] Receiving Cytology Specimens in Glass Containers Morning Histonetters, we have come across a little problem. We are performing some work from local small hospitals. Our concern is that we are receiving many large volumes of cytology fluids in glass containers. I checked the JCAHO website and could not find anything relating to transporting specimens in glass containers. Does anyone know any regulations concerning transporting specimens in glass containers? Once again, I thank you for your efforts and vast knowledge. Joe Nocito Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Thu Feb 3 16:24:16 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] 37% formaldehyde In-Reply-To: References: Message-ID: <4202A490.8080200@umdnj.edu> The surgeon probably does not know that formaldehyde is diluted. Geoff Linda Blazek wrote: >Does anyone know why a surgeon would want to send a liver biopsy >specimen in 37% formaldehyde? Is there some test or procedure that you >would ever want to put a specimen in 37%? >Thanks > > >Linda Blazek, HT (ASCP) >Department of Pathology >Children's Medical Center >Dayton, Ohio 45404 >(937) 641-3358 >fax (937)641-5482 >blazekl@childrensdayton.org > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > -- -- ********************************************** 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 brett_connolly <@t> merck.com Thu Feb 3 13:25:40 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Same Host Ab(fluorescence IHC) HELP...!!! Message-ID: Swaram, What results are you getting? Assuming that you have worked out each antibody individually first on your tissue why not try Zenon (Molecular Probes) primary antibody labeling using different Alexafluors? You can skip all those blocking headaches. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Swaram Sent: Thursday, February 03, 2005 2:00 PM To: HISTONET Cc: histonet@pathology.swmed.edu Subject: [Histonet] Same Host Ab(fluorescence IHC) HELP...!!! Dear Histonetters, Here is the problem that is giving me a lot of grey hairs even though I am only 26. Its making me crazy and I am in deep despair. I cant go on like this.. Please help..! :'( I have to do Fluorescent based IHC on two proteins on old paraffin fixed skin tissue. The first antibody telomerase is from Mouse. I use goat antimouse Fab2 fragments of Alexa488 for the secondary detection. The second Ab that I have to stain is melanoma cells and I use Pan Melanoma cocktail from Biocare Medicals for this purpose. This is again from Mouse and has several IgG subtypes like IgG1K, IgG2a, IgG2b etc. I use Goat antimouse TRITC as my secondary for this step. Before the application of my second Primary(Pan Melanoma), I use Mouse (H+L) and incubate for 30 minutes, and then add (Mouse Fab(H+L) for blocking any epitopes on the first primary Ab (telomerase) so that my second secondary (TRITC) will not bind to that. However, I have been far from successful. I have run out of ideas. There are no good Ab for using against Melanoma (especially basal melanocytes) from any other hosts that work well. Here is a scheme that I use from Jacksonlab website. [1]http://www.jacksonimmuno.com/technical/examplec.asp Please help... ! Thank you all.. SWARAM References 1. http://www.jacksonimmuno.com/technical/examplec.asp _______________________________________________ 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 Rachael_Emerson <@t> URMC.Rochester.edu Thu Feb 3 13:31:51 2005 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] avian/biotin block Message-ID: Let me try to sum up a long story and get to my question...... Basically I am staining mouse embryos-frozen sections, for megakaryocytes. I had trouble with endogenous AP and 15% acetic acid seemed to help. I am having trouble with endogenous biotin, as well. I tried Vector's Avidin/Biotin Blocking Kit and lost my signal all together. I am looking for a surface antigen and I am afraid the kit blocked it all. I would really appreciate any suggestions on blocking endogenous biotin on frozen sections? Also, I tried to us an AP secondary, but my signal was dimished. Thanks Rachael Rachael L. Emerson Center for Human Genetics and Molecular Pediatric Diseases University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 Tel (585) 275-5073 Fax (585) 276-0232 From settembr <@t> umdnj.edu Thu Feb 3 13:28:07 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] 37% formaldehyde Message-ID: Linda, It is my understanding that 37% formaldehyde is Formalin? We put livers in formalin every day and produce most of our stains with that fixative. Dana Settembre Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ >>> Linda Blazek 2/3/2005 1:48:13 PM >>> Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Thu Feb 3 13:35:57 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] Receiving Cytology Specimens in Glass Containers In-Reply-To: Message-ID: see, this is why I love you all. People responded to and told me to contact the US Department of Transportation. Now I feel like "Duh?", but hey, it's no different than any other day. Thanks netters. Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Thursday, February 03, 2005 9:37 AM To: Histonet Subject: [Histonet] Receiving Cytology Specimens in Glass Containers Morning Histonetters, we have come across a little problem. We are performing some work from local small hospitals. Our concern is that we are receiving many large volumes of cytology fluids in glass containers. I checked the JCAHO website and could not find anything relating to transporting specimens in glass containers. Does anyone know any regulations concerning transporting specimens in glass containers? Once again, I thank you for your efforts and vast knowledge. Joe Nocito Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Feb 3 13:39:49 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:34 2005 Subject: [Histonet] avian/biotin block In-Reply-To: Message-ID: <200502031939.j13JdiG7025839@chip.viawest.net> I would consider using a non-biotin detection system such as Hrp Labelled Polymer, there are several out there now, the one I use is from DAKO called Envision. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Emerson, Rachael Sent: Thursday, February 03, 2005 12:32 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] avian/biotin block Let me try to sum up a long story and get to my question...... Basically I am staining mouse embryos-frozen sections, for megakaryocytes. I had trouble with endogenous AP and 15% acetic acid seemed to help. I am having trouble with endogenous biotin, as well. I tried Vector's Avidin/Biotin Blocking Kit and lost my signal all together. I am looking for a surface antigen and I am afraid the kit blocked it all. I would really appreciate any suggestions on blocking endogenous biotin on frozen sections? Also, I tried to us an AP secondary, but my signal was dimished. Thanks Rachael Rachael L. Emerson Center for Human Genetics and Molecular Pediatric Diseases University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 Tel (585) 275-5073 Fax (585) 276-0232 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Thu Feb 3 13:40:22 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: I think Geoff hit the nail on the head. The surgeon doesn't realize or remember that 'formalin' is supposed to be 3.7%. I feel bad for the patient. Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] 37% formaldehyde Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ 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 bmcmahill <@t> incytepathology.com Thu Feb 3 13:29:15 2005 From: bmcmahill <@t> incytepathology.com (Bonnie J. McMahill) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Rubber cement (for in situ) Message-ID: We use the smaller coverslips (we have a large supply of 18 mm round ones that work really well), and apply the rubber cement around the edge with a 5 ml syringe. It works very well and comes off easily after hybridization with a tweezer. Bonnie McMahill Histology Supervisor InCyte Pathology Spokane, WA > -----Original Message----- > From: Connolly, Brett M [SMTP:brett_connolly@merck.com] > Sent: Thursday, February 03, 2005 9:46 AM > To: '- -'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Rubber cement (for in situ) > > Don't use the brush. Use 22 mm wide coverslips instead of 24 mm. With a > plastic transfer pipet lay a thin line of rubber cement at the junction of > the coverslip and the slide. After hybridization the solidified rubber > cement easily peels away. Actually, your temps are relatively low for ISH, > we've hybridized at >65C. > > Brett > > Brett M. Connolly, Ph.D. > Merck & Co., Inc. > MRL, Imaging Research > WP26A-3000 > PO Box 4 > West Point, PA 19486 > PH 215-652-2501 > fax. 215-652-2075 > e-mail. brett_connolly@merck.com > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of - - > Sent: Thursday, February 03, 2005 12:21 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Rubber cement (for in situ) > > > Hello, > > I am starting to work up a protocol for in situ hybridization and discovered > I need to seal my slides because of the high temps and long incubation. Can > anyone help me with the following questions: > > a. I noticed the rubber cement is quite fluid. Using the brush that it > comes with the can (or another method), how does one go about applying it > after your sample is coverslipped? > > b. I've heard about fumes ... what concerns should I have; safety > precautions should I take? (my normal range of temperature will be between > 43 and 55 degrees Celcius within a Boekel Slide Moat or all purpose > incubator) > > c. Are there better methods to use when sealing slides? > > d. What is the best way to remove the rubber cement/coverslip after > performing hybridization? > > THANK YOU ALL VERY MUCH!!!!! > > > --------------------------------- > Do you Yahoo!? > Yahoo! Mail - 250MB free storage. Do more. Manage less. > _______________________________________________ > 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. > ------------------------------------------------------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SHargrove <@t> urhcs.org Thu Feb 3 13:46:28 2005 From: SHargrove <@t> urhcs.org (SHargrove@urhcs.org) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Dissolving fat around lymphnodes of breast tissue Message-ID: Carnoys solution will not dissolve the fat but will highlight the nodes. Strip the specimen first though. Susie Hargrove Histology United Regional Health Care Systems 940-764-3198 The documents inside this electronic transmission contain confidential information belonging to the sender that is legally privileged. This information is intended only for the use of the individual or entity named above. The authorized recipient of this information is prohibited from disclosing this information to any other party and is required to destroy the information after its stated need has been fulfilled, unless otherwise required by law. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or action taken in reliance on the contents of these documents is strictly prohibited. If you received this electronic transmission in error, please notify the sender immediately to arrange for return. From gu.lang <@t> gmx.at Thu Feb 3 13:48:03 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:35 2005 Subject: AW: [Histonet] Nomenclature: HT equivalent in France In-Reply-To: <20050203173839.5528gmx1@mx067.gmx.net> Message-ID: In Austria we are called "medizinisch-technische Analytiker" (medical-technical Analysist). The course lasts three years and you can work in every medical laboratory. There is no special education for histotechnology. It is compareable with "a bachelor-degree", but it is not accredited as a university-course. If someone wants to study medicine after, you have to start from the beginning. In near future our name will change in something like "biomedical analysist", and the course should be upgraded and taught in academies. Hope this helps Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Johnson, Teri Gesendet: Donnerstag, 03. Februar 2005 18:38 An: Histonet Betreff: [Histonet] Nomenclature: HT equivalent in France For those histologists working in France (and other European countries), what is the nomenclature used for your position? In the UK we are biomedical scientists? And what are the education requirements for such? Any info (or links to helpful websites) is appreciated. Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Thu Feb 3 13:53:44 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] avian/biotin block Message-ID: Rachael, I've switched to the polymer-based secondaries (DAKO Envision, Vector Impress, etc.)....biotin-free systems. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Emerson, Rachael Sent: Thursday, February 03, 2005 2:32 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] avian/biotin block Let me try to sum up a long story and get to my question...... Basically I am staining mouse embryos-frozen sections, for megakaryocytes. I had trouble with endogenous AP and 15% acetic acid seemed to help. I am having trouble with endogenous biotin, as well. I tried Vector's Avidin/Biotin Blocking Kit and lost my signal all together. I am looking for a surface antigen and I am afraid the kit blocked it all. I would really appreciate any suggestions on blocking endogenous biotin on frozen sections? Also, I tried to us an AP secondary, but my signal was dimished. Thanks Rachael Rachael L. Emerson Center for Human Genetics and Molecular Pediatric Diseases University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 Tel (585) 275-5073 Fax (585) 276-0232 _______________________________________________ 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 BlazekL <@t> childrensdayton.org Thu Feb 3 13:53:05 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: There isn't any mistake about what he wanted. One of our techs was here late last night because one of our more "cranky" surgeons was doing a procedure in OR. He called to the lab for "formaldehyde". He was sent formalin. He preceded to yell (very loudly) that he did not want formalin he wanted "formaldehyde". Linda >>> "Connolly, Brett M" 2/3/2005 2:40:22 PM >>> I think Geoff hit the nail on the head. The surgeon doesn't realize or remember that 'formalin' is supposed to be 3.7%. I feel bad for the patient. Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] 37% formaldehyde Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ 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. ------------------------------------------------------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RossS <@t> BaylorHealth.edu Thu Feb 3 14:04:45 2005 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: <17A8DA5A05167C4EA20BFA750167736B0BB6E2@BHDAEXCH13.bhcs.pvt> So he wanted a pure formaldehyde gas? I wish you had some around to give him. :) Ah the confusion between formaldehyde, formalin, and 10% NBF. It can make for some great "Who's on first" type moments. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:53 PM To: brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde There isn't any mistake about what he wanted. One of our techs was here late last night because one of our more "cranky" surgeons was doing a procedure in OR. He called to the lab for "formaldehyde". He was sent formalin. He preceded to yell (very loudly) that he did not want formalin he wanted "formaldehyde". Linda >>> "Connolly, Brett M" 2/3/2005 2:40:22 PM >>> I think Geoff hit the nail on the head. The surgeon doesn't realize or remember that 'formalin' is supposed to be 3.7%. I feel bad for the patient. Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] 37% formaldehyde Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ 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. ------------------------------------------------------------------------ ------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From BlazekL <@t> childrensdayton.org Thu Feb 3 14:07:30 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: It's probably a good thing I wasn't here to deal with it last night. Sometimes my mouth can get me in trouble. I would have loved to give him the gas. >>> "Stapf, Ross" 02/03/2005 3:04:45 PM >>> So he wanted a pure formaldehyde gas? I wish you had some around to give him. :) Ah the confusion between formaldehyde, formalin, and 10% NBF. It can make for some great "Who's on first" type moments. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:53 PM To: brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde There isn't any mistake about what he wanted. One of our techs was here late last night because one of our more "cranky" surgeons was doing a procedure in OR. He called to the lab for "formaldehyde". He was sent formalin. He preceded to yell (very loudly) that he did not want formalin he wanted "formaldehyde". Linda >>> "Connolly, Brett M" 2/3/2005 2:40:22 PM >>> I think Geoff hit the nail on the head. The surgeon doesn't realize or remember that 'formalin' is supposed to be 3.7%. I feel bad for the patient. Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] 37% formaldehyde Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ 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. ------------------------------------------------------------------------ ------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From mucram11 <@t> comcast.net Thu Feb 3 14:17:44 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: <020320052017.7261.420286E7000D234C00001C5D2205886360CECE030E9D0C9A03@comcast.net> Linda, WOW!! Can you ask him if he had a special reason for requiring the strongest formaldehyde he could get? Was it a rare infectious case? Should we ask how the tissue sectioned or just be glad we did not have to section it today? Pam Marcum -------------- Original message -------------- > There isn't any mistake about what he wanted. One of our techs was here > late last night because one of our more "cranky" surgeons was doing a > procedure in OR. He called to the lab for "formaldehyde". He was sent > formalin. He preceded to yell (very loudly) that he did not want > formalin he wanted "formaldehyde". > Linda > > >>> "Connolly, Brett M" 2/3/2005 2:40:22 PM > >>> > I think Geoff hit the nail on the head. The surgeon doesn't realize or > remember that 'formalin' is supposed to be 3.7%. I feel bad for the > patient. > > Brett > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda > Blazek > Sent: Thursday, February 03, 2005 1:48 PM > To: histonet@pathology.swmed.edu > Subject: [Histonet] 37% formaldehyde > > > Does anyone know why a surgeon would want to send a liver biopsy > specimen in 37% formaldehyde? Is there some test or procedure that > you > would ever want to put a specimen in 37%? > Thanks > > > Linda Blazek, HT (ASCP) > Department of Pathology > Children's Medical Center > Dayton, Ohio 45404 > (937) 641-3358 > fax (937)641-5482 > blazekl@childrensdayton.org > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------------------------------------------------------ > 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. > ------------------------------------------------------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@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 brett_connolly <@t> merck.com Thu Feb 3 14:22:45 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: So let us know what tests he wants that need the unbuffered 37%. Thanks, Brett -----Original Message----- From: Linda Blazek [mailto:BlazekL@childrensdayton.org] Sent: Thursday, February 03, 2005 3:08 PM To: RossS@BaylorHealth.edu; brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde It's probably a good thing I wasn't here to deal with it last night. Sometimes my mouth can get me in trouble. I would have loved to give him the gas. >>> "Stapf, Ross" 02/03/2005 3:04:45 PM >>> So he wanted a pure formaldehyde gas? I wish you had some around to give him. :) Ah the confusion between formaldehyde, formalin, and 10% NBF. It can make for some great "Who's on first" type moments. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:53 PM To: brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde There isn't any mistake about what he wanted. One of our techs was here late last night because one of our more "cranky" surgeons was doing a procedure in OR. He called to the lab for "formaldehyde". He was sent formalin. He preceded to yell (very loudly) that he did not want formalin he wanted "formaldehyde". Linda >>> "Connolly, Brett M" 2/3/2005 2:40:22 PM >>> I think Geoff hit the nail on the head. The surgeon doesn't realize or remember that 'formalin' is supposed to be 3.7%. I feel bad for the patient. Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] 37% formaldehyde Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ 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. ------------------------------------------------------------------------ ------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ------------------------------------------------------------------------------ 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 Lynne.Bell <@t> hitchcock.org Thu Feb 3 14:28:00 2005 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: As one of our pathologists always says, "You can always tell a surgeon - but you can't tell him much!" Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From timothy.m.coskran <@t> pfizer.com Thu Feb 3 14:32:10 2005 From: timothy.m.coskran <@t> pfizer.com (Coskran, Timothy M) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Same Host Ab flour problems Message-ID: <599089EF29304C44AF1D84B1C223AC6313EAF0@groamrexm03.amer.pfizer.com> If you know the Ig concentration of the telomerase mouse monoclonal, you could try to biotinylate it using the Dako ARK kit. Then follow this with streptavidin Alexa 488 from Molecular Probes. This technique was presented by Chris van der Loos at an NSH a while back. The ARK kit provides an easy way to biotinylate a mouse antibody so that you can base the detection of this antibody on biotin and then use an anti-mouse IgG secondary (HRP, fluor, etc) for the second mouse antibody. You may have to titrate the telomerase antibody. In my experience, you have to increase your primary antibody concentration when using the ARK kit. Good Luck Tim Coskran Pfizer LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From BlazekL <@t> childrensdayton.org Thu Feb 3 14:31:41 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: I will let you all know how the specimen turns out tomorrow. As an added note he also wanted glutaraldehyde which he placed very large chunks of liver tissue in. We retrieved several pieces of that and put it into 10% NBF. The patient specimen will be salvaged. The specimen that was in the 37% was changed to 10% NBF this morning. We will have results tomorrow. So far he has not asked for any other than the routine testing. Our pathologist has agreed to talk to him. But I doubt it will happen. Thanks for all the input. I was afraid that after 30 years in histology I was missing something basic. Linda >>> "Connolly, Brett M" 02/03/2005 3:22:45 PM >>> So let us know what tests he wants that need the unbuffered 37%. Thanks, Brett -----Original Message----- From: Linda Blazek [mailto:BlazekL@childrensdayton.org] Sent: Thursday, February 03, 2005 3:08 PM To: RossS@BaylorHealth.edu; brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde It's probably a good thing I wasn't here to deal with it last night. Sometimes my mouth can get me in trouble. I would have loved to give him the gas. >>> "Stapf, Ross" 02/03/2005 3:04:45 PM >>> So he wanted a pure formaldehyde gas? I wish you had some around to give him. :) Ah the confusion between formaldehyde, formalin, and 10% NBF. It can make for some great "Who's on first" type moments. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:53 PM To: brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde There isn't any mistake about what he wanted. One of our techs was here late last night because one of our more "cranky" surgeons was doing a procedure in OR. He called to the lab for "formaldehyde". He was sent formalin. He preceded to yell (very loudly) that he did not want formalin he wanted "formaldehyde". Linda >>> "Connolly, Brett M" 2/3/2005 2:40:22 PM >>> I think Geoff hit the nail on the head. The surgeon doesn't realize or remember that 'formalin' is supposed to be 3.7%. I feel bad for the patient. Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] 37% formaldehyde Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ 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. ------------------------------------------------------------------------ ------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ------------------------------------------------------------------------------ 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 mcauliff <@t> umdnj.edu Thu Feb 3 17:38:09 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde In-Reply-To: References: Message-ID: <4202B5E1.7050005@umdnj.edu> As I wrote, he does not understand that "formalin" is 3.7% formaldehyde and than nobody fixes tissue in 37% formaldehyde. I suggest getting the pathologist to explain it to him. Geoff Linda Blazek wrote: >There isn't any mistake about what he wanted. One of our techs was here >late last night because one of our more "cranky" surgeons was doing a >procedure in OR. He called to the lab for "formaldehyde". He was sent >formalin. He preceded to yell (very loudly) that he did not want >formalin he wanted "formaldehyde". >Linda > > > >>>>"Connolly, Brett M" 2/3/2005 2:40:22 PM >>>> >>>> >>>> >I think Geoff hit the nail on the head. The surgeon doesn't realize or >remember that 'formalin' is supposed to be 3.7%. I feel bad for the >patient. > >Brett > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda >Blazek >Sent: Thursday, February 03, 2005 1:48 PM >To: histonet@pathology.swmed.edu >Subject: [Histonet] 37% formaldehyde > > >Does anyone know why a surgeon would want to send a liver biopsy >specimen in 37% formaldehyde? Is there some test or procedure that >you >would ever want to put a specimen in 37%? >Thanks > > >Linda Blazek, HT (ASCP) >Department of Pathology >Children's Medical Center >Dayton, Ohio 45404 >(937) 641-3358 >fax (937)641-5482 >blazekl@childrensdayton.org > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >------------------------------------------------------------------------------ >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. >------------------------------------------------------------------------------ > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From swaram <@t> myrealbox.com Thu Feb 3 14:49:05 2005 From: swaram <@t> myrealbox.com (Swaram) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Same Host Ab(fluorescence IHC) HELP...!!! In-Reply-To: References: Message-ID: <42028E41.8060205@myrealbox.com> Dear Histonetters, I did try the Alexa Zenon labelling kit for labelling my Second Secondary for MART-1 (from Upstate). But unfortunately, even at concentrations of 1:5 of the labelling complex, it hardly stained anything. I think, the stearic hinderance is too much for the complex to attach efficiently to epitopes unmasked after citrate buffer cooking on the paraffin fixed slides. I am now thinking, if reducing the labelling ratios of (1:3) {that Molecular Probes recommends} to 1:1 along with not adding the blocking reagent will help me in increasing the binding and overall signal intensity? Does anyone think this is tryable..? even logical? I mean if the labeled complx is too big with addition of several Fab fragments, its reasonable to assume that the complx cant bind to the epitope well. (I find the edge staining effect, which probably is the washed off Ab complx). So if I reduce the amount of Zenon labelling reagent (which is Fab fragments bound to Alexa dye) to almost 2-3 per primary Ab, then the complex has better access to the epitope and the maze of the paraffin world.. ? Thanks for the suggestions on direct labelling of the Ab to a fluorochrome, but I find that to do that.. most of the Abs have to be free from Glycine/NaCl buffer and unfortunately mfgrs dont sell them. Thanks to you who responded and sparked my interests in these solutions.. I wish I can get some more tips.. Regards, Swaram, UCSF From swaram <@t> myrealbox.com Thu Feb 3 14:49:05 2005 From: swaram <@t> myrealbox.com (Swaram) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Same Host Ab(fluorescence IHC) HELP...!!! In-Reply-To: References: Message-ID: <42028E41.8060205@myrealbox.com> Dear Histonetters, I did try the Alexa Zenon labelling kit for labelling my Second Secondary for MART-1 (from Upstate). But unfortunately, even at concentrations of 1:5 of the labelling complex, it hardly stained anything. I think, the stearic hinderance is too much for the complex to attach efficiently to epitopes unmasked after citrate buffer cooking on the paraffin fixed slides. I am now thinking, if reducing the labelling ratios of (1:3) {that Molecular Probes recommends} to 1:1 along with not adding the blocking reagent will help me in increasing the binding and overall signal intensity? Does anyone think this is tryable..? even logical? I mean if the labeled complx is too big with addition of several Fab fragments, its reasonable to assume that the complx cant bind to the epitope well. (I find the edge staining effect, which probably is the washed off Ab complx). So if I reduce the amount of Zenon labelling reagent (which is Fab fragments bound to Alexa dye) to almost 2-3 per primary Ab, then the complex has better access to the epitope and the maze of the paraffin world.. ? Thanks for the suggestions on direct labelling of the Ab to a fluorochrome, but I find that to do that.. most of the Abs have to be free from Glycine/NaCl buffer and unfortunately mfgrs dont sell them. Thanks to you who responded and sparked my interests in these solutions.. I wish I can get some more tips.. Regards, Swaram, UCSF From akbitting <@t> geisinger.edu Thu Feb 3 14:49:59 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Wheat germ agglutinin Message-ID: Does anyone have a good procedure for Cardiac Myocyte staining for cell size measurement on FFPE tissue? I have one that a research doc gave me, but it is very vague. Also, where can I buy DRAQ-5 and WGA-TRITC? 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 gcallis <@t> montana.edu Thu Feb 3 14:55:38 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:35 2005 Subject: amused as Re: [Histonet] 37% formaldehyde Message-ID: <6.0.0.22.1.20050203134031.01b57c08@gemini.msu.montana.edu> Linda and everyone, I have been rolling across the floor with laughter from replies. Maybe Linda should forward all these replies to him - but look out, Linda et al, the computer might come flying out his office door!!! Seriously though, and Brett said it - feel bad for that poor patient!! Thank folks! I appreciated your senses of humor. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From brett_connolly <@t> merck.com Thu Feb 3 13:25:40 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Same Host Ab(fluorescence IHC) HELP...!!! Message-ID: Swaram, What results are you getting? Assuming that you have worked out each antibody individually first on your tissue why not try Zenon (Molecular Probes) primary antibody labeling using different Alexafluors? You can skip all those blocking headaches. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Swaram Sent: Thursday, February 03, 2005 2:00 PM To: HISTONET Cc: histonet@pathology.swmed.edu Subject: [Histonet] Same Host Ab(fluorescence IHC) HELP...!!! Dear Histonetters, Here is the problem that is giving me a lot of grey hairs even though I am only 26. Its making me crazy and I am in deep despair. I cant go on like this.. Please help..! :'( I have to do Fluorescent based IHC on two proteins on old paraffin fixed skin tissue. The first antibody telomerase is from Mouse. I use goat antimouse Fab2 fragments of Alexa488 for the secondary detection. The second Ab that I have to stain is melanoma cells and I use Pan Melanoma cocktail from Biocare Medicals for this purpose. This is again from Mouse and has several IgG subtypes like IgG1K, IgG2a, IgG2b etc. I use Goat antimouse TRITC as my secondary for this step. Before the application of my second Primary(Pan Melanoma), I use Mouse (H+L) and incubate for 30 minutes, and then add (Mouse Fab(H+L) for blocking any epitopes on the first primary Ab (telomerase) so that my second secondary (TRITC) will not bind to that. However, I have been far from successful. I have run out of ideas. There are no good Ab for using against Melanoma (especially basal melanocytes) from any other hosts that work well. Here is a scheme that I use from Jacksonlab website. [1]http://www.jacksonimmuno.com/technical/examplec.asp Please help... ! Thank you all.. SWARAM References 1. http://www.jacksonimmuno.com/technical/examplec.asp _______________________________________________ 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 vazquezr <@t> ohsu.edu Thu Feb 3 15:05:52 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: Lynne, That is a good one, I love it... Robyn >>> "Bell, Lynne" 2/3/2005 12:28:00 PM >>> As one of our pathologists always says, "You can always tell a surgeon - but you can't tell him much!" Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Thu Feb 3 15:16:04 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: <020320052116.21053.4202949400054EA10000523D2200758942CECE030E9D0C9A03@comcast.net> Ain't it the truth. If they do listen it is only to tell you are wrong. -------------- Original message -------------- > Lynne, > That is a good one, I love it... > > Robyn > > >>> "Bell, Lynne" 2/3/2005 12:28:00 PM >>> > > > As one of our pathologists always says, "You can always tell a surgeon > - > but you can't tell him much!" > > Lynne A. Bell, HT (ASCP) > Central Vermont Hospital > P. O. Box 547 > Barre, VT 05641 > 802-371-4122 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From froyer <@t> bitstream.net Thu Feb 3 15:43:45 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde In-Reply-To: References: Message-ID: <42029B11.7050304@bitstream.net> My dear 'ol pathologist, God rest his soul, would always say... "If you want to hide a $10.00 bill from a surgeon, put it in a textbook!" ~ Ford ;-) Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Bell, Lynne wrote: >As one of our pathologists always says, "You can always tell a surgeon - >but you can't tell him much!" > >Lynne A. Bell, HT (ASCP) >Central Vermont Hospital >P. O. Box 547 >Barre, VT 05641 >802-371-4122 > > From Jackie.O'Connor <@t> abbott.com Thu Feb 3 15:52:47 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: What's the difference between God and a surgeon? God knows he's not a surgeon. Ford Royer Sent by: histonet-bounces@lists.utsouthwestern.edu 02/03/2005 03:43 PM To: histonet@pathology.swmed.edu cc: Subject: Re: [Histonet] 37% formaldehyde My dear 'ol pathologist, God rest his soul, would always say... "If you want to hide a $10.00 bill from a surgeon, put it in a textbook!" ~ Ford ;-) Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Bell, Lynne wrote: >As one of our pathologists always says, "You can always tell a surgeon - >but you can't tell him much!" > >Lynne A. Bell, HT (ASCP) >Central Vermont Hospital >P. O. Box 547 >Barre, VT 05641 >802-371-4122 > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Feb 3 16:34:13 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] avian/biotin block In-Reply-To: <43119.209.246.116.5.1107463229.squirrel@webmail> Message-ID: <200502032234.j13MY8G7026755@chip.viawest.net> Has anyone tried the Signet Acuity Mouse-on-Mouse Detection System "Biotin Free"? This might be a good alternative to an ABC detection for MonM here. Signet sent me a sample but I have not yet had time to try it. It uses an hrplabelled polymer link which is anti-ms and anti-rabbit with special blocking reagents before the primary and another block before the link. Patsy -----Original Message----- From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu] Sent: Thursday, February 03, 2005 1:40 PM To: Patsy Ruegg Cc: 'Emerson, Rachael'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] avian/biotin block Good idea - the polymers are superb - but EnVision is not suitable for mouse embryos but you could look for one that was suitable. I use DAKO Biotin Blocking System to routinely stain cell surface antigens in mouse tissue even finicky hematopoietic tissues. IMO, it is likely though that if your signal disappeared after blocking that it was non-specific - do you have a positive control tissue where it remained after blocking? > I would consider using a non-biotin detection system such as Hrp > Labelled Polymer, there are several out there now, the one I use is > from DAKO called Envision. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Emerson, Rachael > Sent: Thursday, February 03, 2005 12:32 PM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] avian/biotin block > > Let me try to sum up a long story and get to my question...... > Basically I am staining mouse embryos-frozen sections, for megakaryocytes. > I had trouble with endogenous AP and 15% acetic acid seemed to help. I > am having trouble with endogenous biotin, as well. I tried Vector's > Avidin/Biotin Blocking Kit and lost my signal all together. I am > looking for a surface antigen and I am afraid the kit blocked it all. > > I would really appreciate any suggestions on blocking endogenous > biotin on frozen sections? > > Also, I tried to us an AP secondary, but my signal was dimished. > Thanks > Rachael > > Rachael L. Emerson > Center for Human Genetics and Molecular Pediatric Diseases University > of Rochester Medical Center > 575 Elmwood Avenue MRBX 1.11301 > Rochester, NY 14642 > > Tel (585) 275-5073 > Fax (585) 276-0232 > > From haldana <@t> unimoron.edu.ar Thu Feb 3 16:31:29 2005 From: haldana <@t> unimoron.edu.ar (Hernan Aldana Marcos) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Help with paraformadehide and PCR Message-ID: <00d501c50a40$1b04a0c0$a904a8c0@um.edu> Dear Histonetters I need to make PCR in tissues fixed in formaldehyde or Bouin. Does anyone know some technique to do that? Or how can extract the excesses of formaldehyde of the tissues. I read that the sodium borohidrure can help in Immunohistochemistry. Thanks in advance Dr. Hern?n J. Aldana Marcos Facultad de Medicina. Universidad de Mor?n Machado 914. B1708JPD. Buenos Aires. Argentina e-mail alternativo hernanjavier@yahoo.com web: http://hjaldanamarcos.bravepages.com http://histologia.bigthicketdirectory.net/main.html From slimwillie <@t> cox.net Thu Feb 3 22:36:21 2005 From: slimwillie <@t> cox.net (Jerry Wilson) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Productivity Message-ID: <000901c50a73$13d26700$432e6044@no.cox.net> I think this question may have been addressed previously, but could I get a general idea from HIsto Tech's that process, embed and section a general mix of specimens (hospital surgicals, skins, gi's, etc.) of what volumes an individual tech would be expected to complete in one hour? For instance: How many blocks could one tech be reasonably expected to embed in one hour? How many blocks would one expect to section in one hour? I understand that everyone is different in the way they perform certain functions, but a general average of what would be expected would be appreciated. Thanks Jerry Wilson From Kemlo.Rogerson <@t> elht.nhs.uk Fri Feb 4 02:42:10 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Paraffin wax.[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF21@bhrv-nt-11.bhrv.nwest.nhs.uk> Surgipath I expect. I have no links to that Company other than knowing Stan for 30 years. There is also Solmedia and CellPath too, plus loads of others you can access on the net. -----Original Message----- From: Ian Montgomery [mailto:ian.montgomery@bio.gla.ac.uk] Sent: 03 February 2005 17:13 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin wax.[Scanned] After several years my stock of paraffin wax needs replacing. Currently, in the UK, who sells good reliable wax. Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mark.lewis <@t> thermo.com Fri Feb 4 02:53:28 2005 From: mark.lewis <@t> thermo.com (LEWIS, MARK A.) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde & the UK & Europe Message-ID: Concerning Formaldehyde & Formalin.. I would like to hear from our colleagues from around the world concerning the terminology and meaning of the differences between Formaldehyde and Formalin. I was always under the assumption that Formaldehyde was a gas "put" into water at it's highest concentration of 37 - 40 %. This 37-40% Formaldehyde when it is diluted with water then becomes or receives the terminology called Formalin whether it is 10 %, 20% etc... I am over in the UK and have been having some controversy with my colleagues here in that they want to use "concentrated formalin" for a particular procedure. If I take the formaldehyde purchased from my vendor and make up what I call a concentrated solution ( 37% solution), I will have 37% formalin, not the same thing. I feel that is confusing especially for those of us across the ocean, because I will want to ask How concentrated? If I hear the word formaldehyde, then I think of the 37-40% that we use to make up our 10% NBF and I associate that in my mind as concentrate. I especially would like to hear from those in other countries across the world. I even looked at a bottle of Formaldehyde that they have and it says Formaldehyde (formalin). I'm going to search some texts and talk to some chemists just because I want to be saying the correct thing without confusing everyone when it comes to the procedure being done. Thanks ! Mark Lewis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Linda Blazek Sent: Thu 2/3/2005 3:07 PM To: RossS@BaylorHealth.edu; brett_connolly@merck.com; histonet@pathology.swmed.edu Cc: Subject: RE: [Histonet] 37% formaldehyde It's probably a good thing I wasn't here to deal with it last night. Sometimes my mouth can get me in trouble. I would have loved to give him the gas. >>> "Stapf, Ross" 02/03/2005 3:04:45 PM >>> So he wanted a pure formaldehyde gas? I wish you had some around to give him. :) Ah the confusion between formaldehyde, formalin, and 10% NBF. It can make for some great "Who's on first" type moments. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:53 PM To: brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde There isn't any mistake about what he wanted. One of our techs was here late last night because one of our more "cranky" surgeons was doing a procedure in OR. He called to the lab for "formaldehyde". He was sent formalin. He preceded to yell (very loudly) that he did not want formalin he wanted "formaldehyde". Linda >>> "Connolly, Brett M" 2/3/2005 2:40:22 PM >>> I think Geoff hit the nail on the head. The surgeon doesn't realize or remember that 'formalin' is supposed to be 3.7%. I feel bad for the patient. Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] 37% formaldehyde Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ 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. ------------------------------------------------------------------------ ------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Fri Feb 4 03:18:11 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Marker for skeletal muscle satellite cells Message-ID: <001501c50a9a$729cfee0$e8345c9f@Carlos> If anyone has any experience of such antibodies, I would be most grateful for details. Ideally, I want to demonstrate them in frozen sections and FFPWs. At the moment I can think of MCAD, which is present at the membrane interface of these cells and the fibres. From Kemlo.Rogerson <@t> elht.nhs.uk Fri Feb 4 03:27:23 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde & the UK & Europe[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF23@bhrv-nt-11.bhrv.nwest.nhs.uk> 37% formaldehyde gas in water is concentrated formalin. 10% formalin is 10mls of that solution added to 90mls of water including buffer, isn't it? IMHO -----Original Message----- From: LEWIS, MARK A. [mailto:mark.lewis@thermo.com] Sent: 04 February 2005 08:53 To: Linda Blazek; RossS@BaylorHealth.edu; brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde & the UK & Europe[Scanned] Concerning Formaldehyde & Formalin.. I would like to hear from our colleagues from around the world concerning the terminology and meaning of the differences between Formaldehyde and Formalin. I was always under the assumption that Formaldehyde was a gas "put" into water at it's highest concentration of 37 - 40 %. This 37-40% Formaldehyde when it is diluted with water then becomes or receives the terminology called Formalin whether it is 10 %, 20% etc... I am over in the UK and have been having some controversy with my colleagues here in that they want to use "concentrated formalin" for a particular procedure. If I take the formaldehyde purchased from my vendor and make up what I call a concentrated solution ( 37% solution), I will have 37% formalin, not the same thing. I feel that is confusing especially for those of us across the ocean, because I will want to ask How concentrated? If I hear the word formaldehyde, then I think of the 37-40% that we use to make up our 10% NBF and I associate that in my mind as concentrate. I especially would like to hear from those in other countries across the world. I even looked at a bottle of Formaldehyde that they have and it says Formaldehyde (formalin). I'm going to search some texts and talk to some chemists just because I want to be saying the correct thing without confusing everyone when it comes to the procedure being done. Thanks ! Mark Lewis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Linda Blazek Sent: Thu 2/3/2005 3:07 PM To: RossS@BaylorHealth.edu; brett_connolly@merck.com; histonet@pathology.swmed.edu Cc: Subject: RE: [Histonet] 37% formaldehyde It's probably a good thing I wasn't here to deal with it last night. Sometimes my mouth can get me in trouble. I would have loved to give him the gas. >>> "Stapf, Ross" 02/03/2005 3:04:45 PM >>> So he wanted a pure formaldehyde gas? I wish you had some around to give him. :) Ah the confusion between formaldehyde, formalin, and 10% NBF. It can make for some great "Who's on first" type moments. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:53 PM To: brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde There isn't any mistake about what he wanted. One of our techs was here late last night because one of our more "cranky" surgeons was doing a procedure in OR. He called to the lab for "formaldehyde". He was sent formalin. He preceded to yell (very loudly) that he did not want formalin he wanted "formaldehyde". Linda >>> "Connolly, Brett M" 2/3/2005 2:40:22 PM >>> I think Geoff hit the nail on the head. The surgeon doesn't realize or remember that 'formalin' is supposed to be 3.7%. I feel bad for the patient. Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] 37% formaldehyde Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ 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. ------------------------------------------------------------------------ ------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From John.Auld <@t> whnt.nhs.uk Fri Feb 4 03:33:56 2005 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Re: Nomenclature: HT equivalent in France Message-ID: Hi Teri You are correct about the UK. All path lab technical staff are biomedical scientists, requiring BSc plus 1 years OTJ traing before becoming registered (mandatory). This is changing slightly as new degree courses which will incorporate some of the OTJ training are starting up. These will allow the graduate to be registered at graduation, hopefully improving recruitment. For progression to more senior levels MSc is required. The Institute of Biomedical Scientists website has a lot of info www.ibms.org Hope this is helpful John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral Internal extn 2560 External Tel 0151 604 7025 Message: 2 Date: Thu, 3 Feb 2005 11:37:47 -0600 From: "Johnson, Teri" Subject: [Histonet] Nomenclature: HT equivalent in France To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" For those histologists working in France (and other European countries), what is the nomenclature used for your position? In the UK we are biomedical scientists? And what are the education requirements for such? Any info (or links to helpful websites) is appreciated. Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From John.Auld <@t> whnt.nhs.uk Fri Feb 4 03:56:04 2005 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Re: 37% formaldehyde in the UK Message-ID: Mark In the UK formaldehyde is a gas the concentrate bought is 37 - 40% gas dissolved in water. This is then diluted to 4% gas in water, or 10% of the commercial concentrate. Unfortunately there is no formal (sorry for the pun) nomeclature for what is what. It is comonly accepted that formaldehyde is the 37 - 40% concentrate and formalin (4% gas) is the diluted form. While this may seem confusing I would be surpised at any histologist attempting to use concentrated formaldehyde for histology. It is sometimes used to fumigate saftey cabinets and cryostats though. John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral Internal extn 2560 External Tel 0151 604 7025 From: LEWIS, MARK A. [mailto:mark.lewis@thermo.com] Sent: 04 February 2005 08:53 To: Linda Blazek; RossS@BaylorHealth.edu; brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde & the UK & Europe[Scanned] Concerning Formaldehyde & Formalin.. I would like to hear from our colleagues from around the world concerning the terminology and meaning of the differences between Formaldehyde and Formalin. I was always under the assumption that Formaldehyde was a gas "put" into water at it's highest concentration of 37 - 40 %. This 37-40% Formaldehyde when it is diluted with water then becomes or receives the terminology called Formalin whether it is 10 %, 20% etc... I am over in the UK and have been having some controversy with my colleagues here in that they want to use "concentrated formalin" for a particular procedure. If I take the formaldehyde purchased from my vendor and make up what I call a concentrated solution ( 37% solution), I will have 37% formalin, not the same thing. I feel that is confusing especially for those of us across the ocean, because I will want to ask How concentrated? If I hear the word formaldehyde, then I think of the 37-40% that we use to make up our 10% NBF and I associate that in my mind as concentrate. I especially would like to hear from those in other countries across the world. I even looked at a bottle of Formaldehyde that they have and it says Formaldehyde (formalin). I'm going to search some texts and talk to some chemists just because I want to be saying the correct thing without confusing everyone when it comes to the procedure being done. Thanks ! Mark Lewis From c.m.vanderloos <@t> amc.uva.nl Fri Feb 4 04:30:29 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] RE: Same Host Ab(fluorescence IHC) HELP...!!! Message-ID: <9a6d169a6ff9.9a6ff99a6d16@amc.uva.nl> Hello Swaram, I have been testing this Jackon protocol once and could not figure out a good staining either. I presume that this type of protocol only works under "ideal" conditions. That means if you have exactly titrated all your reagents perfectly. Just a little too much of one reagent will result into non-specific, or better to say "unwanted" staining. As mentioned on Histonet, the approach to biotinylate one primary via the Dako ARKit (or even both primaries via the Zenon kit), is much more safer. 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 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From Swaram Date Thu, 03 Feb 2005 10:59:32 -0800 To HISTONET Cc histonet@pathology.swmed.edu Subject [Histonet] Same Host Ab(fluorescence IHC) HELP...!!! Dear Histonetters, Here is the problem that is giving me a lot of grey hairs even though I am only 26. Its making me crazy and I am in deep despair. I cant go on like this.. Please help..! :'( I have to do Fluorescent based IHC on two proteins on old paraffin fixed skin tissue. The first antibody telomerase is from Mouse. I use goat antimouse Fab2 fragments of Alexa488 for the secondary detection. The second Ab that I have to stain is melanoma cells and I use Pan Melanoma cocktail from Biocare Medicals for this purpose. This is again from Mouse and has several IgG subtypes like IgG1K, IgG2a, IgG2b etc. I use Goat antimouse TRITC as my secondary for this step. Before ! ; t References 1. mailto:c.m.vanderloos@amc.uva.nl From Kemlo.Rogerson <@t> elht.nhs.uk Fri Feb 4 05:20:01 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Re: Nomenclature: HT equivalent in France[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF25@bhrv-nt-11.bhrv.nwest.nhs.uk> One slight point. I think technical staff also include MLA's (Medical Laboratory Assistants) who despite some having an 'inappropriate degree' are perfectly trainable to ultimately become BMS's; in fact some carry out very responsible tasks. We also have Phlebotomists and Cytoscreeners too. I am also confused as to whether BMS's are 'technical staff' or 'scientific staff'. When we were state registered under the Medical Technicians Board it could have been argued so, but now we have a protected title and some have chartered status, then I don't know. But I do know that all Lab Staff are NOT Biomedical Scientists that I do know. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: John Auld [mailto:John.Auld@whnt.nhs.uk] Sent: 04 February 2005 09:34 To: histonet@lists.utsouthwestern.edu Cc: TJJ@Stowers-Institute.org Subject: [Histonet] Re: Nomenclature: HT equivalent in France[Scanned] Hi Teri You are correct about the UK. All path lab technical staff are biomedical scientists, requiring BSc plus 1 years OTJ traing before becoming registered (mandatory). This is changing slightly as new degree courses which will incorporate some of the OTJ training are starting up. These will allow the graduate to be registered at graduation, hopefully improving recruitment. For progression to more senior levels MSc is required. The Institute of Biomedical Scientists website has a lot of info www.ibms.org Hope this is helpful John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral Internal extn 2560 External Tel 0151 604 7025 Message: 2 Date: Thu, 3 Feb 2005 11:37:47 -0600 From: "Johnson, Teri" Subject: [Histonet] Nomenclature: HT equivalent in France To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" For those histologists working in France (and other European countries), what is the nomenclature used for your position? In the UK we are biomedical scientists? And what are the education requirements for such? Any info (or links to helpful websites) is appreciated. Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From John.Auld <@t> whnt.nhs.uk Fri Feb 4 05:33:43 2005 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Re: Nomenclature: HT equivalent in France[Scanned] Message-ID: Hi Kemlo I was trying to keep our somewhat complex situation as simple as possible for our overseas colleagues. Also it seems to me that biomedical scientist equivalents in some other countries are called technicians, technologists or similar. John PS I have an MLA in the second year of a BMS BSc course and doing very well, hope that's not tempting fate. Kemlo Rogerson , histonet@lists.utsouthwestern.edu cc: TJJ@Stowers-Institute.org 02/04/2005 11:20 Subject: RE: [Histonet] Re: Nomenclature: HT equivalent in AM France[Scanned] One slight point. I think technical staff also include MLA's (Medical Laboratory Assistants) who despite some having an 'inappropriate degree' are perfectly trainable to ultimately become BMS's; in fact some carry out very responsible tasks. We also have Phlebotomists and Cytoscreeners too. I am also confused as to whether BMS's are 'technical staff' or 'scientific staff'. When we were state registered under the Medical Technicians Board it could have been argued so, but now we have a protected title and some have chartered status, then I don't know. But I do know that all Lab Staff are NOT Biomedical Scientists that I do know. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: John Auld [mailto:John.Auld@whnt.nhs.uk] Sent: 04 February 2005 09:34 To: histonet@lists.utsouthwestern.edu Cc: TJJ@Stowers-Institute.org Subject: [Histonet] Re: Nomenclature: HT equivalent in France[Scanned] Hi Teri You are correct about the UK. All path lab technical staff are biomedical scientists, requiring BSc plus 1 years OTJ traing before becoming registered (mandatory). This is changing slightly as new degree courses which will incorporate some of the OTJ training are starting up. These will allow the graduate to be registered at graduation, hopefully improving recruitment. For progression to more senior levels MSc is required. The Institute of Biomedical Scientists website has a lot of info www.ibms.org Hope this is helpful John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral Internal extn 2560 External Tel 0151 604 7025 Message: 2 Date: Thu, 3 Feb 2005 11:37:47 -0600 From: "Johnson, Teri" Subject: [Histonet] Nomenclature: HT equivalent in France To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" For those histologists working in France (and other European countries), what is the nomenclature used for your position? In the UK we are biomedical scientists? And what are the education requirements for such? Any info (or links to helpful websites) is appreciated. Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Fri Feb 4 05:43:58 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde Message-ID: And a surgeon once said of pathologists that the reason they always seem to know everything is that their elbows are closer to the books than other people's are. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Ford Royer [mailto:froyer@bitstream.net] Sent: 03 February 2005 21:44 To: histonet@pathology.swmed.edu Subject: Re: [Histonet] 37% formaldehyde My dear 'ol pathologist, God rest his soul, would always say... "If you want to hide a $10.00 bill from a surgeon, put it in a textbook!" ~ Ford ;-) Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Bell, Lynne wrote: >As one of our pathologists always says, "You can always tell a surgeon - >but you can't tell him much!" > >Lynne A. Bell, HT (ASCP) >Central Vermont Hospital >P. O. Box 547 >Barre, VT 05641 >802-371-4122 > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Shawnster73 <@t> aol.com Fri Feb 4 07:06:27 2005 From: Shawnster73 <@t> aol.com (Shawnster73@aol.com) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] peptide staining Message-ID: <55AA0799.48828F46.3F5C7C89@aol.com> Trying to stain a peptide using IHC methods. ?Looking for anyone who has some pointers reguarding "secondaries." Michelle Schwab HT(ASCP) From asmith <@t> mail.barry.edu Fri Feb 4 09:08:09 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] formalin vs formaldehyde Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3C89@exchsrv01.barrynet.barry.edu> In the U.S. I have always heard the 37% w/v solution of formaldehyde gas in water called formalin. This is the usage in the 3rd edition of Kiernan, the 4th edition of Lillie, and the 2nd edition of Sheehan & Hrapchak. Thus, a 1 to 9 (or 1 in 10) dilution of this is 10% formalin (3.7% formaldehyde). Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 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 gcallis <@t> montana.edu Fri Feb 4 09:34:47 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Productivity In-Reply-To: <000901c50a73$13d26700$432e6044@no.cox.net> References: <000901c50a73$13d26700$432e6044@no.cox.net> Message-ID: <6.0.0.22.1.20050204082739.01b0cca0@gemini.msu.montana.edu> Jerry, There is a superb publication "A report from the National Society for Histotechnology Productiviey Task Force", LaFreiniere M, Sheppard B and Carson F. 27(4):293-295 J Histotechnology Dec 2004. It will answer a lot of your questions, including times to section blocks etc. If you have an NSH member or are a member yourself, you will have journal in hand. If not, you can obtain an electronic copy from NSH, just email them at histo@nsh.org and request this special report by title, issue. At 09:36 PM 2/3/2005, you wrote: >I think this question may have been addressed previously, but could I get >a general idea from HIsto Tech's that process, embed and section a general >mix of specimens (hospital surgicals, skins, gi's, etc.) of what volumes >an individual tech would be expected to complete in one hour? >For instance: How many blocks could one tech be reasonably expected to >embed in one hour? How many blocks would one expect to section in one hour? >I understand that everyone is different in the way they perform certain >functions, but a general average of what would be expected would be >appreciated. > >Thanks >Jerry Wilson >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From tpmorken <@t> labvision.com Fri Feb 4 10:59:27 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde & the UK & Europe Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F95A@usca0082k08.labvision.apogent.com> Ah...the annual formaldehyde / formalin discussion, destined to recur ad infinitum. >From my reading this is what I have been able to deduce: Technically, formaldehyde is the gas only (paraformaldehyde being the polymerized form). ANY solution of formaldehyde in water is technically termed "formalin". This gas in water is a saturated solution at 37% (by weight) or 40% (by volume). Any dilution, then, is a percentage of the saturated mix. So 10% formalin is roughly 4% formaldehyde in water. Unfortunately, manufacturers ( for example, Fisher) have decided to call the saturated solution "formaldehyde" thereby forever confusing the issue. So we have to use terms like "10% formalin (4% formaldehyde)" or "concentrated formaldehyde (37%)" to clarify every time we write a procedure. Here are the Fisher Scientifc products in my catalog. Formaldehyde, 37% by weight Formaldehyde, 40% by volume Formaldehyde, 10% w/w Formalin, neutral buffered, 10% Formalin, 10% solution Note that Fisher uses "Formaldehyde" for the saturated solution as well as a solution of 10% by weight (that's 10% formaldehyde in water), and "Formalin" for the dilution of the saturated solution (4% formaldehyde). Sigma calls it "Formaldehyde Solution (Formalin) 37% in water." It doesn't seem to me that anything will change, and for clarity, we'll all have to keep indicating the actual percentage of formaldehyde our soluions contain. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LEWIS, MARK A. Sent: Friday, February 04, 2005 12:53 AM To: Linda Blazek; RossS@BaylorHealth.edu; brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde & the UK & Europe Concerning Formaldehyde & Formalin.. I would like to hear from our colleagues from around the world concerning the terminology and meaning of the differences between Formaldehyde and Formalin. I was always under the assumption that Formaldehyde was a gas "put" into water at it's highest concentration of 37 - 40 %. This 37-40% Formaldehyde when it is diluted with water then becomes or receives the terminology called Formalin whether it is 10 %, 20% etc... I am over in the UK and have been having some controversy with my colleagues here in that they want to use "concentrated formalin" for a particular procedure. If I take the formaldehyde purchased from my vendor and make up what I call a concentrated solution ( 37% solution), I will have 37% formalin, not the same thing. I feel that is confusing especially for those of us across the ocean, because I will want to ask How concentrated? If I hear the word formaldehyde, then I think of the 37-40% that we use to make up our 10% NBF and I associate that in my mind as concentrate. I especially would like to hear from those in other countries across the world. I even looked at a bottle of Formaldehyde that they have and it says Formaldehyde (formalin). I'm going to search some texts and talk to some chemists just because I want to be saying the correct thing without confusing everyone when it comes to the procedure being done. Thanks ! Mark Lewis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Linda Blazek Sent: Thu 2/3/2005 3:07 PM To: RossS@BaylorHealth.edu; brett_connolly@merck.com; histonet@pathology.swmed.edu Cc: Subject: RE: [Histonet] 37% formaldehyde It's probably a good thing I wasn't here to deal with it last night. Sometimes my mouth can get me in trouble. I would have loved to give him the gas. >>> "Stapf, Ross" 02/03/2005 3:04:45 PM >>> So he wanted a pure formaldehyde gas? I wish you had some around to give him. :) Ah the confusion between formaldehyde, formalin, and 10% NBF. It can make for some great "Who's on first" type moments. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:53 PM To: brett_connolly@merck.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] 37% formaldehyde There isn't any mistake about what he wanted. One of our techs was here late last night because one of our more "cranky" surgeons was doing a procedure in OR. He called to the lab for "formaldehyde". He was sent formalin. He preceded to yell (very loudly) that he did not want formalin he wanted "formaldehyde". Linda >>> "Connolly, Brett M" 2/3/2005 2:40:22 PM >>> I think Geoff hit the nail on the head. The surgeon doesn't realize or remember that 'formalin' is supposed to be 3.7%. I feel bad for the patient. Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, February 03, 2005 1:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] 37% formaldehyde Does anyone know why a surgeon would want to send a liver biopsy specimen in 37% formaldehyde? Is there some test or procedure that you would ever want to put a specimen in 37%? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ 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, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From scoop <@t> mail.nih.gov Fri Feb 4 11:31:56 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] 37% formaldehyde & the UK & Europe Message-ID: I asked a question about formalin/formaldehyde nomenclature about a year ago and received several excellent replies. The question is answered in the following paper (which a very nice and patient histonetter sent me the reference for): Proper nomenclature of formaldehyde and paraformaldehyde fixatives for histochemistry. Manoonkitiwongsa, P.S. The Histochemical Journal 34: 365-367. 2002. Even though I got such good and informative replies to my question I cannot really reproduce them here in a way that would be clear and articulate enough for anyone to understand, so I suggest people read the article. The article is very clear and informative and after reading it carefully I understand formalin/formaldehyde for about 30 minutes before I am hopelessly confused again. If anyone needs it I can send a pdf. Sharon -- Sharon Cooperman M.D., Ph.D. NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From zhuxx003 <@t> umn.edu Fri Feb 4 11:42:01 2005 From: zhuxx003 <@t> umn.edu (Danielle Jin) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] CLC1 Ab labeling Message-ID: <4203B3E9.15382017@umn.edu> Hello! Histonetters, I have been trying to use every commercially availabled chloride channel antibodys (CLC1) to label frosen human muscle sections without success. Would anyone give me any suggesions what to do/try? I tried different dilutions of the CLC1, include straight, with vector ABC kit and plus TSA amplification kit from PerkinElmer. And I tried use Cy3-goat anti-rabbit IgG as secondary Ab. None worked, the signals are very fade with high backgrounds. Any suggestions will be appreciated. Danielle Jin From dpahisto <@t> yahoo.com Fri Feb 4 11:56:42 2005 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] HPV/DAKO Stainer Message-ID: <20050204175643.77421.qmail@web41315.mail.yahoo.com> Anyone out there using the Dako Autostainer to perform HPV testing? Can you send a synopsis of the Hybridizer procedure and what CPT code are you using to bill for the procedure? Cindy DuBois Delta Pathology Stockton, CA --------------------------------- Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. From csawrenk <@t> bccancer.bc.ca Fri Feb 4 12:22:09 2005 From: csawrenk <@t> bccancer.bc.ca (Sawrenko, Christina) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Double Staining kits Message-ID: <57AD92D1EDD1854BAAC02DD7DDAF0E3601700551@srvex10.phsabc.ehcnet.ca> Good Morning! Does anyone out there have any experience with the Double staining cocktails from Biocare? We are interested in the Kappa + Lambda cocktail, Ki-67 + Caspase-3, L26 + CD3 and L26 + Ki-67. How about their Double Stain polymer detection kits? In addition, has anyone used DakoCytomation Double stain kits? Thanks in advance for any help. Chris Sawrenko BC Cancer Agency-VCC Vancouver, British Columbia, Canada From vbaker60 <@t> yahoo.com Fri Feb 4 13:12:39 2005 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Did the Histonet search page move? Message-ID: <20050204191240.86497.qmail@web50101.mail.yahoo.com> Hi Does anyone know if we still have the Histonet search page working? I've tried to access it twice and I keep coming up with "unable to find page". have a nice weekend. Vikki Baker __________________________________ Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. http://info.mail.yahoo.com/mail_250 From bhewlett <@t> cogeco.ca Fri Feb 4 13:23:58 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde (lengthy) Message-ID: <008e01c50aef$13e603b0$6400a8c0@mainbox> I was hoping that John Kiernan would jump in reply to this issue with his usual eloquence! However, here goes. Confusion in terminology has been common since Blum introduced this agent as a fixative in 1893! It never ceases to amaze me that this should be so, the issue has been repeatedly addressed in all major Histotechnology texts since before the early fifties ( my student days in the UK). The following information is from the 10th Edition(1981) of the Condensed Chemical Dictionary and the10th Edition(1983) of the Merck Index (the only ones at hand). Formaldehyde is a gas. It is readily soluble in water up to 55% and is commercially available to us as 37%, 44% and 50% aqueous solutions which may contain up to 15% methanol. These commercial grades are called Formalin. Formaldehyde solution (Merck Index) The USP grade is about 37% (37-40%) w/v formaldehyde gas in water, usually with 10-15% methanol added to prevent polymerization. This solution is considered to be full strength and is also known as Formalin 100% or Formalin 40 which signifies that it contains 40 grams of formaldehyde within 100mL of the solution. It is this solution that produces most of the confusion since it is referred to and thought of as 100% Formalin. Paraformaldehyde (Merck Index) A white crystalline powder of polymerized formaldehyde, obtained by concentrating formaldehyde solution. Upon solution in water depolymerization and evolution of formaldehyde occurs. Thus an aqueous solution containing 4 grams of paraformaldehyde is essentially the same as a solution of 4% formaldehyde. There is NO such thing as a solution of paraformaldehyde. Right John? The concentration of formaldehyde used for fixation has been the subject of much confusion (see above). The concentration of formaldehyde in compound fixatives varies widely - ranging from 0.5 to 15% w/v. The majority of fixatives, using formaldehyde as the sole fixative agent, have a concentration of formaldehyde between 2.5 and 4% w/v. The concentration of formaldehyde in a fixative should be stated as the percentage by weight of the gas, rather than as a percentage of the formalin(sic) or paraformaldehyde(sic) used to prepare it. Thus: "4% formaldehyde" - not 10% formalin. "4% formaldehyde, from paraformaldehyde" - not 4% paraformaldehyde. "NBF means Neutral buffered formaldehyde" - (not formalin) and is 4% w/v formaldehyde in phosphate buffer pH 7.0- 7.2. Bryan Hewlett Consultant Technologist QMP-LS From tpmorken <@t> labvision.com Fri Feb 4 13:33:49 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde ( lengthy) Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328C9C6@usca0082k08.labvision.apogent.com> I think "Formalin" and "Formal" were trade names from a company and just went into common usage. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Hewlett Sent: Friday, February 04, 2005 11:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde (lengthy) I was hoping that John Kiernan would jump in reply to this issue with his usual eloquence! However, here goes. Confusion in terminology has been common since Blum introduced this agent as a fixative in 1893! It never ceases to amaze me that this should be so, the issue has been repeatedly addressed in all major Histotechnology texts since before the early fifties ( my student days in the UK). The following information is from the 10th Edition(1981) of the Condensed Chemical Dictionary and the10th Edition(1983) of the Merck Index (the only ones at hand). Formaldehyde is a gas. It is readily soluble in water up to 55% and is commercially available to us as 37%, 44% and 50% aqueous solutions which may contain up to 15% methanol. These commercial grades are called Formalin. Formaldehyde solution (Merck Index) The USP grade is about 37% (37-40%) w/v formaldehyde gas in water, usually with 10-15% methanol added to prevent polymerization. This solution is considered to be full strength and is also known as Formalin 100% or Formalin 40 which signifies that it contains 40 grams of formaldehyde within 100mL of the solution. It is this solution that produces most of the confusion since it is referred to and thought of as 100% Formalin. Paraformaldehyde (Merck Index) A white crystalline powder of polymerized formaldehyde, obtained by concentrating formaldehyde solution. Upon solution in water depolymerization and evolution of formaldehyde occurs. Thus an aqueous solution containing 4 grams of paraformaldehyde is essentially the same as a solution of 4% formaldehyde. There is NO such thing as a solution of paraformaldehyde. Right John? The concentration of formaldehyde used for fixation has been the subject of much confusion (see above). The concentration of formaldehyde in compound fixatives varies widely - ranging from 0.5 to 15% w/v. The majority of fixatives, using formaldehyde as the sole fixative agent, have a concentration of formaldehyde between 2.5 and 4% w/v. The concentration of formaldehyde in a fixative should be stated as the percentage by weight of the gas, rather than as a percentage of the formalin(sic) or paraformaldehyde(sic) used to prepare it. Thus: "4% formaldehyde" - not 10% formalin. "4% formaldehyde, from paraformaldehyde" - not 4% paraformaldehyde. "NBF means Neutral buffered formaldehyde" - (not formalin) and is 4% w/v formaldehyde in phosphate buffer pH 7.0- 7.2. Bryan Hewlett Consultant Technologist QMP-LS _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Fri Feb 4 13:45:44 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:35 2005 Subject: [BULK] - RE: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde ( lengthy) Message-ID: I thought formalin was a five star motel with a tux only lounge. >>> "Morken, Tim - Labvision" 02/04/05 02:33PM >>> I think "Formalin" and "Formal" were trade names from a company and just went into common usage. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Hewlett Sent: Friday, February 04, 2005 11:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde (lengthy) I was hoping that John Kiernan would jump in reply to this issue with his usual eloquence! However, here goes. Confusion in terminology has been common since Blum introduced this agent as a fixative in 1893! It never ceases to amaze me that this should be so, the issue has been repeatedly addressed in all major Histotechnology texts since before the early fifties ( my student days in the UK). The following information is from the 10th Edition(1981) of the Condensed Chemical Dictionary and the10th Edition(1983) of the Merck Index (the only ones at hand). Formaldehyde is a gas. It is readily soluble in water up to 55% and is commercially available to us as 37%, 44% and 50% aqueous solutions which may contain up to 15% methanol. These commercial grades are called Formalin. Formaldehyde solution (Merck Index) The USP grade is about 37% (37-40%) w/v formaldehyde gas in water, usually with 10-15% methanol added to prevent polymerization. This solution is considered to be full strength and is also known as Formalin 100% or Formalin 40 which signifies that it contains 40 grams of formaldehyde within 100mL of the solution. It is this solution that produces most of the confusion since it is referred to and thought of as 100% Formalin. Paraformaldehyde (Merck Index) A white crystalline powder of polymerized formaldehyde, obtained by concentrating formaldehyde solution. Upon solution in water depolymerization and evolution of formaldehyde occurs. Thus an aqueous solution containing 4 grams of paraformaldehyde is essentially the same as a solution of 4% formaldehyde. There is NO such thing as a solution of paraformaldehyde. Right John? The concentration of formaldehyde used for fixation has been the subject of much confusion (see above). The concentration of formaldehyde in compound fixatives varies widely - ranging from 0.5 to 15% w/v. The majority of fixatives, using formaldehyde as the sole fixative agent, have a concentration of formaldehyde between 2.5 and 4% w/v. The concentration of formaldehyde in a fixative should be stated as the percentage by weight of the gas, rather than as a percentage of the formalin(sic) or paraformaldehyde(sic) used to prepare it. Thus: "4% formaldehyde" - not 10% formalin. "4% formaldehyde, from paraformaldehyde" - not 4% paraformaldehyde. "NBF means Neutral buffered formaldehyde" - (not formalin) and is 4% w/v formaldehyde in phosphate buffer pH 7.0- 7.2. Bryan Hewlett Consultant Technologist QMP-LS _______________________________________________ Histonet mailing list Histonet@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 bhewlett <@t> cogeco.ca Fri Feb 4 13:55:02 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde (lengthy) References: <0556BE8AC5551E4E8AF6BB9E42509BA20328C9C6@usca0082k08.labvision.apogent.com> Message-ID: <00a701c50af3$6c335000$6400a8c0@mainbox> Tim, "Formal" is quite different, it is another name for Methylal (dimethoxymethane) a flammable colourless, volatile liquid used as a solvent. Bryan ----- Original Message ----- From: "Morken, Tim - Labvision" To: Sent: Friday, February 04, 2005 2:33 PM Subject: RE: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde (lengthy) > I think "Formalin" and "Formal" were trade names from a company and just > went into common usage. > > > Tim Morken > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan > Hewlett > Sent: Friday, February 04, 2005 11:24 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde > (lengthy) > > > I was hoping that John Kiernan would jump in reply to this issue with his > usual eloquence! However, here goes. > > Confusion in terminology has been common since Blum introduced this agent as > a fixative in 1893! It never ceases to amaze me that this should be so, the > issue has been repeatedly addressed in all major Histotechnology texts since > before the early fifties ( my student days in the UK). > > The following information is from the 10th Edition(1981) of the Condensed > Chemical Dictionary and the10th Edition(1983) of the Merck Index (the only > ones at hand). > > Formaldehyde is a gas. > It is readily soluble in water up to 55% and is commercially available to us > as 37%, 44% and 50% aqueous solutions which may contain up to 15% methanol. > These commercial grades are called Formalin. > > Formaldehyde solution (Merck Index) > The USP grade is about 37% (37-40%) w/v formaldehyde gas in water, usually > with 10-15% methanol added to prevent polymerization. This solution is > considered to be full strength and is also known as Formalin 100% or > Formalin 40 which signifies that it contains 40 grams of formaldehyde within > 100mL of the solution. It is this solution that produces most of the > confusion since it is referred to and thought of as 100% Formalin. > > Paraformaldehyde (Merck Index) > A white crystalline powder of polymerized formaldehyde, obtained by > concentrating formaldehyde solution. Upon solution in water depolymerization > and evolution of formaldehyde occurs. Thus an aqueous solution containing 4 > grams of paraformaldehyde is essentially the same as a solution of 4% > formaldehyde. There is NO such thing as a solution of paraformaldehyde. > Right John? > > The concentration of formaldehyde used for fixation has been the subject of > much confusion (see above). The concentration of formaldehyde in compound > fixatives varies widely - ranging from 0.5 to 15% w/v. The majority of > fixatives, using formaldehyde as the sole fixative agent, have a > concentration of formaldehyde between 2.5 and 4% w/v. The concentration of > formaldehyde in a fixative should be stated as the percentage by weight of > the gas, rather than as a percentage of the > formalin(sic) or paraformaldehyde(sic) used to prepare it. > > Thus: > "4% formaldehyde" - not 10% formalin. > "4% formaldehyde, from paraformaldehyde" - not 4% paraformaldehyde. "NBF > means Neutral buffered formaldehyde" - (not formalin) and is 4% w/v > formaldehyde in phosphate buffer pH 7.0- 7.2. > > Bryan Hewlett > > Consultant Technologist > QMP-LS > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From JWEEMS <@t> sjha.org Fri Feb 4 14:00:42 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde(lengthy) Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507A80@sjhaexc02.sjha.org> And all this time I thought it was how to dress for a party. I could have blown up! Have a good weekend everyone... j:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bryan Hewlett Sent: Friday, February 04, 2005 2:55 PM To: Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde(lengthy) Tim, "Formal" is quite different, it is another name for Methylal (dimethoxymethane) a flammable colourless, volatile liquid used as a solvent. Bryan ----- Original Message ----- From: "Morken, Tim - Labvision" To: Sent: Friday, February 04, 2005 2:33 PM Subject: RE: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde (lengthy) > I think "Formalin" and "Formal" were trade names from a company and just > went into common usage. > > > Tim Morken > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan > Hewlett > Sent: Friday, February 04, 2005 11:24 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde > (lengthy) > > > I was hoping that John Kiernan would jump in reply to this issue with his > usual eloquence! However, here goes. > > Confusion in terminology has been common since Blum introduced this agent as > a fixative in 1893! It never ceases to amaze me that this should be so, the > issue has been repeatedly addressed in all major Histotechnology texts since > before the early fifties ( my student days in the UK). > > The following information is from the 10th Edition(1981) of the Condensed > Chemical Dictionary and the10th Edition(1983) of the Merck Index (the only > ones at hand). > > Formaldehyde is a gas. > It is readily soluble in water up to 55% and is commercially available to us > as 37%, 44% and 50% aqueous solutions which may contain up to 15% methanol. > These commercial grades are called Formalin. > > Formaldehyde solution (Merck Index) > The USP grade is about 37% (37-40%) w/v formaldehyde gas in water, usually > with 10-15% methanol added to prevent polymerization. This solution is > considered to be full strength and is also known as Formalin 100% or > Formalin 40 which signifies that it contains 40 grams of formaldehyde within > 100mL of the solution. It is this solution that produces most of the > confusion since it is referred to and thought of as 100% Formalin. > > Paraformaldehyde (Merck Index) > A white crystalline powder of polymerized formaldehyde, obtained by > concentrating formaldehyde solution. Upon solution in water depolymerization > and evolution of formaldehyde occurs. Thus an aqueous solution containing 4 > grams of paraformaldehyde is essentially the same as a solution of 4% > formaldehyde. There is NO such thing as a solution of paraformaldehyde. > Right John? > > The concentration of formaldehyde used for fixation has been the subject of > much confusion (see above). The concentration of formaldehyde in compound > fixatives varies widely - ranging from 0.5 to 15% w/v. The majority of > fixatives, using formaldehyde as the sole fixative agent, have a > concentration of formaldehyde between 2.5 and 4% w/v. The concentration of > formaldehyde in a fixative should be stated as the percentage by weight of > the gas, rather than as a percentage of the > formalin(sic) or paraformaldehyde(sic) used to prepare it. > > Thus: > "4% formaldehyde" - not 10% formalin. > "4% formaldehyde, from paraformaldehyde" - not 4% paraformaldehyde. "NBF > means Neutral buffered formaldehyde" - (not formalin) and is 4% w/v > formaldehyde in phosphate buffer pH 7.0- 7.2. > > Bryan Hewlett > > Consultant Technologist > QMP-LS > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From nmuvarak <@t> facstaff.wisc.edu Fri Feb 4 14:17:12 2005 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL E MUVARAK) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Masson's Trichrome Stain Message-ID: <4541b24591cb.4591cb4541b2@wiscmail.wisc.edu> Hello. I want to do a Masson's trichrome stain on some rat arteries. I never did this before. All I know are the purpose and the principle. So I googled up a protocol from a website from which I had gotten a protocol for VVG, which worked pretty well on our frozen tissue. But this protocol mentions only paraffin sections. Would it work on frozen, aceton-fixed tissue? Here's a brief description of the protocol: Bouin's solution is used as a fixative (contains saturated picric acid, formaldehyde, and acetic acid). Hematoxylin for 10 min. Biebrich scarlet for 5 min. Phosphotungstic/phosphomolybdic acid for 10 min. Aniline blue for 5 min. 1% acetic acid for 1 min. Of course, all steps have washes. Then dehydrate, clear, and coverslip. Does that sound right for frozen sections? Thank you for your time. Regards, Nidal Muvarak Dept. of Biomedical Engineering University of Wisconsin Madison, WI 53706 From BlazekL <@t> childrensdayton.org Fri Feb 4 14:21:04 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Resolution of 37% case Message-ID: Dear all, Thanks to all that responded to my question about a surgeon putting a liver biopsy in "formaldehyde". All of the responses lightened the day. And now the results are in. 1. The "meek" (formerly cranky) surgeon admitted there must have been a misunderstanding between himself and his surgery tech team, but the histo tech. should have known that there was a misunderstanding and should have not sent the 37%. 2. The specimen survived abet some formalin pigment. Thanks again to all! Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org From ploykasek <@t> phenopath.com Fri Feb 4 14:36:59 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Double Staining kits In-Reply-To: <57AD92D1EDD1854BAAC02DD7DDAF0E3601700551@srvex10.phsabc.ehcnet.ca> Message-ID: Good afternoon Chris. I do not have any experience with the Biocare, but do have experience with the Dakocytomation. I have had very good results with the Dako kit. I have even done some triple labeling. There are times on research projects when I use the main components from the dako kit with chromogens from Vector. Patti Loykasek PhenoPath Laboratories Seattle, WA> Good Morning! > Does anyone out there have any experience with the Double staining > cocktails from Biocare? We are interested in the Kappa + Lambda > cocktail, Ki-67 + Caspase-3, L26 + CD3 and L26 + Ki-67. How about > their Double Stain polymer detection kits? > In addition, has anyone used DakoCytomation Double stain kits? > > Thanks in advance for any help. > Chris Sawrenko > BC Cancer Agency-VCC > Vancouver, British Columbia, Canada > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Fri Feb 4 14:35:36 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Clinicians surgeons and pathologists Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328C9C9@usca0082k08.labvision.apogent.com> While clinicians know something about everything and surgeons know everything about something, pathologists know everything about everything - but it's too late. Tim Morken And a surgeon once said of pathologists that the reason they always seem to know everything is that their elbows are closer to the books than other people's are. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Ford Royer [mailto:froyer@bitstream.net] Sent: 03 February 2005 21:44 To: histonet@pathology.swmed.edu Subject: Re: [Histonet] 37% formaldehyde My dear 'ol pathologist, God rest his soul, would always say... "If you want to hide a $10.00 bill from a surgeon, put it in a textbook!" ~ Ford ;-) Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Bell, Lynne wrote: >As one of our pathologists always says, "You can always tell a surgeon >- but you can't tell him much!" > >Lynne A. Bell, HT (ASCP) >Central Vermont Hospital >P. O. Box 547 >Barre, VT 05641 >802-371-4122 > From pruegg <@t> ihctech.net Fri Feb 4 14:58:05 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Region VII/CSH Meeting Message-ID: <200502042057.j14KvwG7028890@chip.viawest.net> This is posted as an invitation to attendees and vendors to join Region VII and the Colorado Society for Histotechnology at our Symposium Convention in April 2005. Please help us get the word out. Patsy Ruegg 2005 Colorado Society of Histotechnology & NSH Region VII Meeting ( www.coloradohisto.org/2005) Dates: April 22 & 23, 2005 Location: ----------------- Beaver Run Resort 620 Village Rd. Breckenridge, CO 80424-2115 Ph: (800) 525-2253 (970) 453-6000 Fx: (970) 453-4284 www.beaverrun.com Registration: -------------------- Friday: 12:00 - 1:00 Saturday: 7:00 - 8:00 Exhibitors: -------------------- Friday: 9:30 - 6:00 Saturday: 9:30 - 5:00 Schedule: Friday, April 23rd ----------------------------- 12:00 - 1:00 Registration 1:00 - 2:30 Class 1-A or 1-B 2:30 - 3:00 Break 3:00 - 4:30 Class 2-A or 2-B 4:30 - 6:00 Social Saturday, April 24th -------------------- 7:00 - 8:00 Registration and Breakfast 8:00 - 9:30 Class 3-A or 3-B 9:30 - 10:00 Break 10:00 -11:30 Class 4-A or 4-C 11:30 - 1:00 Lunch and CSH Business Meeting 1:00 - 2:30 Class 5-A or 5-B 2:30 - 3:00 Break 3:00 - 4:30 Class 6-A or 6-B 4:30 - 5:00 Vendo drawing Deadlines: ---------- Hotel Reservations - March 8, 2005 CSH Pre-registration - April 2, 2005 **************************************************************************** **** PROGRAM Note: Some of the speaker abstracts were not available at the time of this posting. However, all abstracts are being posted on the CSH website as they become available. We apologize for any inconvenience. **************************************************************************** **** SESSION 1: FRIDAY AFTERNOON, APRIL 22, 2005 1:00 PM - 2:30 PM, Presentations 1-A. The History of Automation in Histology Mr. H. Skip Brown, B.A., HT (ASCP) Lab Management Consultants - St. Louis, MO Abstract Pending CEU credit hours: 1.5 ---------------------------------------------------------------------------- ---- 1-B. Update for Markers in Breast Cancer Dr. Meenakshi Singh, M.D. Associate Director of Surgical Pathology, UCHSC - Aurora, CO This presentation will cover the clinical relevance of estrogen receptor, progesterone receptor and Her 2neu analysis by immunohistochemistry, image analysis and fluorescent in situ hybridization studies in invasive breast cancer. The expression profile of a patient's tumor for these markers is used in tailoring treatment decisions and assessing prognosis for individual patients. Recently the scope of ER/PR IHC has been expanded to also being incorporate these into the clinical work up of cases of non-invasive breast cancer (ductal carcinoma in situ). The potential uses of newer markers in breast cancer and metastatic breast cancer will also be discussed; these shall include clinical research done in Dr. Singh's laboratory at the University of Colorado Health Sciences Center in recent years. CEU credit hours: 1.5 **************************************************************************** **** 2:30 PM - 3:00 PM, Break (Breckenridge Ballroom) **************************************************************************** **** 3:00 PM - 4:30 PM, Presentations 2-A. The Challenges of Change Mr. H. Skip Brown, B.A., HT (ASCP) Lab Management Consultants - St. Louis, MO Abstract Pending CEU credit hours: 1.5 ---------------------------------------------------------------------------- ---- 2-B. Changes in the QIHC Ms. Patsy Ruegg, HT(ASCP) QIHC IHCtech, FitzSimmons Bioscience Park - Aurora, CO., www.ihctech.net This presentation can help prepare potential examinees for the new written format for the QIHC exam by going over the projects required in the past and their application to the written questions anticipated on the new exam. The subject categories asked about on this written exam will be covered in this presentation. A list of IHC theory books and handbooks to use to prepare for the exam will be provided. CEU credit hours: 1.5 **************************************************************************** **** SESSION 2: SATURDAY MORNING, APRIL 23, 2005 =========================================== 8:00 AM - 9:30 AM, Presentations 3-A. Contrasting Sharp and Blunt Force Trauma Dr. Stephen Cina, M.D. Weld County Coroner Pathologist, Mckee Medical Center - Loveland, CO Following this presentation, the participant will be able to: 1. Discriminate between sharp force and blunt force injuries. 2. Recognize the significance of "pattern injuries." 3. Associate the types of injuries caused by various weapons types. A cutaneous injury may be the result of blunt or sharp force trauma. Even to the trained eye, it may be difficult to discriminate between lacerations, cuts, stabs or gunshot wounds. This presentation will focus on the diagnostic features of blunt force and sharp force injuries. Mechanisms of these types of injuries will be discussed. Due to the nature of the topic, this presentation will utilize many graphic images to illustrate salient points. CEU credit hours: 1.5 ---------------------------------------------------------------------------- ---- 3-B. Localization of Mycobacterium tuberculosis in diseased tissue utilizing in situ hybridization in combination with immunohistochemistry Ms. Liz Chlipala, B.S., HTL (ASCP) QIHC, Premier Lab, LLC. - Boulder, CO, www.premierhistology.com Ms. Allison L. St. Amand, M.S., University of Colorado at Boulder Diagnosis of tuberculosis generally relies on a combination of acid fast staining of histological specimens, and culture methods. AFB staining is unable to speciate different mycobacteria. In addition mycobacterial acid fastness can be partly or completely lost at some stages of growth of the organisms. Alternatively in situ hybridization can be used to detect bacterial distribution and morphology in the context of histopathology of the tissue. In situ hybridization utilizing 16S ribosomal RNA oligonulceotide probes allows for rapid speciation of mycobacteria. This technique used in combination with immunohistochemistry allows for better understanding of the disease process. This lecture will demonstrate the use of ISH, IHC, and IF in animal models of Mycobacterium tuberculosis infections. CEU credit hours: 1.5 **************************************************************************** **** 9:30 AM - 10:00 AM, Break (Breckenridge Ballroom) **************************************************************************** **** SESSION 2: SATURDAY MORNING, APRIL 23, 2005 =========================================== 10:00 AM - 11:30 AM, Presentations 4-A. Theory and Practice of Microwave Technology Mr. H. Skip Brown, B.A., HT (ASCP) Lab Management Consultants - St. Louis, MO Abstract Pending CEU credit hours: 1.5 ---------------------------------------------------------------------------- ---- 4-B. The Human Genome Project Ms. Donna Lawson, HT (ASCP) QIHC Technical Trainer, Ventana Medical Systems - Tuscon, AZ, www.ventanamed.com The Human Genome Project was officially started in 1990; the involvement became a world wide project of scientists and organizations. The goal was to generate a high-quality reference DNA sequence for the human genome's 3 billion base pairs and to identify all human genes. The project completed in 2003. Even though it was a federally funded project there was great effort to make the information discovered available to the private sector. Grants were funded for innovative research which in turn created a multibillion dollar biotechnology industry. What does this all mean to the medical profession? Because of the project a new world of drug discovery, genetic testing, and bioethics were created. This workshop will cover the inception of the Project and the terminology of genetics in a simple format. It will also discuss the areas the project has touch in medicine, agriculture and industry. The questions of bioethics that arose during the project will be covered along with some possible answers. CEU credit hours: 1.5 **************************************************************************** **** 11:30 AM - 1:00 PM Lunch & CSH Business Meeting (Breckenridge Ballroom) **************************************************************************** **** SESSION 3: SATURDAY AFTERNOON, APRIL 23, 2005 ============================================= 1:00 PM - 2:30 PM, Presentations 5-A. Great Cases from Small Places (I) Dr. Thomas Haas, D.O. Pathologist, Mercy Health System - Janesville, WI In many hospitals, there is no communication or connection between the work done by histotechnologists, the interpretations and recommendations made by pathologists, and the final diagnosis and treatment of the patient by clinicians. Oftentimes, the histotechnologist is not encouraged to attend "Tumor Boards" or other peer-review situations involving patient care. This can be frustrating to those involved in the "behind the scenes" work of the laboratory, and can tend to squelch understanding, interest, and learning on the part of the histotechnologist, when no feedback or integration of "why" certain stains and investigative laboratory techniques were ordered. This seminar will explore a number of different pathology cases and their final diagnoses, beginning with patient information, progressing through the reasoning behind ordering special stains, immunohistochemistry, flow cytometry, and other pertinent laboratory methods, with an explanation of the disease process, diagnostic methods, and final treatment of the patient. These cases range from those seen everyday in the hospital pathology laboratory, to those requiring outside expert interpretation. CEU credit hours: 1.5 ---------------------------------------------------------------------------- ---- 5-B. Laser Capture Microdissection (LCM) - HT Perspective Ms. Ann Bennett Lossing, Field Application Scientist, Arcturus, Inc. - Mountain View, CA, www.arctur.com Gene Expression profiling of specific cell types in both frozen and formalin-fixed breast cancer biopsy samples. Microarrays are valuable tools for studying normal and induced variations in gene expression. Microgram amounts of total RNA are required for target preparation for most microarray platforms. Consequently, whole tissue biopsies are typically used for these studies. However, distinct differences have been shown between gene expression data obtained from whole tissue biopsies, which are essentially mixed cell populations, and that obtained from homogenous populations of cells. Microdissection enables the profiling of native expression levels of thousands of genes, in a selected cell population. In this presentation we will show how microdissection, combined with RNA amplification to produce the amounts of aRNA needed for microarray analysis, have allowed us to generate expression profiles in specific cell populations obtained from breast cancer biopsy samples. As a result of recent technological advances, we are able to isolate RNA from both frozen and formalin fixed tissue samples, and used this RNA to obtain expression profiles. These highly reproducible expression profiles have been used to generate molecular signatures for different stages of breast cancer using frozen biopsy tissues and microarray analysis. Our presentation will focus on the importance of developing histological methods for providing excellent visualization of cell types, while at the same time, preserving preserve RNA quality. Topics to be discussed:- Laser Capture Microdissection Technology Microgenomics Reagent Systems Paradise T Formalin-Fixed Tissue - Expression Analysis System Application Review - Arcturus LCM-based Publications (over 550 available to date!) CEU credit hours: 1.5 **************************************************************************** **** 2:30 PM - 3:00 PM Break (Breckenridge Ballroom) **************************************************************************** **** 3:00 PM - 4:30 PM, Presentations 6-A. Great Cases from Small Places (II) Dr. Thomas Haas, D.O. Pathologist, Mercy Health System - Janesville, WI See presentation 5-A for abstract CEU credit hours: 1.5 ---------------------------------------------------------------------------- ---- 6-B. Dewax, Rehydration, and Antigen Retrieval the Easy Way Ms. Patricia Piatti & Mr. Allen Younger Biogenex, San Ramon, CA How do you conduct Dewax, Rehydration, and Antigen Retrieval for your formalin-fixed, paraffin-embedded tissue sections? Do you baby-sit every tedious step of toxic xylene and alcohol dewax and rehydration pretreatment? Do you have difficulty in preserving tissue sample morphology and obtaining consistent results when you using traditional pressure cookers, water baths, and microwaves for antigen retrieval? Do you need a lot of guess work and training? Now, with BioGenex's EZ Retrieval System, the sample pretreatment has never been as easy as today. EZ RetrieverT allows you to shorten multiple steps of traditional tissue sample pretreatment from 85min to less than 30 mins. With a throughput as high as 96 slides/run, you may even save more than 4 hours of work and you can almost walk away from the whole process. With the ease of use digital interface and temperature probe, you can standardize the procedure with little training. By using eco-friendly ZEN-AR solutions that have been optimized for each antibody, you can preserves the tissue morphology and gain high quality and reproducible results. To learn more about the ZEN Retrieval System, please join the ZEN Retrieval System work shop that will be held in the following locations or visit our website at www.biogenex.com. CEU credit hours: 1.5 **************************************************************************** **** 4:30 PM - 5:00 PM, Vendo drawing (Breckenridge Ballroom) **************************************************************************** **** Online registration and additonal information regarding the meeting can be found at www.coloradohisto.org/2005 From pmarcum <@t> polysciences.com Fri Feb 4 15:45:09 2005 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Resolution of 37% case In-Reply-To: Message-ID: <000001c50b02$cc5cc880$7f00a8c0@PMARCUM2K> I knew he (the surgeon) couldn't be wrong and some one from histology would get blamed somehow. Glad to hear the specimen was salvagable and the patient did not suffer. Thanks, Pam Marcum > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda > Blazek > Sent: Friday, February 04, 2005 3:21 PM > To: histonet@pathology.swmed.edu > Subject: [Histonet] Resolution of 37% case > > > Dear all, > Thanks to all that responded to my question about a surgeon putting a > liver biopsy in "formaldehyde". All of the responses lightened the day. > And now the results are in. > > 1. The "meek" (formerly cranky) surgeon admitted there must have been > a misunderstanding between himself and his surgery tech team, but the > histo tech. should have known that there was a misunderstanding and > should have not sent the 37%. > 2. The specimen survived abet some formalin pigment. > Thanks again to all! > > > > > Linda Blazek, HT (ASCP) > Department of Pathology > Children's Medical Center > Dayton, Ohio 45404 > (937) 641-3358 > fax (937)641-5482 > blazekl@childrensdayton.org > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From froyer <@t> bitstream.net Fri Feb 4 16:08:06 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Clinicians surgeons and pathologists In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328C9C9@usca0082k08.labvision.apogent.com> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328C9C9@usca0082k08.labvision.apogent.com> Message-ID: <4203F246.10401@bitstream.net> AND of course... What's the difference between a Urologist and a Pathologist? A Urologist washes his hands BEFORE he goes to the bathroom..... Morken, Tim - Labvision wrote: >While clinicians know something about everything and surgeons know >everything about something, pathologists know everything about everything - >but it's too late. > >Tim Morken > > > >And a surgeon once said of pathologists that the reason they always seem to >know everything is that their elbows are closer to the books than other >people's are. > >Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant >Pathologist Rotherham General Hospital South Yorkshire England > terry.marshall@rothgen.nhs.uk > >-----Original Message----- >From: Ford Royer [mailto:froyer@bitstream.net] >Sent: 03 February 2005 21:44 >To: histonet@pathology.swmed.edu >Subject: Re: [Histonet] 37% formaldehyde > > >My dear 'ol pathologist, God rest his soul, would always say... "If you >want to hide a $10.00 bill from a surgeon, put it in a textbook!" > >~ Ford ;-) > >Ford M. Royer, MT(ASCP) >Midwest Science Biocenter >Minneapolis, MN > > >Bell, Lynne wrote: > > > >>As one of our pathologists always says, "You can always tell a surgeon >>- but you can't tell him much!" >> >>Lynne A. Bell, HT (ASCP) >>Central Vermont Hospital >>P. O. Box 547 >>Barre, VT 05641 >>802-371-4122 >> >> >> > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From froyer <@t> bitstream.net Fri Feb 4 16:15:43 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde(lengthy) In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E507A80@sjhaexc02.sjha.org> References: <83AACDB0810528418AA106F9AE9B7F7E507A80@sjhaexc02.sjha.org> Message-ID: <4203F40F.5040203@bitstream.net> No, no, Joyce. You are all confused. A "Formal" is a bright red farm tractor. My uncle had three of them, and he wasn't any kind of doctor at all. ~ Ford ;-) Weems, Joyce wrote: >And all this time I thought it was how to dress for a party. I could have blown up! > >Have a good weekend everyone... j:>) > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bryan >Hewlett >Sent: Friday, February 04, 2005 2:55 PM >To: Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] RE: Formaldehyde vs Formalin vs >paraformaldehyde(lengthy) > > >Tim, > >"Formal" is quite different, it is another name for Methylal >(dimethoxymethane) a flammable colourless, volatile liquid used as a >solvent. > >Bryan > >----- Original Message ----- >From: "Morken, Tim - Labvision" >To: >Sent: Friday, February 04, 2005 2:33 PM >Subject: RE: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde >(lengthy) > > > > >>I think "Formalin" and "Formal" were trade names from a company and just >>went into common usage. >> >> >>Tim Morken >> >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan >>Hewlett >>Sent: Friday, February 04, 2005 11:24 AM >>To: histonet@lists.utsouthwestern.edu >>Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde >>(lengthy) >> >> >>I was hoping that John Kiernan would jump in reply to this issue with his >>usual eloquence! However, here goes. >> >>Confusion in terminology has been common since Blum introduced this agent >> >> >as > > >>a fixative in 1893! It never ceases to amaze me that this should be so, >> >> >the > > >>issue has been repeatedly addressed in all major Histotechnology texts >> >> >since > > >>before the early fifties ( my student days in the UK). >> >>The following information is from the 10th Edition(1981) of the Condensed >>Chemical Dictionary and the10th Edition(1983) of the Merck Index (the only >>ones at hand). >> >>Formaldehyde is a gas. >>It is readily soluble in water up to 55% and is commercially available to >> >> >us > > >>as 37%, 44% and 50% aqueous solutions which may contain up to 15% >> >> >methanol. > > >>These commercial grades are called Formalin. >> >>Formaldehyde solution (Merck Index) >>The USP grade is about 37% (37-40%) w/v formaldehyde gas in water, >> >> >usually > > >>with 10-15% methanol added to prevent polymerization. This solution is >>considered to be full strength and is also known as Formalin 100% or >>Formalin 40 which signifies that it contains 40 grams of formaldehyde >> >> >within > > >>100mL of the solution. It is this solution that produces most of the >>confusion since it is referred to and thought of as 100% Formalin. >> >>Paraformaldehyde (Merck Index) >>A white crystalline powder of polymerized formaldehyde, obtained by >>concentrating formaldehyde solution. Upon solution in water >> >> >depolymerization > > >>and evolution of formaldehyde occurs. Thus an aqueous solution containing >> >> >4 > > >>grams of paraformaldehyde is essentially the same as a solution of 4% >>formaldehyde. There is NO such thing as a solution of paraformaldehyde. >>Right John? >> >>The concentration of formaldehyde used for fixation has been the subject >> >> >of > > >>much confusion (see above). The concentration of formaldehyde in compound >>fixatives varies widely - ranging from 0.5 to 15% w/v. The majority of >>fixatives, using formaldehyde as the sole fixative agent, have a >>concentration of formaldehyde between 2.5 and 4% w/v. The concentration of >>formaldehyde in a fixative should be stated as the percentage by weight of >>the gas, rather than as a percentage of the >>formalin(sic) or paraformaldehyde(sic) used to prepare it. >> >>Thus: >>"4% formaldehyde" - not 10% formalin. >>"4% formaldehyde, from paraformaldehyde" - not 4% paraformaldehyde. "NBF >>means Neutral buffered formaldehyde" - (not formalin) and is 4% w/v >>formaldehyde in phosphate buffer pH 7.0- 7.2. >> >>Bryan Hewlett >> >>Consultant Technologist >>QMP-LS >> >> >> >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. >Thank you. Saint Josephs Health System, Inc. > > > >------------------------------------------------------------------------ > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From JWEEMS <@t> sjha.org Fri Feb 4 16:09:19 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] RE: Formaldehyde vs Formalinvs paraformaldehyde(lengthy) Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507A8E@sjhaexc02.sjha.org> That's right. I plum forgot. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ford Royer Sent: Friday, February 04, 2005 5:16 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Formaldehyde vs Formalinvs paraformaldehyde(lengthy) No, no, Joyce. You are all confused. A "Formal" is a bright red farm tractor. My uncle had three of them, and he wasn't any kind of doctor at all. ~ Ford ;-) Weems, Joyce wrote: >And all this time I thought it was how to dress for a party. I could have blown up! > >Have a good weekend everyone... j:>) > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bryan >Hewlett >Sent: Friday, February 04, 2005 2:55 PM >To: Morken, Tim - Labvision; histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] RE: Formaldehyde vs Formalin vs >paraformaldehyde(lengthy) > > >Tim, > >"Formal" is quite different, it is another name for Methylal >(dimethoxymethane) a flammable colourless, volatile liquid used as a >solvent. > >Bryan > >----- Original Message ----- >From: "Morken, Tim - Labvision" >To: >Sent: Friday, February 04, 2005 2:33 PM >Subject: RE: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde >(lengthy) > > > > >>I think "Formalin" and "Formal" were trade names from a company and just >>went into common usage. >> >> >>Tim Morken >> >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan >>Hewlett >>Sent: Friday, February 04, 2005 11:24 AM >>To: histonet@lists.utsouthwestern.edu >>Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde >>(lengthy) >> >> >>I was hoping that John Kiernan would jump in reply to this issue with his >>usual eloquence! However, here goes. >> >>Confusion in terminology has been common since Blum introduced this agent >> >> >as > > >>a fixative in 1893! It never ceases to amaze me that this should be so, >> >> >the > > >>issue has been repeatedly addressed in all major Histotechnology texts >> >> >since > > >>before the early fifties ( my student days in the UK). >> >>The following information is from the 10th Edition(1981) of the Condensed >>Chemical Dictionary and the10th Edition(1983) of the Merck Index (the only >>ones at hand). >> >>Formaldehyde is a gas. >>It is readily soluble in water up to 55% and is commercially available to >> >> >us > > >>as 37%, 44% and 50% aqueous solutions which may contain up to 15% >> >> >methanol. > > >>These commercial grades are called Formalin. >> >>Formaldehyde solution (Merck Index) >>The USP grade is about 37% (37-40%) w/v formaldehyde gas in water, >> >> >usually > > >>with 10-15% methanol added to prevent polymerization. This solution is >>considered to be full strength and is also known as Formalin 100% or >>Formalin 40 which signifies that it contains 40 grams of formaldehyde >> >> >within > > >>100mL of the solution. It is this solution that produces most of the >>confusion since it is referred to and thought of as 100% Formalin. >> >>Paraformaldehyde (Merck Index) >>A white crystalline powder of polymerized formaldehyde, obtained by >>concentrating formaldehyde solution. Upon solution in water >> >> >depolymerization > > >>and evolution of formaldehyde occurs. Thus an aqueous solution containing >> >> >4 > > >>grams of paraformaldehyde is essentially the same as a solution of 4% >>formaldehyde. There is NO such thing as a solution of paraformaldehyde. >>Right John? >> >>The concentration of formaldehyde used for fixation has been the subject >> >> >of > > >>much confusion (see above). The concentration of formaldehyde in compound >>fixatives varies widely - ranging from 0.5 to 15% w/v. The majority of >>fixatives, using formaldehyde as the sole fixative agent, have a >>concentration of formaldehyde between 2.5 and 4% w/v. The concentration of >>formaldehyde in a fixative should be stated as the percentage by weight of >>the gas, rather than as a percentage of the >>formalin(sic) or paraformaldehyde(sic) used to prepare it. >> >>Thus: >>"4% formaldehyde" - not 10% formalin. >>"4% formaldehyde, from paraformaldehyde" - not 4% paraformaldehyde. "NBF >>means Neutral buffered formaldehyde" - (not formalin) and is 4% w/v >>formaldehyde in phosphate buffer pH 7.0- 7.2. >> >>Bryan Hewlett >> >>Consultant Technologist >>QMP-LS >> >> >> >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. >Thank you. Saint Josephs Health System, Inc. > > > >------------------------------------------------------------------------ > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From carl.hobbs <@t> kcl.ac.uk Sat Feb 5 12:19:53 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Surgeons and their ways Message-ID: <001f01c50baf$49cb9bf0$0201a8c0@lynne> They are a breed of people who desperately want to change from being called "Mr" to "Dr" and then.......back to "Mr"!!!! Many times have I been on standby for receiving surgical specimens in the early evening when, not hearing anything, I have strolled down to Theatre to find out what the delay is......only to discover it in darkness ( no, they were not working in the dark, altho some may say they usually do lol).......the Surgeon cancelled and couldn't be bothered to inform us! -------------- next part -------------- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.300 / Virus Database: 265.8.5 - Release Date: 03/02/2005 From Kathy.Paton <@t> waitematadhb.govt.nz Sun Feb 6 14:40:45 2005 From: Kathy.Paton <@t> waitematadhb.govt.nz (Kathy Paton (WDHB)) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde (lengthy) Message-ID: According to The Microtomists Vade-Mecum 1937 you are absolutely correct. Formol, Formalin and Formalose are commercial names for the saturated solution. Cor lummy....you learn something new each day Kathy Paton Auckland New Zealand -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Saturday, 5 February 2005 08:34 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde (lengthy) I think "Formalin" and "Formal" were trade names from a company and just went into common usage. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Hewlett Sent: Friday, February 04, 2005 11:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde (lengthy) I was hoping that John Kiernan would jump in reply to this issue with his usual eloquence! However, here goes. Confusion in terminology has been common since Blum introduced this agent as a fixative in 1893! It never ceases to amaze me that this should be so, the issue has been repeatedly addressed in all major Histotechnology texts since before the early fifties ( my student days in the UK). The following information is from the 10th Edition(1981) of the Condensed Chemical Dictionary and the10th Edition(1983) of the Merck Index (the only ones at hand). Formaldehyde is a gas. It is readily soluble in water up to 55% and is commercially available to us as 37%, 44% and 50% aqueous solutions which may contain up to 15% methanol. These commercial grades are called Formalin. Formaldehyde solution (Merck Index) The USP grade is about 37% (37-40%) w/v formaldehyde gas in water, usually with 10-15% methanol added to prevent polymerization. This solution is considered to be full strength and is also known as Formalin 100% or Formalin 40 which signifies that it contains 40 grams of formaldehyde within 100mL of the solution. It is this solution that produces most of the confusion since it is referred to and thought of as 100% Formalin. Paraformaldehyde (Merck Index) A white crystalline powder of polymerized formaldehyde, obtained by concentrating formaldehyde solution. Upon solution in water depolymerization and evolution of formaldehyde occurs. Thus an aqueous solution containing 4 grams of paraformaldehyde is essentially the same as a solution of 4% formaldehyde. There is NO such thing as a solution of paraformaldehyde. Right John? The concentration of formaldehyde used for fixation has been the subject of much confusion (see above). The concentration of formaldehyde in compound fixatives varies widely - ranging from 0.5 to 15% w/v. The majority of fixatives, using formaldehyde as the sole fixative agent, have a concentration of formaldehyde between 2.5 and 4% w/v. The concentration of formaldehyde in a fixative should be stated as the percentage by weight of the gas, rather than as a percentage of the formalin(sic) or paraformaldehyde(sic) used to prepare it. Thus: "4% formaldehyde" - not 10% formalin. "4% formaldehyde, from paraformaldehyde" - not 4% paraformaldehyde. "NBF means Neutral buffered formaldehyde" - (not formalin) and is 4% w/v formaldehyde in phosphate buffer pH 7.0- 7.2. Bryan Hewlett Consultant Technologist QMP-LS _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Sun Feb 6 14:54:48 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Re: Formaldehyde vs Formalin (Is it formol, not formal?) References: <0556BE8AC5551E4E8AF6BB9E42509BA20328C9C6@usca0082k08.labvision.apogent.com> <00a701c50af3$6c335000$6400a8c0@mainbox> Message-ID: <42068418.1CD25A5D@uwo.ca> Dear Bryan, As always, you're right. Formal is methylal. I'd never noticed that use of "formal" before. You're quite right of course; it's all there in the Merck Index and elsewhere, for all to read. One implication of this is that we probably should not write "formal-saline." This was advocated by J.R.Baker (Principles of Biological Microtechnique) on the grounds that formaldehyde was an ALdehyde, and not and alcohOL as implied by "formol." A little methylal (= "formal") is present in commercial formalin (= aqueous formaldehyde, 37%w/w = 40%w/v) because of the small amount of methanol that's added to retard polymerization, which results in precipitation of insoluble paraformaldehyde (= polyoxymethylene). There is an inconsistency in the Merck Index (I'm looking at the 12th edition, 1996) that amounts to an error [Gasp!] in the "Formaldehyde" entry. There it states that cooling or evaporation of an aqueous formaldehyde solution yields solid trioxymethylene. That's wrong; the solid stuff is paraformaldehyde, a much higher polymer. Trioxymethylene is a cyclic trimer - 3 formaldehyde molecules joined in a small ring - more often called trioxane. The Merck Index entries for "Paraformaldehyde" and "s-Trioxane" are up to date and correct. The former states that the name trioxymethylene has, in the past, been wrongly applied to paraformaldehyde. Less than 30 years ago I had at least one jar of paraformaldehyde from a major N. American supplier with "(trioxymethylene)" also on the label, as if it were a synonym. Trioxane (real trioxymethylene) is very different from paraformaldehyde. It dissolves in water without much depolymerization - just enough to yield safely antiseptic amounts of formaldehyde. It does not depolymerize at all at pH >7 and could not simply be used as a solid source of formaldehyde for making neutral buffered fixatives. My source of chemistry for all the above is JF Walker's book, "Formaldehyde" (3rd ed 1964; reprinted by Krieger, NY in 1975). I was lucky enough to find one in a 2nd hand bookshop in Britain about 15 years ago, quite by chance. John. ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 ______________________________________________________ Bryan Hewlett wrote: > > Tim, > > "Formal" is quite different, it is another name for Methylal > (dimethoxymethane) a flammable colourless, volatile liquid used as a > solvent. > > Bryan > > ----- Original Message ----- > From: "Morken, Tim - Labvision" > To: > Sent: Friday, February 04, 2005 2:33 PM > Subject: RE: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde > (lengthy) > > > I think "Formalin" and "Formal" were trade names from a company and just > > went into common usage. > > > > > > Tim Morken > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan > > Hewlett > > Sent: Friday, February 04, 2005 11:24 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] RE: Formaldehyde vs Formalin vs paraformaldehyde > > (lengthy) > > > > > > I was hoping that John Kiernan would jump in reply to this issue with his > > usual eloquence! However, here goes. > > > > Confusion in terminology has been common since Blum introduced this agent > as > > a fixative in 1893! It never ceases to amaze me that this should be so, > the > > issue has been repeatedly addressed in all major Histotechnology texts > since > > before the early fifties ( my student days in the UK). > > > > The following information is from the 10th Edition(1981) of the Condensed > > Chemical Dictionary and the10th Edition(1983) of the Merck Index (the only > > ones at hand). > > > > Formaldehyde is a gas. > > It is readily soluble in water up to 55% and is commercially available to > us > > as 37%, 44% and 50% aqueous solutions which may contain up to 15% > methanol. > > These commercial grades are called Formalin. > > > > Formaldehyde solution (Merck Index) > > The USP grade is about 37% (37-40%) w/v formaldehyde gas in water, > usually > > with 10-15% methanol added to prevent polymerization. This solution is > > considered to be full strength and is also known as Formalin 100% or > > Formalin 40 which signifies that it contains 40 grams of formaldehyde > within > > 100mL of the solution. It is this solution that produces most of the > > confusion since it is referred to and thought of as 100% Formalin. > > > > Paraformaldehyde (Merck Index) > > A white crystalline powder of polymerized formaldehyde, obtained by > > concentrating formaldehyde solution. Upon solution in water > depolymerization > > and evolution of formaldehyde occurs. Thus an aqueous solution containing > 4 > > grams of paraformaldehyde is essentially the same as a solution of 4% > > formaldehyde. There is NO such thing as a solution of paraformaldehyde. > > Right John? > > > > The concentration of formaldehyde used for fixation has been the subject > of > > much confusion (see above). The concentration of formaldehyde in compound > > fixatives varies widely - ranging from 0.5 to 15% w/v. The majority of > > fixatives, using formaldehyde as the sole fixative agent, have a > > concentration of formaldehyde between 2.5 and 4% w/v. The concentration of > > formaldehyde in a fixative should be stated as the percentage by weight of > > the gas, rather than as a percentage of the > > formalin(sic) or paraformaldehyde(sic) used to prepare it. > > > > Thus: > > "4% formaldehyde" - not 10% formalin. > > "4% formaldehyde, from paraformaldehyde" - not 4% paraformaldehyde. "NBF > > means Neutral buffered formaldehyde" - (not formalin) and is 4% w/v > > formaldehyde in phosphate buffer pH 7.0- 7.2. > > > > Bryan Hewlett > > > > Consultant Technologist > > QMP-LS > > > > _______________________________ From bhewlett <@t> cogeco.ca Sun Feb 6 17:57:32 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] RE: Formaldehyde vs Formalin (Is it formol, not formal?) Message-ID: <001f01c50ca7$9faa5860$6400a8c0@mainbox> Dear John, Thanks for the confirmation. In spite of J.R.Baker's advocacy , perhaps a better name for 'formal-saline' would be '4% formaldehyde in saline'? As to the small amount of methylal present in commercial formaldehyde solutions, I thought that mentioning that fact might only further add to the confusion! The same inconsistency in the "Formaldehyde" entry you mention is present in my earlier edition of the Merck Index. I am on the lookout for JF Walker's book, I hope to have the same luck as you. Best regards, Bryan P.S. Great short article in 'The Cutting Edge', I look forward to part two. From cytologer <@t> msn.com Sun Feb 6 22:24:31 2005 From: cytologer <@t> msn.com (Choi Ul Soo) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] PCNA on methanol fixed cell smears along with Ki67.. Message-ID: Dear histonetters! Hello, I am a graduate student, DVM, studying on immunocytochemical evaluation of some proliferation markers on methanol fixed cell smears from canine mammary tumors. Markers are PCNA, Ki67 antibodies, and some others. Smears have been stained with Diff Quik, a modified Wright stain using methanol fixation. As it turned out, I have only a few cases PCNA positive i.e. 7 / 47 cases. Correponding tissue sections have also been immunostained and all are positive with PCNA with variable positivities. I have used Oncogene Cat # NA03 PCNA antibody. The reagents, and immunostaining procedures are same both for smears and tissue sections except that the smears are stored ones and transferred to adhesive slides using liquid mounting medium. I have successfully retrieved Ki67 antigens on methanol fixed smears after trasferring to adhesive slides using antigen retrieval. My question is that 'Is there anybody who has ever tried PCNA on methanol fixed smears, and got similar experiences Or, who has ever heard of similar issues of PCNA like this?' Any information of any kind regarding this issue would be appreciated. I would gratefully receive any guidances or advices on this problem, because I should write up a 'discussion' section of the article. Lots of thanks in advance! Ul Soo Choi DVM, graduate student, VMTH of CVM, Seoul National Univ., Seoul, Korea _________________________________________________________________ ??? ???? ??? ????, ???... ??? ?? http://www.msn.co.kr/money/interlotto/ From ASelf <@t> gmhsc.com Mon Feb 7 05:07:17 2005 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Histotech Job Opportunity Message-ID: <39836CD6DB61654E8F95A35898C92186D70DA1@exchange.gmhpost.com> We have an opening for a histotechnician at Georgetown Memorial Hospital in Georgetown, SC. Short drive to Myrtle Beach and Charleston. This is a full time day shift position. If interested, or for more details you can e-mail me directly or give me a call @ 843-527-7179. Thank You, Amy Self Georgetown Memorial Hospital Georgetown, SC Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or an agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. From Diane.Gladney <@t> se.amedd.army.mil Mon Feb 7 05:44:56 2005 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Resolution of 37% case Message-ID: <4D55B2E997EFAE4DA6081DDE100B83022B0F7B@amedmlsermc133.amed.ds.army.mil> As always, the Histotech gets the blame. The surgeon is never wrong and refuses to listen to the one person that has the expertise to know that he is "way off base" to ask for 37% formaldehyde. Thankfully, the patient's specimen wasn't totally ruined since the Histotech did the right thing when it arrived at the lab. It gets very frustrating to be treated like you are an idiot. The Histotechs are still the "red-headed step children" of the lab and our profession is not given the respect that it deserves. I have to deal with this prejudice each and every day. It doesn't help when your pathologist tells you to do whatever the surgeon wants knowing full well that the surgeon is wrong. Now that I have that off my chest, keep doing the great job that you do for your patients. Stay strong and stand your ground. This is the only way that I have been able to make a difference. I have had the rare opportunity to have a pathologist that would not let the surgeons "walk all over" his histotechs. Hang in there! Happy Cutting, Diane Diane C. Gladney, HT (ASCP) Supervisor, Histology/Cytology Moncrief Army Community Hospital Dept. of Pathology 4500 Stuart Avenue P.O. Box 484 Ft. Jackson, SC 29207 Email: diane.gladney@se.amedd.army.mil Phone: 803-751-2530 FAX: 803-751-7829 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Friday, February 04, 2005 3:21 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Resolution of 37% case Dear all, Thanks to all that responded to my question about a surgeon putting a liver biopsy in "formaldehyde". All of the responses lightened the day. And now the results are in. 1. The "meek" (formerly cranky) surgeon admitted there must have been a misunderstanding between himself and his surgery tech team, but the histo tech. should have known that there was a misunderstanding and should have not sent the 37%. 2. The specimen survived abet some formalin pigment. Thanks again to all! Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BlazekL <@t> childrensdayton.org Mon Feb 7 08:31:43 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Clinicians surgeons and pathologists Message-ID: Then there were the five doctors that went duck hunting. They were sitting in the duck blind when a duck flew over. The obstetrician stood up to shoot, and said "wait a minute, I think that's a male and I don't deal with males." Another duck flew over and the family practitioner stood up to shoot and said "Oh wait a minute, I'm not sure if it is a duck. It might be a duck, but I think I'll wait and see." Then a third duck flew over. The psychiatrist stood up and said "That is a duck! But wait, does he know he's a duck?" and sat back down. When the fourth duck flew over the surgeon jumped up and shot it and turned to the pathologist and said "Go see if that's a duck". Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Morken, Tim - Labvision" 02/04/2005 3:35:36 PM >>> While clinicians know something about everything and surgeons know everything about something, pathologists know everything about everything - but it's too late. Tim Morken And a surgeon once said of pathologists that the reason they always seem to know everything is that their elbows are closer to the books than other people's are. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Ford Royer [mailto:froyer@bitstream.net] Sent: 03 February 2005 21:44 To: histonet@pathology.swmed.edu Subject: Re: [Histonet] 37% formaldehyde My dear 'ol pathologist, God rest his soul, would always say... "If you want to hide a $10.00 bill from a surgeon, put it in a textbook!" ~ Ford ;-) Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Bell, Lynne wrote: >As one of our pathologists always says, "You can always tell a surgeon >- but you can't tell him much!" > >Lynne A. Bell, HT (ASCP) >Central Vermont Hospital >P. O. Box 547 >Barre, VT 05641 >802-371-4122 > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Mon Feb 7 08:50:34 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Clinicians surgeons and pathologists Message-ID: <020720051450.7903.4207803A0009667600001EDF2200751150CECE030E9D0C9A03@comcast.net> I think I know these guys!! Good One. Pam -------------- Original message -------------- > Then there were the five doctors that went duck hunting. They were > sitting in the duck blind when a duck flew over. The obstetrician stood > up to shoot, and said "wait a minute, I think that's a male and I don't > deal with males." Another duck flew over and the family practitioner > stood up to shoot and said "Oh wait a minute, I'm not sure if it is a > duck. It might be a duck, but I think I'll wait and see." Then a third > duck flew over. The psychiatrist stood up and said "That is a duck! > But wait, does he know he's a duck?" and sat back down. When the > fourth duck flew over the surgeon jumped up and shot it and turned to > the pathologist and said "Go see if that's a duck". > > Linda Blazek, HT (ASCP) > Department of Pathology > Children's Medical Center > Dayton, Ohio 45404 > (937) 641-3358 > fax (937)641-5482 > blazekl@childrensdayton.org > > > >>> "Morken, Tim - Labvision" 02/04/2005 > 3:35:36 PM >>> > > While clinicians know something about everything and surgeons know > everything about something, pathologists know everything about > everything - > but it's too late. > > Tim Morken > > > > And a surgeon once said of pathologists that the reason they always > seem to > know everything is that their elbows are closer to the books than > other > people's are. > > Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant > Pathologist Rotherham General Hospital South Yorkshire England > terry.marshall@rothgen.nhs.uk > > -----Original Message----- > From: Ford Royer [mailto:froyer@bitstream.net] > Sent: 03 February 2005 21:44 > To: histonet@pathology.swmed.edu > Subject: Re: [Histonet] 37% formaldehyde > > > My dear 'ol pathologist, God rest his soul, would always say... "If you > > want to hide a $10.00 bill from a surgeon, put it in a textbook!" > > ~ Ford ;-) > > Ford M. Royer, MT(ASCP) > Midwest Science Biocenter > Minneapolis, MN > > > Bell, Lynne wrote: > > >As one of our pathologists always says, "You can always tell a surgeon > > >- but you can't tell him much!" > > > >Lynne A. Bell, HT (ASCP) > >Central Vermont Hospital > >P. O. Box 547 > >Barre, VT 05641 > >802-371-4122 > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Mon Feb 7 09:22:37 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Clinicians surgeons and pathologists Message-ID: My version is that the surgeon washes his hand before he pees, the physician after he pees, and the pathologist in his pee, 'cos he knows that it's sterile. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Ford Royer [mailto:froyer@bitstream.net] Sent: 04 February 2005 22:08 To: Morken, Tim - Labvision Cc: histonet@pathology.swmed.edu Subject: Re: [Histonet] Clinicians surgeons and pathologists AND of course... What's the difference between a Urologist and a Pathologist? A Urologist washes his hands BEFORE he goes to the bathroom..... Morken, Tim - Labvision wrote: >While clinicians know something about everything and surgeons know >everything about something, pathologists know everything about everything - >but it's too late. > >Tim Morken > > > >And a surgeon once said of pathologists that the reason they always seem to >know everything is that their elbows are closer to the books than other >people's are. > >Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant >Pathologist Rotherham General Hospital South Yorkshire England > terry.marshall@rothgen.nhs.uk > >-----Original Message----- >From: Ford Royer [mailto:froyer@bitstream.net] >Sent: 03 February 2005 21:44 >To: histonet@pathology.swmed.edu >Subject: Re: [Histonet] 37% formaldehyde > > >My dear 'ol pathologist, God rest his soul, would always say... "If you >want to hide a $10.00 bill from a surgeon, put it in a textbook!" > >~ Ford ;-) > >Ford M. Royer, MT(ASCP) >Midwest Science Biocenter >Minneapolis, MN > > >Bell, Lynne wrote: > > > >>As one of our pathologists always says, "You can always tell a surgeon >>- but you can't tell him much!" >> >>Lynne A. Bell, HT (ASCP) >>Central Vermont Hospital >>P. O. Box 547 >>Barre, VT 05641 >>802-371-4122 >> >> >> > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ASelf <@t> gmhsc.com Mon Feb 7 10:06:58 2005 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Histotech Job Opportunity Message-ID: <39836CD6DB61654E8F95A35898C92186D70DA2@exchange.gmhpost.com> If this is a repeat I apologize - we are having computer problems this morning..... We have an opening for a histotechnician at Georgetown Memorial Hospital in Georgetown, SC. Short drive to Myrtle Beach and Charleston. This is a full time day shift position. If interested, or for more details you can e-mail me directly or give me a call @ 843-527-7179. Thank You, Amy Self Georgetown Memorial Hospital Georgetown, SC Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or an agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. From Bultema.Brent <@t> mayo.edu Mon Feb 7 11:25:45 2005 From: Bultema.Brent <@t> mayo.edu (Bultema, Brent T.) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Histology Technician Career Opportunity - Mayo Clinic Message-ID: <3CE454ED960EEB4387BDE95E401776A801E67B32@excsrv18.mayo.edu> Histology Technician Mayo Clinic in Rochester, MN, invites you to consider a rewarding career in our Department of Laboratory Medicine and Pathology. Due to significant growth, we seek a Histology Technician to perform technical procedures including: accessioning and processing specimens; microtomy; paraffin, frozen, plastics; staining - routine H&E, immunofluorescence, immunohistochemical, and in situ hybridization. Available openings include one day shift with a variable schedule and one evening shift from 3-11pm. Prefer Associate's Degree with a combination of 12 semester hours of biology and chemistry. HT (ASCP) required. Must also have QIHC (ASCP) certification, or attainment within two years of hire for Immunostaining Laboratory. Laboratory equipment operated includes: balance, microtome, cryostat, tissue processor, autostainer, coverslipper, pH meter, pipette, and waterbath. Additional opportunities in the clinical labs are available, including careers as Lab Assistants, Technical Specialists, Development Technologists and Quality/Education Specialists. As one of the largest clinical laboratories in the world, a highly skilled team of professionals throughout our 40 specialty laboratories perform more than 11 million tests per year. Mayo Clinic also serves as a reference laboratory for clinics and hospitals in the United States and abroad. To apply, please visit www.mayoclinic.org/jobs-rst and reference job posting #2880. For more information, please contact: Mayo Clinic Tina Cronk, Human Resources Phone: 800-562-7984 Mayo Clinic is an affirmative action and equal opportunity employer. Post-offer/pre-employment drug screening is required. From JNocito <@t> Pathreflab.com Mon Feb 7 11:48:10 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Testing Water for Resistively? Message-ID: Hey Histoland, CAP is putting me through the fire. How does one check for resistively in water? Do you send it out for testing or do you have a meter. CAP didn't like my answers the first time. Thanks in advance. Joe From tpmorken <@t> labvision.com Mon Feb 7 12:23:53 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Testing Water for Resistively? Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328C9D5@usca0082k08.labvision.apogent.com> Joe, Whoever supplies your distilled water should be able to supply this number for you. Otherwise, you can either get a resistivity probe for your pH meter (to do occasional checks), or you can get a resistivity meter that attaches directly to your distilled water supply pipe. It measures the ion level of the water, so if it is deionized, it will have high resistivity (will resist electric current, will not conduct electricity well). Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Monday, February 07, 2005 9:48 AM To: Histonet Subject: [Histonet] Testing Water for Resistively? Hey Histoland, CAP is putting me through the fire. How does one check for resistively in water? Do you send it out for testing or do you have a meter. CAP didn't like my answers the first time. Thanks in advance. Joe _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From miffordclark <@t> optonline.net Mon Feb 7 14:33:09 2005 From: miffordclark <@t> optonline.net (clifford berger) Date: Fri Sep 16 15:24:35 2005 Subject: [Histonet] Testing Water for Resistively? References: <0556BE8AC5551E4E8AF6BB9E42509BA20328C9D5@usca0082k08.labvision.apogent.com> Message-ID: <000601c50d54$3c8f5ed0$0300a8c0@dellovo0ll7kuk> It should be noted that once Dl water leaves the piping, its resistivity will drop because the water absorbs dissolved carbon dioxide from the atmosphere. -Cliff Berger Decal Chemical Corp www.decal-bone.com ----- Original Message ----- From: "Morken, Tim - Labvision" To: "'Joe Nocito'" ; Sent: Monday, February 07, 2005 1:23 PM Subject: RE: [Histonet] Testing Water for Resistively? > Joe, > > Whoever supplies your distilled water should be able to supply this number > for you. Otherwise, you can either get a resistivity probe for your pH > meter > (to do occasional checks), or you can get a resistivity meter that > attaches > directly to your distilled water supply pipe. It measures the ion level of > the water, so if it is deionized, it will have high resistivity (will > resist > electric current, will not conduct electricity well). > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito > Sent: Monday, February 07, 2005 9:48 AM > To: Histonet > Subject: [Histonet] Testing Water for Resistively? > > > Hey Histoland, > CAP is putting me through the fire. How does one check for resistively in > water? Do you send it out for testing or do you have a meter. CAP didn't > like my answers the first time. Thanks in advance. > > > > Joe > _______________________________________________ > Histonet mailing list > Histonet@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 Mon Feb 7 14:34:52 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Testing Water for Resistively? In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328C9D5@usca0082k08.labvision.apogent.com> Message-ID: <200502072034.j17KYre7016638@chip.viawest.net> Joe, We had a meter and passed it around the whole department and checked resistivity ? Once per month I believe. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Monday, February 07, 2005 11:24 AM To: 'Joe Nocito'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Testing Water for Resistively? Joe, Whoever supplies your distilled water should be able to supply this number for you. Otherwise, you can either get a resistivity probe for your pH meter (to do occasional checks), or you can get a resistivity meter that attaches directly to your distilled water supply pipe. It measures the ion level of the water, so if it is deionized, it will have high resistivity (will resist electric current, will not conduct electricity well). Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Monday, February 07, 2005 9:48 AM To: Histonet Subject: [Histonet] Testing Water for Resistively? Hey Histoland, CAP is putting me through the fire. How does one check for resistively in water? Do you send it out for testing or do you have a meter. CAP didn't like my answers the first time. Thanks in advance. Joe _______________________________________________ Histonet mailing list Histonet@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 MTitford <@t> aol.com Mon Feb 7 14:35:33 2005 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Happy Mardi Gras! Message-ID: <22222366.279C938F.00762DB1@aol.com> Happy Mardi Gras to everyone from Alabama's Gulf Coast! Don't call me to-morrow - it's a day off work!! Mike Titford USA Pathology Mobile AL USA From cbass <@t> bidmc.harvard.edu Mon Feb 7 15:25:03 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] marking liver in vivo for later collection Message-ID: Hello everyone, This may be more of a surgery question than histology, but as the knowledge on this forum seems rather diverse I was hoping someone could help me. I am injecting various viral vectors directly into the liver (i.e. intrahepatically) of mice. As you can imagine, the incision for this injection is rather small compared to the entire liver. When I collect the tissue, it is sometimes difficult to determine where the injection was made. As my virus doesn't seem to diffuse very far from the injection site, I would like to mark the injection site in some way. Right now when I collect the tissue I can usually narrow it down to the correct lobe. Is there any way to mark the liver at the time of injection so that collection is easy later? I have heard of surgeons burning their initials into tissue that they were removing for later analysis. Could this be possible, perhaps just a small prick with a heated probe? I don't want to do anything that is unethical, could compromise the health of the animal, or cause undue stress or pain. Any advice would be appreciated. It is so much easier with the brain where you can see the site of injection and follow a needle track. Thanks, Caroline Bass From juan.gutierrez <@t> christushealth.org Mon Feb 7 17:26:01 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Testing Water for Resistively? Message-ID: We have a meter on the line. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Monday, February 07, 2005 11:48 AM To: Histonet Subject: [Histonet] Testing Water for Resistively? Hey Histoland, CAP is putting me through the fire. How does one check for resistively in water? Do you send it out for testing or do you have a meter. CAP didn't like my answers the first time. Thanks in advance. Joe _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From slimwillie <@t> cox.net Tue Feb 8 10:49:45 2005 From: slimwillie <@t> cox.net (Jerry Wilson) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Productivity References: <000901c50a73$13d26700$432e6044@no.cox.net> <6.0.0.22.1.20050204082739.01b0cca0@gemini.msu.montana.edu> Message-ID: <001f01c50dfe$31bdbe20$432e6044@no.cox.net> Thanks to everyone who replied to this question. Big help. I received the report from NSH today. Looks very helpful. Thanks again Jerry Wilson ----- Original Message ----- From: "Gayle Callis" To: "Jerry Wilson" ; Sent: Friday, February 04, 2005 9:34 AM Subject: Re: [Histonet] Productivity > Jerry, > > There is a superb publication "A report from the National Society for > Histotechnology Productiviey Task Force", LaFreiniere M, Sheppard B and > Carson F. 27(4):293-295 J Histotechnology Dec 2004. It will answer a lot > of your questions, including times to section blocks etc. > > If you have an NSH member or are a member yourself, you will have journal > in hand. If not, you can obtain an electronic copy from NSH, just email > them at histo@nsh.org and request this special report by title, issue. > > > At 09:36 PM 2/3/2005, you wrote: > >I think this question may have been addressed previously, but could I get > >a general idea from HIsto Tech's that process, embed and section a general > >mix of specimens (hospital surgicals, skins, gi's, etc.) of what volumes > >an individual tech would be expected to complete in one hour? > >For instance: How many blocks could one tech be reasonably expected to > >embed in one hour? How many blocks would one expect to section in one hour? > >I understand that everyone is different in the way they perform certain > >functions, but a general average of what would be expected would be > >appreciated. > > > >Thanks > >Jerry Wilson > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > From pathrm35 <@t> adelphia.net Tue Feb 8 12:22:40 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] automatic fire extinguishing system Message-ID: <28069694.1107886960732.JavaMail.root@web3.mail.adelphia.net> We are in the process of building a new lab. Does anyone know if an automatic fire extinguishing system (overhead sprinkler sytem type) is required for private histology laboratories? Thanks, Ron From jengirl1014 <@t> yahoo.com Tue Feb 8 12:26:08 2005 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] (no subject) Message-ID: <20050208182608.94549.qmail@web60604.mail.yahoo.com> Happy Mardi Gras and Happy Chinese New Year, too! May you all eat, drink, and be merry! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From CCollins <@t> propathlab.com Tue Feb 8 12:51:05 2005 From: CCollins <@t> propathlab.com (Cory Collins) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Respiratory Syncytial Virus (RSV) Message-ID: Hey Histonetters, I'm the supervisor of an IHC Lab in Dallas (ProPath) and my pathologist is looking for a known positive control for Respiratory Syncytial Virus (RSV). We have numerous other postive control and are willing to trade if someone has some extra RSV. Thanks a bunch, Cory Cory Collins, HT (ASCP) QIHC Immunohistochemistry Supervisor ProPath 8267 Elmbrook Drive Suite 100 Dallas, TX 75247 214-638-2000 ext 2027 214-237-1730 Fax To learn more about ProPath, please visit http://www.ProPathLab.com ______________________________________________________________________________ This e-mail may contain confidential or privileged information. If you think you have received this e-mail in error, please advise the sender by reply e-mail and then delete this e-mail immediately. From POWELL_SA <@t> Mercer.edu Tue Feb 8 15:47:11 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Georgia Society for Histotechnology Meeting Message-ID: The annual GSH meeting will be held in Atlanta this year. I have added the program, hotel information and registration form below. Once again in the year 2005 the Georgia Society of Cytology and the Georgia Society for Histotechnology have joined to present a combined annual symposium. This year we are not only repeating the joint venture but we are also returning to a previous location ? the Hyatt Regency Suites Perimeter Northwest ? Atlanta. The Hyatt features luxuriously decorated rooms, indoor swimming pool and health club as well as an excellent on site restaurant. The hotel offers complimentary shuttle service within a 5 mile radius and covers a wide variety of restaurant and evening entertainment opportunities. A block of rooms is reserved until February 18, 2005. Mention the Georgia Society of Histotechnology to receive the single/double rate of $84.00, triple/quad rate of $94.00 and Regency Suite rate of $375.00. Call 770-956-1234 or fax 770-916-1120 to make your reservations. Joint Sessions of histology and cytology will be held featuring current topics of common interest. Registration for your respective program gives you access to the other society?s program at no additional charge. A special treat this year will be the dinner theater with a scientific flavor. Many scientific companies will be present to exhibit laboratory products and services along with the most current products and technologies for both cytology and histology. Following the Dinner Theater an opportunity of mix and mingle with venders and colleagues in the exhibit area will be available. Drawings for completed Vender Bingo as well as other drawings and prizes will be held at this time. You must be present to win. For more information on the area/events visit www.accessatlanta.com. Room Reservations: The host hotel is the Hyatt Regency Suites Perimeter Northwest ? Atlanta. Room reservations should be made directly with the Hyatt by calling 770-956-1234 or faxing 770-916-1120. When making your reservation, identify yourself as an attendee of the Georgia Society of Histotechnology/Cytology meeting. The following rates are guaranteed until Feb. 18, 2005 Single/double occupancy $ 84.00 Triple/quad occupancy $ 94.00 Regency Suite $ 375.00 Check-in time 3:00 p.m. Check-out time 12:00 p.m. Directions: From I-285 take I-75 North. Take Exit 260 Windy Hill Road; go east ? miles. The Hyatt Regency suites will be on the left, before the Powers Ferry intersection. Program Histology Cytology Saturday, March 5, 2005 7:30 ? 8:30 a.m. Registration 8:00 ? 9:00 a.m. Registration 8:30 a.m. ? 12:00 p.m. 9:00 ? 10:00 a.m. Lymphoma Panel Workups Atypical Glandular Cells of Undetermined Significance: Laura Hughs Dr. Stephen Lau, Grady Health System Sponsored by Cell Marque Corp. 10:00 ?10:30 a.m. 9:30 ? 10:00 a.m. Break ? View Exhibits Break ? View Exhibits 10:30 ? 11:30 Lung Fine Needle Aspiration Dr. Roger Lane, Southeastern Pathology 11:30 ? 12:00 Pap Smear Litigation Update Dr. Roger Lan 12:00 ? 1:30 p.m. Lunch ? View Exhibits 12:00 ? 1:30 p.m. Lunch ? View Exhibits Joint Cytology/Histology Sessions 1:30 - 2:30 p.m. Obesity: What?s Happening in Our Society? Annie B. Carr, MS, RD, CDC 2:30 ? 3:00 p.m. Break ? View Exhibits 3:00 ? 4:30 p.m Case Studies in Forensic Pathology Geoffry Smith M.D. Fulton Cty Med Examiner 4:30 ? 5:45 p.m Nipple Duct Lavage; Cytology and Histology Joanne Piratzky, M.D. 5:45 p.m. GSH General Membership Meeting 5:45 p.m. GSC General Membership Meeting 6:30 ? 10:30 p.m. Dinner Theater/Exhibits/Cash Bar Sunday, March 6, 2004 7:30-8:00 a.m. Registration 8:00 ? 9:00 a.m. Registration 8:00 ? 10:30 a.m. 9:00- 10:00 a.m. The Whole Enchilada Urocyt Donna Willis HT/HTL(ASCP) Brian Hancock, Cytyc Corp 10:30 ? 11:00 a.m. 10:00 ? 10:30 a.m. Break Break 11:00 a.m. ? 1:00 p.m. 10:30 ? 11:30 a.m. The Fun Side of Presenting a Workshop FNA, Preparation and Fundamentals Linda Jenkins HT(ASCP) Dr. Julie Baird, Grady Health System 1:30 ? GSH BOD meeting Symposium Registration: To register, complete the Registration form using with either the Histology or Cytology portion only. Registrations postmarked after February 21, 2005 must include a late fee of $10.00. Histology: Full symposium registration fee includes Saturday and Sunday symposium sessions (including the ability to attend Cytology lectures at no additional charge), Saturday lunch break, Saturday Dinner Theater and Exhibit viewing and all morning and afternoon breaks. One day only fees are available for either Saturday or Sunday and include only activities and sessions scheduled for that day. Additional Saturday Dinner Theater tickets are $30.00 and MUST be ordered in advance. Registrations postmarked after February 21, 2005 must include a late fee of $10.00. Your annual GSH dues may be paid with your registration, entitling you to the member discount. Annual dues are $10.00 for the calendar year 2005. Make check payable to: Georgia Society for Histotechnology. Cytology: Full symposium registration fee includes Saturday and Sunday symposium sessions (including the ability to attend Histology lectures at no additional charge), Saturday lunch break, Saturday Dinner theater and Exhibit viewing and all morning and afternoon breaks and GSC annual dues. Additional Saturday Dinner Theater tickets are $30.00 and MUST be ordered in advance. Registrations postmarked after February 21, 2005 must include a late fee of $10.00. Make check payable to: Georgia Society of Cytology. Program Cancellation Policies: Refunds can only be issued for cancellation requests received by February 21, 2005. NO refunds will be made after that date for any reason. Attendance by a substitute person is encouraged in lieu of cancellation. Please return registration form and fees (for both cytology and histology) to: Connie Wavrin GSC/GSH Symposium Chairperson 2667 Meadow Court Chamblee, GA 30341 We are unable to accept credit card registrations. Additional information is available from: Histology Cytology Venders Connie Wavrin Shirley VanDuzer Ruby Huggins-Rodriquez w. ph. 678-443-2332 w. ph. 404-321-6111 ext 4081 h. ph. 770-513-3290 fax: 678-443-2339 fax: 404-235-3007 email: Rodriquezr@bellsouth.net email: cwavrin@labmd.org email: Shirley.vanduzer@med.va.gov GSH NEWS UPDATE The good news continues! Dues were free in 2003; half price in 2004 and due to good financial health dues will remain half price in 2005. That?s just $10.00! Name: _________________________________________________________________ Home Mailing Address: __________________________________________________ City, State, Zip Code: _____________________________________________________ Organization/Employer: ____________________________________________________ Work Mailing Address: ____________________________________________________ City, State, Zip Code: ______________________________________________________ W.Ph: ( )__________________H.Ph.: ( )_______________ Fax: ( )________________ Home e-mail:____________________________________________Work email__________________________________ Dues must be current and paid in full to be eligible for member discount. Include with symposium registration or send to: Shirley Powell 156 Oakridge Ave Macon, GA 31204 From NMargaryan <@t> childrensmemorial.org Tue Feb 8 16:48:57 2005 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Monoclonal Anti-cytokeratin CK5 Message-ID: <63B8B599DE283148B92E83C78B32C15D4C89B6@cmhexbe2.childrensmemorial.org> Hello everyone, Is anybody of you used the Ab Monoclonal Anti-cytokeratin CK5, clone CK5 from SIGMA for immunostaining on FFPE tissue? If so, please, send me a protocol. Thanks a lot, Naira Naira V. Margaryan, Ph.D, D.V.M. Research Associate II Children's Memorial Research Center 2300 Children's Plaza, Box 222 Chicago, IL 60614-3394 Tel: 773-880-4000/5-6740 Fax: 773-755-6594 nmargaryan@childrensmemorial.org For Express Mail: CMRC, Room C.473 2430 N. Halsted Street Chicago, IL 60614-4314 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 shive003 <@t> umn.edu Tue Feb 8 16:54:26 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] FCV IHC Message-ID: <004c01c50e31$241b9160$41065486@auxs.umn.edu> Does anybody out there have a successful IHC protocol for Feline Calicivirus? If so, would you share it with me (along with the name of your supplier)? Thanks much in advance. Jan Shivers U of MN Vet Diag Lab From frosscis <@t> usc.edu Tue Feb 8 17:05:58 2005 From: frosscis <@t> usc.edu (Fred Ross-Cisneros) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Microglia Message-ID: <000901c50e32$c02ff810$7f607d80@sadunlab> Dear All, What would be a specific antibody (that's commercially availble) to CNS micoglia? I've heard that CD-68 and ED-1 can be used. From your experience, what is the best for human and/or murine/rodent tissues. Thanks. Fred Fred N. Ross-Cisneros Sadun Neuro-Ophthalmology Lab USC Keck School of Medicine and Doheny Eye Institute 1355 San Pablo Street, DVRC 311 Los Angeles, CA 90033-1026 Tel.: (323) 442-6667 Fax: (323) 442-6688 From bjackson <@t> unipathllc.com Wed Feb 9 11:39:56 2005 From: bjackson <@t> unipathllc.com (Brianna Jackson) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Decloaking chamber problem Message-ID: Hello Everyone, This question is for those of you using the Biocare Medical decloaking chamber or the Dako Pascal decloaking chamber. We're having trouble with the coplin jars boiling over during retrieval during the depressurizing portion of retrieval. After the alarm sounds we turn the decloaker off and don't disturb the chamber until it has completely depressurized. We've started using the taller coplin jars from Scienceware and it still happens. It doesn't seem to be dependant on the type of retrieval solution, the amount of retrieval solution in the coplin jar, barometric pressure, the number of slides or whether or not we use the little perforated disk (heat shield) in the bottom of the decloaker. Our patient specimens are always OK, being at the bottom of the slide, but we have to repeat cases that boiled over because the control tissue at the top of the slide no longer stains by IHC. If anyone has figured out a solution to this problem any help would be appreciated. Thanks, Brianna Jackson, BS, HTL(ASCP), QIHC UniPath Denver, CO 303-512-2220 From JMyers1 <@t> aol.com Wed Feb 9 11:56:51 2005 From: JMyers1 <@t> aol.com (JMyers1@aol.com) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Would a lab equipment 'refurbisher' want to buy a used IHC stainer? Message-ID: <36.6bfd7fa8.2f3ba8e3@aol.com> Dear Histonetters: I was wondering if anyone has had any experience selling a used IHC slide stainer to one of those lab equipment refurbishing/reselling companies? I consult with laboratories on acquiring new IHC stainers, and have often been asked about this possibility. I don't know if 'refurbishers' would be interested in doing this, and hoped that someone out there in cyberspace might be able to give me a little guidance. Any feedback is appreciated. J.D. Myers From bgoldman <@t> wam.umd.edu Wed Feb 9 12:03:46 2005 From: bgoldman <@t> wam.umd.edu (Beth A. Goldman) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] (no subject) Message-ID: Hi everyone. I was wondering if any of you knew where I could purchase a staining net. I'm not sure if this is actually what the product is called. It is basically a round dish about 2 inches high, with netting on the bottom. It is divided into sections by glass. I've been using some borrowed ones for staining brain tissue, with an acetylcholinesterse reaction. I'd like to buy some of my own, but I have no idea of where I can buy some. Thanks, Beth University of Maryland, College Park From shive003 <@t> umn.edu Wed Feb 9 12:04:04 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Decloaking chamber problem References: Message-ID: <004e01c50ed1$bdf12b60$41065486@auxs.umn.edu> I use Tissue Tek rectangular plastic containers and slide racks. No problem with overheating/overboiling, and I decloak in Dako Target Retrieval Solution for 20 minutes, followed by a 25 minute cooldown inside the cooker. Jan Shivers U of MN Vet Diag Lab ----- Original Message ----- From: "Brianna Jackson" To: Sent: Wednesday, February 09, 2005 11:39 AM Subject: [Histonet] Decloaking chamber problem > Hello Everyone, > > > > This question is for those of you using the Biocare Medical decloaking > chamber or the Dako Pascal decloaking chamber. We're having trouble with > the coplin jars boiling over during retrieval during the depressurizing > portion of retrieval. After the alarm sounds we turn the decloaker off and > don't disturb the chamber until it has completely depressurized. We've > started using the taller coplin jars from Scienceware and it still happens. > It doesn't seem to be dependant on the type of retrieval solution, the > amount of retrieval solution in the coplin jar, barometric pressure, the > number of slides or whether or not we use the little perforated disk (heat > shield) in the bottom of the decloaker. Our patient specimens are always > OK, being at the bottom of the slide, but we have to repeat cases that > boiled over because the control tissue at the top of the slide no longer > stains by IHC. > > If anyone has figured out a solution to this problem any help > would be appreciated. > > > > Thanks, > > > > Brianna Jackson, BS, HTL(ASCP), QIHC > > UniPath > > Denver, CO > > 303-512-2220 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From haldana <@t> unimoron.edu.ar Wed Feb 9 13:44:30 2005 From: haldana <@t> unimoron.edu.ar (Hernan Aldana Marcos) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Pineal research antibody Message-ID: <005501c50edf$c5bb27c0$a904a8c0@um.edu> Dear Histonet mailing list Does anyone know some antibody specific for pineal gland (melatonin, HIOMT, etc)? Thanks in advance Dr. Hern?n J. Aldana Marcos Facultad de Medicina. Universidad de Mor?n Machado 914. B1708JPD. Buenos Aires. Argentina e-mail alternativo hernanjavier@yahoo.com web: http://hjaldanamarcos.bravepages.com http://histologia.bigthicketdirectory.net/main.html From MAUGER <@t> email.chop.edu Wed Feb 9 13:36:31 2005 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Decloaking chamber problem Message-ID: Hi Brianna, We use the larger, rectangular 'boats' that hold a plastic slide rack(24 slide capacity) We have never had boiling over. More retrieval soln is used, but it saves you from repeating,and is more standardized. Jo >>> "Brianna Jackson" 02/09/05 12:39 PM >>> Hello Everyone, This question is for those of you using the Biocare Medical decloaking chamber or the Dako Pascal decloaking chamber. We're having trouble with the coplin jars boiling over during retrieval during the depressurizing portion of retrieval. After the alarm sounds we turn the decloaker off and don't disturb the chamber until it has completely depressurized. We've started using the taller coplin jars from Scienceware and it still happens. It doesn't seem to be dependant on the type of retrieval solution, the amount of retrieval solution in the coplin jar, barometric pressure, the number of slides or whether or not we use the little perforated disk (heat shield) in the bottom of the decloaker. Our patient specimens are always OK, being at the bottom of the slide, but we have to repeat cases that boiled over because the control tissue at the top of the slide no longer stains by IHC. If anyone has figured out a solution to this problem any help would be appreciated. Thanks, Brianna Jackson, BS, HTL(ASCP), QIHC UniPath Denver, CO 303-512-2220 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Don.Birgerson <@t> leica-microsystems.com Wed Feb 9 14:49:19 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] (no subject) Message-ID: Hi Beth, I think you want to contact a company called "Brain Research Laboratories" (www.brainsearchlab.com ) 617-965-5544 in Newton Mass. They list staining nets. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Beth A. Goldman" To: histonet@lists.utsouthwestern.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] (no subject) western.edu 02/09/2005 12:03 PM Hi everyone. I was wondering if any of you knew where I could purchase a staining net. I'm not sure if this is actually what the product is called. It is basically a round dish about 2 inches high, with netting on the bottom. It is divided into sections by glass. I've been using some borrowed ones for staining brain tissue, with an acetylcholinesterse reaction. I'd like to buy some of my own, but I have no idea of where I can buy some. Thanks, Beth University of Maryland, College Park _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Beth A. Goldman" To: histonet@lists.utsouthwestern.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] (no subject) western.edu 02/09/2005 12:03 PM Hi everyone. I was wondering if any of you knew where I could purchase a staining net. I'm not sure if this is actually what the product is called. It is basically a round dish about 2 inches high, with netting on the bottom. It is divided into sections by glass. I've been using some borrowed ones for staining brain tissue, with an acetylcholinesterse reaction. I'd like to buy some of my own, but I have no idea of where I can buy some. Thanks, Beth University of Maryland, College Park _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From bosborn <@t> hsc.usf.edu Wed Feb 9 14:57:50 2005 From: bosborn <@t> hsc.usf.edu (Barbara Osborn) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] blue flourescent nuclear dyes Message-ID: <000a01c50eea$042fe130$9d46f783@hscnet.hsc.usf.edu> We are just beginning flourescent staining for confocal microscopy in our lab, and our confocal microscope did not come with a UV laser, so we cannot detect DAPI as a nuclear stain. We've been told to use Draque 5 (sp) which emits blue in visible light instead of DAPI to counterstain nuclei. I cannot for the life of me find any info on this dye. Molecular Probes offers a blue flourescent nuclear dye called BOBO-1 iodide. Does anyone out there in histoland have any information about Draque 5, where to purchase it, or has anyone tried the BOBO-1 from Moleculare Probes? Or does anyone use an alternative nuclear dye? We plan to triple stain using nuclear dye/ Alexa Flour 488/ Alexa Flour 568 so I need something that does not overlap too much with the 488. Thanks. From leahcox27 <@t> yahoo.com Wed Feb 9 14:51:00 2005 From: leahcox27 <@t> yahoo.com (Leah Cox) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Blood smear for parasites Message-ID: <20050209205101.8997.qmail@web50206.mail.yahoo.com> I am staining a thick blood smear for parasites, probably malaria. Does anyone know a good procedure for it? --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From bgoldman <@t> wam.umd.edu Wed Feb 9 14:51:56 2005 From: bgoldman <@t> wam.umd.edu (Beth A. Goldman) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] (no subject) In-Reply-To: References: Message-ID: Thanks! On Wed, 9 Feb 2005 Don.Birgerson@leica-microsystems.com wrote: > > Hi Beth, > I think you want to contact a company called "Brain Research > Laboratories" (www.brainsearchlab.com ) 617-965-5544 in Newton Mass. They > list staining nets. > > Don Birgerson > Leica Microsystems > Technical Assistance Center > Don.Birgerson@Leica-Microsystems.Com > 1-800-248-0123 ext 5918 > > > > "Beth A. Goldman" > To: histonet@lists.utsouthwestern.edu > Sent by: cc: > histonet-bounces@lists.utsouth Subject: [Histonet] (no subject) > western.edu > > > 02/09/2005 12:03 PM > > > > > > > Hi everyone. > I was wondering if any of you knew where I could purchase a staining > net. I'm not sure if this is actually what the product is called. It is > basically a round dish about 2 inches high, with netting on the bottom. It > is divided into sections by glass. I've been using some borrowed ones for > staining brain tissue, with an acetylcholinesterse reaction. I'd like to > buy some of my own, but I have no idea of where I can buy some. > > Thanks, > > Beth > University of Maryland, College Park > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > Don Birgerson > Leica Microsystems > Technical Assistance Center > Don.Birgerson@Leica-Microsystems.Com > 1-800-248-0123 ext 5918 > > > > "Beth A. Goldman" > To: histonet@lists.utsouthwestern.edu > Sent by: cc: > histonet-bounces@lists.utsouth Subject: [Histonet] (no subject) > western.edu > > > 02/09/2005 12:03 PM > > > > > > > Hi everyone. > I was wondering if any of you knew where I could purchase a staining > net. I'm not sure if this is actually what the product is called. It is > basically a round dish about 2 inches high, with netting on the bottom. It > is divided into sections by glass. I've been using some borrowed ones for > staining brain tissue, with an acetylcholinesterse reaction. I'd like to > buy some of my own, but I have no idea of where I can buy some. > > Thanks, > > Beth > University of Maryland, College Park > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ______________________________________________________________________ > This email has been scanned by the MessageLabs Email Security System. > For more information please visit http://www.messagelabs.com/email > ______________________________________________________________________ > From leahcox27 <@t> yahoo.com Wed Feb 9 16:03:28 2005 From: leahcox27 <@t> yahoo.com (Leah Cox) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Blood smear for parasites In-Reply-To: <20050209205101.8997.qmail@web50206.mail.yahoo.com> Message-ID: <20050209220328.44639.qmail@web50207.mail.yahoo.com> Thanks to everyone who has responded so far! I originally tried the Wrights stain but it turned out way too dark, maybe because the smear is very thick. Has anyone tried to stain for less time on thicker tissue? Leah Cox wrote:I am staining a thick blood smear for parasites, probably malaria. Does anyone know a good procedure for it? --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From katherine-walters <@t> uiowa.edu Wed Feb 9 16:32:23 2005 From: katherine-walters <@t> uiowa.edu (Walters, Katherine S) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] blue flourescent nuclear dyes Message-ID: Hi Barbara, We use Draq-5 and we get it from Biostatus in the UK. Their website is http://www.biostatus.co.uk/. It is supposed to work in the far-red range, but some of our researchers have reported an excitation in 488nm and 568 nm as well. I personally have not witnessed this. Hope this helps, Kathy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barbara Osborn Sent: Wednesday, February 09, 2005 2:58 PM To: Histonet Subject: [Histonet] blue flourescent nuclear dyes We are just beginning flourescent staining for confocal microscopy in our lab, and our confocal microscope did not come with a UV laser, so we cannot detect DAPI as a nuclear stain. We've been told to use Draque 5 (sp) which emits blue in visible light instead of DAPI to counterstain nuclei. I cannot for the life of me find any info on this dye. Molecular Probes offers a blue flourescent nuclear dye called BOBO-1 iodide. Does anyone out there in histoland have any information about Draque 5, where to purchase it, or has anyone tried the BOBO-1 from Moleculare Probes? Or does anyone use an alternative nuclear dye? We plan to triple stain using nuclear dye/ Alexa Flour 488/ Alexa Flour 568 so I need something that does not overlap too much with the 488. Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Feb 9 17:33:06 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] blue flourescent nuclear dyes In-Reply-To: References: Message-ID: <6.0.0.22.1.20050209163003.01b24e60@gemini.msu.montana.edu> Interesting fluorescent dye. It says for use in live cells, but have you tried it with fixed cells. DAPI will work on fixed cells, and we have the same problem with our CLSM - did not have the laser for exciting the DAPI. Thanks for the information on Biostatus, it has been filed away!! At 03:32 PM 2/9/2005, you wrote: >Hi Barbara, > > >We use Draq-5 and we get it from Biostatus in the UK. Their website is >http://www.biostatus.co.uk/. It is supposed to work in the far-red >range, but some of our researchers have reported an excitation in 488nm >and 568 nm as well. I personally have not witnessed this. > >Hope this helps, >Kathy > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barbara >Osborn >Sent: Wednesday, February 09, 2005 2:58 PM >To: Histonet >Subject: [Histonet] blue flourescent nuclear dyes > >We are just beginning flourescent staining for confocal microscopy in >our lab, and our confocal microscope did not come with a UV laser, so we >cannot detect DAPI as a nuclear stain. We've been told to use Draque 5 >(sp) which emits blue in visible light instead of DAPI to counterstain >nuclei. I cannot for the life of me find any info on this dye. >Molecular Probes offers a blue flourescent nuclear dye called BOBO-1 >iodide. Does anyone out there in histoland have any information about >Draque 5, where to purchase it, or has anyone tried the BOBO-1 from >Moleculare Probes? Or does anyone use an alternative nuclear dye? We >plan to triple stain using nuclear dye/ Alexa Flour 488/ Alexa Flour 568 >so I need something that does not overlap too much with the 488. >Thanks. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Feb 9 17:43:51 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] blue flourescent nuclear dyes References: <000a01c50eea$042fe130$9d46f783@hscnet.hsc.usf.edu> Message-ID: <420AA037.1E7733E3@uwo.ca> If you want a blue emission you'll almost certainly need near-UV excitation. The excitation wavelength of a fluorochrome is always shorter than the wavelength of the emitted light. -- ------------------------------- 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 _______________________________ Barbara Osborn wrote: > > We are just beginning flourescent staining for confocal microscopy in our lab, and our confocal microscope did not come with a UV laser, so we cannot detect DAPI as a nuclear stain. We've been told to use Draque 5 (sp) which emits blue in visible light instead of DAPI to counterstain nuclei. I cannot for the life of me find any info on this dye. Molecular Probes offers a blue flourescent nuclear dye called BOBO-1 iodide. Does anyone out there in histoland have any information about Draque 5, where to purchase it, or has anyone tried the BOBO-1 from Moleculare Probes? Or does anyone use an alternative nuclear dye? We plan to triple stain using nuclear dye/ Alexa Flour 488/ Alexa Flour 568 so I need something that does not overlap too much with the 488. Thanks. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CrochiereSteve <@t> aol.com Wed Feb 9 21:42:01 2005 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] DeCloaker Message-ID: <6b.3e9cd94b.2f3c3209@aol.com> In a message dated 2/9/2005 1:07:23 PM Eastern Standard Time, histonet-request@lists.utsouthwestern.edu writes: bjackson@unipathllc.com Do you leave the covers off the coplin jars. I found that pressure builds up in the coplin jar if covered and this could cause your problem with boiling over. I use plastic coplin jars or the 250 ml tissue tek staining "boxes" without covers and have not experienced a boiling over incident. Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 From lpwenk <@t> sbcglobal.net Thu Feb 10 04:19:32 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Wage and Salary Survey Message-ID: <004501c50f5a$04479300$ba30d445@domainnotset.invalid> The 2003 Wage and Salary Survey from ASCP Board of Registry is now available on line. http://www.ascp.org/bor/center/center_research.asp Click on "2003 Wage and Vacancy Survey of Medical Laboratories". You will need Adobe to open the PDF file. If you don't have it, there is a link below the survey link. I couldn't find in the article exactly WHEN in 2003 the survey was taken, but if memory serves me, it was in the fall of 2003, so the information is a little over 1 year old. ASCP BOR was doing the survey every other year, and reporting the results 1 year later. Because of the market changing so quickly, they are now doing it every year. The survey breaks down the information into: - tech position (HT, HTL, supervisor, etc.) - type of lab (hospital, private, etc.) - number of beds - city size - annual testing volume - region of US Number of vacancies are not as high as the previous surveys found - but the last paragraph was interesting - "Nearly every laboratory profiled described a scenario of disappearing vacancies. After budget cuts or lengthy recruitment searches, budgeted vacancies were cut. Students are not attracted to the long hours of laboratory work. Not enough certified staff entering the workforce to replace the number of laboratory staff expected to retire in the next 3 to 5 years." Looking at only hospital vacancies, regardless of location, size, etc: MT = 4.3% CT = 4.1% HTL = 3.1% MLT = 6.1% HT = 5.3% HT/HTL Supervisor = 4.9% MT Supervisor = 3.3% MLT Supervisor = 1.2% CT Supervisor = 1% PBT = 6.3% PBT Supervisor = 3.4% Good reading. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From linhines <@t> yahoo.com Thu Feb 10 05:42:20 2005 From: linhines <@t> yahoo.com (Linda Hines) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] RE: Histoprocessing with microwave Message-ID: <20050210114220.6913.qmail@web61207.mail.yahoo.com> I have just accepted an employment offer in a lab, that uses a microwave for histoprocessing. I have never worked with tissue that is processed in the microwave. Do you use the same paraffin for infiltration and embedding? If not what paraffin is recommended? How does the tissue cut? What are some of the problems to look out for? Are you also drying slides in the micrwave? What are your procedures? Thanx in advance for your answers and help. Linda Hines HT --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From katherine-walters <@t> uiowa.edu Thu Feb 10 08:03:18 2005 From: katherine-walters <@t> uiowa.edu (Walters, Katherine S) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] blue flourescent nuclear dyes Message-ID: Hi Gayle, It is supposed to work on live and dead cells, I've only used it on fixed cells, but others in our lab have used live ones. Kathy -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Wednesday, February 09, 2005 5:33 PM To: Walters, Katherine S; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] blue flourescent nuclear dyes Interesting fluorescent dye. It says for use in live cells, but have you tried it with fixed cells. DAPI will work on fixed cells, and we have the same problem with our CLSM - did not have the laser for exciting the DAPI. Thanks for the information on Biostatus, it has been filed away!! At 03:32 PM 2/9/2005, you wrote: >Hi Barbara, > > >We use Draq-5 and we get it from Biostatus in the UK. Their website is >http://www.biostatus.co.uk/. It is supposed to work in the far-red >range, but some of our researchers have reported an excitation in 488nm >and 568 nm as well. I personally have not witnessed this. > >Hope this helps, >Kathy > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barbara >Osborn >Sent: Wednesday, February 09, 2005 2:58 PM >To: Histonet >Subject: [Histonet] blue flourescent nuclear dyes > >We are just beginning flourescent staining for confocal microscopy in >our lab, and our confocal microscope did not come with a UV laser, so we >cannot detect DAPI as a nuclear stain. We've been told to use Draque 5 >(sp) which emits blue in visible light instead of DAPI to counterstain >nuclei. I cannot for the life of me find any info on this dye. >Molecular Probes offers a blue flourescent nuclear dye called BOBO-1 >iodide. Does anyone out there in histoland have any information about >Draque 5, where to purchase it, or has anyone tried the BOBO-1 from >Moleculare Probes? Or does anyone use an alternative nuclear dye? We >plan to triple stain using nuclear dye/ Alexa Flour 488/ Alexa Flour 568 >so I need something that does not overlap too much with the 488. >Thanks. >_______________________________________________ >Histonet mailing list >Histonet@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 lyonm <@t> upstate.edu Thu Feb 10 08:42:11 2005 From: lyonm <@t> upstate.edu (Michael J. Lyon) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Developmental Myosin Control Tissue Message-ID: Hi: We are in need of a positive control for human developmental myosin antibody. Does anyone have some that they would be willing to sell? Thanks Michael J. Lyon, Ph.D. Otolaryngology Research Labs SUNY Upstate Medical University 750 East Adams Street Syracuse, NY 13210 Voice 315-464-7253 Fax 315-464-5572 From bosborn <@t> hsc.usf.edu Thu Feb 10 08:52:17 2005 From: bosborn <@t> hsc.usf.edu (Barbara Osborn) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] blue flourescent nuclear dyes References: Message-ID: <005501c50f80$1dba3420$9d46f783@hscnet.hsc.usf.edu> Thankyou for your replies. I found the info that I needed and I apparently had totally misunderstood the nature of Draq5 and it's excitation and emmission spectra. Now it all makes much more sense! Thanks! ----- Original Message ----- From: "Walters, Katherine S" To: "Barbara Osborn" ; "Histonet" Sent: Wednesday, February 09, 2005 5:32 PM Subject: RE: [Histonet] blue flourescent nuclear dyes > Hi Barbara, > > > We use Draq-5 and we get it from Biostatus in the UK. Their website is > http://www.biostatus.co.uk/. It is supposed to work in the far-red > range, but some of our researchers have reported an excitation in 488nm > and 568 nm as well. I personally have not witnessed this. > > Hope this helps, > Kathy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barbara > Osborn > Sent: Wednesday, February 09, 2005 2:58 PM > To: Histonet > Subject: [Histonet] blue flourescent nuclear dyes > > We are just beginning flourescent staining for confocal microscopy in > our lab, and our confocal microscope did not come with a UV laser, so we > cannot detect DAPI as a nuclear stain. We've been told to use Draque 5 > (sp) which emits blue in visible light instead of DAPI to counterstain > nuclei. I cannot for the life of me find any info on this dye. > Molecular Probes offers a blue flourescent nuclear dye called BOBO-1 > iodide. Does anyone out there in histoland have any information about > Draque 5, where to purchase it, or has anyone tried the BOBO-1 from > Moleculare Probes? Or does anyone use an alternative nuclear dye? We > plan to triple stain using nuclear dye/ Alexa Flour 488/ Alexa Flour 568 > so I need something that does not overlap too much with the 488. > Thanks. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From JWEEMS <@t> sjha.org Thu Feb 10 12:01:32 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Pathology Consults and Path Contracts Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507AE5@sjhaexc02.sjha.org> Hello Everyone, We are going in circles trying to obtain referral numbers for path cases being sent for consultation. We are told that for any HMO we need a referral number from the primary care physician. We don't usually know who the primary is and have to make several calls to make people understand what we are asking for. How are you handling this? Also many consultants have stopped billing patient insurance and are only billing the facility. If your pathologists are contracted by the hospital, does their private billing service handle this issue or is it a part of their contract for the hospital to handle it? Thanks for your help, 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 Instaken <@t> aol.com Thu Feb 10 12:16:58 2005 From: Instaken <@t> aol.com (Instaken@aol.com) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] automatic fire extinguishing system Message-ID: <1f6.4f76afc.2f3cff1a@aol.com> I would be interested in this also. I notice CAP has a checklist item regarding fire supression systems. How have independent labs who are CAP accredited addressed this? Thanks, Ken From Virginia.Ross <@t> nrc-cnrc.gc.ca Thu Feb 10 12:48:53 2005 From: Virginia.Ross <@t> nrc-cnrc.gc.ca (Ross, Virginia) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Raynaud's Syndrome Message-ID: <10C94843061E094A98C02EB77CFC32870A73DDCA@nrcmrdex1d.imsb.nrc.ca> I have developed Raynaud's and I am awaiting diagnosis for suspected autoimmune diseases and I have a collegue who worked with the some of the same chemicals who is disabled with mixed connective tissue disease. Raynaud's has been linked to vinyl chloride, cryostats and vibrating machines (vibrotomes) but I have not worked with any of these. I have worked with the traditional histology chemicals, solvents, and methymethacrylate. I am interested in seeing if there is a link between histologists and autoimmune diseases, and which chemicals may be implicated. I would greatly appreciate any information anyone has on any of these topics. Thank you, Virginia From BriggsK <@t> drmc.drhsi.org Thu Feb 10 13:23:33 2005 From: BriggsK <@t> drmc.drhsi.org (Kevin Briggs) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] ASR Disclaimer Message-ID: <7C1D5BE1ADF06E488AEB7A6247214A860D260D@HORNET.drmc.drhsi.org> Hi Netters! I am curious to know how other laboratories are handling the CAP Checklist item ANP.12425: "If patient testing is performed using Class I analyte-specific reagents (ASR's) obtained or purchased from an outside vendor, does the patient report include the disclaimer required by federal regulations?" I am specifically searching for examples of canned text comments or disclaimers suitable for reporting any of our antibodies simultaneously. Thanks, Kevin D. Briggs, MS, CT(ASCP) Team Leader- Cytopathology/ Histopathology Services Danville Regional Medical Center 142 South Main Street Danville, VA 24541 Telephone: (434) 799-4470 ext.5451 Fax: (434) 799-2118 E-mail: briggsk@drhsi.org From sharon.osborn <@t> dnax.org Thu Feb 10 13:51:04 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] RE: sorted cells from spleen to stain Message-ID: <29B25753F6B1D51196110002A589D44402397ED6@PALMSG30.us.schp.com> Histoland experts..... I have a researcher who is macerating spleen, sorting the cells, lysing out the red cells and wants a good stain to identify the cells, primarily B cells with some T cells. These are on cytospin samples. He has used T-Blue, Wright-Giemsa and just Giemsa. It seems the staining could be better for it is not showing nuclear detail that the PI thinks it should be. Does anyone have suggestions for a different histochemical technic or ways to tweak what he is currently doing? Thanks. Sharon Osborn DNAX Palo Alto, CA ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From akbitting <@t> geisinger.edu Thu Feb 10 14:13:46 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] ASR Disclaimer Message-ID: We are currently involved in adding this to our reports as well. One example that I think is very good comes from University of Pittsburgh Medical System. It reads: "The following statement applies to all Immunohistochemistry, in-situ Hybridization Assays (ISH & FISH), and Immunofluorescent Testing: The testing was developed and its performance characteristics determined by the University of Pittsburgh, Department of Pathology, as required by CLIA '88 regulations. The testing has not been cleared or approved for the specific use by the U.S. Food and Drug Administration, but the FDA has determined such approval is not necessary for clinical use." We tried putting a disclaimer on our Molecular Path reports last year, and one payor kept rejecting the billing because that disclaimer used the word "Research". So, I suggest it is best to avoid the use of that word if possible. 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 >>> "Kevin Briggs" 02/10/05 02:23PM >>> Hi Netters! I am curious to know how other laboratories are handling the CAP Checklist item ANP.12425: "If patient testing is performed using Class I analyte-specific reagents (ASR's) obtained or purchased from an outside vendor, does the patient report include the disclaimer required by federal regulations?" I am specifically searching for examples of canned text comments or disclaimers suitable for reporting any of our antibodies simultaneously. Thanks, Kevin D. Briggs, MS, CT(ASCP) Team Leader- Cytopathology/ Histopathology Services Danville Regional Medical Center 142 South Main Street Danville, VA 24541 Telephone: (434) 799-4470 ext.5451 Fax: (434) 799-2118 E-mail: briggsk@drhsi.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From catherine.scott <@t> mpiresearch.com Thu Feb 10 15:00:28 2005 From: catherine.scott <@t> mpiresearch.com (Catherine Scott) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Elastin Stain for GMA/MMA Message-ID: Hello Everyone, I have been trying to find an elastin stain for GMA and MMA stented arteries. Does anyone know of a procedure or publication that may help? Thanks, Cathy Catherine L. Scott, B.A., HT (ASCP), ALAT Associate Director, Pathology Services 54943 North Main Street Mattawan, Michigan 49071-9366 USA Telephone: 269.668.3336 ext. 1218 Fax: 269.668.4151 email: catherine.scott@mpiresearch.com ****************************************************************************** Visit the MPI Website at http://www.mpiresearch.com Confidentiality Notice: "By accessing this e-mail communication, the intended recipient hereby consents to the transmittal and receipt of confidential information via electronicmedium. This e-mail is strictly confidential and intended solely for the addressee. If you are not the intended recipient you must not use, disclose, or copy this transmission. If you have received this message in error, please contact the sender and delete the material from any computer, disc drive, diskette, or other storage device or media. 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MPI Research, Inc. hereby expressly disclaims any and all liability for any viruses or data corrupting contents which may result from recipient's access or use of this e-mail or its contents." ******************************************************************************* MPI Research, Inc. - (269) 668-3336 ****************************************************************************** Visit the MPI Website at http://www.mpiresearch.com Confidentiality Notice: "By accessing this e-mail communication, the intended recipient hereby consents to the transmittal and receipt of confidential information via electronicmedium. This e-mail is strictly confidential and intended solely for the addressee. If you are not the intended recipient you must not use, disclose, or copy this transmission. If you have received this message in error, please contact the sender and delete the material from any computer, disc drive, diskette, or other storage device or media. This e-mail is not intended to impose, nor shall it be construed as imposing, any legally binding obligation upon MPI Research, Inc., or any of its subsidiaries or affiliated companies. Neither MPI Research, Inc. nor any of its subsidiaries or affiliated companies gives any representation or warranty as to the accuracy or completeness of the contents of this e-mail. MPI Research, Inc. shall not be held liable to any person resulting from the use of any information contained in this e-mail, and shall not be liable to any person who acts or omits to do anything in reliance upon receipt of this e-mail. MPI Research, Inc. hereby expressly disclaims any and all liability for any viruses or data corrupting contents which may result from recipient's access or use of this e-mail or its contents." ******************************************************************************* MPI Research, Inc. - (269) 668-3336 ****************************************************************************** Visit the MPI Website at http://www.mpiresearch.com Confidentiality Notice: "By accessing this e-mail communication, the intended recipient hereby consents to the transmittal and receipt of confidential information via electronicmedium. This e-mail is strictly confidential and intended solely for the addressee. If you are not the intended recipient you must not use, disclose, or copy this transmission. If you have received this message in error, please contact the sender and delete the material from any computer, disc drive, diskette, or other storage device or media. This e-mail is not intended to impose, nor shall it be construed as imposing, any legally binding obligation upon MPI Research, Inc., or any of its subsidiaries or affiliated companies. Neither MPI Research, Inc. nor any of its subsidiaries or affiliated companies gives any representation or warranty as to the accuracy or completeness of the contents of this e-mail. MPI Research, Inc. shall not be held liable to any person resulting from the use of any information contained in this e-mail, and shall not be liable to any person who acts or omits to do anything in reliance upon receipt of this e-mail. MPI Research, Inc. hereby expressly disclaims any and all liability for any viruses or data corrupting contents which may result from recipient?s access or use of this e-mail or its contents." ******************************************************************************* MPI Research, Inc. - (269) 668-3336 From ynwang <@t> u.washington.edu Thu Feb 10 15:16:08 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Elastin Stain for GMA/MMA (fwd) Message-ID: Hello Catherine, When we did our elastin staining of rat and tissue enineered arteries we used Hart's mofified method counterstained with tartrazine, as recommended by Barry Starcher (we tried an elasting staining kit which didn't work veyr well for us). We only stained arteries alone (no stents) and we are not experts in histology. I'm sure someone else on the histonet would be a better source for a protocol. However, we do have the protocol we used if you would like it. Best regards Yak-Nam Wang > Hello Everyone, > > > > I have been trying to find an elastin stain for GMA and MMA stented arteries. > Does anyone know of a procedure or publication that may help? > > > > Thanks, > > Cathy > > > > Catherine L. Scott, B.A., HT (ASCP), ALAT > > Associate Director, Pathology Services > > 54943 North Main Street > > Mattawan, Michigan 49071-9366 USA > > Telephone: 269.668.3336 ext. 1218 > > Fax: 269.668.4151 > > email: catherine.scott@mpiresearch.com > > > > ****************************************************************************** > Visit the MPI Website at http://www.mpiresearch.com > > Confidentiality Notice: > "By accessing this e-mail communication, the intended recipient hereby > consents > to the transmittal and receipt of confidential information via > electronicmedium. > This e-mail is strictly confidential and intended solely for the addressee. > > If you are not the intended recipient you must not use, disclose, or copy > this > transmission. If you have received this message in error, please contact the > sender and delete the material from any computer, disc drive, diskette, or > other > storage device or media. > > This e-mail is not intended to impose, nor shall it be construed as imposing, > any legally binding obligation upon MPI Research, Inc., or any of its > subsidiaries > or affiliated companies. > > Neither MPI Research, Inc. nor any of its subsidiaries or affiliated > companies > gives any representation or warranty as to the accuracy or completeness of > the > contents of this e-mail. > > MPI Research, Inc. shall not be held liable to any person resulting from the > use > of any information contained in this e-mail, and shall not be liable to any > person who acts or omits to do anything in reliance upon receipt of this > e-mail. > MPI Research, Inc. hereby expressly disclaims any and all liability for any > viruses or data corrupting contents which may result from recipient's access > or > use of this e-mail or its contents." > ******************************************************************************* > MPI Research, Inc. - (269) 668-3336 > > ****************************************************************************** > Visit the MPI Website at http://www.mpiresearch.com > > Confidentiality Notice: > "By accessing this e-mail communication, the intended recipient hereby > consents > to the transmittal and receipt of confidential information via > electronicmedium. > This e-mail is strictly confidential and intended solely for the addressee. > > If you are not the intended recipient you must not use, disclose, or copy > this > transmission. If you have received this message in error, please contact the > sender and delete the material from any computer, disc drive, diskette, or > other > storage device or media. > > This e-mail is not intended to impose, nor shall it be construed as imposing, > any legally binding obligation upon MPI Research, Inc., or any of its > subsidiaries > or affiliated companies. > > Neither MPI Research, Inc. nor any of its subsidiaries or affiliated > companies > gives any representation or warranty as to the accuracy or completeness of > the > contents of this e-mail. > > MPI Research, Inc. shall not be held liable to any person resulting from the > use > of any information contained in this e-mail, and shall not be liable to any > person who acts or omits to do anything in reliance upon receipt of this > e-mail. > MPI Research, Inc. hereby expressly disclaims any and all liability for any > viruses or data corrupting contents which may result from recipient's access > or > use of this e-mail or its contents." > ******************************************************************************* > MPI Research, Inc. - (269) 668-3336 > > > ****************************************************************************** > Visit the MPI Website at http://www.mpiresearch.com > > Confidentiality Notice: > "By accessing this e-mail communication, the intended recipient hereby > consents > to the transmittal and receipt of confidential information via > electronicmedium. > This e-mail is strictly confidential and intended solely for the addressee. > > If you are not the intended recipient you must not use, disclose, or copy > this > transmission. If you have received this message in error, please contact the > sender and delete the material from any computer, disc drive, diskette, or > other > storage device or media. > > This e-mail is not intended to impose, nor shall it be construed as imposing, > any legally binding obligation upon MPI Research, Inc., or any of its > subsidiaries > or affiliated companies. > > Neither MPI Research, Inc. nor any of its subsidiaries or affiliated > companies > gives any representation or warranty as to the accuracy or completeness of > the > contents of this e-mail. > > MPI Research, Inc. shall not be held liable to any person resulting from the > use > of any information contained in this e-mail, and shall not be liable to any > person who acts or omits to do anything in reliance upon receipt of this > e-mail. > MPI Research, Inc. hereby expressly disclaims any and all liability for any > viruses or data corrupting contents which may result from recipient’s access > or > use of this e-mail or its contents." > ******************************************************************************* > MPI Research, Inc. - (269) 668-3336 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emerald_lake77 <@t> yahoo.com Thu Feb 10 15:49:52 2005 From: emerald_lake77 <@t> yahoo.com (- -) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Silicon Tubing Message-ID: <20050210214952.61088.qmail@web42301.mail.yahoo.com> Hello, I was just approached regarding processing and sectioning silicon tubing that may be used to cover certain arteries of murine models. The tubing is very soft and flexible, and a sharp blade slices through as if going through air. However, I have never processed such samples and do not know what is the best way to do so... if it's at all possible. Does anyone have any suggestions (routine processing, frozen, etc.)?? Thank you all in advance. --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From scoop <@t> mail.nih.gov Thu Feb 10 17:51:31 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] mouse perfusion Message-ID: Hi, Does anyone know of a place I can buy or have a suggestion of how to make an apparatus for positioning mice during perfusion and catching the perfusate that runs off? We have an old home made thing consisting of a metal grate and a plastic square tank that is gigantic and awkward. We tape the mice down on it by the extremities. Any suggestions on something better would be appreciated. Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From aetechserv <@t> hotmail.com Fri Feb 11 04:35:54 2005 From: aetechserv <@t> hotmail.com (awdaf asdfadf) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Dako autostainer Message-ID: Hello, Just a quick plug for the Dako autostainer plus. It is a very good instrument - labs actually buy it ! It doesn't have to be allegedly given away like a competitors' autostainer : HINT: "the name's _ _ _ _ , James _ _ _ _ " . I've heard that this machine has more bugs than a rotten redwood - allegedly. Stick with what you know - Dako is , in many people's opinion , a stable, robust and reliable platform. _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From juan.gutierrez <@t> christushealth.org Fri Feb 11 05:52:28 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Dako autostainer Message-ID: Ventana's Benchmark XT can still run circles around the DAKO. 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 awdaf asdfadf Sent: Friday, February 11, 2005 4:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dako autostainer Hello, Just a quick plug for the Dako autostainer plus. It is a very good instrument - labs actually buy it ! It doesn't have to be allegedly given away like a competitors' autostainer : HINT: "the name's _ _ _ _ , James _ _ _ _ " . I've heard that this machine has more bugs than a rotten redwood - allegedly. Stick with what you know - Dako is , in many people's opinion , a stable, robust and reliable platform. _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jhofecker <@t> yahoo.com Fri Feb 11 07:38:45 2005 From: jhofecker <@t> yahoo.com (Jennifer Hofecker) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Tennessee Meeting Message-ID: <20050211133845.22828.qmail@web21005.mail.yahoo.com> Hi, I just wanted to let everyone know that the annual Tennessee Society for Histotechnology meeting will be held in Memphis this year. The dates are April 7-9. Location is the Radisson - Downtown. Workshops start on Friday morning and continue through Saturday afternoon. Topics for workshops include: Tissue Microarrays Interpretation of Special Stains Tolerance in the Workplace Today's artifact- Tomorrow's Facts IHC Roundtable discussion Is Your Tissue Processor Fighting you? Simply Silver Grossing for Histotechs There is also a scientific session free with paid registration: Laser Capture Microdissection: Principles, Techniques, and Applications. If you would like a complete program or more information, please contact: President: Rhonda Schalk tshpresident@juno.com (865)481-1172 (work) Pres. Elect: Jennifer Hofecker jhofecker@yahoo.com (615)343-0083 work Hope to see y'all there! __________________________________ Do you Yahoo!? The all-new My Yahoo! - What will yours do? http://my.yahoo.com From AFeatherstone <@t> KaleidaHealth.Org Fri Feb 11 07:42:06 2005 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] stat lab marker Message-ID: A while ago people were talking about a superiour slide marker. Could some one tell me the name and vendor? Thanks Annette Featherstone HT/MLT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GUTIERREZ, JUAN Sent: Friday, February 11, 2005 06:52 To: awdaf asdfadf; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Dako autostainer Ventana's Benchmark XT can still run circles around the DAKO. 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 awdaf asdfadf Sent: Friday, February 11, 2005 4:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dako autostainer Hello, Just a quick plug for the Dako autostainer plus. It is a very good instrument - labs actually buy it ! It doesn't have to be allegedly given away like a competitors' autostainer : HINT: "the name's _ _ _ _ , James _ _ _ _ " . I've heard that this machine has more bugs than a rotten redwood - allegedly. Stick with what you know - Dako is , in many people's opinion , a stable, robust and reliable platform. _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 JWEEMS <@t> sjha.org Fri Feb 11 08:06:02 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] stat lab marker Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507AFF@sjhaexc02.sjha.org> It is StatLab - 800-442-3573. It really is a good product. j:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Featherstone, Annette Sent: Friday, February 11, 2005 8:42 AM To: 'GUTIERREZ, JUAN'; awdaf asdfadf; histonet@lists.utsouthwestern.edu Subject: [Histonet] stat lab marker A while ago people were talking about a superiour slide marker. Could some one tell me the name and vendor? Thanks Annette Featherstone HT/MLT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GUTIERREZ, JUAN Sent: Friday, February 11, 2005 06:52 To: awdaf asdfadf; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Dako autostainer Ventana's Benchmark XT can still run circles around the DAKO. 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 awdaf asdfadf Sent: Friday, February 11, 2005 4:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dako autostainer Hello, Just a quick plug for the Dako autostainer plus. It is a very good instrument - labs actually buy it ! It doesn't have to be allegedly given away like a competitors' autostainer : HINT: "the name's _ _ _ _ , James _ _ _ _ " . I've heard that this machine has more bugs than a rotten redwood - allegedly. Stick with what you know - Dako is , in many people's opinion , a stable, robust and reliable platform. _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** 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 wood <@t> dcpah.msu.edu Fri Feb 11 08:54:19 2005 From: wood <@t> dcpah.msu.edu (Thomas Wood) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Warthin-Starry protocol needed Message-ID: Can anyone give me a good Warthin-Starry protocol for spirochetes, ours no longer does the job. Tom Wood BS HT Michigan State University Veterinary Medicine From JQB7 <@t> CDC.GOV Fri Feb 11 08:56:21 2005 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Warthin-Starry protocol needed Message-ID: Good W-S? Isn't that an oxymoron? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Thomas Wood Sent: Friday, February 11, 2005 9:54 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Warthin-Starry protocol needed Can anyone give me a good Warthin-Starry protocol for spirochetes, ours no longer does the job. Tom Wood BS HT Michigan State University Veterinary Medicine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Fri Feb 11 08:58:37 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:36 2005 Subject: AW: [Histonet] Raynaud's Syndrome In-Reply-To: <10C94843061E094A98C02EB77CFC32870A73DDCA@nrcmrdex1d.imsb.nrc.ca> Message-ID: Hi I am sorry to hear about your illness. What I can tell you ist, that I had a workmate, who has been working for thirty years in histology. She is retarded in 1999. She has also Mb. Raynaud (that's the disease with the "white fingers"?). And latetly I was told, that she is consulting the doctors because of another autoimmun-disease, where the esophagus is affected. I don't know the details, but I think there was no examination about connections between the diseases and the chemical exposition during her livetime. I can remember, that she told us about airdrying xylol-wet slides on the radiatior .... Just one story of a histotech Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Ross, Virginia Gesendet: Donnerstag, 10. Februar 2005 19:49 An: 'histonet@lists.utsouthwestern.edu' Betreff: [Histonet] Raynaud's Syndrome I have developed Raynaud's and I am awaiting diagnosis for suspected autoimmune diseases and I have a collegue who worked with the some of the same chemicals who is disabled with mixed connective tissue disease. Raynaud's has been linked to vinyl chloride, cryostats and vibrating machines (vibrotomes) but I have not worked with any of these. I have worked with the traditional histology chemicals, solvents, and methymethacrylate. I am interested in seeing if there is a link between histologists and autoimmune diseases, and which chemicals may be implicated. I would greatly appreciate any information anyone has on any of these topics. Thank you, Virginia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vbaker60 <@t> yahoo.com Fri Feb 11 09:08:47 2005 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Warthin-Starry protocol needed In-Reply-To: Message-ID: <20050211150847.69139.qmail@web50103.mail.yahoo.com> What particular protocol are you using? Lab made or commercial? Is it possible that your control tissue has been depleted? Back when I still in clinical, no one else would do them and I always agreed with the saying that they were 'worthless and sorry'. Once in a very, very blue moon I got results that were worth something. Vikki Baker Mt. Sinai School of Medicine New York, NY 10029 --- "Bartlett, Jeanine" wrote: > Good W-S? Isn't that an oxymoron? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Thomas > Wood > Sent: Friday, February 11, 2005 9:54 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Warthin-Starry protocol needed > > > Can anyone give me a good Warthin-Starry protocol > for spirochetes, ours > no longer does the job. > > Tom Wood BS HT > Michigan State University > Veterinary Medicine > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. http://promotions.yahoo.com/new_mail From Charles.Embrey <@t> carle.com Fri Feb 11 09:11:49 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Raynaud's Syndrome Message-ID: Retarded or retired? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Friday, February 11, 2005 8:59 AM To: Histonetliste (Histonetliste) Subject: AW: [Histonet] Raynaud's Syndrome Hi I am sorry to hear about your illness. What I can tell you ist, that I had a workmate, who has been working for thirty years in histology. She is retarded in 1999. She has also Mb. Raynaud (that's the disease with the "white fingers"?). And latetly I was told, that she is consulting the doctors because of another autoimmun-disease, where the esophagus is affected. I don't know the details, but I think there was no examination about connections between the diseases and the chemical exposition during her livetime. I can remember, that she told us about airdrying xylol-wet slides on the radiatior .... Just one story of a histotech Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Ross, Virginia Gesendet: Donnerstag, 10. Februar 2005 19:49 An: 'histonet@lists.utsouthwestern.edu' Betreff: [Histonet] Raynaud's Syndrome I have developed Raynaud's and I am awaiting diagnosis for suspected autoimmune diseases and I have a collegue who worked with the some of the same chemicals who is disabled with mixed connective tissue disease. Raynaud's has been linked to vinyl chloride, cryostats and vibrating machines (vibrotomes) but I have not worked with any of these. I have worked with the traditional histology chemicals, solvents, and methymethacrylate. I am interested in seeing if there is a link between histologists and autoimmune diseases, and which chemicals may be implicated. I would greatly appreciate any information anyone has on any of these topics. Thank you, Virginia _______________________________________________ Histonet mailing list Histonet@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 vazquezr <@t> ohsu.edu Fri Feb 11 09:15:33 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Raynaud's Syndrome Message-ID: Thank you for asking that...I almost started looking for a different line of work....;0) Robyn OHSU >>> "Charles.Embrey" 02/11/05 7:11 AM >>> Retarded or retired? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Friday, February 11, 2005 8:59 AM To: Histonetliste (Histonetliste) Subject: AW: [Histonet] Raynaud's Syndrome Hi I am sorry to hear about your illness. What I can tell you ist, that I had a workmate, who has been working for thirty years in histology. She is retarded in 1999. She has also Mb. Raynaud (that's the disease with the "white fingers"?). And latetly I was told, that she is consulting the doctors because of another autoimmun-disease, where the esophagus is affected. I don't know the details, but I think there was no examination about connections between the diseases and the chemical exposition during her livetime. I can remember, that she told us about airdrying xylol-wet slides on the radiatior .... Just one story of a histotech Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Ross, Virginia Gesendet: Donnerstag, 10. Februar 2005 19:49 An: 'histonet@lists.utsouthwestern.edu' Betreff: [Histonet] Raynaud's Syndrome I have developed Raynaud's and I am awaiting diagnosis for suspected autoimmune diseases and I have a collegue who worked with the some of the same chemicals who is disabled with mixed connective tissue disease. Raynaud's has been linked to vinyl chloride, cryostats and vibrating machines (vibrotomes) but I have not worked with any of these. I have worked with the traditional histology chemicals, solvents, and methymethacrylate. I am interested in seeing if there is a link between histologists and autoimmune diseases, and which chemicals may be implicated. I would greatly appreciate any information anyone has on any of these topics. Thank you, Virginia _______________________________________________ Histonet mailing list Histonet@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 NMargaryan <@t> childrensmemorial.org Fri Feb 11 09:17:41 2005 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Raynaud's Syndrome Message-ID: <63B8B599DE283148B92E83C78B32C15D4C8AB5@cmhexbe2.childrensmemorial.org> Dark joke and correction.......... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Friday, February 11, 2005 9:12 AM To: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Raynaud's Syndrome Retarded or retired? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Friday, February 11, 2005 8:59 AM To: Histonetliste (Histonetliste) Subject: AW: [Histonet] Raynaud's Syndrome Hi I am sorry to hear about your illness. What I can tell you ist, that I had a workmate, who has been working for thirty years in histology. She is retarded in 1999. She has also Mb. Raynaud (that's the disease with the "white fingers"?). And latetly I was told, that she is consulting the doctors because of another autoimmun-disease, where the esophagus is affected. I don't know the details, but I think there was no examination about connections between the diseases and the chemical exposition during her livetime. I can remember, that she told us about airdrying xylol-wet slides on the radiatior .... Just one story of a histotech Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Ross, Virginia Gesendet: Donnerstag, 10. Februar 2005 19:49 An: 'histonet@lists.utsouthwestern.edu' Betreff: [Histonet] Raynaud's Syndrome I have developed Raynaud's and I am awaiting diagnosis for suspected autoimmune diseases and I have a collegue who worked with the some of the same chemicals who is disabled with mixed connective tissue disease. Raynaud's has been linked to vinyl chloride, cryostats and vibrating machines (vibrotomes) but I have not worked with any of these. I have worked with the traditional histology chemicals, solvents, and methymethacrylate. I am interested in seeing if there is a link between histologists and autoimmune diseases, and which chemicals may be implicated. I would greatly appreciate any information anyone has on any of these topics. Thank you, Virginia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The 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 gcallis <@t> montana.edu Fri Feb 11 10:15:30 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Abandoned Warthin-Starry protocol in favor of Steiner In-Reply-To: <20050211150847.69139.qmail@web50103.mail.yahoo.com> References: <20050211150847.69139.qmail@web50103.mail.yahoo.com> Message-ID: <6.0.0.22.1.20050211090828.01b9ac18@gemini.msu.montana.edu> Dear all, After dinking with WS, we went with Steiner (in house made reagents) and used a superb protocol from Peggy Wenk's laboratory. We could do several racks of slides in a day, mass production with some organization, but there were no headaches with her method - it never failed to work well. Key was timing and watching that control develop. I fully agree, a good positive control is needed. We are forever grateful to Peggy on this one. At 08:08 AM 2/11/2005, you wrote: >What particular protocol are you using? Lab made or >commercial? Is it possible that your control tissue >has been depleted? >Back when I still in clinical, no one else would do >them and I always agreed with the saying that they >were 'worthless and sorry'. Once in a very, very blue >moon I got results that were worth something. > >Vikki Baker >Mt. Sinai School of Medicine >New York, NY 10029 > > >--- "Bartlett, Jeanine" wrote: > > > Good W-S? Isn't that an oxymoron? > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] > > On Behalf Of Thomas > > Wood > > Sent: Friday, February 11, 2005 9:54 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] Warthin-Starry protocol needed > > > > > > Can anyone give me a good Warthin-Starry protocol > > for spirochetes, ours > > no longer does the job. > > > > Tom Wood BS HT > > Michigan State University > > Veterinary Medicine > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > >__________________________________ >Do you Yahoo!? >Yahoo! Mail - Helps protect you from nasty viruses. >http://promotions.yahoo.com/new_mail > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Terry.Marshall <@t> rothgen.nhs.uk Fri Feb 11 10:28:36 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Abandoned Warthin-Starry protocol in favor of Steiner Message-ID: Dinking? What's that? Not in any dictionary on the web. Good word though:-) Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 11 February 2005 16:16 To: Victoria Baker; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Abandoned Warthin-Starry protocol in favor of Steiner Dear all, After dinking with WS, we went with Steiner (in house made reagents) and used a superb protocol from Peggy Wenk's laboratory. We could do several racks of slides in a day, mass production with some organization, but there were no headaches with her method - it never failed to work well. Key was timing and watching that control develop. I fully agree, a good positive control is needed. We are forever grateful to Peggy on this one. At 08:08 AM 2/11/2005, you wrote: >What particular protocol are you using? Lab made or >commercial? Is it possible that your control tissue >has been depleted? >Back when I still in clinical, no one else would do >them and I always agreed with the saying that they >were 'worthless and sorry'. Once in a very, very blue >moon I got results that were worth something. > >Vikki Baker >Mt. Sinai School of Medicine >New York, NY 10029 > > >--- "Bartlett, Jeanine" wrote: > > > Good W-S? Isn't that an oxymoron? > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] > > On Behalf Of Thomas > > Wood > > Sent: Friday, February 11, 2005 9:54 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] Warthin-Starry protocol needed > > > > > > Can anyone give me a good Warthin-Starry protocol > > for spirochetes, ours > > no longer does the job. > > > > Tom Wood BS HT > > Michigan State University > > Veterinary Medicine > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > >__________________________________ >Do you Yahoo!? >Yahoo! Mail - Helps protect you from nasty viruses. >http://promotions.yahoo.com/new_mail > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From POWELL_SA <@t> Mercer.edu Fri Feb 11 10:38:27 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Warthin-Starry protocol needed In-Reply-To: Message-ID: Tom, I have used a very good modification of Gabriel-Steiner's original technique that works well. It was in an old issue of Histo-Logic, January 1988. It can be found at http://www.sakura-americas.com/histologic/pdf/88_jan.pdf and requires no developing with almost not background staining. Spirochetes are easily seen. You can also do IHC which demonstrates them nicely, I have gotten the antibody from Biocare in the past. Shirley Powell -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Thomas Wood Sent: Friday, February 11, 2005 9:54 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Warthin-Starry protocol needed Can anyone give me a good Warthin-Starry protocol for spirochetes, ours no longer does the job. Tom Wood BS HT Michigan State University Veterinary Medicine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hgpardo <@t> uniovi.es Fri Feb 11 10:40:34 2005 From: hgpardo <@t> uniovi.es (Hector Gonzalez Pardo) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] cresyl violet staining on unfixed frozen brain sections Message-ID: <000c01c51058$68a64220$7a47239c@p48> Hi everyone! I am trying to perform Nissl staining using cresyl violet acetate on 30 micrometer-tick cryostat sections from rat brain (I usually don't need gelatinized slides). After re-hydriting the sections in alcohols (70%, 80%, 96% and 100%), I immerse the sections for 10-15 min in a solution of cresyl violet acetate (Sigma, 0,5 % at pH 4). I differentiate the sections using 70% ethanol with a few drops of glacial acetic acid (for abut 15-30 secs.), but when I try to deydrate the sections using 80%, 96% and 100 % ethanol, I always lose the staing. Finally, I get a blueish pale-looking Nissl staining on my sections, and I cannot see anything. Does anyone know how to solve it? I've tried to first de-fat the sections using ethanol for a few minutes but I did'nt work. Thank you! From hgpardo <@t> uniovi.es Fri Feb 11 10:50:38 2005 From: hgpardo <@t> uniovi.es (Hector Gonzalez Pardo) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] isopentane freezing of rat brain Message-ID: <001501c51059$d0cdd010$7a47239c@p48> Hi, I am using for the first time isopentane (2-methylbutane) to freeze my rat brains for histochemistry. However, when I do the histochemical staining, the tissue looks like "granulated" under the microscope, especially in the center. We usually immerse slowly (30 secs.) the entire brain on a weighing plastic boat in a glass vase containing isopentane chilled at -40 ?C. Then we wait for another 30 secs until it looks white on the surface, and we store it at -40?C until sectioning using a cryostate. We used before a time-consuming procedure that worked, using cryoprotection with sucrose overnight, covering the whole brain with a cryogel (OCT compound / Tissue-Tek) and using a kind of freon-22 cooled by liquid nitrogen. Since we would like to continue using the easier isopentane procedure, does anyone know what are we doing wrong? Thanks! From kbroomal <@t> NEMOURS.ORG Fri Feb 11 11:11:29 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Raynaud's Syndrome Message-ID: <9BCAC308B27CAD4196D8AC9ADF037AAA110A792F@wlmmsx01.nemours.org> Yeah, I was staring to get a little worried there! Kristen -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: Friday, February 11, 2005 10:16 AM To: Charles.Embrey@carle.com; gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Raynaud's Syndrome Thank you for asking that...I almost started looking for a different line of work....;0) Robyn OHSU >>> "Charles.Embrey" 02/11/05 7:11 AM >>> Retarded or retired? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Friday, February 11, 2005 8:59 AM To: Histonetliste (Histonetliste) Subject: AW: [Histonet] Raynaud's Syndrome Hi I am sorry to hear about your illness. What I can tell you ist, that I had a workmate, who has been working for thirty years in histology. She is retarded in 1999. She has also Mb. Raynaud (that's the disease with the "white fingers"?). And latetly I was told, that she is consulting the doctors because of another autoimmun-disease, where the esophagus is affected. I don't know the details, but I think there was no examination about connections between the diseases and the chemical exposition during her livetime. I can remember, that she told us about airdrying xylol-wet slides on the radiatior .... Just one story of a histotech Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Ross, Virginia Gesendet: Donnerstag, 10. Februar 2005 19:49 An: 'histonet@lists.utsouthwestern.edu' Betreff: [Histonet] Raynaud's Syndrome I have developed Raynaud's and I am awaiting diagnosis for suspected autoimmune diseases and I have a collegue who worked with the some of the same chemicals who is disabled with mixed connective tissue disease. Raynaud's has been linked to vinyl chloride, cryostats and vibrating machines (vibrotomes) but I have not worked with any of these. I have worked with the traditional histology chemicals, solvents, and methymethacrylate. I am interested in seeing if there is a link between histologists and autoimmune diseases, and which chemicals may be implicated. I would greatly appreciate any information anyone has on any of these topics. Thank you, Virginia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Feb 11 11:35:09 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] dinking defined in terms of Warthin Starry and amused!! In-Reply-To: References: Message-ID: <6.0.0.22.1.20050211101601.01b4cd18@gemini.msu.montana.edu> Terry, You have me laughing. "Dinking" - One can dink with - an object, procedure and probably thoughts. I have used fiddly dink, twiddle or fiddle with, or tweak(ing). In terms of the Warthin Starry, we tweaked, fiddled, twiddled and "dinked" this method, but not the Steiner - our chosen method. The term came from our electron microscopist's vernacular, year ago and he was always "fiddly dinking" with things, usually to make whatever he was doing work better. A clever person with lots of creativeness. Where he got the term, I don't know. Well, I am off to "dink" around in my the lab, it is a slow day and a Friday. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Fri Feb 11 11:46:31 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Problem with cresyl violet staining on unfixed frozen brain sections In-Reply-To: <000c01c51058$68a64220$7a47239c@p48> References: <000c01c51058$68a64220$7a47239c@p48> Message-ID: <6.0.0.22.1.20050211103811.01b55c40@gemini.msu.montana.edu> Hector, You may be taking out too much stain with your alcohol rinsing steps or the acetic acid step (this may be too long and acid too concentrated). Fat is probably not your problem but too much stain removal. See B-2 steps for that comment. Your sections are thick, so staining them may take a bit more patience. You say unfixed frozen sections, so you are letting the absolute alcohol fix the fresh, unfixed brain frozen section, then rehydrate? These are some staining hints we have with our method: Variations to Cresyl violet A. If staining is too dark, staining time can be increased. If too dark, acetic acid water can be used to differentiate out stain (1 - 2 ml acetic acid in 250 ml distilled water) with 1 to 2 very quick dips, rinse with water, dehydrate, clear and coverslip. Use acid water step carefully, you can overdo removal of stain. B. To lighten staining 1) Staining time can be decreased to 45 sec, rinsed with 70% ethanol 3 dips then into 95% ethanol. 2) Water rinses, acetic acid water dips, and alcohols will remove stain Good luck At 09:40 AM 2/11/2005, you wrote: >Hi everyone! >I am trying to perform Nissl staining using cresyl violet acetate on 30 >micrometer-tick cryostat sections from rat brain (I usually don't need >gelatinized slides). After re-hydriting the sections in alcohols (70%, >80%, 96% and 100%), I immerse the sections for 10-15 min in a solution of >cresyl violet acetate (Sigma, 0,5 % at pH 4). I differentiate the sections >using 70% ethanol with a few drops of glacial acetic acid (for abut 15-30 >secs.), but when I try to deydrate the sections using 80%, 96% and 100 % >ethanol, I always lose the staing. Finally, I get a blueish pale-looking >Nissl staining on my sections, and I cannot see anything. Does anyone know >how to solve it? I've tried to first de-fat the sections using ethanol for >a few minutes but I did'nt work. Thank you! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Matthew_Frank <@t> URMC.Rochester.edu Fri Feb 11 11:51:28 2005 From: Matthew_Frank <@t> URMC.Rochester.edu (Frank, Matthew) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] cresyl violet staining on unfixed frozen brain sec tions Message-ID: Sounds like you may be over-differentiating the sections. I would try going straight from the staining solution to a water rinse and dehydrate quickly 95% and 100% ethanol. The staining procedure is simple but you can over differentiate. In "Troubleshooting Histology Stains" by Richard Horobin & John Bancroft 1998 it list differentiation as the cause of weak staining. -----Original Message----- From: Hector Gonzalez Pardo [mailto:hgpardo@uniovi.es] Sent: Friday, February 11, 2005 11:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cresyl violet staining on unfixed frozen brain sections Hi everyone! I am trying to perform Nissl staining using cresyl violet acetate on 30 micrometer-tick cryostat sections from rat brain (I usually don't need gelatinized slides). After re-hydriting the sections in alcohols (70%, 80%, 96% and 100%), I immerse the sections for 10-15 min in a solution of cresyl violet acetate (Sigma, 0,5 % at pH 4). I differentiate the sections using 70% ethanol with a few drops of glacial acetic acid (for abut 15-30 secs.), but when I try to deydrate the sections using 80%, 96% and 100 % ethanol, I always lose the staing. Finally, I get a blueish pale-looking Nissl staining on my sections, and I cannot see anything. Does anyone know how to solve it? I've tried to first de-fat the sections using ethanol for a few minutes but I did'nt work. Thank you! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jfish <@t> gladstone.ucsf.edu Fri Feb 11 11:57:54 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] cresyl violet staining on unfixed frozen brain sections In-Reply-To: <000c01c51058$68a64220$7a47239c@p48> References: <000c01c51058$68a64220$7a47239c@p48> Message-ID: Hi Hector, I differentiate in 95% Ethanol (can add 0.1% acetic acid, but not necessary), nothing less. It seems that going into a lower concentration of ethanol destains the slides. After 95%, I continue dehydration in 100% (3x) 10 dips each, then 2 minutes in xylene, 20 dips in fresh xylene, and coverslip out of another fresh xylene. Works great! Jo Dee At 5:40 PM +0100 2/11/05, Hector Gonzalez Pardo wrote: >Hi everyone! >I am trying to perform Nissl staining using cresyl violet acetate on >30 micrometer-tick cryostat sections from rat brain (I usually don't >need gelatinized slides). After re-hydriting the sections in >alcohols (70%, 80%, 96% and 100%), I immerse the sections for 10-15 >min in a solution of cresyl violet acetate (Sigma, 0,5 % at pH 4). I >differentiate the sections using 70% ethanol with a few drops of >glacial acetic acid (for abut 15-30 secs.), but when I try to >deydrate the sections using 80%, 96% and 100 % ethanol, I always >lose the staing. Finally, I get a blueish pale-looking Nissl >staining on my sections, and I cannot see anything. Does anyone know >how to solve it? I've tried to first de-fat the sections using >ethanol for a few minutes but I did'nt work. Thank you! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** ********** Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 734-5766 Fax: (415) 355-0230 E-mail: jfish@gladstone.ucsf.edu Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158 From JBurrill <@t> criver.com Fri Feb 11 12:08:56 2005 From: JBurrill <@t> criver.com (JBurrill@criver.com) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Warthin-Starry protocol needed Message-ID: Tom, We routinely use this protocol with excellent reproducibility and results. http://www.nottingham.ac.uk/pathology/protocols/warthin.html Jason D. Burrill Manager, Histology Charles River Laboratories 251 Ballardvale Street Wilmington, MA 01887 ph: 978-658-6000 ext 1652 fax: 978-988-8793 E-mail: jburrill@criver.com From tpmorken <@t> labvision.com Fri Feb 11 12:14:19 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] dinking defined ... and amused!! Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CA15@usca0082k08.labvision.apogent.com> I understand "dinking" as takng the time to explore all the purmutaions of a given endevour. For me, it came to full fruition when I (along with a group of like-minded souls) took a long, long bicycle trip, during which we refined "dinking" to a fine art, spending quality time at cafes, pubs, roadside views. It was the life. Tim Morken 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 Gayle Callis Sent: Friday, February 11, 2005 9:35 AM To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: [Histonet] dinking defined in terms of Warthin Starry and amused!! Terry, You have me laughing. "Dinking" - One can dink with - an object, procedure and probably thoughts. I have used fiddly dink, twiddle or fiddle with, or tweak(ing). In terms of the Warthin Starry, we tweaked, fiddled, twiddled and "dinked" this method, but not the Steiner - our chosen method. The term came from our electron microscopist's vernacular, year ago and he was always "fiddly dinking" with things, usually to make whatever he was doing work better. A clever person with lots of creativeness. Where he got the term, I don't know. Well, I am off to "dink" around in my the lab, it is a slow day and a Friday. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JQB7 <@t> CDC.GOV Fri Feb 11 12:30:55 2005 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] dinking defined ... and amused!! Message-ID: What he said. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Friday, February 11, 2005 1:14 PM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] dinking defined ... and amused!! I understand "dinking" as takng the time to explore all the purmutaions of a given endevour. For me, it came to full fruition when I (along with a group of like-minded souls) took a long, long bicycle trip, during which we refined "dinking" to a fine art, spending quality time at cafes, pubs, roadside views. It was the life. Tim Morken 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 Gayle Callis Sent: Friday, February 11, 2005 9:35 AM To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: [Histonet] dinking defined in terms of Warthin Starry and amused!! Terry, You have me laughing. "Dinking" - One can dink with - an object, procedure and probably thoughts. I have used fiddly dink, twiddle or fiddle with, or tweak(ing). In terms of the Warthin Starry, we tweaked, fiddled, twiddled and "dinked" this method, but not the Steiner - our chosen method. The term came from our electron microscopist's vernacular, year ago and he was always "fiddly dinking" with things, usually to make whatever he was doing work better. A clever person with lots of creativeness. Where he got the term, I don't know. Well, I am off to "dink" around in my the lab, it is a slow day and a Friday. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Fri Feb 11 12:37:07 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] isopentane freezing of rat brain Message-ID: <5784D843593D874C93E9BADCB87342AB102796@tpiserver03.Coretech-holdings.com> Trying to slow down is all wrong. Chill the isopentane to -80, as cold as you can get it with dry ice, and plunge instantly to the bottom, full immersion of the tissue in cold isopentane. The poor appearance, especially in the middle, is freezing artifact due to freezing too slowly. The tissue will freeze vitreous, without expansion and resulting tissue damage, but gradually over time the ice will crystalize, even while stored at -40, and expand. Thus, the sooner after freezing you can get to the sectioning, the better the tissue quality will be. 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 Hector Gonzalez Pardo Sent: Friday, February 11, 2005 10:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] isopentane freezing of rat brain Hi, I am using for the first time isopentane (2-methylbutane) to freeze my rat brains for histochemistry. However, when I do the histochemical staining, the tissue looks like "granulated" under the microscope, especially in the center. We usually immerse slowly (30 secs.) the entire brain on a weighing plastic boat in a glass vase containing isopentane chilled at -40 ?C. Then we wait for another 30 secs until it looks white on the surface, and we store it at -40?C until sectioning using a cryostate. We used before a time-consuming procedure that worked, using cryoprotection with sucrose overnight, covering the whole brain with a cryogel (OCT compound / Tissue-Tek) and using a kind of freon-22 cooled by liquid nitrogen. Since we would like to continue using the easier isopentane procedure, does anyone know what are we doing wrong? Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Fri Feb 11 12:41:10 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] cresyl violet staining on unfixed frozen brain sections References: <000c01c51058$68a64220$7a47239c@p48> Message-ID: <420CFC46.F85C2C41@uwo.ca> 15-30 seconds is rather fast for a critical differentiation, unless you are handling individual slides. Consider omitting the acetic acid or reducing its concentration, especially if you are staining several slides in a Coplin jar or rack. When the neurons look right, or when they are slightly overstained, quickly move the slides into the first of three changes of 100% alcohol, agitate vigorously and then go into the second 100%. Alcohol-water mixtures extract most dyes more rapidly than either water or 100% alcohol used alone. For what it's worth, I prefer toluidine blue or neutral red to cresyl violet for Nissl staining, partly because the dye solutions can be kept and re-used for years. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Hector Gonzalez Pardo wrote: > > Hi everyone! > I am trying to perform Nissl staining using cresyl violet acetate on 30 micrometer-tick cryostat sections from rat brain (I usually don't need gelatinized slides). After re-hydriting the sections in alcohols (70%, 80%, 96% and 100%), I immerse the sections for 10-15 min in a solution of cresyl violet acetate (Sigma, 0,5 % at pH 4). I differentiate the sections using 70% ethanol with a few drops of glacial acetic acid (for abut 15-30 secs.), but when I try to deydrate the sections using 80%, 96% and 100 % ethanol, I always lose the staing. Finally, I get a blueish pale-looking Nissl staining on my sections, and I cannot see anything. Does anyone know how to solve it? I've tried to first de-fat the sections using ethanol for a few minutes but I did'nt work. Thank you! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Fri Feb 11 12:42:35 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] dinking defined ... and amused!! OT Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3C8D@exchsrv01.barrynet.barry.edu> The more common phrase is "puttering around." Kenneth Grahame(The Wind in the Willows) used "messing about." "Believe me, my young friend," Water Rat said, "there is nothing in life half so important as messing about in boats." C.P. Snow (Science and Government) warmly commended Water Rat, suggesting that it would be a good thing if scientists spent more time messing about. 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 Morken, Tim - Labvision Sent: Friday, February 11, 2005 1:14 PM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] dinking defined ... and amused!! I understand "dinking" as takng the time to explore all the purmutaions of a given endevour. For me, it came to full fruition when I (along with a group of like-minded souls) took a long, long bicycle trip, during which we refined "dinking" to a fine art, spending quality time at cafes, pubs, roadside views. It was the life. Tim Morken 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 Gayle Callis Sent: Friday, February 11, 2005 9:35 AM To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: [Histonet] dinking defined in terms of Warthin Starry and amused!! Terry, You have me laughing. "Dinking" - One can dink with - an object, procedure and probably thoughts. I have used fiddly dink, twiddle or fiddle with, or tweak(ing). In terms of the Warthin Starry, we tweaked, fiddled, twiddled and "dinked" this method, but not the Steiner - our chosen method. The term came from our electron microscopist's vernacular, year ago and he was always "fiddly dinking" with things, usually to make whatever he was doing work better. A clever person with lots of creativeness. Where he got the term, I don't know. Well, I am off to "dink" around in my the lab, it is a slow day and a Friday. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ 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 TJJ <@t> Stowers-Institute.org Fri Feb 11 12:55:41 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Re: Warthin-Starry protocol needed Message-ID: Tom, I agree with Gayle, we gleefully discarded our W-S stain in favor of a Modified Steiner. For ease, Sigma sells a staining kit (HT-1-1A). It's expensive, but has everything you need to do the technique, and it works very well. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From llewllew <@t> shaw.ca Fri Feb 11 13:17:01 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] cresyl violet staining on unfixed frozen brain sections References: <000c01c51058$68a64220$7a47239c@p48> <420CFC46.F85C2C41@uwo.ca> Message-ID: <001401c5106e$44117950$80004246@yourlk4rlmsu> When I did this method (admittedly on paraffin sections) years ago, I used only a few drops of acetic acid in a staining dish (200-300 mL) of 95% ethanol. However, integral to the method was dehydrating and clearing with xylene before differentiation. The method I used specified to stain, rinse with water, dehydrate in ethanols and clear with xylene, then leave overnight in xylene before differentiating. This took too long, so I used to leave the slides about 30 minutes. I found that it did help to remove dye from the background faster than from the cells and improved contrast. Since the sections have already been dehydrated, the post-differentiation dehydration can be gentler. Bryan Llewellyn From RBARNHART <@t> summithealth.org Fri Feb 11 13:24:10 2005 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Water bath temperature Message-ID: We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? From brett_connolly <@t> merck.com Fri Feb 11 13:27:14 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] dinking defined Message-ID: I haven't used that term in a long time, but apparently the police do it too... Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com Deputies altered ID tags in WTO set-to, report says Just 'dinking around,' says sheriff spokesman By KERY MURAKAMI SEATTLE POST-INTELLIGENCER REPORTER As King County sheriff's deputies engaged in a standoff with protesters during the World Trade Organization conference last fall, a "handful" of deputies removed or rearranged identifying letters on their riot helmets, according to a draft of a Sheriff's Office report on the handling of the protests. The admission has led to speculation that the deputies were attempting to avoid being held accountable for their actions, but sheriff's spokesman John Urquhart strongly denied that was the case. He said deputies, forced to stand for long stretches of time, were just "dinking around" when they switched stick-on letters spelling out their "nicknames." According to the draft report obtained by the Seattle Post-Intelligencer, the Sheriff's Office decided the deputies should be identifiable to supervisors and to the public. Names were stitched on their shirts. In addition, riot officers were provided with 'stick-on' letters to be applied to the backs of their helmets so that unit leaders could identify and direct them as individuals. However, the names were covered at times by rain gear. And "some individual team members rearranged/removed the 'stick-on' letters from their helmets, making them identifiable only to those who knew them or knew their 'new' name as created on their helmets," the report says. That's a concern for some observers, including the American Civil Liberties Union and a former Seattle police sergeant who serves on a Seattle City Council citizens panel reviewing the WTO. Protesters have complained they weren't able to identify officers they accuse of using excessive force because their names were not visible. Seattle Police spokesman Clem Benton said he did not know whether any Seattle police officers changed lettering on their helmets. When asked about the issue, Urquhart said "you're trying to make a mountain out of a molehill," saying that deputies acted with discipline. In one case, however, the Sheriff's Office was initially unable to identify a riot-gear clad deputy who was videotaped pepper-spraying two art students sitting in a car on Capitol Hill during the protests. Urquhart said the students refused to provide the sheriff's office with a copy of the videotape, and even if they had the roughly 15-second incident showed the deputy only from the front and with a gas mask on. Sheriff's officials called on the unidentified deputy to come forward, and Deputy John Vanderwalker, a 19-year patrolman, identified himself about two weeks later. Urquhart said a "handful" of deputies from one 15-member squad changed their letters. Urquhart said the Sheriff's Office, reviewing videotapes of the protest, identified one deputy from the letters on his helmet as possibly using excessive force, and is investigating. Urquhart said changing the letters to avoid identification would be considered a violation of policy. However, he said the office was not investigating because "there was no indication of any nefarious motive." Nevertheless, ACLU spokesman Doug Honig said the office may not have received more complaints because protesters couldn't identify the offending deputies. And Urquhart's explanation that the deputies were just "dinking around," Honig said, "was hard to believe . . . People shouldn't be fooling around with their identification." "If it's true, it's troubling," said Timothy Burgess, a former police sergeant, and currently head of Seattle Ethics and Elections Commission. Burgess also heads the City Council citizens panel reviewing the events during the conference, and brought up the issue of the stick-on letters at a meeting this week. Burgess yesterday said he was even more troubled by other aspects of the draft Sheriff's Office report. Even so, Burgess said, "police officers in that kind of a situation shouldn't be 'dinking' around with their identification. Anything that identifies them should be clear. Anyway, it's kind of juvenile." -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu ] On Behalf Of Morken, Tim - Labvision Sent: Friday, February 11, 2005 1:14 PM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] dinking defined ... and amused!! I understand "dinking" as takng the time to explore all the purmutaions of a given endevour. For me, it came to full fruition when I (along with a group of like-minded souls) took a long, long bicycle trip, during which we refined "dinking" to a fine art, spending quality time at cafes, pubs, roadside views. It was the life. Tim Morken 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 Gayle Callis Sent: Friday, February 11, 2005 9:35 AM To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: [Histonet] dinking defined in terms of Warthin Starry and amused!! Terry, You have me laughing. "Dinking" - One can dink with - an object, procedure and probably thoughts. I have used fiddly dink, twiddle or fiddle with, or tweak(ing). In terms of the Warthin Starry, we tweaked, fiddled, twiddled and "dinked" this method, but not the Steiner - our chosen method. The term came from our electron microscopist's vernacular, year ago and he was always "fiddly dinking" with things, usually to make whatever he was doing work better. A clever person with lots of creativeness. Where he got the term, I don't know. Well, I am off to "dink" around in my the lab, it is a slow day and a Friday. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ 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 gu.lang <@t> gmx.at Fri Feb 11 14:30:08 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:36 2005 Subject: AW: [Histonet] Raynaud's Syndrome Message-ID: Oh, my mistake! I think I have to emprove my English. She is retired and enjoys it. Gudrun -----Urspr?ngliche Nachricht----- Von: Charles.Embrey [mailto:Charles.Embrey@carle.com] Gesendet: Freitag, 11. Februar 2005 16:12 An: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Betreff: RE: [Histonet] Raynaud's Syndrome Retarded or retired? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Friday, February 11, 2005 8:59 AM To: Histonetliste (Histonetliste) Subject: AW: [Histonet] Raynaud's Syndrome Hi I am sorry to hear about your illness. What I can tell you ist, that I had a workmate, who has been working for thirty years in histology. She is retarded in 1999. She has also Mb. Raynaud (that's the disease with the "white fingers"?). And latetly I was told, that she is consulting the doctors because of another autoimmun-disease, where the esophagus is affected. I don't know the details, but I think there was no examination about connections between the diseases and the chemical exposition during her livetime. I can remember, that she told us about airdrying xylol-wet slides on the radiatior .... Just one story of a histotech Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Ross, Virginia Gesendet: Donnerstag, 10. Februar 2005 19:49 An: 'histonet@lists.utsouthwestern.edu' Betreff: [Histonet] Raynaud's Syndrome I have developed Raynaud's and I am awaiting diagnosis for suspected autoimmune diseases and I have a collegue who worked with the some of the same chemicals who is disabled with mixed connective tissue disease. Raynaud's has been linked to vinyl chloride, cryostats and vibrating machines (vibrotomes) but I have not worked with any of these. I have worked with the traditional histology chemicals, solvents, and methymethacrylate. I am interested in seeing if there is a link between histologists and autoimmune diseases, and which chemicals may be implicated. I would greatly appreciate any information anyone has on any of these topics. Thank you, Virginia _______________________________________________ Histonet mailing list Histonet@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 Pantur <@t> aol.com Fri Feb 11 14:42:05 2005 From: Pantur <@t> aol.com (Pantur@aol.com) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Wanted: Technicon Tissue Processors, Model 2A Message-ID: We are currently looking for some Technicon tissue processors, model 2A, monos or duos. We can use either complete or incomplete units, working or for parts. Best regards, Angelos Panagopoulos Manager Pantur, Inc. 5126 East 5th Street..........P O Box 6471 Austin, TX 78702...............Austin, TX 78762 USA.................................USA Telephone: (512) 385-6232 Fax: (512) 385-6253 e-mail: pantur@aol.com From funderwood <@t> mcohio.org Fri Feb 11 14:43:58 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:36 2005 Subject: [BULK] - [Histonet] Water bath temperature Message-ID: Hi Rebecca, This is just a dart at the old dart board. Could you be getting bubbles trapped under your sections? This is also more likely to happen if you're using alcohol in the water bath. ...ahhh, darts and a cold one on a Friday. Anyone care to join me? Fred >>> "Rebecca Barnhart" 02/11/05 02:24PM >>> We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rbarnhart <@t> summithealth.org Fri Feb 11 15:05:30 2005 From: rbarnhart <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:36 2005 Subject: [BULK] - [Histonet] Water bath temperature Message-ID: We do not use alcohol in the water bath. We use a product called Halt and I tested it at 45 with and with out Halt, both had polka dots. >>> "Fred Underwood" 2/11/2005 3:43:58 PM >>> Hi Rebecca, This is just a dart at the old dart board. Could you be getting bubbles trapped under your sections? This is also more likely to happen if you're using alcohol in the water bath. ...ahhh, darts and a cold one on a Friday. Anyone care to join me? Fred >>> "Rebecca Barnhart" 02/11/05 02:24PM >>> We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Feb 11 15:19:34 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Archived Material Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507B14@sjhaexc02.sjha.org> Can someone direct me to written documentation for when to create a new billing episode for archived pathology material? Is it after 30 days? Thanks for your help. j Have a good weekend everyone. 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 victor <@t> pathology.washington.edu Fri Feb 11 15:22:15 2005 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Archived Material In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E507B14@sjhaexc02.sjha.org> References: <83AACDB0810528418AA106F9AE9B7F7E507B14@sjhaexc02.sjha.org> Message-ID: <420D2207.1020801@pathology.washington.edu> Joyce, I don't know the source of the documentation, but we have been instructed to create a new billing episode for anything greater than 60 days. Victor Weems, Joyce wrote: >Can someone direct me to written documentation for when to create a new billing episode for archived pathology material? Is it after 30 days? > >Thanks for your help. j >Have a good weekend everyone. > >Joyce Weems >Pathology Manager >Saint Joseph's Hospital >5665 Peachtree Dunwoody Rd NE >Atlanta, GA 30342 >404-851-7376 - Phone >404-851-7831 - Fax > > > >Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. >Thank you. Saint Josephs Health System, Inc. > > > >------------------------------------------------------------------------ > >Confidentiality Notice ** The information contained in this message may >be privileged and is confidential information intended for the use of >the addressee listed above. If you are neither the intended recipient >nor the employee or agent responsible for delivering this message to the >intended recipient, you are hereby notified that any disclosure, >copying, distribution or the taking of any action in reliance on the >contents of this information is strictly prohibited. If you have >received this communication in error, please notify us immediately by >replying to the message and deleting it from your computer. >Thank you. Saint Joseph?s Health System, Inc. > > > >------------------------------------------------------------------------ > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. From eva.alstromer <@t> histolab.se Fri Feb 11 15:31:37 2005 From: eva.alstromer <@t> histolab.se (eva alstromer) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] unsubscribe In-Reply-To: <20050208182608.94549.qmail@web60604.mail.yahoo.com> References: <20050208182608.94549.qmail@web60604.mail.yahoo.com> Message-ID: -----Original Message----- From: Jennifer Sipes To: histonet@lists.utsouthwestern.edu Date: Tue, 8 Feb 2005 10:26:08 -0800 (PST) Subject: [Histonet] (no subject) > Happy Mardi Gras and Happy Chinese New Year, too! May you all eat, > drink, and be merry! > > > Jennifer K. Sipes, RALAT > Sr. Laboratory Technician > Johns Hopkins University > Ross 929 > 720 Rutland Avenue > Baltimore, MD 21205 > phone: 410-614-0131 > cell: 443-413-0853 > e-mail: jengirl1014@yahoo.com > > > > > > > > > > > > --------------------------------- > Do you Yahoo!? > Yahoo! Search presents - Jib Jab's 'Second Term' > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ttruscot <@t> vetmed.wsu.edu Fri Feb 11 15:27:56 2005 From: ttruscot <@t> vetmed.wsu.edu (Truscott, Tom) Date: Fri Sep 16 15:24:36 2005 Subject: [Histonet] Water bath temperature Message-ID: <95111571D1C6B5498C64F68DD0769BB506BECD@cvm36.vetmed.wsu.edu> Rebecca, Just a guess. Perhaps there's a reaction from the heat or water or both that separates a component of the paraplast and makes an greasy or insoluble bubble that blocks staining but disappears in the final dehydration. Good Luck! Tom Truscott -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca Barnhart Sent: Friday, February 11, 2005 11:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Feb 11 15:30:15 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] B-Plus Fixative Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507B16@sjhaexc02.sjha.org> My pathologists are unhappy with B-Plus fixed bone marrows, especially immunos. They have pulled previous cases on patient that were fixed in B5 and of couse see quite a difference side by side. Does anyone else hear this complaint? 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 BlazekL <@t> childrensdayton.org Fri Feb 11 15:34:03 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] B-Plus Fixative Message-ID: Joyce, We see a difference in the B5 and B-Plus but it is coming down to the wire that there will be no Mercury used in the hospital at all. Linda Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Weems, Joyce" 02/11 4:30 PM >>> My pathologists are unhappy with B-Plus fixed bone marrows, especially immunos. They have pulled previous cases on patient that were fixed in B5 and of couse see quite a difference side by side. Does anyone else hear this complaint? 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. From froyer <@t> bitstream.net Fri Feb 11 16:00:10 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Wanted: In-Reply-To: References: Message-ID: <420D2AEA.2010009@bitstream.net> We are looking for used Leica 2125 microtomes. Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Pantur@aol.com wrote: >We are currently looking for some Technicon tissue processors, model 2A, >monos or duos. We can use either complete or incomplete units, working or for >parts. > >Best regards, > >Angelos Panagopoulos >Manager > >Pantur, Inc. >5126 East 5th Street..........P O Box 6471 >Austin, TX 78702...............Austin, TX 78762 >USA.................................USA > >Telephone: (512) 385-6232 >Fax: (512) 385-6253 >e-mail: pantur@aol.com >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From mcclistd <@t> musc.edu Fri Feb 11 15:59:09 2005 From: mcclistd <@t> musc.edu (David McClister) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] blue nuclear stain Message-ID: With the use of confocal, you can do sequential imaging that will allow the machine to detect to dyes independent from a third one. As far as a good dye to use, To-Pro-3 from Molecular Probes is wonderful. You might want to think about changing your Alexa 543 dye to a 633 or 647. Hope this Helps!! From mcauliff <@t> umdnj.edu Fri Feb 11 19:32:53 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Microglia In-Reply-To: <000901c50e32$c02ff810$7f607d80@sadunlab> References: <000901c50e32$c02ff810$7f607d80@sadunlab> Message-ID: <420D5CC5.9070201@umdnj.edu> Hi Fred: For mouse, Mac1 or F4/80 work well on frozen sections of lightly fixed (buffered formalin) material. For rat, OX-42 is the equivalent of Mac1. ED-1 is another marker for rat microglia. I don't have experience with human tissues. While it is possible to get some of these markers to work on paraffin embedded material, frozen sections of fixed material are a better choice. Geoff Fred Ross-Cisneros wrote: >Dear All, > >What would be a specific antibody (that's commercially availble) to CNS >micoglia? I've heard that CD-68 and ED-1 can be used. From your >experience, what is the best for human and/or murine/rodent tissues. > >Thanks. > >Fred > > > >Fred N. Ross-Cisneros > >Sadun Neuro-Ophthalmology Lab > >USC Keck School of Medicine and > >Doheny Eye Institute > >1355 San Pablo Street, DVRC 311 > >Los Angeles, CA 90033-1026 > >Tel.: (323) 442-6667 > >Fax: (323) 442-6688 > > > > >_______________________________________________ >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 chesarato <@t> hotmail.com Fri Feb 11 23:01:54 2005 From: chesarato <@t> hotmail.com (Cesar Francisco Romero) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Santa Cruz Technologies Antibodies Message-ID: Dear People from Histonet I have purchased two antibodies from the Company Santa Cruz Tech. ( California ). These antibodies are FAK and Pax5. I can not make them stain anything. They do not provide a Technical Datasheet ( a real shame ! ). I have tried Low pH - High pH - Pepsin and Trypsin but nothing works at all. Does anybody have any experience with these antibodies or other antibodies from this company ? Thank You in advance Dr. C?sar Romero Pathologist Buenos Aires Argentina _________________________________________________________________ MSN Amor: busca tu ? naranja http://latam.msn.com/amor/ From rmeyers <@t> ctcsurge.com Fri Feb 11 23:12:32 2005 From: rmeyers <@t> ctcsurge.com (Roger Meyers) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] automated response Message-ID: <10502120012.AA12477972@mail.ctcsurge.com> I will be traveling and unavailable after 2:00pm on Friday 2/11/05. If you need immediate assistance, please call Computer Trust Corporation at 617-557-9264. From herme013 <@t> umn.edu Sat Feb 12 09:18:47 2005 From: herme013 <@t> umn.edu (Yves Heremans) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] peroxidase/biotin block before or after permeabilization ? Message-ID: <643D5B77-7D09-11D9-BB19-000393BA7938@umn.edu> Dear All, I am performing peroxidase-DAB staining on cultured cells and was wondering whether peroxidase and biotin blocking should be performed before or after the permeabilization step with PBS + Tween ? Does permeabilization before blocking improve blocking due to better penetration ? My protocol is : peroxidase block with 0.3% H202 in methanol for 30 min. at RT, followed by avidin-biotin block from Dako, followed by PBS + Tween for 15 min. , followed by 30 min. block with fish skin gelatin in PBS. Molly From histo007 <@t> hotmail.com Sat Feb 12 13:04:51 2005 From: histo007 <@t> hotmail.com (Jim Ball) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Histology Schools Message-ID: Is there an accredited histology school in the state of Alabama, that may have been assosiated with the University of Alabama. I was told there use to be one, but it lost its accreidation, if this is true do they plan on reapplying for accreidation. I have a friend that would like this information, but does not have access to the inter-net. I told him I would ask the histonet and relay all respones to him. From katri <@t> cogeco.ca Sat Feb 12 13:46:30 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Santa Cruz Technologies Antibodies References: Message-ID: <001801c5113b$8cb5e240$6a9a9618@Katri> Hi Cesar, Have you tried to talk to their (Santa Cruz) Techical support Tel: 1-800-457-3801? They should be able to tell you how any particular antibody works. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Cesar Francisco Romero" To: Sent: Saturday, February 12, 2005 12:01 AM Subject: [Histonet] Santa Cruz Technologies Antibodies > Dear People from Histonet > > I have purchased two antibodies from the Company Santa Cruz Tech. ( > California ). > These antibodies are FAK and Pax5. > I can not make them stain anything. > They do not provide a Technical Datasheet ( a real shame ! ). > I have tried Low pH - High pH - Pepsin and Trypsin but nothing works at > all. > > Does anybody have any experience with these antibodies or other antibodies > from this company ? > > Thank You in advance > > Dr. C?sar Romero > Pathologist > Buenos Aires > Argentina > > _________________________________________________________________ > MSN Amor: busca tu ? naranja http://latam.msn.com/amor/ > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From georgecole <@t> ev1.net Sun Feb 13 02:42:50 2005 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] " Dinking" Message-ID: <000001c511a8$00c5ad20$074dbad0@hppav> Hey Techs out there--- there are possible, highly interesting useages for the Term "dinking" around. There are a couple of entries in Webster's 9th New Collegiate Dictionary: 1. Dink: to shorten and 2. to make a drop shot---(for you basketball fans). Histotechs, said to be dinking about, I suppose, could 1: be described as shortening the confusion by chugging about finding answers and or 2 Making a drop shot with a good answer. But the next line under these two entries are the terms: Dinky or Dinkey----a small locomotive used esp. for hauling freight, logging and shunting----where "dinking" refers to hauling freight, especially in movements of goods about the yard----also used in the slang version ---"dinking about"-like a small; busy locomotive busy about the train yards that picks up and moves things----but in this usage, it is a person picking up questions, picking away at them, as one who picks away at a problem until it is dropped next to its answer. I hope that both Merriam and Webster publishing since 1831, will be beyond caring if their entries are chugged, ,juggled and dinked about the Great Yard of Usage. georgecole@ev1.net From pruegg <@t> ihctech.net Sun Feb 13 12:06:30 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] mouse perfusion In-Reply-To: Message-ID: <200502131806.j1DI6Ylk088884@pro12.abac.com> We used to have a devise which was a clear plastic tube for holding rats while we took blood from their tails, there may be something like that smaller for mice, don't know where it came from, I could ask the animal care lab. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Cooperman Sent: Thursday, February 10, 2005 4:52 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] mouse perfusion Hi, Does anyone know of a place I can buy or have a suggestion of how to make an apparatus for positioning mice during perfusion and catching the perfusate that runs off? We have an old home made thing consisting of a metal grate and a plastic square tank that is gigantic and awkward. We tape the mice down on it by the extremities. Any suggestions on something better would be appreciated. Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Sun Feb 13 12:09:17 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Elastin Stain for GMA/MMA (fwd) In-Reply-To: Message-ID: <200502131809.j1DI9Lfb089883@pro12.abac.com> I have done Pentachrome stain for elastic fibers on rat pulmonary arteries with good results, I use the Pentachrome staining kit from Polyscientific. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Y. Wang Sent: Thursday, February 10, 2005 2:16 PM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Elastin Stain for GMA/MMA (fwd) Hello Catherine, When we did our elastin staining of rat and tissue enineered arteries we used Hart's mofified method counterstained with tartrazine, as recommended by Barry Starcher (we tried an elasting staining kit which didn't work veyr well for us). We only stained arteries alone (no stents) and we are not experts in histology. I'm sure someone else on the histonet would be a better source for a protocol. However, we do have the protocol we used if you would like it. Best regards Yak-Nam Wang > Hello Everyone, > > > > I have been trying to find an elastin stain for GMA and MMA stented arteries. > Does anyone know of a procedure or publication that may help? > > > > Thanks, > > Cathy > > > > Catherine L. Scott, B.A., HT (ASCP), ALAT > > Associate Director, Pathology Services > > 54943 North Main Street > > Mattawan, Michigan 49071-9366 USA > > Telephone: 269.668.3336 ext. 1218 > > Fax: 269.668.4151 > > email: catherine.scott@mpiresearch.com > > > > **************************************************************************** ** > Visit the MPI Website at http://www.mpiresearch.com > > Confidentiality Notice: > "By accessing this e-mail communication, the intended recipient hereby > consents > to the transmittal and receipt of confidential information via > electronicmedium. > This e-mail is strictly confidential and intended solely for the addressee. > > If you are not the intended recipient you must not use, disclose, or copy > this > transmission. If you have received this message in error, please contact the > sender and delete the material from any computer, disc drive, diskette, or > other > storage device or media. > > This e-mail is not intended to impose, nor shall it be construed as imposing, > any legally binding obligation upon MPI Research, Inc., or any of its > subsidiaries > or affiliated companies. > > Neither MPI Research, Inc. nor any of its subsidiaries or affiliated > companies > gives any representation or warranty as to the accuracy or completeness of > the > contents of this e-mail. > > MPI Research, Inc. shall not be held liable to any person resulting from the > use > of any information contained in this e-mail, and shall not be liable to any > person who acts or omits to do anything in reliance upon receipt of this > e-mail. > MPI Research, Inc. hereby expressly disclaims any and all liability for any > viruses or data corrupting contents which may result from recipient's access > or > use of this e-mail or its contents." > **************************************************************************** *** > MPI Research, Inc. - (269) 668-3336 > > **************************************************************************** ** > Visit the MPI Website at http://www.mpiresearch.com > > Confidentiality Notice: > "By accessing this e-mail communication, the intended recipient hereby > consents > to the transmittal and receipt of confidential information via > electronicmedium. > This e-mail is strictly confidential and intended solely for the addressee. > > If you are not the intended recipient you must not use, disclose, or copy > this > transmission. If you have received this message in error, please contact the > sender and delete the material from any computer, disc drive, diskette, or > other > storage device or media. > > This e-mail is not intended to impose, nor shall it be construed as imposing, > any legally binding obligation upon MPI Research, Inc., or any of its > subsidiaries > or affiliated companies. > > Neither MPI Research, Inc. nor any of its subsidiaries or affiliated > companies > gives any representation or warranty as to the accuracy or completeness of > the > contents of this e-mail. > > MPI Research, Inc. shall not be held liable to any person resulting from the > use > of any information contained in this e-mail, and shall not be liable to any > person who acts or omits to do anything in reliance upon receipt of this > e-mail. > MPI Research, Inc. hereby expressly disclaims any and all liability for any > viruses or data corrupting contents which may result from recipient's access > or > use of this e-mail or its contents." > **************************************************************************** *** > MPI Research, Inc. - (269) 668-3336 > > > **************************************************************************** ** > Visit the MPI Website at http://www.mpiresearch.com > > Confidentiality Notice: > "By accessing this e-mail communication, the intended recipient hereby > consents > to the transmittal and receipt of confidential information via > electronicmedium. > This e-mail is strictly confidential and intended solely for the addressee. > > If you are not the intended recipient you must not use, disclose, or copy > this > transmission. If you have received this message in error, please contact the > sender and delete the material from any computer, disc drive, diskette, or > other > storage device or media. > > This e-mail is not intended to impose, nor shall it be construed as imposing, > any legally binding obligation upon MPI Research, Inc., or any of its > subsidiaries > or affiliated companies. > > Neither MPI Research, Inc. nor any of its subsidiaries or affiliated > companies > gives any representation or warranty as to the accuracy or completeness of > the > contents of this e-mail. > > MPI Research, Inc. shall not be held liable to any person resulting from the > use > of any information contained in this e-mail, and shall not be liable to any > person who acts or omits to do anything in reliance upon receipt of this > e-mail. > MPI Research, Inc. hereby expressly disclaims any and all liability for any > viruses or data corrupting contents which may result from recipients access > or > use of this e-mail or its contents." > **************************************************************************** *** > MPI Research, Inc. - (269) 668-3336 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pathrm35 <@t> adelphia.net Sun Feb 13 12:44:13 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] dermpath histotech position in SE Florida Message-ID: <30833902.1108320253219.JavaMail.root@web2.mail.adelphia.net> We are currently seeking a full time first shift histotech for our small but growing dermpath lab in southeast Florida. Position requires ASCP as either a HT or HTL and Florida license as a technician or technologist or eligible. We offer: -- a new lab currently being built in a professional business park -- a smaller yet professional atmosphere -- lighter volume but growing -- a sign on bonus -- close proximity to the ocean for outdoor activities -- no micro managment If interested please contact: Ron Martin fax 561-721-1249 phone 561-721-2400 From monika.sternesjo <@t> akademiska.se Mon Feb 14 02:59:23 2005 From: monika.sternesjo <@t> akademiska.se (monika.sternesjo@akademiska.se) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] unsubscibe Message-ID: From JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk Mon Feb 14 03:42:42 2005 From: JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk (JOHN PHILLIPS) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature Message-ID: <166A1E642B5B644DA694C08FD29D0ADC3E135F@ztroy.new-tr.wales.nhs.uk> The problem with staining may be due to unsatisfactory removal of the wax prior to staining for H&E. I think you are looking to the wrong place for the solution to your problem. Try extending your dewax time in xylene before you hydrate your sections on the staining run. If the problem goes away PERMANANTLY then you have your answer.Good luck. John, Wrexham,Wales.U.K. -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 11 February 2005 19:24 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r rheolwr systemau wybod drwy ddefnyddio'r manylion isod. Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain Cymru. Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau?r Ddeddf Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru ar www.newalesnhstrust.org.uk This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager using the details below. The contents of this email represent the views of the individual(s) named above and do not necessarily represent the views of the North East Wales NHS Trust. Please be aware that, under the terms of the Freedom of Information Act 2000, the North East Wales NHS Trust may be required to make public the content of any emails or correspondence received. For futher information on Freedom of Information, please refer to the North East Wales NHS Trust website at www.newalesnhstrust.org.uk For further assistance, please contact system.administrator@new-tr.wales.nhs.uk. From aweimer <@t> clinomicsbio.com Mon Feb 14 06:13:15 2005 From: aweimer <@t> clinomicsbio.com (Aleasha Weimer) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Histology Schools Message-ID: <145F33B84793CF488578F86355B32E691D8A7F@mail.clinomicsbio.com> I believe that the only accredited histology school is in New York. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jim Ball Sent: Saturday, February 12, 2005 2:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology Schools Is there an accredited histology school in the state of Alabama, that may have been assosiated with the University of Alabama. I was told there use to be one, but it lost its accreidation, if this is true do they plan on reapplying for accreidation. I have a friend that would like this information, but does not have access to the inter-net. I told him I would ask the histonet and relay all respones to him. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RBARNHART <@t> summithealth.org Mon Feb 14 06:20:48 2005 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature Message-ID: I am sure the paraffin is completely removed. I know this because when I stain a slide cut with the water bath at a temperature of 45 and at the same time a slide cut with the water bath set at 38, the slide cut at 45 will show polka dots and the slide cut at 38 will not. >>> "George Cole" 2/13/2005 2:34:30 AM >>> Rebecca; I will only add a quick swiping question to the wiser replies you should get---the fact that your tissue would not take up the hematoxylin is interesting, because I stained thousands of my frozen muscle and nerve section UNFIXED in hematoxylin and eosin--with the eosin differentiated in 95% alcohol--- and got beautiful, consistent results. May I ask, have you removed the paraffin completely from the tissues before you tried to stain them? georgecole@ev1.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca Barnhart Sent: Friday, February 11, 2005 11:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Feb 14 06:37:12 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF75@bhrv-nt-11.bhrv.nwest.nhs.uk> After 'floating out' on the waterbath, do you dry in an oven afterwards and, if so, what is the temperature of the oven and do both sets (from the 45 and 38 waterbaths) go in the same oven for drying? For the same time? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 14 February 2005 12:21 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Water bath temperature[Scanned] I am sure the paraffin is completely removed. I know this because when I stain a slide cut with the water bath at a temperature of 45 and at the same time a slide cut with the water bath set at 38, the slide cut at 45 will show polka dots and the slide cut at 38 will not. >>> "George Cole" 2/13/2005 2:34:30 AM >>> Rebecca; I will only add a quick swiping question to the wiser replies you should get---the fact that your tissue would not take up the hematoxylin is interesting, because I stained thousands of my frozen muscle and nerve section UNFIXED in hematoxylin and eosin--with the eosin differentiated in 95% alcohol--- and got beautiful, consistent results. May I ask, have you removed the paraffin completely from the tissues before you tried to stain them? georgecole@ev1.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca Barnhart Sent: Friday, February 11, 2005 11:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RBARNHART <@t> summithealth.org Mon Feb 14 06:43:31 2005 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature[Scanned] Message-ID: Our stainer has an oven on it and it is set at 70. And yes both slides do go in the same oven. To make sure everything was equal I stain both slides at the same time. >>> Kemlo Rogerson 2/14/2005 7:37:12 AM >>> After 'floating out' on the waterbath, do you dry in an oven afterwards and, if so, what is the temperature of the oven and do both sets (from the 45 and 38 waterbaths) go in the same oven for drying? For the same time? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 14 February 2005 12:21 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Water bath temperature[Scanned] I am sure the paraffin is completely removed. I know this because when I stain a slide cut with the water bath at a temperature of 45 and at the same time a slide cut with the water bath set at 38, the slide cut at 45 will show polka dots and the slide cut at 38 will not. >>> "George Cole" 2/13/2005 2:34:30 AM >>> Rebecca; I will only add a quick swiping question to the wiser replies you should get---the fact that your tissue would not take up the hematoxylin is interesting, because I stained thousands of my frozen muscle and nerve section UNFIXED in hematoxylin and eosin--with the eosin differentiated in 95% alcohol--- and got beautiful, consistent results. May I ask, have you removed the paraffin completely from the tissues before you tried to stain them? georgecole@ev1.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca Barnhart Sent: Friday, February 11, 2005 11:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pathrm35 <@t> adelphia.net Mon Feb 14 07:13:05 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Histology Schools Message-ID: <8327720.1108386785714.JavaMail.root@web3.mail.adelphia.net> Check the NSH website for a listing of schools. Go under Education then Schools for the listings. ---- Aleasha Weimer wrote: > I believe that the only accredited histology school is in New York. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jim Ball > Sent: Saturday, February 12, 2005 2:05 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histology Schools > > Is there an accredited histology school in the state of Alabama, that > may > have been assosiated with the University of Alabama. I was told there > use > to be one, but it lost its accreidation, if this is true do they plan on > > reapplying for accreidation. I have a friend that would like this > information, but does not have access to the inter-net. I told him I > would > ask the histonet and relay all respones to him. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From HornHV <@t> archildrens.org Mon Feb 14 08:33:17 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Histology Schools Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322BA7A@EMAIL.archildrens.org> No, there is an accredited histology school here in Little Rock, Ar. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Aleasha Weimer Sent: Monday, February 14, 2005 6:13 AM To: Jim Ball; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histology Schools I believe that the only accredited histology school is in New York. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jim Ball Sent: Saturday, February 12, 2005 2:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology Schools Is there an accredited histology school in the state of Alabama, that may have been assosiated with the University of Alabama. I was told there use to be one, but it lost its accreidation, if this is true do they plan on reapplying for accreidation. I have a friend that would like this information, but does not have access to the inter-net. I told him I would ask the histonet and relay all respones to him. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From gcallis <@t> montana.edu Mon Feb 14 09:42:35 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Polka dots and RE: Water bath temperature In-Reply-To: <166A1E642B5B644DA694C08FD29D0ADC3E135F@ztroy.new-tr.wales. nhs.uk> References: <166A1E642B5B644DA694C08FD29D0ADC3E135F@ztroy.new-tr.wales.nhs.uk> Message-ID: <6.0.0.22.1.20050214083128.01b51de8@gemini.msu.montana.edu> I agree with John on this polka dot problem. Also, stirring your paraffin redistributes polymer and other additives. This should be done just before embedding EACH day insure these polymers have not settled to the bottom into little blobs or solid polka dots that may not be totally removed by xylenes. In addition to increasing time in xylene, you can also add one more container of xylene or xylene substitute and change the first absolute alcohol after xylene more often to prevent excessive carry over of paraffin residues. A quick test for rehydration solvents. Take a pasteur pipette and sample each alcohol change after xylene. Squirt this sample into water. If it turns milky white, then you have carry over of xylene and paraffin into the alcohols, and need to rotate out all changes more frequently. If this happens in the 95% just before 70% and water, you really have trouble with paraffin residues removal. At 02:42 AM 2/14/2005, you wrote: >The problem with staining may be due to unsatisfactory removal of the wax >prior to staining for H&E. I think you are looking to the wrong place for >the solution to your problem. Try extending your dewax time in xylene >before you hydrate your sections on the staining run. If the problem goes >away PERMANANTLY then you have your answer. Good luck. > >John, >Wrexham,Wales.U.K. > >-----Original Message----- >From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] >Sent: 11 February 2005 19:24 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Water bath temperature > > >We were having a problem that the tissue would not take up the >hematoxlyin but would take up the eosin but only in some areas, so it >looked like polka dots. We started one by one changing solution trying >to figure out where it was coming from. We had all fresh solutions and >still had polka dots. I tried the only other thing left to change, I >lowered the temperature of the water bath from 45 to 38 and this worked. > We currently are using ParaPlast Xtra with a melting point of 52. We >have all tried figure out why the temperature in the water bath will >make the difference. Several times I have cut the same tissue at >different temperatures ranging from 38 - 45 and every time there are >polka dots at the higher temperatures. Does anyone have any idea why >this would be? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn >gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu >atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r >rheolwr systemau wybod drwy ddefnyddio'r manylion isod. > >Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, >felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain >Cymru. > >Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i >Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys >unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau?r Ddeddf >Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, >cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru >ar www.newalesnhstrust.org.uk > > >This email and any files transmitted with it are confidential and >intended solely for the use of the individual or entity to whom they >are addressed. If you have received this email in error please notify >the system manager using the details below. > >The contents of this email represent the views of the individual(s) >named above and do not necessarily represent the views of the >North East Wales NHS Trust. > >Please be aware that, under the terms of the Freedom of Information Act >2000, the North East Wales NHS Trust may be required to make public the >content of any emails or correspondence received. For futher >information on Freedom of Information, please refer to the North East >Wales NHS Trust website at www.newalesnhstrust.org.uk > >For further assistance, please contact >system.administrator@new-tr.wales.nhs.uk. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From POWELL_SA <@t> Mercer.edu Mon Feb 14 09:54:35 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Histology Schools In-Reply-To: <145F33B84793CF488578F86355B32E691D8A7F@mail.clinomicsbio.com> Message-ID: We have a NAACLS approved histology school in Georgia, it is at Darton College. www.darton.edu Shirley Powell -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Aleasha Weimer Sent: Monday, February 14, 2005 7:13 AM To: Jim Ball; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histology Schools I believe that the only accredited histology school is in New York. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jim Ball Sent: Saturday, February 12, 2005 2:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology Schools Is there an accredited histology school in the state of Alabama, that may have been assosiated with the University of Alabama. I was told there use to be one, but it lost its accreidation, if this is true do they plan on reapplying for accreidation. I have a friend that would like this information, but does not have access to the inter-net. I told him I would ask the histonet and relay all respones to him. _______________________________________________ Histonet mailing list Histonet@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 Mon Feb 14 10:01:29 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Polka dots and RE: Water bath temperature[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF80@bhrv-nt-11.bhrv.nwest.nhs.uk> But why does it happen only with the 'hotter' bath? Surely the wax would not be removed from the sections floated out on the 'cooler' bath as well? You would assume that it is either the effect of the 'extra' heat or that there is something odd about that waterbath. I assume that the bath is clean and we are not dealing with wax melting in the 'hotter' bath and thereby contaminating the sections? -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 14 February 2005 15:43 To: JOHN PHILLIPS; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Polka dots and RE: Water bath temperature[Scanned] I agree with John on this polka dot problem. Also, stirring your paraffin redistributes polymer and other additives. This should be done just before embedding EACH day insure these polymers have not settled to the bottom into little blobs or solid polka dots that may not be totally removed by xylenes. In addition to increasing time in xylene, you can also add one more container of xylene or xylene substitute and change the first absolute alcohol after xylene more often to prevent excessive carry over of paraffin residues. A quick test for rehydration solvents. Take a pasteur pipette and sample each alcohol change after xylene. Squirt this sample into water. If it turns milky white, then you have carry over of xylene and paraffin into the alcohols, and need to rotate out all changes more frequently. If this happens in the 95% just before 70% and water, you really have trouble with paraffin residues removal. At 02:42 AM 2/14/2005, you wrote: >The problem with staining may be due to unsatisfactory removal of the wax >prior to staining for H&E. I think you are looking to the wrong place for >the solution to your problem. Try extending your dewax time in xylene >before you hydrate your sections on the staining run. If the problem goes >away PERMANANTLY then you have your answer. Good luck. > >John, >Wrexham,Wales.U.K. > >-----Original Message----- >From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] >Sent: 11 February 2005 19:24 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Water bath temperature > > >We were having a problem that the tissue would not take up the >hematoxlyin but would take up the eosin but only in some areas, so it >looked like polka dots. We started one by one changing solution trying >to figure out where it was coming from. We had all fresh solutions and >still had polka dots. I tried the only other thing left to change, I >lowered the temperature of the water bath from 45 to 38 and this worked. > We currently are using ParaPlast Xtra with a melting point of 52. We >have all tried figure out why the temperature in the water bath will >make the difference. Several times I have cut the same tissue at >different temperatures ranging from 38 - 45 and every time there are >polka dots at the higher temperatures. Does anyone have any idea why >this would be? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn >gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu >atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r >rheolwr systemau wybod drwy ddefnyddio'r manylion isod. > >Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, >felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain >Cymru. > >Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i >Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys >unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau?r Ddeddf >Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, >cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru >ar www.newalesnhstrust.org.uk > > >This email and any files transmitted with it are confidential and >intended solely for the use of the individual or entity to whom they >are addressed. If you have received this email in error please notify >the system manager using the details below. > >The contents of this email represent the views of the individual(s) >named above and do not necessarily represent the views of the >North East Wales NHS Trust. > >Please be aware that, under the terms of the Freedom of Information Act >2000, the North East Wales NHS Trust may be required to make public the >content of any emails or correspondence received. For futher >information on Freedom of Information, please refer to the North East >Wales NHS Trust website at www.newalesnhstrust.org.uk > >For further assistance, please contact >system.administrator@new-tr.wales.nhs.uk. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RBARNHART <@t> summithealth.org Mon Feb 14 10:06:12 2005 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:37 2005 Subject: [Possible SPAM] - RE: [Histonet] Polka dots and RE: Water bath temperature[Scanned] - Found word(s) Message-ID: Yes the waterbath is cleaned everyday and there is no wax left on it. I am sure that all the wax is being removed or we would see it on both section (from the waterbath at 38 and 45). I have done test time after time staining several slides (some cut at 38 and others at 45) at the same time and the slides cut at 45 will always show polka dots, even with fresh americlear (we do not use xylene) and alcohols. >>> Kemlo Rogerson 2/14/2005 11:01:29 AM >>> But why does it happen only with the 'hotter' bath? Surely the wax would not be removed from the sections floated out on the 'cooler' bath as well? You would assume that it is either the effect of the 'extra' heat or that there is something odd about that waterbath. I assume that the bath is clean and we are not dealing with wax melting in the 'hotter' bath and thereby contaminating the sections? -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 14 February 2005 15:43 To: JOHN PHILLIPS; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Polka dots and RE: Water bath temperature[Scanned] I agree with John on this polka dot problem. Also, stirring your paraffin redistributes polymer and other additives. This should be done just before embedding EACH day insure these polymers have not settled to the bottom into little blobs or solid polka dots that may not be totally removed by xylenes. In addition to increasing time in xylene, you can also add one more container of xylene or xylene substitute and change the first absolute alcohol after xylene more often to prevent excessive carry over of paraffin residues. A quick test for rehydration solvents. Take a pasteur pipette and sample each alcohol change after xylene. Squirt this sample into water. If it turns milky white, then you have carry over of xylene and paraffin into the alcohols, and need to rotate out all changes more frequently. If this happens in the 95% just before 70% and water, you really have trouble with paraffin residues removal. At 02:42 AM 2/14/2005, you wrote: >The problem with staining may be due to unsatisfactory removal of the wax >prior to staining for H&E. I think you are looking to the wrong place for >the solution to your problem. Try extending your dewax time in xylene >before you hydrate your sections on the staining run. If the problem goes >away PERMANANTLY then you have your answer. Good luck. > >John, >Wrexham,Wales.U.K. > >-----Original Message----- >From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] >Sent: 11 February 2005 19:24 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Water bath temperature > > >We were having a problem that the tissue would not take up the >hematoxlyin but would take up the eosin but only in some areas, so it >looked like polka dots. We started one by one changing solution trying >to figure out where it was coming from. We had all fresh solutions and >still had polka dots. I tried the only other thing left to change, I >lowered the temperature of the water bath from 45 to 38 and this worked. > We currently are using ParaPlast Xtra with a melting point of 52. We >have all tried figure out why the temperature in the water bath will >make the difference. Several times I have cut the same tissue at >different temperatures ranging from 38 - 45 and every time there are >polka dots at the higher temperatures. Does anyone have any idea why >this would be? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn >gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu >atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r >rheolwr systemau wybod drwy ddefnyddio'r manylion isod. > >Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, >felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain >Cymru. > >Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i >Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys >unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau?r Ddeddf >Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, >cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru >ar www.newalesnhstrust.org.uk > > >This email and any files transmitted with it are confidential and >intended solely for the use of the individual or entity to whom they >are addressed. If you have received this email in error please notify >the system manager using the details below. > >The contents of this email represent the views of the individual(s) >named above and do not necessarily represent the views of the >North East Wales NHS Trust. > >Please be aware that, under the terms of the Freedom of Information Act >2000, the North East Wales NHS Trust may be required to make public the >content of any emails or correspondence received. For futher >information on Freedom of Information, please refer to the North East >Wales NHS Trust website at www.newalesnhstrust.org.uk > >For further assistance, please contact >system.administrator@new-tr.wales.nhs.uk. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Feb 14 10:26:10 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:37 2005 Subject: [Possible SPAM] - RE: [Histonet] Polka dots and RE: Water bathtemperature[Scanned] - Found word(s)[Scanned][Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF81@bhrv-nt-11.bhrv.nwest.nhs.uk> Could you try something for me? Could you try post fixing the 45 degree section is something? Take two 45 degree sections treated exactly the same and post fix one in maybe mercury. Then stain both of them; if the formalin one has polka dots and the mercury one doesn't you have some answer. If both have polka dots, then I'm stumped. I'm just assuming it is the effect of extra heat on poorly fixed tissue that is stopping the mordanting effect of aluminium ions on the nucleic acids; a touch of mercury may be the solution. If it works, then fix in formalin longer; sound sense? -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 14 February 2005 16:06 To: histonet@lists.utsouthwestern.edu Subject: [Possible SPAM] - RE: [Histonet] Polka dots and RE: Water bathtemperature[Scanned] - Found word(s)[Scanned] Yes the waterbath is cleaned everyday and there is no wax left on it. I am sure that all the wax is being removed or we would see it on both section (from the waterbath at 38 and 45). I have done test time after time staining several slides (some cut at 38 and others at 45) at the same time and the slides cut at 45 will always show polka dots, even with fresh americlear (we do not use xylene) and alcohols. >>> Kemlo Rogerson 2/14/2005 11:01:29 AM >>> But why does it happen only with the 'hotter' bath? Surely the wax would not be removed from the sections floated out on the 'cooler' bath as well? You would assume that it is either the effect of the 'extra' heat or that there is something odd about that waterbath. I assume that the bath is clean and we are not dealing with wax melting in the 'hotter' bath and thereby contaminating the sections? -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 14 February 2005 15:43 To: JOHN PHILLIPS; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Polka dots and RE: Water bath temperature[Scanned] I agree with John on this polka dot problem. Also, stirring your paraffin redistributes polymer and other additives. This should be done just before embedding EACH day insure these polymers have not settled to the bottom into little blobs or solid polka dots that may not be totally removed by xylenes. In addition to increasing time in xylene, you can also add one more container of xylene or xylene substitute and change the first absolute alcohol after xylene more often to prevent excessive carry over of paraffin residues. A quick test for rehydration solvents. Take a pasteur pipette and sample each alcohol change after xylene. Squirt this sample into water. If it turns milky white, then you have carry over of xylene and paraffin into the alcohols, and need to rotate out all changes more frequently. If this happens in the 95% just before 70% and water, you really have trouble with paraffin residues removal. At 02:42 AM 2/14/2005, you wrote: >The problem with staining may be due to unsatisfactory removal of the wax >prior to staining for H&E. I think you are looking to the wrong place for >the solution to your problem. Try extending your dewax time in xylene >before you hydrate your sections on the staining run. If the problem goes >away PERMANANTLY then you have your answer. Good luck. > >John, >Wrexham,Wales.U.K. > >-----Original Message----- >From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] >Sent: 11 February 2005 19:24 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Water bath temperature > > >We were having a problem that the tissue would not take up the >hematoxlyin but would take up the eosin but only in some areas, so it >looked like polka dots. We started one by one changing solution trying >to figure out where it was coming from. We had all fresh solutions and >still had polka dots. I tried the only other thing left to change, I >lowered the temperature of the water bath from 45 to 38 and this worked. > We currently are using ParaPlast Xtra with a melting point of 52. We >have all tried figure out why the temperature in the water bath will >make the difference. Several times I have cut the same tissue at >different temperatures ranging from 38 - 45 and every time there are >polka dots at the higher temperatures. Does anyone have any idea why >this would be? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn >gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu >atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r >rheolwr systemau wybod drwy ddefnyddio'r manylion isod. > >Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, >felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain >Cymru. > >Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i >Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys >unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau?r Ddeddf >Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, >cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru >ar www.newalesnhstrust.org.uk > > >This email and any files transmitted with it are confidential and >intended solely for the use of the individual or entity to whom they >are addressed. If you have received this email in error please notify >the system manager using the details below. > >The contents of this email represent the views of the individual(s) >named above and do not necessarily represent the views of the >North East Wales NHS Trust. > >Please be aware that, under the terms of the Freedom of Information Act >2000, the North East Wales NHS Trust may be required to make public the >content of any emails or correspondence received. For futher >information on Freedom of Information, please refer to the North East >Wales NHS Trust website at www.newalesnhstrust.org.uk > >For further assistance, please contact >system.administrator@new-tr.wales.nhs.uk. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@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 nena.dimaano <@t> stryker.com Mon Feb 14 10:36:59 2005 From: nena.dimaano <@t> stryker.com (Dimaano, Nena) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Job Description: Scientist Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD13F349@HOS2KEXCHCL.howost.strykercorp.com> My boss and I need your help in setting up a job description. We wanted to find out if there were someone doing histology work with an associate scientist or scientist job title and is willing to share a sample of his or her job description. I appreciate your help. Sincerely, Nena Dimaano,MT/HT(ASCP) Advanced Technology Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com From xuesong.chen <@t> yale.edu Mon Feb 14 10:43:03 2005 From: xuesong.chen <@t> yale.edu (Xuesong Chen) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Loss if LacZ staining after mounting slides, Message-ID: <1108399383.4210d5173f8df@webmail.med.yale.edu> Hi histonetters, I need help on lacZ counter staining. I?m trying to stain bone/cartilage for beta galactosidase. H&E does not seem a good choice because lacZ blue is not accentrated. I have been using Nuclear Fast Red (CI 60760) as a nuclear counterstain, but after dehydrating and mounting I lose the the entire beta gal over the course of two or three weeks. My protocol: Cryojane sectioning femur/tibia and air dry the slides 10 min in 1XPBS 5 min in 2%PFA in 1XPBS 10 min washing in 1XPBS Beta galactosidase staining overnight at 37C in dark 10 min washing in 1XPBS 15 min counterstaining in 0.1% nuclear fast red 10 min washing in 1XPBS 5 min in 70% ethanol 5 min in 95% ethanol 5 min in 100% ethanol 5 min in Histoclear 5 min in Histoclear Cytoseal 60 (low viscosity) to cover the slip. I'm wondering is there any people using nuclear fast red for beta-gal counterstain and whether you have experienced the same problem, or alternatively, advice any possibilities that cause this. Thanks Xuesong From Kemlo.Rogerson <@t> elht.nhs.uk Mon Feb 14 10:51:02 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] SurgiPath MacroPath[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF85@bhrv-nt-11.bhrv.nwest.nhs.uk> Has anyone any experience of using this analogue video camera set up with computer and image handling software? It's sold to assist with recording evidence at the cut up (?grossing). From Terry.Marshall <@t> rothgen.nhs.uk Mon Feb 14 11:16:44 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature Message-ID: Why from the description is it not just our old friend pink disease? (BayleyJH {1949}J.Path.BAct., 61,448) Are the polka dots too small or too many or too invariable, or any combination thereof? As usual, a picture would be worth a thousand words. She wore an itsy bitsy teeny weeny polka dot bikini ....... I was never sure if it was the polka dots that were teeny weeny. Probably not. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 11 February 2005 19:24 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Mon Feb 14 11:18:06 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] " Dinking" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB41AC@fh2k093.fhmis.net> you mean...fooling around with...? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of George Cole Sent: Sunday, February 13, 2005 3:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] " Dinking" Hey Techs out there--- there are possible, highly interesting useages for the Term "dinking" around. There are a couple of entries in Webster's 9th New Collegiate Dictionary: 1. Dink: to shorten and 2. to make a drop shot---(for you basketball fans). Histotechs, said to be dinking about, I suppose, could 1: be described as shortening the confusion by chugging about finding answers and or 2 Making a drop shot with a good answer. But the next line under these two entries are the terms: Dinky or Dinkey----a small locomotive used esp. for hauling freight, logging and shunting----where "dinking" refers to hauling freight, especially in movements of goods about the yard----also used in the slang version ---"dinking about"-like a small; busy locomotive busy about the train yards that picks up and moves things----but in this usage, it is a person picking up questions, picking away at them, as one who picks away at a problem until it is dropped next to its answer. I hope that both Merriam and Webster publishing since 1831, will be beyond caring if their entries are chugged, ,juggled and dinked about the Great Yard of Usage. georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From conniemoss <@t> relia.net Mon Feb 14 12:00:37 2005 From: conniemoss <@t> relia.net (Connie McManus) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Equipment needed Message-ID: <1199.207.173.181.11.1108404037.squirrel@email.relia.net> Hi everyone. We are looking for histology equipment for our labs. We need... 1 tissue processor 2 microtome 3 stainer 4 IHC stainer If you have any of these you would like to sell, please contact me privately. Thanks bunches! -- Connie McManus Mt Ogden Scientific Services 950 W Kershaw, Suite E Ogden UT 8401 tel: 866/933-6677 fax: 435/514-1781 conniemoss@relia.net www.mtogdensci.com From conniemoss <@t> relia.net Mon Feb 14 12:06:01 2005 From: conniemoss <@t> relia.net (Connie McManus) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Equipment needed 2 Message-ID: <1209.207.173.181.11.1108404361.squirrel@email.relia.net> I forgot, we also need... 1 embedder 2 slide and cassette labelers thanks again. -- Connie McManus Mt Ogden Scientific Services 950 W Kershaw, Suite E Ogden UT 8401 tel: 866/933-6677 fax: 435/514-1781 conniemoss@relia.net www.mtogdensci.com From rmeyers <@t> ctcsurge.com Mon Feb 14 12:08:04 2005 From: rmeyers <@t> ctcsurge.com (Roger Meyers) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] automated response Message-ID: <10502141308.AA11303842@mail.ctcsurge.com> I will be out of the office from Monday, February 14 through Thursday, February 17, 2005. If you need immediate assistance, please call Computer Trust Corporation at 617-557-9264. From KMH.02 <@t> ex.uchs.org Mon Feb 14 12:14:47 2005 From: KMH.02 <@t> ex.uchs.org (Hopkins, Karen) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Dako Autostainer Message-ID: <640B23A5DC4B234BB065E56F2DB30596D9625B@uchex2ucmc.uchs.org> Hi We have had a problem with our IHC autostainer and I was wondering if anyone else has experienced the same problem. We had a Dako Autostainer and over time (about three years) found that we had intermittent problems with slides not being stained. It started with slides in the first and second slide position (the positive control was not staining) when we had a large run but eventually we noticed it at other positions, spiratically. We have another instrument now, since Nov and the other day experienced the same problem. We ran three sets of known controls (we send one to each of the facilities we stain for) and one of the three was positive, the other two were not. We were using a RTU ER antibody. ???? Also, the spec sheet from Dako states we should run the ER with Env or Lsab2. We get better results with ENV+. The stain with ENV is extremely pale. Any comments or suggestions? From ploykasek <@t> phenopath.com Mon Feb 14 12:37:36 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] LAB solution Message-ID: Hi. I was wondering if anyone has tried this room temperature antigen unmasking solution called L.A.B. From Polysciences? Thanks for the info. I am receiving a sample, and am willing to share my experience. Patti Loykasek PhenoPath Laboratories From jkiernan <@t> uwo.ca Mon Feb 14 12:40:20 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Wax removal incomplete Message-ID: <4210F094.55088792@uwo.ca> A recent publication from Ireland shows (by laser confocal Raman microspectroscopy, no less) that wax removal from sections, as usually practised, is always incomplete. The authors were able to remove all the paraffin, however, with hexane. The other things they tried were xylene, "histoclear" followed by hot antigen retrieval, and "trilogy" (a product that combines dewaxing and retrieval - ? a solvent-detergent mixture). All three failed to remove all the wax, even with repeated, prolonged treatments. For hexane, 18 hours were needed for complete was removal. The investigators found that the residual paraffin seriously interfered with the Raman spectra of rehydrated sections. They are investigating the Raman technique (which they explain fairly simply) for early diagnosis of tumours. They also claim stronger immunostaining after dewaxing in hexane (18h) than after xylene (18h). They tested only one antibody (to cytokeratin), and the pair of photos doesn't make the point very impressively. They authors also state that hexane is less toxic than xylene. I can add that the least expensive n-hexane is a little cheaper than xylenes (mixed isomers) in one catalogue checked. Both solvents are flammable, and n-hexane (BP 69C) is more volatile than xylene. Both solvents are miscible with 100% alcohol. The reference is: O'Faolain E et 6 al. (2005) Raman spectroscopic evaluation of efficacy of current paraffin wax section dewaxing agents. J. Histochem. Cytochem. 53(1): 121-129. -- ------------------------------- 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 _______________________________ From JQB7 <@t> CDC.GOV Mon Feb 14 13:11:00 2005 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Histology Schools Message-ID: There are many schools, but not in Alabama: Arkansas Arizona California Connecticutt DC Delaware Florida Georgia Illinois Indiana Maryland Michigan Minnesota New Mexico New Jersey New York Ohio Pennsylvania Texas Wisconsin -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jim Ball Sent: Sat 2/12/2005 2:04 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Histology Schools Is there an accredited histology school in the state of Alabama, that may have been assosiated with the University of Alabama. I was told there use to be one, but it lost its accreidation, if this is true do they plan on reapplying for accreidation. I have a friend that would like this information, but does not have access to the inter-net. I told him I would ask the histonet and relay all respones to him. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RossS <@t> BaylorHealth.edu Mon Feb 14 13:32:27 2005 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Dako Autostainer Message-ID: <17A8DA5A05167C4EA20BFA750167736B0BB6F7@BHDAEXCH13.bhcs.pvt> At my former institution we were doing a large volume of immuno's and would every 3 to 6 months start having inconsistant staining and have to get parts replaced on the probe assembly. That usually fixed it, so we got the the point where every 3 months we had them in for a PM. The components that were problematic were part of the normal PM. One time the problem continued and multiple things were replaced (this was on a 4 or 5 year old machine). It turned out that it was the power supply transformer in the instrument. It would occassionally kick out an odd voltage and thus the pipet would pick up too little reagent. In spite of that problem I personally still prefer the Dako. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hopkins, Karen Sent: Monday, February 14, 2005 12:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dako Autostainer Hi We have had a problem with our IHC autostainer and I was wondering if anyone else has experienced the same problem. We had a Dako Autostainer and over time (about three years) found that we had intermittent problems with slides not being stained. It started with slides in the first and second slide position (the positive control was not staining) when we had a large run but eventually we noticed it at other positions, spiratically. We have another instrument now, since Nov and the other day experienced the same problem. We ran three sets of known controls (we send one to each of the facilities we stain for) and one of the three was positive, the other two were not. We were using a RTU ER antibody. ???? Also, the spec sheet from Dako states we should run the ER with Env or Lsab2. We get better results with ENV+. The stain with ENV is extremely pale. Any comments or suggestions? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From rcharles <@t> state.pa.us Mon Feb 14 13:59:45 2005 From: rcharles <@t> state.pa.us (Charles, Roger) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] shirley powell Spirochete IHC Message-ID: I am looking for Shirley Powell to contact me about her spirochete ihc. I would like to know if this ab also works on leptospirosis? Roger Charles Microbiologist Pennsylvania Veterinary Laboratory 2305 N Cameron St Harrisburg, PA 17110 717-787-8808 From akbitting <@t> geisinger.edu Mon Feb 14 15:42:54 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Cutting sections using Supercassettes Message-ID: Hi All, I am, once again, ona quest for knowledge.... I need to adapt my rotary microtomes, Microm 315s and R-J 2030s, to be able to cut whole prostate sections using Surgipath's super cassettes. Can anyone lead me to a clamp that will work? 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 Kemlo.Rogerson <@t> elht.nhs.uk Tue Feb 15 02:18:34 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Polka dots and RE: Water bath temperature[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF89@bhrv-nt-11.bhrv.nwest.nhs.uk> I am honoured to act as Gayle's secretary, despite her getting my name wrong . -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 14 February 2005 18:58 To: Kemlo Rogerson Subject: RE: [Histonet] Polka dots and RE: Water bath temperature[Scanned] Kemalo , I unsubscribed from Histonet this a.m. for travel. You can put my reply on Histonet if you wish. If the waterbath is not the problem and all rehydration agents have been tested or changed with timing reevaluated for good paraffin removal AND the paraffin is stirred before embedding, maybe the culprit is the paraffin itself. Age of paraffin lot or possibly defective paraffin during manufacturing. Then manufacturer could be contacted and asked about this problem. At 09:01 AM 2/14/2005, you wrote: >But why does it happen only with the 'hotter' bath? Surely the wax would not >be removed from the sections floated out on the 'cooler' bath as well? You >would assume that it is either the effect of the 'extra' heat or that there >is something odd about that waterbath. I assume that the bath is clean and >we are not dealing with wax melting in the 'hotter' bath and thereby >contaminating the sections? > >-----Original Message----- >From: Gayle Callis [mailto:gcallis@montana.edu] >Sent: 14 February 2005 15:43 >To: JOHN PHILLIPS; Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Polka dots and RE: Water bath temperature[Scanned] > >I agree with John on this polka dot problem. Also, stirring your paraffin >redistributes polymer and other additives. This should be done just before >embedding EACH day insure these polymers have not settled to the bottom >into little blobs or solid polka dots that may not be totally removed by >xylenes. In addition to increasing time in xylene, you can also add one >more container of xylene or xylene substitute and change the first absolute >alcohol after xylene more often to prevent excessive carry over of paraffin >residues. > >A quick test for rehydration solvents. Take a pasteur pipette and sample >each alcohol change after xylene. Squirt this sample into water. If it >turns milky white, then you have carry over of xylene and paraffin into the >alcohols, and need to rotate out all changes more frequently. If this >happens in the 95% just before 70% and water, you really have trouble with >paraffin residues removal. > > > >At 02:42 AM 2/14/2005, you wrote: > >The problem with staining may be due to unsatisfactory removal of the wax > >prior to staining for H&E. I think you are looking to the wrong place for > >the solution to your problem. Try extending your dewax time in xylene > >before you hydrate your sections on the staining run. If the problem goes > >away PERMANANTLY then you have your answer. Good luck. > > > >John, > >Wrexham,Wales.U.K. > > > >-----Original Message----- > >From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] > >Sent: 11 February 2005 19:24 > >To: histonet@lists.utsouthwestern.edu > >Subject: [Histonet] Water bath temperature > > > > > >We were having a problem that the tissue would not take up the > >hematoxlyin but would take up the eosin but only in some areas, so it > >looked like polka dots. We started one by one changing solution trying > >to figure out where it was coming from. We had all fresh solutions and > >still had polka dots. I tried the only other thing left to change, I > >lowered the temperature of the water bath from 45 to 38 and this worked. > > We currently are using ParaPlast Xtra with a melting point of 52. We > >have all tried figure out why the temperature in the water bath will > >make the difference. Several times I have cut the same tissue at > >different temperatures ranging from 38 - 45 and every time there are > >polka dots at the higher temperatures. Does anyone have any idea why > >this would be? > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn > >gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu > >atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r > >rheolwr systemau wybod drwy ddefnyddio'r manylion isod. > > > >Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, > >felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain > >Cymru. > > > >Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i > >Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys > >unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau?r Ddeddf > >Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, > >cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru > >ar www.newalesnhstrust.org.uk > > > > > >This email and any files transmitted with it are confidential and > >intended solely for the use of the individual or entity to whom they > >are addressed. If you have received this email in error please notify > >the system manager using the details below. > > > >The contents of this email represent the views of the individual(s) > >named above and do not necessarily represent the views of the > >North East Wales NHS Trust. > > > >Please be aware that, under the terms of the Freedom of Information Act > >2000, the North East Wales NHS Trust may be required to make public the > >content of any emails or correspondence received. For futher > >information on Freedom of Information, please refer to the North East > >Wales NHS Trust website at www.newalesnhstrust.org.uk > > > >For further assistance, please contact > >system.administrator@new-tr.wales.nhs.uk. > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Kemlo.Rogerson <@t> elht.nhs.uk Tue Feb 15 02:28:28 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF8B@bhrv-nt-11.bhrv.nwest.nhs.uk> Doesn't the adjective affect the noun? So 'polka dot bikini' being the noun it implies that the 'polka dot bikini' was 'itsy bitsy'. You would have to say 'itsy bitsy bikini with teeny weeny polka dots' to mean what you have suggested. In order for this not to be frivolous please follow the link for an English lesson http://www-users.cs.york.ac.uk/~susan/cyc/a/adj.htm. -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 February 2005 17:17 To: Rebecca Barnhart; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Water bath temperature[Scanned] Why from the description is it not just our old friend pink disease? (BayleyJH {1949}J.Path.BAct., 61,448) Are the polka dots too small or too many or too invariable, or any combination thereof? As usual, a picture would be worth a thousand words. She wore an itsy bitsy teeny weeny polka dot bikini ....... I was never sure if it was the polka dots that were teeny weeny. Probably not. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 11 February 2005 19:24 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Tue Feb 15 02:30:16 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EF8C@bhrv-nt-11.bhrv.nwest.nhs.uk> Because, if you read the thread properly, it doesn't affect sibling H&E's cut from the same block but floated out on a hotter water bath. Maybe technical ought to be left to the 'technicians'. IMHO. -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 February 2005 17:17 To: Rebecca Barnhart; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Water bath temperature[Scanned] Why from the description is it not just our old friend pink disease? (BayleyJH {1949}J.Path.BAct., 61,448) Are the polka dots too small or too many or too invariable, or any combination thereof? As usual, a picture would be worth a thousand words. She wore an itsy bitsy teeny weeny polka dot bikini ....... I was never sure if it was the polka dots that were teeny weeny. Probably not. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 11 February 2005 19:24 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Tue Feb 15 03:59:51 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Histology Schools References: Message-ID: <007c01c51345$18d65ce0$752ed445@domainnotset.invalid> NAACLS (National Accrediting Agency for Clinical Laboratory Sciences), which accredits/approves HT/HTL, MT/MLT, Pathologist Assistant, Cytogenetics Technologists, Phlebotomy and Clinical Assistant programs - lists 25 accredited HT programs and 2 accredited HTL programs. http://www.naacls.org/search/programs.asp Under "TYPE", click on either HT or HTL, leave the "STATES" as ALL, and click on "SEARCH". I found no programs in Alabama. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Jim Ball" To: Sent: Saturday, February 12, 2005 2:04 PM Subject: [Histonet] Histology Schools > Is there an accredited histology school in the state of Alabama, that may > have been assosiated with the University of Alabama. I was told there use > to be one, but it lost its accreidation, if this is true do they plan on > reapplying for accreidation. I have a friend that would like this > information, but does not have access to the inter-net. I told him I would > ask the histonet and relay all respones to him. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dsnider <@t> shrinenet.org Tue Feb 15 07:07:28 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] non xylene based dehydrants Message-ID: Ok everyone, What non xylene based products are being used by everyone? What are their health risks, hazards, etc. compared to xylene? Do they work as well as it does? Is it easily disposed of? The lab here uses xylene, which is my choice, but the restrictions on storing, pouring on/off, disposal, etc. borders on paranoia...:) So I am exploring the alternatives. The only exposure I've had to this type of product was years ago. Hemo-D, which gave me tremendous headaches.... I appreciate all your imput, and thanks in advance! Deanna Snider HT Shriners Hospital for Children Research Dept. Cincinnati, OH 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 Terry.Marshall <@t> rothgen.nhs.uk Tue Feb 15 07:14:33 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature[Scanned] Message-ID: Follow your own link. Dot *is* a noun. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 15 February 2005 08:28 To: Marshall Terry Dr, Consultant Histopathologist; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Water bath temperature[Scanned] Doesn't the adjective affect the noun? So 'polka dot bikini' being the noun it implies that the 'polka dot bikini' was 'itsy bitsy'. You would have to say 'itsy bitsy bikini with teeny weeny polka dots' to mean what you have suggested. In order for this not to be frivolous please follow the link for an English lesson http://www-users.cs.york.ac.uk/~susan/cyc/a/adj.htm. -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 February 2005 17:17 To: Rebecca Barnhart; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Water bath temperature[Scanned] Why from the description is it not just our old friend pink disease? (BayleyJH {1949}J.Path.BAct., 61,448) Are the polka dots too small or too many or too invariable, or any combination thereof? As usual, a picture would be worth a thousand words. She wore an itsy bitsy teeny weeny polka dot bikini ....... I was never sure if it was the polka dots that were teeny weeny. Probably not. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 11 February 2005 19:24 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Tue Feb 15 07:36:43 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature[Scanned] Message-ID: Thank you Kemlo for your insightful comments. However, though necessarily amateurish about things technical, I can't help feeling that:- "I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked.", means the exact opposite of what you said, viz. "Because, if you read the thread properly, it doesn't affect sibling H&E's cut from the same block but floated out on a hotter water bath." Perhaps, if you read the thread properly, you may prevent making a dick of yourself with these gratuitously rude comments. Furthermore, until the cause of pink disease is known, your arguement that it couldn't have been that merely because schedule A prevented it and schedule B didn't (however befuddled you were about which was which) does not follow. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk From: Kemlo Rogerson Because, if you read the thread properly, it doesn't affect sibling H&E's cut from the same block but floated out on a hotter water bath. Maybe technical ought to be left to the 'technicians'. IMHO. -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist Why from the description is it not just our old friend pink disease? (BayleyJH {1949}J.Path.BAct., 61,448) Are the polka dots too small or too many or too invariable, or any combination thereof? As usual, a picture would be worth a thousand words. She wore an itsy bitsy teeny weeny polka dot bikini ....... I was never sure if it was the polka dots that were teeny weeny. Probably not. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 11 February 2005 19:24 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@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 browning <@t> HHSC.CA Tue Feb 15 07:45:20 2005 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] B-Plus Fixative Message-ID: <3AADFB88753AD31189C100902786B91C0E278702@hch_nt_exchange.hhsc.ca> We are currently using B5 fixative for bone marrows, and as much as we don't like using it, and it gives problems with some of the antibodies, after extensive testing of B5 substitutes, I found them all inferior to B5. The only fixative that gave consistent staining with all antibodies was 10% NBF, but only after 24 hours fixation, and this time limitation was not acceptable for some clinical cases. Our biggest problem with 10% NBF was producing a H&E section that demonstrated the same crispness of nuclear detail that is achieved with B5. -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Friday, February 11, 2005 4:30 PM To: Histonet Subject: [Histonet] B-Plus Fixative My pathologists are unhappy with B-Plus fixed bone marrows, especially immunos. They have pulled previous cases on patient that were fixed in B5 and of couse see quite a difference side by side. Does anyone else hear this complaint? 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. 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 lchung <@t> ppmh.org Tue Feb 15 08:02:20 2005 From: lchung <@t> ppmh.org (Chung, Luong) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] B-Plus Fixative Message-ID: All, Since we are on the B-5 topic, does anyone still using B-5 in the state of Georgia? Is it a mandatory or a voluntary to stop using B-5? Thanks for any information. Bruce Chung, MSM, CT(ASCP) Anatomic Pathology Manager Phoebe Putney Memorial Hospital lchung@ppmh.org From mark.lewis <@t> thermo.com Tue Feb 15 08:08:50 2005 From: mark.lewis <@t> thermo.com (LEWIS, MARK A.) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Wax removal incomplete Message-ID: I often wondered about this due to the "plastic "polymers added to the different types of paraffins. Perhaps it's these "polymers" that are not being removed with the typical solvents. Interesting ... Thanks John ! Mark ? Mark A. Lewis B.A., H.T.(ASCP) Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation 171 Industry Drive Pittsburgh, Pa. 15275 Phone:? 412-747-4013 Fax?????? 412-788-6557 E-mail: mark.lewis@thermo.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Monday, February 14, 2005 1:40 PM To: Histonet Subject: [Histonet] Wax removal incomplete A recent publication from Ireland shows (by laser confocal Raman microspectroscopy, no less) that wax removal from sections, as usually practised, is always incomplete. The authors were able to remove all the paraffin, however, with hexane. The other things they tried were xylene, "histoclear" followed by hot antigen retrieval, and "trilogy" (a product that combines dewaxing and retrieval - ? a solvent-detergent mixture). All three failed to remove all the wax, even with repeated, prolonged treatments. For hexane, 18 hours were needed for complete was removal. The investigators found that the residual paraffin seriously interfered with the Raman spectra of rehydrated sections. They are investigating the Raman technique (which they explain fairly simply) for early diagnosis of tumours. They also claim stronger immunostaining after dewaxing in hexane (18h) than after xylene (18h). They tested only one antibody (to cytokeratin), and the pair of photos doesn't make the point very impressively. They authors also state that hexane is less toxic than xylene. I can add that the least expensive n-hexane is a little cheaper than xylenes (mixed isomers) in one catalogue checked. Both solvents are flammable, and n-hexane (BP 69C) is more volatile than xylene. Both solvents are miscible with 100% alcohol. The reference is: O'Faolain E et 6 al. (2005) Raman spectroscopic evaluation of efficacy of current paraffin wax section dewaxing agents. J. Histochem. Cytochem. 53(1): 121-129. -- ------------------------------- 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 _______________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Tue Feb 15 08:49:27 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EFA8@bhrv-nt-11.bhrv.nwest.nhs.uk> I'm not too proud to admit that you have me on that. Can't win all the time, I apologise. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 15 February 2005 13:37 To: Kemlo Rogerson; Rebecca Barnhart; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Water bath temperature[Scanned] Thank you Kemlo for your insightful comments. However, though necessarily amateurish about things technical, I can't help feeling that:- "I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked.", means the exact opposite of what you said, viz. "Because, if you read the thread properly, it doesn't affect sibling H&E's cut from the same block but floated out on a hotter water bath." Perhaps, if you read the thread properly, you may prevent making a dick of yourself with these gratuitously rude comments. Furthermore, until the cause of pink disease is known, your arguement that it couldn't have been that merely because schedule A prevented it and schedule B didn't (however befuddled you were about which was which) does not follow. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk From: Kemlo Rogerson Because, if you read the thread properly, it doesn't affect sibling H&E's cut from the same block but floated out on a hotter water bath. Maybe technical ought to be left to the 'technicians'. IMHO. -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist Why from the description is it not just our old friend pink disease? (BayleyJH {1949}J.Path.BAct., 61,448) Are the polka dots too small or too many or too invariable, or any combination thereof? As usual, a picture would be worth a thousand words. She wore an itsy bitsy teeny weeny polka dot bikini ....... I was never sure if it was the polka dots that were teeny weeny. Probably not. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 11 February 2005 19:24 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From julien_lambreydesouza <@t> uqar.qc.ca Tue Feb 15 08:51:01 2005 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien Lambrey De Souza) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] plant samara Message-ID: <005501c5136d$c44a3dd0$cd11d784@rcloutier06> Hello all histoneters, I am having a hard time cutting samara (birch fruit) using the classical paraffin method. I am also having no success in finding information for this technique on the net and in relevant publications. It seems that plant histology is not as popular as animal or human histology... and not as well documented. I have no problem cutting the samara up to a certain date, after which the pericarp seems to harden. I must find a way to soften the pericarp of the samara prior to cutting for the more developed fruits. I have tried soaking the blocks for a few days in a warm mixture of acetic acid and ethanol (1:9), but this yielded no success. I then have tried to soften the samara prior to infiltration with the same mixture. This doesn't seem to soften them at all. I would appreciate any advice. Is their anyone out there doing routine histology on plant material? Thanks, Julien Lambrey de Souza Biologie ?volutive, Universit? du Qu?bec ? Rimouski, (418) 723-1986 #1438 From Luis.Chiriboga <@t> med.nyu.edu Tue Feb 15 09:11:54 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Wax removal incomplete In-Reply-To: Message-ID: Likewise, I have also wondered about these "plasticizers" added to paraffin. There really is no literature (that I can find) discussing this issue. We recently were studying Pten expression in skin samples dating as far back as the late 70's. Interestingly, samples collected in the mid 1980's did not deparaffinize as well as more recent (and older) samples. It turned out that these samples required substantial heating (upwards of 80C, normally we do at 60C) to generate IHC labeling. In regards to the Raman paper, I have done quite a bit of similar work with FTIR in FFPE tissue and would like to point out some critical points. Both Raman and FTIR techniques are very sensitive to molecular structure. Unfortunately the authors do not describe or show from where (histologically) they collected their spectra. This can influence the resulting spectra quite dramatically (remember they are using a 2um spot size). Although both techniques are considered "fingerprinting" methods in that they can identify compounds based on their unique vibrational signatures, they also suffer from the averaging effect. Thus, other compounds (many compounds will have C-C stretching and CH2 bending and deformation modes) that produce similar spectral signatures will be averaged into the overall spectrum. 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 LEWIS, MARK A. Sent: Tuesday, February 15, 2005 9:09 AM To: John Kiernan; Histonet Subject: RE: [Histonet] Wax removal incomplete I often wondered about this due to the "plastic "polymers added to the different types of paraffins. Perhaps it's these "polymers" that are not being removed with the typical solvents. Interesting ... Thanks John ! Mark ? Mark A. Lewis B.A., H.T.(ASCP) Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation 171 Industry Drive Pittsburgh, Pa. 15275 Phone:? 412-747-4013 Fax?????? 412-788-6557 E-mail: mark.lewis@thermo.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Monday, February 14, 2005 1:40 PM To: Histonet Subject: [Histonet] Wax removal incomplete A recent publication from Ireland shows (by laser confocal Raman microspectroscopy, no less) that wax removal from sections, as usually practised, is always incomplete. The authors were able to remove all the paraffin, however, with hexane. The other things they tried were xylene, "histoclear" followed by hot antigen retrieval, and "trilogy" (a product that combines dewaxing and retrieval - ? a solvent-detergent mixture). All three failed to remove all the wax, even with repeated, prolonged treatments. For hexane, 18 hours were needed for complete was removal. The investigators found that the residual paraffin seriously interfered with the Raman spectra of rehydrated sections. They are investigating the Raman technique (which they explain fairly simply) for early diagnosis of tumours. They also claim stronger immunostaining after dewaxing in hexane (18h) than after xylene (18h). They tested only one antibody (to cytokeratin), and the pair of photos doesn't make the point very impressively. They authors also state that hexane is less toxic than xylene. I can add that the least expensive n-hexane is a little cheaper than xylenes (mixed isomers) in one catalogue checked. Both solvents are flammable, and n-hexane (BP 69C) is more volatile than xylene. Both solvents are miscible with 100% alcohol. The reference is: O'Faolain E et 6 al. (2005) Raman spectroscopic evaluation of efficacy of current paraffin wax section dewaxing agents. J. Histochem. Cytochem. 53(1): 121-129. -- ------------------------------- 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 _______________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bosborn <@t> hsc.usf.edu Tue Feb 15 09:36:56 2005 From: bosborn <@t> hsc.usf.edu (Barbara Osborn) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Re: LAB solution Message-ID: <004501c51374$2ec00f40$9d46f783@hscnet.hsc.usf.edu> I have tried LAB with multiple antibodies and have had pretty good success. It has worked with various cytokeratins, vimentin, aSMA, and several other "easy to retreive" antigens by incubating for 20 min at RT. However, I had no success with Ki-67, not even by heating to 60 deg. The problem I had, however, is that the solution started to form a precipitate after a short time, and then failed to work. This was just after a couple of months. I called Polysciences and they were happy to replace the bottle. So far the new bottle has also formed the precipitate, but appears to still be working. They claimed the shelf life should be atleast a year. There are no special storage requirements, RT and keep out of direct sunlight. I store it in a closed cabinet to keep any light away because it comes in a clear plastic bottle. Hope this helps. From P.Krawczyk <@t> amc.uva.nl Tue Feb 15 09:38:22 2005 From: P.Krawczyk <@t> amc.uva.nl (P. Krawczyk) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Immunofluorescence staining artefact? Message-ID: <000c01c51374$61da55c0$08337591@Przemek> Hallo, I'm performing a double fluorescent staining (Cy3 + FITC) of adherent cells fixed with 2%paraformaldehyde. In my stained cells, apart of the typical staining pattern i see small, very bright Cy3 dots, moving fast with a kind of Brownian motion. Their size is in the microscope resolution area (viewed with 100x magn.) Nothing can be spotted with the phase contract, so it's not bacteria. I tried filtering, centrifuging, nothing... It seems, that a long washing reduces this artefact. Are these Cy3 clumps? Anyone knows what it is and how to get rid of it? Any help appreciated, Regards, P. Krawczyk _____________________________ P. M. Krawczyk Dept. of Cell Biology and Histology Academic Medical Centre University of Amsterdam Meibergdreef 15, room M3-107 1105 AZ, Amsterdam The Netherlands Email: P.Krawczyk@amc.uva.nl Tel: +31 20 5668746 Fax: +31 20 6974156 From pathrm35 <@t> adelphia.net Tue Feb 15 09:45:27 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] CAP and proficiency testing Message-ID: <27588852.1108482327958.JavaMail.root@web1.mail.adelphia.net> Does anyone know what the requirements are for proficiency testing for CAP accredited private histology laboratories? Thanks, Ron Martin From TJJ <@t> Stowers-Institute.org Tue Feb 15 09:49:26 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Universal antigen retrieval method Message-ID: Dr. Kiernan brought our attention to a current article on dewaxing agents from the JHC (Vol 53(1): 121-129, 2005) that I read yesterday. In addition to anticipating more discussion about this, I would also like to highlight another article in the same volume. These researchers from Japan are using citraconic anhydride and heat to reverse protein cross-linking in formalin-fixed, paraffin-embedded (FFPE) tissue samples. This may be a very useful tool for those of us doing immunostaining with antibodies that are not characterized for use in FFPE samples. The authors did prospective parallel studies with FFPE and frozen section, although they didn't indicate how the frozen sections were handled, i.e. freshly frozen and acetone fixed post sectioning? At any rate, give it a read and let us know what you think. Is this really as good as it sounds? Namimatsu S et al. (2005) Reversing the Effects of Formalin Fixation with Citraconic Anhydride and Heat: A Universal Antigen Retrieval Method, J Histochem Cytochem. 53(1): 3-11 Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From maxim_71 <@t> mail.ru Tue Feb 15 09:55:34 2005 From: maxim_71 <@t> mail.ru (Maxim) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Checking the endpoint of decalcification Message-ID: <122260515.20050215185534@mail.ru> Dear Histonetters! For checking the endpoint of decalcification is traditionally used 5% solution sodium oxalate or ammonium oxalate. Solubility in 100 ml water for sodium oxalate 3,7v/w, but for ammonium oxalate forms accordingly 4,5 v/w. Thereby, we get the saturated solutions of these salts. Why to this effect do not use potassium oxalate? His solubility in 100 ml water forms 26,4w/v. Can be therefore that in solution is got too high concentration oxalate and this test not will so be sensitive? Or this there is else some explanation? Thank you. Maxim Peshkov, HTL Department of biopsy and cytological research Pathological and anatomical bureau Taganrog Russia, 347942. mailto:maxim_71@mail.ru From jengirl1014 <@t> yahoo.com Tue Feb 15 10:02:06 2005 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Re: non xylene based dehydrates Message-ID: <20050215160207.2800.qmail@web60605.mail.yahoo.com> I use Clear Rite 3 by Richard Allen Scientific. It works great! There is no smell and it is less toxic than the Citirisolv and Xylene. Richard Allen is even nice enough to give you a free sample to try out there product! That's how I got hooked! I even use it for staining, and I get beautiful stains. Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From akbitting <@t> geisinger.edu Tue Feb 15 10:28:19 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] CAP and proficiency testing Message-ID: Our proficiency testing is very simple. We named six general functions and gave an excellent, good or poor rating. Method was direct observation. The tester and techs signed and dated it. Repeat it annually. 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 >>> 02/15/05 10:45AM >>> Does anyone know what the requirements are for proficiency testing for CAP accredited private histology laboratories? Thanks, Ron Martin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From p.j.bergin <@t> qmul.ac.uk Tue Feb 15 10:28:39 2005 From: p.j.bergin <@t> qmul.ac.uk (p.j.bergin@qmul.ac.uk) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Iron staining (cellular) Message-ID: <1108484919.42122337e0844@webapps.qmul.ac.uk> Hi, I was hoping someone could give me advice. I have looked through the archives and have tried a few things already. I am interested in staining cell preps to localize iron. I have tried using mouse spleen cells as a trial, but am mainly interested in human macrophages and monocytes that have been grown on glass slides. I have tried fixing the preps with a methanol/acetone mix and with 4% formalin. I have been trying two different methods; Perls Prussian blue from the stainsfile archive and Turnbull's Blue (for ferric and ferrous iron respectively). Both have been unsuccessful with no staining detected (and the cells are there). Does anyone have a method for these stains that works on cells? do they need to be fixed a specific way? Is there any advice anyone would care to share? Thanks in advance for any help. Phil P.S. The staining protocols I have been using follow..... FERROUS IRON - TURNBULL?S BLUE PURPOSE: To detect ferrous (Fe2+) iron in tissues. PRINCIPLE: Tissue sections are treated with an acidic solution of potassium ferricyanide, any ferrous iron present will react to form an insoluble bright blue pigment called Turnbull?s blue (ferrous ferricyanide). CONTROL: Routine iron control, which includes an negative. TECHNIQUE: Cut paraffin section 4?. EQUIPMENT: Acid clean glassware, non-metallic forceps. REAGENTS: 0.006N Hydrochloric Acid 1% Acetic Acid Hydrochloric acid 2.5 ml Acetic acid, glacial 1.0 ml Distilled water 497.5 ml Distilled water 100.0 ml Mix well. Solution is stable for 1 Mix well. Solution is stable for 1 year. year. CAUTION: Corrosive acid. CAUTION: Avoid contact and inhalation. Potassium Ferricyanide Nuclear-Fast Red: Staining Solution: See Retic Potassium ferricyanide 0.4 gm Hydrochloric acid, 0.006N 40.0 ml Prepare fresh, just before use. CAUTION: Avoid contact and inhalation. SAFETY: Wear gloves, goggles and lab coat. Avoid contact and inhalation. Hydrochloric acid; target organ effects on reproductive system and fetal tissue. Irritant to skin eyes and respiratory system. Potassium ferricyanide; Low toxicity as long as it is not heated, it will release cyanide gas. MINERALS AND PIGMENTS TURNBULL?S FERROUS IRON Page: 2 of 2 Acetic acid: Irritating to respiratory system. Target organ effects on respiratory system by inhalation. Corrosive. PROCEDURE: 1. Deparaffinize and hydrate to distilled water. 2. Place slides in Potassium ferricyanide staining solution for 1 hour. 3. Wash slides in 1% acetic acid. 4. Counterstain slides in nuclear-fast red for 5 minutes. 5. Rinse well in distilled water. 6. Dehydrate, clear and coverslip. RESULTS: Ferrous iron blue Background pink-red REFERENCE: Carson, F, Histotechnology: A Self-Instructional Text, 1st Ed., 1991, pp. 215-16 ASCP Press Perls Prussian Blue Principle The basis for the method is the release of ferric iron from hemosiderin by acid treatment, forming ferric chloride. The ferric iron reacts with potassium ferrocyanide to form ferric ferrocyanide. This is an insoluble, blue compound known as Prussian blue. The intensity of the colour gives some indication as to amount, but it is qualitative only. 4FeCl3 + 3K4Fe(CN)6 = Fe4[Fe(CN)6]3 + 12KCl Stock solution A Hydrochloric acid 2 ml Distilled water 98 ml Stock solution B Potassium ferrocyanide 2 g Distilled water 100 ml Working solution C Solution A 1 part Solution B 1 part Counterstain solution D Neutral red counterstain Fixation & processing Avoid iron containing materials and jars while fixing as these may contaminate the tissue. Acid containing fixatives may remove some of the iron deposits. Otherwise most methods are satisfactory. Method Bring sections to distilled water with xylene and ethanol. Place into working solution C for 15 minutes. Rinse with distilled water, then tap water. Stain with counterstain solution D for one minute Rinse well with tap water. Dehydrate with ethanol. Clear with xylene Expected Results Nuclei - red Notes The working solution C must be made immediately before use. Avoid washing with tap water before treating with solution C, as rust in the water or tap fixtures could cause false positive staining. Wash well at step 3, as traces of iron will form a granular red deposit with neutral red. Iron ores can be demonstrated, but the acid concentration in solution A may need to be increased to 10% or more. From akbitting <@t> geisinger.edu Tue Feb 15 10:30:18 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] non xylene based dehydrants Message-ID: I've used both ClearRite and Histoclear with very good results. 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 >>> "Snider, Deanna" 02/15/05 08:07AM >>> Ok everyone, What non xylene based products are being used by everyone? What are their health risks, hazards, etc. compared to xylene? Do they work as well as it does? Is it easily disposed of? The lab here uses xylene, which is my choice, but the restrictions on storing, pouring on/off, disposal, etc. borders on paranoia...:) So I am exploring the alternatives. The only exposure I've had to this type of product was years ago. Hemo-D, which gave me tremendous headaches.... I appreciate all your imput, and thanks in advance! Deanna Snider HT Shriners Hospital for Children Research Dept. Cincinnati, OH 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From PBugelsk <@t> CNTUS.JNJ.COM Tue Feb 15 10:48:40 2005 From: PBugelsk <@t> CNTUS.JNJ.COM (Bugelski, Peter [CNTUS]) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Iron staining (cellular) Message-ID: <70F83FE9F65318468A612768E7043F8918A068@cntusmaexs9.na.jnj.com> Try the method described in Bugelski PJ. Sequential histochemical staining for resident and recruited macrophages. Journal of Leukocyte Biology. 38(6):687-96, 1985 Dec. It is very sensitive for iron. -----Original Message----- From: p.j.bergin@qmul.ac.uk [mailto:p.j.bergin@qmul.ac.uk] Sent: Tuesday, February 15, 2005 11:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Iron staining (cellular) Hi, I was hoping someone could give me advice. I have looked through the archives and have tried a few things already. I am interested in staining cell preps to localize iron. I have tried using mouse spleen cells as a trial, but am mainly interested in human macrophages and monocytes that have been grown on glass slides. I have tried fixing the preps with a methanol/acetone mix and with 4% formalin. I have been trying two different methods; Perls Prussian blue from the stainsfile archive and Turnbull's Blue (for ferric and ferrous iron respectively). Both have been unsuccessful with no staining detected (and the cells are there). Does anyone have a method for these stains that works on cells? do they need to be fixed a specific way? Is there any advice anyone would care to share? Thanks in advance for any help. Phil P.S. The staining protocols I have been using follow..... FERROUS IRON - TURNBULL'S BLUE PURPOSE: To detect ferrous (Fe2+) iron in tissues. PRINCIPLE: Tissue sections are treated with an acidic solution of potassium ferricyanide, any ferrous iron present will react to form an insoluble bright blue pigment called Turnbull's blue (ferrous ferricyanide). CONTROL: Routine iron control, which includes an negative. TECHNIQUE: Cut paraffin section 4?. EQUIPMENT: Acid clean glassware, non-metallic forceps. REAGENTS: 0.006N Hydrochloric Acid 1% Acetic Acid Hydrochloric acid 2.5 ml Acetic acid, glacial 1.0 ml Distilled water 497.5 ml Distilled water 100.0 ml Mix well. Solution is stable for 1 Mix well. Solution is stable for 1 year. year. CAUTION: Corrosive acid. CAUTION: Avoid contact and inhalation. Potassium Ferricyanide Nuclear-Fast Red: Staining Solution: See Retic Potassium ferricyanide 0.4 gm Hydrochloric acid, 0.006N 40.0 ml Prepare fresh, just before use. CAUTION: Avoid contact and inhalation. SAFETY: Wear gloves, goggles and lab coat. Avoid contact and inhalation. Hydrochloric acid; target organ effects on reproductive system and fetal tissue. Irritant to skin eyes and respiratory system. Potassium ferricyanide; Low toxicity as long as it is not heated, it will release cyanide gas. MINERALS AND PIGMENTS TURNBULL'S FERROUS IRON Page: 2 of 2 Acetic acid: Irritating to respiratory system. Target organ effects on respiratory system by inhalation. Corrosive. PROCEDURE: 1. Deparaffinize and hydrate to distilled water. 2. Place slides in Potassium ferricyanide staining solution for 1 hour. 3. Wash slides in 1% acetic acid. 4. Counterstain slides in nuclear-fast red for 5 minutes. 5. Rinse well in distilled water. 6. Dehydrate, clear and coverslip. RESULTS: Ferrous iron blue Background pink-red REFERENCE: Carson, F, Histotechnology: A Self-Instructional Text, 1st Ed., 1991, pp. 215-16 ASCP Press Perls Prussian Blue Principle The basis for the method is the release of ferric iron from hemosiderin by acid treatment, forming ferric chloride. The ferric iron reacts with potassium ferrocyanide to form ferric ferrocyanide. This is an insoluble, blue compound known as Prussian blue. The intensity of the colour gives some indication as to amount, but it is qualitative only. 4FeCl3 + 3K4Fe(CN)6 = Fe4[Fe(CN)6]3 + 12KCl Stock solution A Hydrochloric acid 2 ml Distilled water 98 ml Stock solution B Potassium ferrocyanide 2 g Distilled water 100 ml Working solution C Solution A 1 part Solution B 1 part Counterstain solution D Neutral red counterstain Fixation & processing Avoid iron containing materials and jars while fixing as these may contaminate the tissue. Acid containing fixatives may remove some of the iron deposits. Otherwise most methods are satisfactory. Method Bring sections to distilled water with xylene and ethanol. Place into working solution C for 15 minutes. Rinse with distilled water, then tap water. Stain with counterstain solution D for one minute Rinse well with tap water. Dehydrate with ethanol. Clear with xylene Expected Results Nuclei - red Notes The working solution C must be made immediately before use. Avoid washing with tap water before treating with solution C, as rust in the water or tap fixtures could cause false positive staining. Wash well at step 3, as traces of iron will form a granular red deposit with neutral red. Iron ores can be demonstrated, but the acid concentration in solution A may need to be increased to 10% or more. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cgin <@t> pen.eiu.edu Tue Feb 15 11:42:56 2005 From: cgin <@t> pen.eiu.edu (ikenwosu) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] RE: Problems with floating sections Message-ID: <421234a0.c.13cb.16323@pen.eiu.edu> Hi Friends, I have been working on BRDU staining of olfactory epithelium. The protocol involves: 30 min 0.1% TX in PBS then 60 min HCl at 60 degrees Celcius. At the end of the second step my tissues are floating off the slide. Could the problem be that I prepared a 700 ml volume of subbing solution and then subbed 900 slides with it? Are there any techniques for refixing tissues to the slide with out destroying or modifying the antigen. When I CV stain the same tissue on the same batch of slides I have no problems with tissues floating. Thanks for your anticpated help. Ike Thedore Nwosu Grad. Student Dept. of Biological Sciences Eastern Illinois University 600 Linclon Ave Charleston, IL 61938 From cgin <@t> pen.eiu.edu Tue Feb 15 11:44:42 2005 From: cgin <@t> pen.eiu.edu (ikenwosu) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] RE: Service Contracts Message-ID: <4212350a.2ae.15bb.15781@pen.eiu.edu> Hi again, Our lab wishes to obtain the services of a good service contractor for our cryostat close (within a few 100 miles)to our location in Charleston, Illinois. Do you know of any reputable and hopefully affordable contractors out there?What would the average service contracts be worth in dollar terms per year? Thanks. Ike Thedore Nwosu Grad. Student Dept. of Biological Sciences Eastern Illinois University 600 Linclon Ave Charleston, IL 61938 From carolb <@t> mail.phys.mcw.edu Tue Feb 15 12:08:01 2005 From: carolb <@t> mail.phys.mcw.edu (Carol Bobrowitz) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Collagen Type IV Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A9635@thor.phys.mcw.edu> Hello everyone, I'm looking for Collagen Type IV specific for rat tissue and to be tested on FFPE sections. Has anyone attempted an anti-human Collagen IV on FFPE rat sections? Either fluorescent or DAB staining I would be interested in. I have found references to Collagen IV on frozen sections but unfortunately my tissue has been used for other studies and is already blocked. Any help in this area would be greatly appreciated. Thank you in advance, Carol Ann Bobrowitz Histology Laboratory Department of Physiology - Room 541 Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, Wisconsin 53226 414-456-8179 FAX 414-456-6546 cbobrowi@mcw.edu From leswes <@t> shaw.ca Tue Feb 15 12:44:37 2005 From: leswes <@t> shaw.ca (Lesley Weston) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Wax removal incomplete In-Reply-To: Message-ID: I could just be displaying my ignorance here, but if the problem is the prescence of polymerised plastics in the wax, then a technique sometimes used for EM might help. To remove polymerised epoxy resins from a block or from thick sections, one exposes them to sodium ethoxide or sodium methoxide for a short time. Either is made very easily by adding sodium hydroxide pellets to absolute ethanol or methanol and leaving it overnight in the fume hood. Since it would be applied to the ex-paraffin sections, you wouldn't have to leave them in for very long, so there shouldn't be too much damage to the tissue, though I've never tried immuno-staining after using this. It would probably work best (if it works at all) after removing the wax with xylene or whatever and rinsing well in 100% alcohol. Lesley Weston. on 15/02/2005 6:08 AM, LEWIS, MARK A. at mark.lewis@thermo.com wrote: > I often wondered about this due to the "plastic "polymers added to the > different types of paraffins. Perhaps it's these "polymers" that are not being > removed with the typical solvents. > > Interesting ... > > Thanks John ! > > Mark > ? > Mark A. Lewis B.A., H.T.(ASCP) > Product Specialist > Anatomical Pathology > Clinical Diagnostics > Thermo Electron Corporation > 171 Industry Drive > Pittsburgh, Pa. 15275 > Phone:? 412-747-4013 > Fax?????? 412-788-6557 > E-mail: mark.lewis@thermo.com > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan > Sent: Monday, February 14, 2005 1:40 PM > To: Histonet > Subject: [Histonet] Wax removal incomplete > > A recent publication from Ireland shows (by laser confocal > Raman microspectroscopy, no less) that wax removal from > sections, as usually practised, is always incomplete. > The authors were able to remove all the paraffin, however, > with hexane. The other things they tried were xylene, > "histoclear" followed by hot antigen retrieval, and > "trilogy" (a product that combines dewaxing and retrieval > - ? a solvent-detergent mixture). All three failed to > remove all the wax, even with repeated, prolonged > treatments. > For hexane, 18 hours were needed for complete was removal. > > The investigators found that the residual paraffin > seriously interfered with the Raman spectra of > rehydrated sections. They are investigating the Raman > technique (which they explain fairly simply) for early > diagnosis of tumours. They also claim stronger > immunostaining after dewaxing in hexane (18h) than after > xylene (18h). They tested only one antibody (to > cytokeratin), > and the pair of photos doesn't make the point very > impressively. > > They authors also state that hexane is less toxic than > xylene. I can add that the least expensive n-hexane is > a little cheaper than xylenes (mixed isomers) in one > catalogue checked. Both solvents are flammable, and > n-hexane (BP 69C) is more volatile than xylene. Both > solvents are miscible with 100% alcohol. > > The reference is: > O'Faolain E et 6 al. (2005) Raman spectroscopic evaluation > of efficacy of current paraffin wax section dewaxing > agents. J. Histochem. Cytochem. 53(1): 121-129. From pruegg <@t> ihctech.net Tue Feb 15 14:35:00 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] FW: Looking for controls Message-ID: <200502152035.j1FKYxFF017438@chip.viawest.net> -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Tuesday, February 15, 2005 1:33 PM To: 'Cara.Mack@UCHSC.edu'; 'Garza-Williams, Sara'; 'carlyne.cool@uchsc.edu' Subject: Looking for controls I was wondering if any of you kind people could help an investigator at the U with this request? Thanks for considering this, Patsy From: Susan Majka [mailto:susan.majka@uchsc.edu] Hi Patsy, Do you know any generous pediatricians who might have paraffin blocks of BPD (bronchopulmonary dysplasia) tissue? SUe From jkiernan <@t> uwo.ca Tue Feb 15 14:56:37 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Wax removal incomplete References: Message-ID: <42126204.10AC7381@uwo.ca> O'Faolain et al did use a wax with added "polymer", from a Dutch supplier. The Raman spectrum of this sectioned wax (without tissue) had characteristic peaks that showed also in the spectra of supposedly dewaxed sections of tissue. They don't discuss the "polymer", but your suggestion could well be right. They should have used pure paraffin as well. -- ------------------------------- 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 _______________________________ "LEWIS, MARK A." wrote: > > I often wondered about this due to the "plastic "polymers added to the different types of paraffins. Perhaps it's these "polymers" that are not being removed with the typical solvents. > > Interesting ... > > Thanks John ! > > Mark > > Mark A. Lewis B.A., H.T.(ASCP) > Product Specialist > Anatomical Pathology > Clinical Diagnostics > Thermo Electron Corporation > 171 Industry Drive > Pittsburgh, Pa. 15275 > Phone: 412-747-4013 > Fax 412-788-6557 > E-mail: mark.lewis@thermo.com > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan > Sent: Monday, February 14, 2005 1:40 PM > To: Histonet > Subject: [Histonet] Wax removal incomplete > > A recent publication from Ireland shows (by laser confocal > Raman microspectroscopy, no less) that wax removal from > sections, as usually practised, is always incomplete. > The authors were able to remove all the paraffin, however, > with hexane. The other things they tried were xylene, > "histoclear" followed by hot antigen retrieval, and > "trilogy" (a product that combines dewaxing and retrieval > - ? a solvent-detergent mixture). All three failed to > remove all the wax, even with repeated, prolonged > treatments. > For hexane, 18 hours were needed for complete was removal. > > The investigators found that the residual paraffin > seriously interfered with the Raman spectra of > rehydrated sections. They are investigating the Raman > technique (which they explain fairly simply) for early > diagnosis of tumours. They also claim stronger > immunostaining after dewaxing in hexane (18h) than after > xylene (18h). They tested only one antibody (to > cytokeratin), > and the pair of photos doesn't make the point very > impressively. > > They authors also state that hexane is less toxic than > xylene. I can add that the least expensive n-hexane is > a little cheaper than xylenes (mixed isomers) in one > catalogue checked. Both solvents are flammable, and > n-hexane (BP 69C) is more volatile than xylene. Both > solvents are miscible with 100% alcohol. > > The reference is: > O'Faolain E et 6 al. (2005) Raman spectroscopic evaluation > of efficacy of current paraffin wax section dewaxing > agents. J. Histochem. Cytochem. 53(1): 121-129. > -- > ------------------------------- > 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 > _______________________________ From Jackie.O'Connor <@t> abbott.com Tue Feb 15 15:12:45 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] RE: Service Contracts Message-ID: Try Ken Hart at Dorn and Hart Microedge, Villa Park, IL 630-832-3843. Ken is the best microtome/cryostat troubleshooter I've ever known. I recommend him highly. My Hospital in Honolulu even flew him to Hawaii once to service our equipment there - he's the best. Jacqueline M. O'Connor HT(ASCP) Assistant Scientist Abbott Laboratories Global Pharmaceutical Research and Development "ikenwosu" Sent by: histonet-bounces@lists.utsouthwestern.edu 02/15/2005 11:44 AM Please respond to cgin To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] RE: Service Contracts Hi again, Our lab wishes to obtain the services of a good service contractor for our cryostat close (within a few 100 miles)to our location in Charleston, Illinois. Do you know of any reputable and hopefully affordable contractors out there?What would the average service contracts be worth in dollar terms per year? Thanks. Ike Thedore Nwosu Grad. Student Dept. of Biological Sciences Eastern Illinois University 600 Linclon Ave Charleston, IL 61938 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Tue Feb 15 15:57:16 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Water bath temperature[Scanned] Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB41AF@fh2k093.fhmis.net> Or perhaps ...itsy bitsy, teeny weeny LITTLE polka-dot bikini -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: 'Marshall Terry Dr,Consultant Histopathologist'; histonet@lists.utsouthwestern.edu Sent: 2/15/2005 3:28 AM Subject: RE: [Histonet] Water bath temperature[Scanned] Doesn't the adjective affect the noun? So 'polka dot bikini' being the noun it implies that the 'polka dot bikini' was 'itsy bitsy'. You would have to say 'itsy bitsy bikini with teeny weeny polka dots' to mean what you have suggested. In order for this not to be frivolous please follow the link for an English lesson http://www-users.cs.york.ac.uk/~susan/cyc/a/adj.htm. -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 February 2005 17:17 To: Rebecca Barnhart; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Water bath temperature[Scanned] Why from the description is it not just our old friend pink disease? (BayleyJH {1949}J.Path.BAct., 61,448) Are the polka dots too small or too many or too invariable, or any combination thereof? As usual, a picture would be worth a thousand words. She wore an itsy bitsy teeny weeny polka dot bikini ....... I was never sure if it was the polka dots that were teeny weeny. Probably not. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: 11 February 2005 19:24 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath temperature We were having a problem that the tissue would not take up the hematoxlyin but would take up the eosin but only in some areas, so it looked like polka dots. We started one by one changing solution trying to figure out where it was coming from. We had all fresh solutions and still had polka dots. I tried the only other thing left to change, I lowered the temperature of the water bath from 45 to 38 and this worked. We currently are using ParaPlast Xtra with a melting point of 52. We have all tried figure out why the temperature in the water bath will make the difference. Several times I have cut the same tissue at different temperatures ranging from 38 - 45 and every time there are polka dots at the higher temperatures. Does anyone have any idea why this would be? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From maria <@t> ski.org Tue Feb 15 17:44:34 2005 From: maria <@t> ski.org (Maria Mejia) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Looking for neurohisto job in San Francisco Message-ID: <42128962.1060103@ski.org> Greetings to everyone, For the past 15 years, I've been involved in the basic science of neurohistology at a small institute here in San Francisco. Over the years, I've enjoyed working on quite a number of very interesting PI projects of varying scale & priority. I have knowledge of CALOSHA regulations because I also happen to also be the safety person. However, due to financial changes the institute has been downsizing personnel in various Core departments including histology. So, I'm searching for employment. I would prefer to stay in San Francisco area because it's my home. I have a total of 26 years in histology. I'm proficient in a variety of routine & specialized neurohistologic techniques using various tissue processing and sectioning, e.g., paraffin & frozen sections, thick whole mounts, small & large tissue gelatin embedding for thick free floating sections using rotary, cryostat & slide microtomes. I've worked on various neural tissue from different species, e.g., cortical & retinal tissue from cat, mouse, turtle, monkey, rat, rabbit extra-ocular muscles and whole butterfly heads. I've performed a variety of IHC techniques, e.g., dopamine beta hydroxylase (DBH), tyrosine hydroxylase, anti-humanBNDF, c-FOS, ZIF-189, GABA, anti-Cam Kinase type II, ChAT, 3CB2 and protein kinase C, anti-recoverin & etc. If you need someone with my skills, please contact me. I just want to say, I consider being a member of the Histonet part of my histology continuing education. regards Maria Bartola Mejia neurohistology Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org or marius2@mindspring.com From billions <@t> public1.sz.js.cn Tue Feb 15 19:22:06 2005 From: billions <@t> public1.sz.js.cn (Sinoera Tech) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Re: BIOLOGICAL STAINS, DYES AND INDICATORS, DIAGNOSTICS References: <000c01c51374$61da55c0$08337591@Przemek> Message-ID: <004f01c513c5$f2d41ec0$a001a8c0@ming> Dear Sirs, We can manufacture following products for scientific research activities. 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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 Zorbutt <@t> aol.com Wed Feb 16 00:34:56 2005 From: Zorbutt <@t> aol.com (Zorbutt@aol.com) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Telomerase IHC antibody Message-ID: <8d.20d8623e.2f444390@aol.com> Hi, I waswondering if anyone uses the telomerase antibody that is supplied by vectorlabs? The normal way of identifying telomerase is through PCR, but have seen that an antibody is available and wanted to know if it worked just as well as the PCR. Am thinking of using it on paraffin sections. Thanx to anyone who replies. From sbindu20002000 <@t> yahoo.co.in Wed Feb 16 01:09:04 2005 From: sbindu20002000 <@t> yahoo.co.in (s bindu) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Immunohistochemistry control slides Message-ID: <20050216070904.98266.qmail@web8504.mail.in.yahoo.com> Hai all, I am new in the field of immunohistochemistry.My doubt is from where i can get positive control slides for different antibodies like ED1, ED2, ED3,IL1,TNF,CD4 and CD8. Bindu.S Research Assistant Histopathology lab SCTIMST,BMT wing Trivandrum Yahoo! India Matrimony: Find your life partneronline. From David.Muskett <@t> RLC.NHS.UK Wed Feb 16 02:10:20 2005 From: David.Muskett <@t> RLC.NHS.UK (Muskett David) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Bone saw and bone protocols[Scanned] Message-ID: <6AB79F0BA7C4914BA8DD8F5E0C21B9752E2928@AHEXMAIL01.xalderhey.com> Dear All I am currently reviewing our protocols for the processing of bone. Would anybody be willing to share protocols and processing schedules? I am also interested in buying a bone saw, preferably one which allows thin sections to be cut easily. Can anybody recommend me a firm or a saw? 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 Kemlo.Rogerson <@t> elht.nhs.uk Wed Feb 16 03:02:47 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Checking the endpoint of decalcification[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EFAF@bhrv-nt-11.bhrv.nwest.nhs.uk> I would assume that if you can use the sodium salt (NaOCOCOONa) then you use the potassium salt (KOCOCOOK.H2O) as the cation has both the same valency and does do anything. The anion appears exactly the same but the potassium salt is attached to a water molecule. The potassium salt is more toxic than the sodium equivalent. IMHO Kemlo -----Original Message----- From: Maxim [mailto:maxim_71@mail.ru] Sent: 15 February 2005 15:56 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Checking the endpoint of decalcification[Scanned] Dear Histonetters! For checking the endpoint of decalcification is traditionally used 5% solution sodium oxalate or ammonium oxalate. Solubility in 100 ml water for sodium oxalate 3,7v/w, but for ammonium oxalate forms accordingly 4,5 v/w. Thereby, we get the saturated solutions of these salts. Why to this effect do not use potassium oxalate? His solubility in 100 ml water forms 26,4w/v. Can be therefore that in solution is got too high concentration oxalate and this test not will so be sensitive? Or this there is else some explanation? Thank you. Maxim Peshkov, HTL Department of biopsy and cytological research Pathological and anatomical bureau Taganrog Russia, 347942. mailto:maxim_71@mail.ru _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Feb 16 03:11:41 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] Iron staining (cellular)[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EFB1@bhrv-nt-11.bhrv.nwest.nhs.uk> All I can remember is never use an acidic fixative, it removes Fe ions, or anything acidic before staining. The protocols you suggest appear fine; how certain are you that the macrophages have 'ingested' iron? Could it be that cloned cells don't act as they ought and that the iron was never 'ingested' in the first place? I assume you have put through an Fe2 and Fe3 control slide to tell you if your procedure was successful? Kemlo -----Original Message----- From: p.j.bergin@qmul.ac.uk [mailto:p.j.bergin@qmul.ac.uk] Sent: 15 February 2005 16:29 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Iron staining (cellular)[Scanned] Hi, I was hoping someone could give me advice. I have looked through the archives and have tried a few things already. I am interested in staining cell preps to localize iron. I have tried using mouse spleen cells as a trial, but am mainly interested in human macrophages and monocytes that have been grown on glass slides. I have tried fixing the preps with a methanol/acetone mix and with 4% formalin. I have been trying two different methods; Perls Prussian blue from the stainsfile archive and Turnbull's Blue (for ferric and ferrous iron respectively). Both have been unsuccessful with no staining detected (and the cells are there). Does anyone have a method for these stains that works on cells? do they need to be fixed a specific way? Is there any advice anyone would care to share? Thanks in advance for any help. Phil P.S. The staining protocols I have been using follow..... FERROUS IRON - TURNBULL'S BLUE PURPOSE: To detect ferrous (Fe2+) iron in tissues. PRINCIPLE: Tissue sections are treated with an acidic solution of potassium ferricyanide, any ferrous iron present will react to form an insoluble bright blue pigment called Turnbull's blue (ferrous ferricyanide). CONTROL: Routine iron control, which includes an negative. TECHNIQUE: Cut paraffin section 4?. EQUIPMENT: Acid clean glassware, non-metallic forceps. REAGENTS: 0.006N Hydrochloric Acid 1% Acetic Acid Hydrochloric acid 2.5 ml Acetic acid, glacial 1.0 ml Distilled water 497.5 ml Distilled water 100.0 ml Mix well. Solution is stable for 1 Mix well. Solution is stable for 1 year. year. CAUTION: Corrosive acid. CAUTION: Avoid contact and inhalation. Potassium Ferricyanide Nuclear-Fast Red: Staining Solution: See Retic Potassium ferricyanide 0.4 gm Hydrochloric acid, 0.006N 40.0 ml Prepare fresh, just before use. CAUTION: Avoid contact and inhalation. SAFETY: Wear gloves, goggles and lab coat. Avoid contact and inhalation. Hydrochloric acid; target organ effects on reproductive system and fetal tissue. Irritant to skin eyes and respiratory system. Potassium ferricyanide; Low toxicity as long as it is not heated, it will release cyanide gas. MINERALS AND PIGMENTS TURNBULL'S FERROUS IRON Page: 2 of 2 Acetic acid: Irritating to respiratory system. Target organ effects on respiratory system by inhalation. Corrosive. PROCEDURE: 1. Deparaffinize and hydrate to distilled water. 2. Place slides in Potassium ferricyanide staining solution for 1 hour. 3. Wash slides in 1% acetic acid. 4. Counterstain slides in nuclear-fast red for 5 minutes. 5. Rinse well in distilled water. 6. Dehydrate, clear and coverslip. RESULTS: Ferrous iron blue Background pink-red REFERENCE: Carson, F, Histotechnology: A Self-Instructional Text, 1st Ed., 1991, pp. 215-16 ASCP Press Perls Prussian Blue Principle The basis for the method is the release of ferric iron from hemosiderin by acid treatment, forming ferric chloride. The ferric iron reacts with potassium ferrocyanide to form ferric ferrocyanide. This is an insoluble, blue compound known as Prussian blue. The intensity of the colour gives some indication as to amount, but it is qualitative only. 4FeCl3 + 3K4Fe(CN)6 = Fe4[Fe(CN)6]3 + 12KCl Stock solution A Hydrochloric acid 2 ml Distilled water 98 ml Stock solution B Potassium ferrocyanide 2 g Distilled water 100 ml Working solution C Solution A 1 part Solution B 1 part Counterstain solution D Neutral red counterstain Fixation & processing Avoid iron containing materials and jars while fixing as these may contaminate the tissue. Acid containing fixatives may remove some of the iron deposits. Otherwise most methods are satisfactory. Method Bring sections to distilled water with xylene and ethanol. Place into working solution C for 15 minutes. Rinse with distilled water, then tap water. Stain with counterstain solution D for one minute Rinse well with tap water. Dehydrate with ethanol. Clear with xylene Expected Results Nuclei - red Notes The working solution C must be made immediately before use. Avoid washing with tap water before treating with solution C, as rust in the water or tap fixtures could cause false positive staining. Wash well at step 3, as traces of iron will form a granular red deposit with neutral red. Iron ores can be demonstrated, but the acid concentration in solution A may need to be increased to 10% or more. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Andrew.Prior <@t> Smith-Nephew.com Wed Feb 16 03:43:00 2005 From: Andrew.Prior <@t> Smith-Nephew.com (Prior, Andrew) Date: Fri Sep 16 15:24:37 2005 Subject: [Histonet] RE: Problems with floating sections Message-ID: We had some success using 2N HCl at 37deg C for 30 minutes. This was enough to uncover the epitope and sections stayed on slides. If you are cutting more sections try using pre-coated (poly-L?) or charged slides (Superfrost Plus) as they help sections stick during IHC Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. From p.j.bergin <@t> qmul.ac.uk Wed Feb 16 05:24:47 2005 From: p.j.bergin <@t> qmul.ac.uk (p.j.bergin@qmul.ac.uk) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Iron staining (cellular)[Scanned] In-Reply-To: <1030B679AD69D6119C3F00080210DD9D05A3EFB1@bhrv-nt-11.bhrv.nwest.nhs.uk> References: <1030B679AD69D6119C3F00080210DD9D05A3EFB1@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: <1108553087.42132d7f6494f@webapps.qmul.ac.uk> I thought so too (about the adic fixing). So far we have fixed in methenol/acetone, NBF, and tried with just air drying. The cells themselves are just cultured macrophages (primary) and should have iron in them. Unfortunaetlywe have no control, but have been using murine spleen cell smears for the positive and to try get the assay working. Still no luck though. Although if you have a better suggestion for a control slide then that'd be great. Cheers, Phil Quoting Kemlo Rogerson : > All I can remember is never use an acidic fixative, it removes Fe ions, or > anything acidic before staining. The protocols you suggest appear fine; how > certain are you that the macrophages have 'ingested' iron? Could it be that > cloned cells don't act as they ought and that the iron was never 'ingested' > in the first place? I assume you have put through an Fe2 and Fe3 control > slide to tell you if your procedure was successful? > > Kemlo > > -----Original Message----- > From: p.j.bergin@qmul.ac.uk [mailto:p.j.bergin@qmul.ac.uk] > Sent: 15 February 2005 16:29 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Iron staining (cellular)[Scanned] > > > > Hi, > > I was hoping someone could give me advice. I have looked through the > archives > and have tried a few things already. I am interested in staining cell preps > to > localize iron. I have tried using mouse spleen cells as a trial, but am > mainly > interested in human macrophages and monocytes that have been grown on glass > slides. I have tried fixing the preps with a methanol/acetone mix and with > 4% > formalin. I have been trying two different methods; Perls Prussian blue > from > the stainsfile archive and Turnbull's Blue (for ferric and ferrous iron > respectively). Both have been unsuccessful with no staining detected (and > the > cells are there). > > Does anyone have a method for these stains that works on cells? do they need > to > be fixed a specific way? Is there any advice anyone would care to share? > > Thanks in advance for any help. > > Phil > > P.S. The staining protocols I have been using follow..... > > FERROUS IRON - TURNBULL'S BLUE > > PURPOSE: To detect ferrous (Fe2+) iron in tissues. > PRINCIPLE: Tissue sections are treated with an acidic solution of potassium > ferricyanide, any ferrous iron present will react to form an insoluble > bright > blue pigment called Turnbull's blue (ferrous ferricyanide). > CONTROL: Routine iron control, which includes an negative. > TECHNIQUE: Cut paraffin section 4?. > EQUIPMENT: Acid clean glassware, non-metallic forceps. > REAGENTS: > 0.006N Hydrochloric Acid 1% Acetic Acid > Hydrochloric acid 2.5 ml Acetic acid, glacial 1.0 ml Distilled water 497.5 > ml > Distilled water 100.0 ml Mix well. Solution is stable for 1 Mix well. > Solution > is stable for 1 year. year. > CAUTION: Corrosive acid. CAUTION: Avoid contact and inhalation. > Potassium Ferricyanide Nuclear-Fast Red: > Staining Solution: See Retic > Potassium ferricyanide 0.4 gm > Hydrochloric acid, 0.006N 40.0 ml > Prepare fresh, just before use. > CAUTION: Avoid contact and inhalation. > SAFETY: Wear gloves, goggles and lab coat. Avoid contact and inhalation. > Hydrochloric acid; target organ effects on reproductive system and fetal > tissue. > Irritant to skin eyes and respiratory system. > Potassium ferricyanide; Low toxicity as long as it is not heated, it will > release cyanide gas. > > MINERALS AND PIGMENTS > TURNBULL'S FERROUS IRON Page: 2 of 2 > Acetic acid: Irritating to respiratory system. Target organ effects on > respiratory system by inhalation. Corrosive. > PROCEDURE: > 1. Deparaffinize and hydrate to distilled water. > 2. Place slides in Potassium ferricyanide staining solution for 1 hour. > 3. Wash slides in 1% acetic acid. > 4. Counterstain slides in nuclear-fast red for 5 minutes. > 5. Rinse well in distilled water. > 6. Dehydrate, clear and coverslip. > RESULTS: > Ferrous iron blue > Background pink-red > REFERENCE: > Carson, F, Histotechnology: A Self-Instructional Text, 1st Ed., 1991, pp. > 215-16 ASCP Press > > Perls Prussian Blue > > Principle > The basis for the method is the release of ferric iron from hemosiderin by > acid > treatment, forming ferric chloride. The ferric iron reacts with potassium > ferrocyanide to form ferric ferrocyanide. This is an insoluble, blue > compound > known as Prussian blue. The intensity of the colour gives some indication as > to > amount, but it is qualitative only. > > 4FeCl3 + 3K4Fe(CN)6 = Fe4[Fe(CN)6]3 + 12KCl > > Stock solution A > Hydrochloric acid 2 ml > Distilled water 98 ml > > > Stock solution B > Potassium ferrocyanide 2 g > Distilled water 100 ml > > > Working solution C > Solution A 1 part > Solution B 1 part > > > Counterstain solution D > Neutral red counterstain > > > > > Fixation & processing > Avoid iron containing materials and jars while fixing as these may > contaminate > the tissue. Acid containing fixatives may remove some of the iron deposits. > Otherwise most methods are satisfactory. > Method > Bring sections to distilled water with xylene and ethanol. > Place into working solution C for 15 minutes. > Rinse with distilled water, then tap water. > Stain with counterstain solution D for one minute Rinse well with tap water. > Dehydrate with ethanol. > Clear with xylene > > > Expected Results > Nuclei - red > > > Notes > The working solution C must be made immediately before use. > Avoid washing with tap water before treating with solution C, as rust in the > water or tap fixtures could cause false positive staining. > Wash well at step 3, as traces of iron will form a granular red deposit with > neutral red. > Iron ores can be demonstrated, but the acid concentration in solution A may > need > to be increased to 10% or more. > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Kemlo.Rogerson <@t> elht.nhs.uk Wed Feb 16 05:57:26 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Iron staining (cellular)[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EFB4@bhrv-nt-11.bhrv.nwest.nhs.uk> You must have a control slide or you will never get to the bottom of it. Why should the macrophages have iron in them? When I cultured blood cells for my MSc I learnt very quickly that what you got may bear no resemblance to what you started with; cells that posses certain characteristics in vivo may not in vitro. I wonder if you can't stain it as it's not there; haemorrhagic placenta or omentum, I'm told, is a good control (Dave Rushworth; verbal). That fellow J.A. Kiernan has a good section in his book 'Histological and Histochemical Methods' Theory & Practice, 3rd Edition, Published by Arnold 1999. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: p.j.bergin@qmul.ac.uk [mailto:p.j.bergin@qmul.ac.uk] Sent: 16 February 2005 11:25 To: Kemlo Rogerson Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Iron staining (cellular)[Scanned] I thought so too (about the adic fixing). So far we have fixed in methenol/acetone, NBF, and tried with just air drying. The cells themselves are just cultured macrophages (primary) and should have iron in them. Unfortunaetlywe have no control, but have been using murine spleen cell smears for the positive and to try get the assay working. Still no luck though. Although if you have a better suggestion for a control slide then that'd be great. Cheers, Phil Quoting Kemlo Rogerson : > All I can remember is never use an acidic fixative, it removes Fe ions, or > anything acidic before staining. The protocols you suggest appear fine; how > certain are you that the macrophages have 'ingested' iron? Could it be that > cloned cells don't act as they ought and that the iron was never 'ingested' > in the first place? I assume you have put through an Fe2 and Fe3 control > slide to tell you if your procedure was successful? > > Kemlo > > -----Original Message----- > From: p.j.bergin@qmul.ac.uk [mailto:p.j.bergin@qmul.ac.uk] > Sent: 15 February 2005 16:29 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Iron staining (cellular)[Scanned] > > > > Hi, > > I was hoping someone could give me advice. I have looked through the > archives > and have tried a few things already. I am interested in staining cell preps > to > localize iron. I have tried using mouse spleen cells as a trial, but am > mainly > interested in human macrophages and monocytes that have been grown on glass > slides. I have tried fixing the preps with a methanol/acetone mix and with > 4% > formalin. I have been trying two different methods; Perls Prussian blue > from > the stainsfile archive and Turnbull's Blue (for ferric and ferrous iron > respectively). Both have been unsuccessful with no staining detected (and > the > cells are there). > > Does anyone have a method for these stains that works on cells? do they need > to > be fixed a specific way? Is there any advice anyone would care to share? > > Thanks in advance for any help. > > Phil > > P.S. The staining protocols I have been using follow..... > > FERROUS IRON - TURNBULL'S BLUE > > PURPOSE: To detect ferrous (Fe2+) iron in tissues. > PRINCIPLE: Tissue sections are treated with an acidic solution of potassium > ferricyanide, any ferrous iron present will react to form an insoluble > bright > blue pigment called Turnbull's blue (ferrous ferricyanide). > CONTROL: Routine iron control, which includes an negative. > TECHNIQUE: Cut paraffin section 4?. > EQUIPMENT: Acid clean glassware, non-metallic forceps. > REAGENTS: > 0.006N Hydrochloric Acid 1% Acetic Acid > Hydrochloric acid 2.5 ml Acetic acid, glacial 1.0 ml Distilled water 497.5 > ml > Distilled water 100.0 ml Mix well. Solution is stable for 1 Mix well. > Solution > is stable for 1 year. year. > CAUTION: Corrosive acid. CAUTION: Avoid contact and inhalation. > Potassium Ferricyanide Nuclear-Fast Red: > Staining Solution: See Retic > Potassium ferricyanide 0.4 gm > Hydrochloric acid, 0.006N 40.0 ml > Prepare fresh, just before use. > CAUTION: Avoid contact and inhalation. > SAFETY: Wear gloves, goggles and lab coat. Avoid contact and inhalation. > Hydrochloric acid; target organ effects on reproductive system and fetal > tissue. > Irritant to skin eyes and respiratory system. > Potassium ferricyanide; Low toxicity as long as it is not heated, it will > release cyanide gas. > > MINERALS AND PIGMENTS > TURNBULL'S FERROUS IRON Page: 2 of 2 > Acetic acid: Irritating to respiratory system. Target organ effects on > respiratory system by inhalation. Corrosive. > PROCEDURE: > 1. Deparaffinize and hydrate to distilled water. > 2. Place slides in Potassium ferricyanide staining solution for 1 hour. > 3. Wash slides in 1% acetic acid. > 4. Counterstain slides in nuclear-fast red for 5 minutes. > 5. Rinse well in distilled water. > 6. Dehydrate, clear and coverslip. > RESULTS: > Ferrous iron blue > Background pink-red > REFERENCE: > Carson, F, Histotechnology: A Self-Instructional Text, 1st Ed., 1991, pp. > 215-16 ASCP Press > > Perls Prussian Blue > > Principle > The basis for the method is the release of ferric iron from hemosiderin by > acid > treatment, forming ferric chloride. The ferric iron reacts with potassium > ferrocyanide to form ferric ferrocyanide. This is an insoluble, blue > compound > known as Prussian blue. The intensity of the colour gives some indication as > to > amount, but it is qualitative only. > > 4FeCl3 + 3K4Fe(CN)6 = Fe4[Fe(CN)6]3 + 12KCl > > Stock solution A > Hydrochloric acid 2 ml > Distilled water 98 ml > > > Stock solution B > Potassium ferrocyanide 2 g > Distilled water 100 ml > > > Working solution C > Solution A 1 part > Solution B 1 part > > > Counterstain solution D > Neutral red counterstain > > > > > Fixation & processing > Avoid iron containing materials and jars while fixing as these may > contaminate > the tissue. Acid containing fixatives may remove some of the iron deposits. > Otherwise most methods are satisfactory. > Method > Bring sections to distilled water with xylene and ethanol. > Place into working solution C for 15 minutes. > Rinse with distilled water, then tap water. > Stain with counterstain solution D for one minute Rinse well with tap water. > Dehydrate with ethanol. > Clear with xylene > > > Expected Results > Nuclei - red > > > Notes > The working solution C must be made immediately before use. > Avoid washing with tap water before treating with solution C, as rust in the > water or tap fixtures could cause false positive staining. > Wash well at step 3, as traces of iron will form a granular red deposit with > neutral red. > Iron ores can be demonstrated, but the acid concentration in solution A may > need > to be increased to 10% or more. > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@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 ian.montgomery <@t> bio.gla.ac.uk Wed Feb 16 06:23:48 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Cresyl violet. Message-ID: <6.2.1.2.2.20050216122123.030188b0@udcf.gla.ac.uk> Ever thought of Gallocyanine? A progressive stain requiring no differentiation. Einarson, I. 1932 Amer.J.Path. 8. 295 Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From Heather.A.Harper <@t> pcola.med.navy.mil Wed Feb 16 07:44:57 2005 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Need info Message-ID: <807FE48C5A7CC940B973B58D32E7014318A75EC3@nhpens-exch1.pcola.med.navy.mil> I work for the Navy as a civilian histo tech. I have been here for 5 yrs. And as of Valentine's Day, I got land basted by a pathologist presenting me with a new and revised work description of my supervisory duties. I got told that as of April 1st I will have to be trained for 30 days to gross in all small specimens. This is at no extra pay and I am a GS employee. I felt like I was done totally wrong, because they went behind my back and threatened me that if I didn't comply to learning how to gross, that administrative action will be taken against me. Now remind you I would've been far more receptive if I had been asked to gross and possibly be upgraded from a GS-7 to a GS-8 or GS-9. But no, I was treated like I was some enlisted navy sailor. I would like to know if anybody has any information on rights that histo techs have when it comes to grossing. Can an employer threaten you after working for 5 yrs and tell you if you don't comply, administrative action will be taken against you. I assume that means...in the board room, and being told "you're fired". I have the Union involved but any information on employee rights, regulations etc...would be of great help. Thank you in advance. Heather A. Harper From Charles.Embrey <@t> carle.com Wed Feb 16 08:10:37 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:24:38 2005 Subject: FW: [Histonet] Need info Message-ID: -----Original Message----- From: Charles.Embrey Sent: Wednesday, February 16, 2005 8:10 AM To: 'Heather.A.Harper@pcola.med.navy.mil' Subject: RE: [Histonet] Need info Heather, I am sorry to see that this is happening to you. Your union is your best help in the matter. As retired active duty Air Force I understand how orders can be given but as a GS employee you have more power in resisting those orders. The biggest problem is that even if you win the battle you still may lose the war. After everything is settled you will still have to work there and the pathologists may try to make things unpleasant if they chose to. Hopefully you can work out a compromise. Do you meet the CLIA requirements for high complexity testing? If not, you have grounds to refuse biased on the regulation. For the record, I feel that any histotech that grosses tissue for the same wage that he cuts blocks is doing himself a disservice. Standard histology is not considered "high complexity testing" and grossing is. If you are working at a higher standard you should be paid at that level. To willingly accept otherwise is foolish. Charles Embrey, PA(ASCP) Urbana, IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Wednesday, February 16, 2005 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need info I work for the Navy as a civilian histo tech. I have been here for 5 yrs. And as of Valentine's Day, I got land basted by a pathologist presenting me with a new and revised work description of my supervisory duties. I got told that as of April 1st I will have to be trained for 30 days to gross in all small specimens. This is at no extra pay and I am a GS employee. I felt like I was done totally wrong, because they went behind my back and threatened me that if I didn't comply to learning how to gross, that administrative action will be taken against me. Now remind you I would've been far more receptive if I had been asked to gross and possibly be upgraded from a GS-7 to a GS-8 or GS-9. But no, I was treated like I was some enlisted navy sailor. I would like to know if anybody has any information on rights that histo techs have when it comes to grossing. Can an employer threaten you after working for 5 yrs and tell you if you don't comply, administrative action will be taken against you. I assume that means...in the board room, and being told "you're fired". I have the Union involved but any information on employee rights, regulations etc...would be of great help. Thank you in advance. Heather A. Harper _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BlazekL <@t> childrensdayton.org Wed Feb 16 08:38:01 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Need info Message-ID: Heather, I agree with Chuck and also sympathize with the way they have handled changing your job classification. I don't know what a GS-7 pays but I do know that in the "outside world" you are worth a lot with the ability to do grossing. I think every one of us should use every opportunity to further our skills and keep learning new things. It's worth it in the long run. The more valuable you make yourself the more marketable you are in this field where there is such a shortage. I think you should grit your teeth and smile and let them foot the expense of learning something new. Then in a year or so if the opportunity presents its self run quickly for someplace that respects you and involves you in the process of making changes. Linda Blazek, HT (ASCP) From: Charles.Embrey Sent: Wednesday, February 16, 2005 8:10 AM To: 'Heather.A.Harper@pcola.med.navy.mil' Subject: RE: [Histonet] Need info Heather, I am sorry to see that this is happening to you. Your union is your best help in the matter. As retired active duty Air Force I understand how orders can be given but as a GS employee you have more power in resisting those orders. The biggest problem is that even if you win the battle you still may lose the war. After everything is settled you will still have to work there and the pathologists may try to make things unpleasant if they chose to. Hopefully you can work out a compromise. Do you meet the CLIA requirements for high complexity testing? If not, you have grounds to refuse biased on the regulation. For the record, I feel that any histotech that grosses tissue for the same wage that he cuts blocks is doing himself a disservice. Standard histology is not considered "high complexity testing" and grossing is. If you are working at a higher standard you should be paid at that level. To willingly accept otherwise is foolish. Charles Embrey, PA(ASCP) Urbana, IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Wednesday, February 16, 2005 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need info I work for the Navy as a civilian histo tech. I have been here for 5 yrs. And as of Valentine's Day, I got land basted by a pathologist presenting me with a new and revised work description of my supervisory duties. I got told that as of April 1st I will have to be trained for 30 days to gross in all small specimens. This is at no extra pay and I am a GS employee. I felt like I was done totally wrong, because they went behind my back and threatened me that if I didn't comply to learning how to gross, that administrative action will be taken against me. Now remind you I would've been far more receptive if I had been asked to gross and possibly be upgraded from a GS-7 to a GS-8 or GS-9. But no, I was treated like I was some enlisted navy sailor. I would like to know if anybody has any information on rights that histo techs have when it comes to grossing. Can an employer threaten you after working for 5 yrs and tell you if you don't comply, administrative action will be taken against you. I assume that means...in the board room, and being told "you're fired". I have the Union involved but any information on employee rights, regulations etc...would be of great help. Thank you in advance. Heather A. Harper From DDDeltour <@t> mar.med.navy.mil Wed Feb 16 08:54:39 2005 From: DDDeltour <@t> mar.med.navy.mil (Deltour, Douglas D. (HM2)) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Need info Message-ID: <080566D001A3D9459FFC0A391A646C9104BEF832@marxchg03.mar.med.navy.mil> I can sympathize with this situation because I am a military Histo tech and I will be getting out of the Navy next year(If anyone needs me, hint hint). Navy Pathologist are a little different to deal with. The good thing is if you get a bad one they always transfer in a few years. If they are re-writing your job description then the person doing it should have the decency to upgrade it to match your grossing peers. Anyway... Show the union CLIA 88 as follows... CLIA '88 lists the requirements for non-pathologists grossing. Grossing is considered high-complexity testing even if it's a punch biopsy. CLIA '88 states "On or before April 24 1995 (I) be a high school graduate or equivalent; and (b) have documentation of training appropriate for the test performed before analyzing patient specimens"................After that date it requires an associate degree in a biological or chemical science or medical laboratory technology -or- qualify as a medical technologist with a bachelor's degree from an accredited institution -or- earned a bachelor's degree in a chemical, physical, biologic or clinical laboratory science. ref. CLIA '88 493.1489 -----Original Message----- From: Linda Blazek [mailto:BlazekL@childrensdayton.org] Sent: Wednesday, February 16, 2005 9:38 AM To: histonet@lists.utsouthwestern.edu; Harper, Heather A., CIV Subject: Re: [Histonet] Need info Heather, I agree with Chuck and also sympathize with the way they have handled changing your job classification. I don't know what a GS-7 pays but I do know that in the "outside world" you are worth a lot with the ability to do grossing. I think every one of us should use every opportunity to further our skills and keep learning new things. It's worth it in the long run. The more valuable you make yourself the more marketable you are in this field where there is such a shortage. I think you should grit your teeth and smile and let them foot the expense of learning something new. Then in a year or so if the opportunity presents its self run quickly for someplace that respects you and involves you in the process of making changes. Linda Blazek, HT (ASCP) From: Charles.Embrey Sent: Wednesday, February 16, 2005 8:10 AM To: 'Heather.A.Harper@pcola.med.navy.mil' Subject: RE: [Histonet] Need info Heather, I am sorry to see that this is happening to you. Your union is your best help in the matter. As retired active duty Air Force I understand how orders can be given but as a GS employee you have more power in resisting those orders. The biggest problem is that even if you win the battle you still may lose the war. After everything is settled you will still have to work there and the pathologists may try to make things unpleasant if they chose to. Hopefully you can work out a compromise. Do you meet the CLIA requirements for high complexity testing? If not, you have grounds to refuse biased on the regulation. For the record, I feel that any histotech that grosses tissue for the same wage that he cuts blocks is doing himself a disservice. Standard histology is not considered "high complexity testing" and grossing is. If you are working at a higher standard you should be paid at that level. To willingly accept otherwise is foolish. Charles Embrey, PA(ASCP) Urbana, IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Wednesday, February 16, 2005 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need info I work for the Navy as a civilian histo tech. I have been here for 5 yrs. And as of Valentine's Day, I got land basted by a pathologist presenting me with a new and revised work description of my supervisory duties. I got told that as of April 1st I will have to be trained for 30 days to gross in all small specimens. This is at no extra pay and I am a GS employee. I felt like I was done totally wrong, because they went behind my back and threatened me that if I didn't comply to learning how to gross, that administrative action will be taken against me. Now remind you I would've been far more receptive if I had been asked to gross and possibly be upgraded from a GS-7 to a GS-8 or GS-9. But no, I was treated like I was some enlisted navy sailor. I would like to know if anybody has any information on rights that histo techs have when it comes to grossing. Can an employer threaten you after working for 5 yrs and tell you if you don't comply, administrative action will be taken against you. I assume that means...in the board room, and being told "you're fired". I have the Union involved but any information on employee rights, regulations etc...would be of great help. Thank you in advance. Heather A. Harper _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From P.Krawczyk <@t> amc.uva.nl Wed Feb 16 09:10:08 2005 From: P.Krawczyk <@t> amc.uva.nl (P. Krawczyk) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Can TUNEL be used for in situ Double Strand Break detection assay? Message-ID: <000001c51439$9a6bbad0$08337591@Przemek> Hallo, I was wondering whether I could use the TUNEL / ISOL assay to label DNA double strand breaks specificaly. I know, that the terminal transferase (TUNEL) also labels single strand breaks and thus the staining would be probably non-specific. But I also heard that the T3 DNA Ligase (ISOL) labels duplex DNA breaks exclusively. Does anybody have experience with this type of labeling? Can it be used to visualize single DNA breaks under fluorescence microscope? Thank you, regards, P. Krawczyk _____________________________ P. M. Krawczyk Dept. of Cell Biology and Histology Academic Medical Centre University of Amsterdam Meibergdreef 15, room M3-107 1105 AZ, Amsterdam The Netherlands Email: P.Krawczyk@amc.uva.nl Tel: +31 20 5668746 Fax: +31 20 6974156 From Andrew.Prior <@t> Smith-Nephew.com Wed Feb 16 09:18:54 2005 From: Andrew.Prior <@t> Smith-Nephew.com (Prior, Andrew) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Bone saw and bone protocols Message-ID: David, Don't have protocols to hand - but is roughly 8 hours for each step of dehydration. Need to know what your final use for the bone is? Are you embedding and sectioning it or just doing gross pathology? As for the saw, we have just acquired an Exakt cutting system, and the band saw is excellent. Cuts thin (200 micron easy) MMA sections with very little damage to tissue, haven't tried fresh tissue yet. website is www.exakt.de The guys are very helpful. They are having a practical workshop at the start of April which you may be interested in so you can see their stuff ( I'm not employed by them, just happy with the new toys/ equipment in the lab). Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Date: Wed, 16 Feb 2005 08:10:20 -0000 From: "Muskett David" Subject: [Histonet] Bone saw and bone protocols[Scanned] To: Message-ID: <6AB79F0BA7C4914BA8DD8F5E0C21B9752E2928@AHEXMAIL01.xalderhey.com> Content-Type: text/plain; charset="US-ASCII" Dear All I am currently reviewing our protocols for the processing of bone. Would anybody be willing to share protocols and processing schedules? I am also interested in buying a bone saw, preferably one which allows thin sections to be cut easily. Can anybody recommend me a firm or a saw? 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 Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. From mcauliff <@t> umdnj.edu Wed Feb 16 13:18:21 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Iron staining (cellular)[Scanned] In-Reply-To: <1030B679AD69D6119C3F00080210DD9D05A3EFB4@bhrv-nt-11.bhrv.nwest.nhs.uk> References: <1030B679AD69D6119C3F00080210DD9D05A3EFB4@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: <42139C7D.8020108@umdnj.edu> Kelmo has a good point here, cultured macrophages may not have iron in them. Not every macrophage in the spleen has iron deposits, some are newly formed from monocytes or cell division. And, cells in culture do what they want which is not always what we hope for. For control, I suggest sections of spleen, not a few dabs of cells on a slide. Prussian and Turnbull blue are easy to do so it is unlikely that the problem lies with you. Do be sure to mix the HCl + ferrocyanide/ferricyanide immediately before use, not just fresh that day. Geoff Kemlo Rogerson wrote: >You must have a control slide or you will never get to the bottom of it. Why >should the macrophages have iron in them? When I cultured blood cells for my >MSc I learnt very quickly that what you got may bear no resemblance to what >you started with; cells that posses certain characteristics in vivo may not >in vitro. I wonder if you can't stain it as it's not there; haemorrhagic >placenta or omentum, I'm told, is a good control (Dave Rushworth; verbal). > > >That fellow J.A. Kiernan has a good section in his book 'Histological and >Histochemical Methods' Theory & Practice, 3rd Edition, Published by Arnold >1999. > >Kemlo Rogerson >Cellular Pathology Manager >East Lancashire Hospitals NHS Trust >DD. 01254-294162 >Mobile 0774-9754194 > > >-----Original Message----- >From: p.j.bergin@qmul.ac.uk [mailto:p.j.bergin@qmul.ac.uk] >Sent: 16 February 2005 11:25 >To: Kemlo Rogerson >Cc: histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Iron staining (cellular)[Scanned] > >I thought so too (about the adic fixing). So far we have fixed in >methenol/acetone, NBF, and tried with just air drying. The cells themselves >are just cultured macrophages (primary) and should have iron in them. >Unfortunaetlywe have no control, but have been using murine spleen cell >smears >for the positive and to try get the assay working. Still no luck though. >Although if you have a better suggestion for a control slide then that'd be >great. > >Cheers, > >Phil > >Quoting Kemlo Rogerson : > > > >>All I can remember is never use an acidic fixative, it removes Fe ions, or >>anything acidic before staining. The protocols you suggest appear fine; >> >> >how > > >>certain are you that the macrophages have 'ingested' iron? Could it be >> >> >that > > >>cloned cells don't act as they ought and that the iron was never >> >> >'ingested' > > >>in the first place? I assume you have put through an Fe2 and Fe3 control >>slide to tell you if your procedure was successful? >> >>Kemlo >> >>-----Original Message----- >>From: p.j.bergin@qmul.ac.uk [mailto:p.j.bergin@qmul.ac.uk] >>Sent: 15 February 2005 16:29 >>To: histonet@lists.utsouthwestern.edu >>Subject: [Histonet] Iron staining (cellular)[Scanned] >> >> >> >>Hi, >> >>I was hoping someone could give me advice. I have looked through the >>archives >>and have tried a few things already. I am interested in staining cell >> >> >preps > > >>to >>localize iron. I have tried using mouse spleen cells as a trial, but am >>mainly >>interested in human macrophages and monocytes that have been grown on >> >> >glass > > >>slides. I have tried fixing the preps with a methanol/acetone mix and >> >> >with > > >>4% >>formalin. I have been trying two different methods; Perls Prussian blue >>from >>the stainsfile archive and Turnbull's Blue (for ferric and ferrous iron >>respectively). Both have been unsuccessful with no staining detected (and >>the >>cells are there). >> >>Does anyone have a method for these stains that works on cells? do they >> >> >need > > >>to >>be fixed a specific way? Is there any advice anyone would care to share? >> >>Thanks in advance for any help. >> >>Phil >> >>P.S. The staining protocols I have been using follow..... >> >>FERROUS IRON - TURNBULL'S BLUE >> >>PURPOSE: To detect ferrous (Fe2+) iron in tissues. >>PRINCIPLE: Tissue sections are treated with an acidic solution of >> >> >potassium > > >>ferricyanide, any ferrous iron present will react to form an insoluble >>bright >>blue pigment called Turnbull's blue (ferrous ferricyanide). >>CONTROL: Routine iron control, which includes an negative. >>TECHNIQUE: Cut paraffin section 4?. >>EQUIPMENT: Acid clean glassware, non-metallic forceps. >>REAGENTS: >>0.006N Hydrochloric Acid 1% Acetic Acid >>Hydrochloric acid 2.5 ml Acetic acid, glacial 1.0 ml Distilled water 497.5 >>ml >>Distilled water 100.0 ml Mix well. Solution is stable for 1 Mix well. >>Solution >>is stable for 1 year. year. >>CAUTION: Corrosive acid. CAUTION: Avoid contact and inhalation. >>Potassium Ferricyanide Nuclear-Fast Red: >>Staining Solution: See Retic >>Potassium ferricyanide 0.4 gm >>Hydrochloric acid, 0.006N 40.0 ml >>Prepare fresh, just before use. >>CAUTION: Avoid contact and inhalation. >>SAFETY: Wear gloves, goggles and lab coat. Avoid contact and inhalation. >>Hydrochloric acid; target organ effects on reproductive system and fetal >>tissue. >>Irritant to skin eyes and respiratory system. >>Potassium ferricyanide; Low toxicity as long as it is not heated, it will >>release cyanide gas. >> >>MINERALS AND PIGMENTS >>TURNBULL'S FERROUS IRON Page: 2 of 2 >>Acetic acid: Irritating to respiratory system. Target organ effects on >>respiratory system by inhalation. Corrosive. >>PROCEDURE: >>1. Deparaffinize and hydrate to distilled water. >>2. Place slides in Potassium ferricyanide staining solution for 1 hour. >>3. Wash slides in 1% acetic acid. >>4. Counterstain slides in nuclear-fast red for 5 minutes. >>5. Rinse well in distilled water. >>6. Dehydrate, clear and coverslip. >>RESULTS: >>Ferrous iron blue >>Background pink-red >>REFERENCE: >>Carson, F, Histotechnology: A Self-Instructional Text, 1st Ed., 1991, pp. >>215-16 ASCP Press >> >>Perls Prussian Blue >> >>Principle >>The basis for the method is the release of ferric iron from hemosiderin by >>acid >>treatment, forming ferric chloride. The ferric iron reacts with potassium >>ferrocyanide to form ferric ferrocyanide. This is an insoluble, blue >>compound >>known as Prussian blue. The intensity of the colour gives some indication >> >> >as > > >>to >>amount, but it is qualitative only. >> >>4FeCl3 + 3K4Fe(CN)6 = Fe4[Fe(CN)6]3 + 12KCl >> >> Stock solution A >> Hydrochloric acid 2 ml >> Distilled water 98 ml >> >> >> Stock solution B >> Potassium ferrocyanide 2 g >> Distilled water 100 ml >> >> >> Working solution C >> Solution A 1 part >> Solution B 1 part >> >> >> Counterstain solution D >> Neutral red counterstain >> >> >> >> >>Fixation & processing >>Avoid iron containing materials and jars while fixing as these may >>contaminate >>the tissue. Acid containing fixatives may remove some of the iron >> >> >deposits. > > >>Otherwise most methods are satisfactory. >>Method >>Bring sections to distilled water with xylene and ethanol. >>Place into working solution C for 15 minutes. >>Rinse with distilled water, then tap water. >>Stain with counterstain solution D for one minute Rinse well with tap >> >> >water. > > >>Dehydrate with ethanol. >>Clear with xylene >> >> >>Expected Results >>Nuclei - red >> >> >>Notes >>The working solution C must be made immediately before use. >>Avoid washing with tap water before treating with solution C, as rust in >> >> >the > > >>water or tap fixtures could cause false positive staining. >>Wash well at step 3, as traces of iron will form a granular red deposit >> >> >with > > >>neutral red. >>Iron ores can be demonstrated, but the acid concentration in solution A >> >> >may > > >>need >>to be increased to 10% or more. >> >> >> >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@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 > > > > -- -- ********************************************** 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 AFeatherstone <@t> KaleidaHealth.Org Wed Feb 16 10:36:51 2005 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Placenta biopsy and storage Message-ID: We are looking into submitting for biopsy, all of our placentas. We are a large merged hospital with many outreach stations. I would like to know if anyone out there is currently dealing with grossing, sectioning, and storing of these specimens. I would like to know if you are using fixed and/or fresh tissue and if you are ultimately providing PCR and genetic testing. Please contact me either by email or phone. Any information will be greatly appreciated. Annette Featherstone HT/MLT Supervisor, Anatomic Pathology Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 716-859-2625 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Prior, Andrew Sent: Wednesday, February 16, 2005 10:19 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Re: Bone saw and bone protocols David, Don't have protocols to hand - but is roughly 8 hours for each step of dehydration. Need to know what your final use for the bone is? Are you embedding and sectioning it or just doing gross pathology? As for the saw, we have just acquired an Exakt cutting system, and the band saw is excellent. Cuts thin (200 micron easy) MMA sections with very little damage to tissue, haven't tried fresh tissue yet. website is www.exakt.de The guys are very helpful. They are having a practical workshop at the start of April which you may be interested in so you can see their stuff ( I'm not employed by them, just happy with the new toys/ equipment in the lab). Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Date: Wed, 16 Feb 2005 08:10:20 -0000 From: "Muskett David" Subject: [Histonet] Bone saw and bone protocols[Scanned] To: Message-ID: <6AB79F0BA7C4914BA8DD8F5E0C21B9752E2928@AHEXMAIL01.xalderhey.com> Content-Type: text/plain; charset="US-ASCII" Dear All I am currently reviewing our protocols for the processing of bone. Would anybody be willing to share protocols and processing schedules? I am also interested in buying a bone saw, preferably one which allows thin sections to be cut easily. Can anybody recommend me a firm or a saw? 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 Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. _______________________________________________ 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 p.j.bergin <@t> qmul.ac.uk Wed Feb 16 10:37:56 2005 From: p.j.bergin <@t> qmul.ac.uk (p.j.bergin@qmul.ac.uk) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Aqueous mountant Message-ID: <1108571876.421376e4709b6@webapps.qmul.ac.uk> Hi again everyone, I have been using mountquick mountant for a while now and it is great. However, I would like to switch to a good mountant that actually sets hard, prevents the cover slip moving, and is (hopefully) less likely to grow fungus etc. I have had a look through our catalogues and was wondering what other people used and recommended. I mainly want one that is compatable with AEC substrate and is easily available in the UK. Any recommendations would be appreciated. Cheers, Phil From AFeatherstone <@t> KaleidaHealth.Org Wed Feb 16 10:46:26 2005 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] CV5000 Robotic Coverslipper Message-ID: Would anyone have any of the plastic slide racks for the CV 5000 for sale? Annette Featherstone HT/MLT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Angela Bitting Sent: Tuesday, February 15, 2005 11:30 To: histonet@lists.utsouthwestern.edu; dsnider@shrinenet.org Subject: Re: [Histonet] non xylene based dehydrants I've used both ClearRite and Histoclear with very good results. 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 >>> "Snider, Deanna" 02/15/05 08:07AM >>> Ok everyone, What non xylene based products are being used by everyone? What are their health risks, hazards, etc. compared to xylene? Do they work as well as it does? Is it easily disposed of? The lab here uses xylene, which is my choice, but the restrictions on storing, pouring on/off, disposal, etc. borders on paranoia...:) So I am exploring the alternatives. The only exposure I've had to this type of product was years ago. Hemo-D, which gave me tremendous headaches.... I appreciate all your imput, and thanks in advance! Deanna Snider HT Shriners Hospital for Children Research Dept. Cincinnati, OH 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ 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 Heather.A.Harper <@t> pcola.med.navy.mil Wed Feb 16 11:02:07 2005 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Thank you Message-ID: <807FE48C5A7CC940B973B58D32E7014318A75EFB@nhpens-exch1.pcola.med.navy.mil> To everyone who responded to my post. Thank you so much. I am willing to learn to gross but if you saw the amount of work I do, I literally spin circles around my military co-worker. Time management is of the essence. I do have an Associates Degree and graduated from Histology School so I am not sure if the CLIA regs would be to my benefit. My beef is how the whole situation was handled and according to the union regs, you can't just change someone's work PD without sitting down with the employee and discussing the changes. Mine was changed behind my back than presented to me. Even though I am and consider myself a good histo tech, I think more money would talk better than the approach that was used on me. All histo techs know that we are in very high demand, and it took where I work a few years to even keep a histo tech...gees I wonder why. But to all of you who don't work for the government, when you deal w/the military, it's like republican vs. democrats. It's not the great job that we were taught to believe. It has its perks but you deal with a lot more political nonsense than at a regular histology civilian job. Thank you again. If you have additional info, please post. Heather From mari.ann.mailhiot <@t> leica-microsystems.com Wed Feb 16 11:30:35 2005 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] CV5000 Robotic Coverslipper Message-ID: Annette We still do have plastic slide racks for the CV5000. You are welcome to give me a call for the prices and part numbers. Best Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From anne.lewin <@t> bms.com Tue Feb 15 09:28:10 2005 From: anne.lewin <@t> bms.com (Anne C Lewin) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Formalin Fixation times for IHC samples Message-ID: <4212150A.9040600@bms.com> Could I get a general concensus from Histoland about the amount of time needed to fix a sample for IHC without over-fixing? My training/journal searches have led me to believe that the longer a sample is in formalin, the stronger the protiens are cross-linked, and the more difficult it is to break those bonds with antigen retreival and get your antibody to the target. I generally tell the scientists in my department to fix overnight (most samples vary from about the size of a pea to a lima bean), and then to transfer to 70% Ethanol for me to process for paraffin. The total time in fixative tends to be around 24 hours. Morphology has always been great, and my IHC's work well. This question is mainly for my information, since I have used the same tissue prep protocol for a few years now, I want to make sure I am keeping up with current opinions. Thanks! -Anne From skouko <@t> po-box.mcgill.ca Wed Feb 16 12:42:41 2005 From: skouko <@t> po-box.mcgill.ca (skouko@po-box.mcgill.ca) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Antibody Purification for Quantum Dots experiment Message-ID: <1108579361.421394218da5f@www.webmail.mcgill.ca> Hi guys, I want to use the Quantum Dots conjugation kit to label my primary antibody for an immunohistochemistry experiment. The QD protocol requires the antibody to be ascites and protein free. Unfortunately my antibody has ascites in it. I've already performed a "www.biocompare.com" search and there is no equivalent antibody on the market that is ascites free. How do I purify my antibody without losing half the content (as would happen if I used dialysis). Any ideas? Cheers, Sophia Koukoui Behavioural Neuroscience McGill University. From la.sebree <@t> hosp.wisc.edu Wed Feb 16 12:44:45 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] HepPar-1 Message-ID: Hi all, Our pathologists want us to get HepPar-1. What have people tried out there that has worked well for them? We have Ventana instruments. 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 TJasper <@t> smdc.org Wed Feb 16 12:49:31 2005 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Need info Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E45098@harrier> Dear Heather, Your situation raises a lot of questions. First of all I'm quite certain since you are an employee of the government (federal) you are bound by conditions that do not exist in other sectors of employment. Perhaps you need to take a look at the contract you work under. You mentioned union involvement. Your statement"...revised work description of my supervisory duties." is confusing. Maybe I don't fully understand, but I am under the impression that if you belong to a union you do not supervise as that is an administrative/managerial responsibility. Perhaps you are viewed as a "lead tech or working charge tech" type of employee so as not to conflict with your union status? Your question about being threatened by an employer...administrative action...being told "you're fired"; if you are indeed a union member you should be protected against any rash or undocumented action by an employer of this nature. You should be discussing this with your union steward. Again this is unclear as you are a government employee, you have supervisory duties and yet you reference the union. What I know as "non-contract" employees are subject to "firing at will", although this is not possible in any real sense without proper "cause". Employers do have to be accountable for their actions vis-a-vis employees and there is the added value placed on good histotechs these days due to staff shortages. Based on the information you've provided it seems possible you are being treated unfairly. However, I certainly don't know enough about your situation to definitely say that is the case. I supervise both union (contract) and non-union employees. In management we would not be able to make any changes to a (union) job description without union knowledge. Having said that, approximately 2 years ago we did make a change to our histotech (union) job description, which pertained to grossing in at the bench. We added what we call "Minor Gross Dictations" for our histotechs. We defined "minor grosses" as small endoscopy and dermal biopsies, the work is still carried out under the direction of our PA or a pathologist. We did this for a few different reasons: we had just merged with another facility and our pathologists thought they might need the help; our PA is also the administrator of our AP LIS system, at times is pulled away from the bench coverage is required to keep the operation running; it is also becoming a standard of care issue as histotechs possess the ability to handle this task in a knowledgeable and responsible manner. The reality for us is that histotechs "occasionally" perform this duty, in most cases when it comes down to it, due to staff schedules, the task defaults to me. It works out for us as there are times when late "rush" cases come in and they can be added to the evening run when the PA or pathologists are gone. Keep in mind that the histotechs are to set cases aside if they believe, due to complexity, the case should be dictated by the PA or pathologist. Please understand that this works for us, everyone is unique and pathology services will vastly differ one to another. I don't know what is expected of you daily as a histotech and what would be expected of you as the grossing person at the bench. These factors are defined by the nature and scope of your service. To my way of thinking actual grossing bench work is one job and being a histotech is one job. Although I don't know your workflow, I wouldn't expect that you would have to do both, it's probably too much work and you would be subject to legal limitations. Sorry for the ramble, I hope this is of some help, please let us know how this turns out. Good luck! Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Heather.A.Harper@pcola.med.navy.mil [mailto:Heather.A.Harper@pcola.med.navy.mil] Sent: Wednesday, February 16, 2005 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need info I work for the Navy as a civilian histo tech. I have been here for 5 yrs. And as of Valentine's Day, I got land basted by a pathologist presenting me with a new and revised work description of my supervisory duties. I got told that as of April 1st I will have to be trained for 30 days to gross in all small specimens. This is at no extra pay and I am a GS employee. I felt like I was done totally wrong, because they went behind my back and threatened me that if I didn't comply to learning how to gross, that administrative action will be taken against me. Now remind you I would've been far more receptive if I had been asked to gross and possibly be upgraded from a GS-7 to a GS-8 or GS-9. But no, I was treated like I was some enlisted navy sailor. I would like to know if anybody has any information on rights that histo techs have when it comes to grossing. Can an employer threaten you after working for 5 yrs and tell you if you don't comply, administrative action will be taken against you. I assume that means...in the board room, and being told "you're fired". I have the Union involved but any information on employee rights, regulations etc...would be of great help. Thank you in advance. Heather A. Harper _______________________________________________ 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 bliven.laura <@t> marshfieldclinic.org Wed Feb 16 13:00:37 2005 From: bliven.laura <@t> marshfieldclinic.org (bliven.laura@marshfieldclinic.org) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] HIER and EIER Message-ID: <2c1ba01c51459$cceb3a60$8e05010a@mfldclinframe.org> When used in combination, is it better to use HIER (Heat Induced Epitope Retrieval) first or EIER (Enzyme Induced Epitope Retrieval) first. Or does it depend upon the antigen of interest? Any references? Thanks, Laura Bliven From sluhisto <@t> yahoo.com Wed Feb 16 13:23:08 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] cd5 and cd23 on B5 fixed tissue Message-ID: <20050216192308.26909.qmail@web51004.mail.yahoo.com> Hello All: Is anyone doing these two antibodies on B5 fixed tissue with good results? If you are, and would be willing to share your secrets, any info would be appreciated. Both of the vendors and a couple of others have not used these antibodies with B5 fixation as part of their QC procedures and can not help. Thanks, in advance. Susan --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From la.sebree <@t> hosp.wisc.edu Wed Feb 16 13:27:21 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] HIER and EIER Message-ID: We've always done our HIER before the EIER but mainly because that's how our instruments proceed. 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 bliven.laura@marshfieldclinic.org Sent: Wednesday, February 16, 2005 1:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HIER and EIER When used in combination, is it better to use HIER (Heat Induced Epitope Retrieval) first or EIER (Enzyme Induced Epitope Retrieval) first. Or does it depend upon the antigen of interest? Any references? Thanks, Laura Bliven _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed Feb 16 16:39:34 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Formalin Fixation times for IHC samples In-Reply-To: <4212150A.9040600@bms.com> References: <4212150A.9040600@bms.com> Message-ID: <4213CBA6.40804@umdnj.edu> Dear Anne: Standard answer, it depends on the antigen in question. Some survive fixation for 24 hours or more, some are rendered unreactive in 4 hours. Formalin works slowly so longer fixation usually reduces immunoreactivity. Also, the method of detection is important. In my hands the Vector ABC Elite kit gives a much stronger signal than the standard kit and often gives good results when the standard kit shows nothing. Geoff Anne C Lewin wrote: > Could I get a general concensus from Histoland about the amount of > time needed to fix a sample for IHC without over-fixing? My > training/journal searches have led me to believe that the longer a > sample is in formalin, the stronger the protiens are cross-linked, and > the more difficult it is to break those bonds with antigen retreival > and get your antibody to the target. I generally tell the scientists > in my department to fix overnight (most samples vary from about the > size of a pea to a lima bean), and then to transfer to 70% Ethanol for > me to process for paraffin. The total time in fixative tends to be > around 24 hours. Morphology has always been great, and my IHC's work > well. This question is mainly for my information, since I have used > the same tissue prep protocol for a few years now, I want to make sure > I am keeping up with current opinions. > Thanks! > -Anne > >------------------------------------------------------------------------ > >_______________________________________________ >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 Luis.Chiriboga <@t> med.nyu.edu Wed Feb 16 14:11:11 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] HepPar-1 In-Reply-To: Message-ID: works extremely well! From dako (m7158:clone OCH1E5), 10 minute HIER (Microwave) 0.01M pH6 citrate, dilute 1:20 and run for 32 minutes on a Nexus LC -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sebree Linda A. Sent: Wednesday, February 16, 2005 1:45 PM To: Histonet (E-mail) Subject: [Histonet] HepPar-1 Hi all, Our pathologists want us to get HepPar-1. What have people tried out there that has worked well for them? We have Ventana instruments. Thanks, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From browning <@t> HHSC.CA Wed Feb 16 14:23:55 2005 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Formalin Fixation times for IHC samples Message-ID: <3AADFB88753AD31189C100902786B91C0E278708@hch_nt_exchange.hhsc.ca> 24 hours works best for us, we have standardized the retrieval protocols based on this period of fixation. -----Original Message----- From: Anne C Lewin [mailto:anne.lewin@bms.com] Sent: Tuesday, February 15, 2005 10:28 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Formalin Fixation times for IHC samples Could I get a general concensus from Histoland about the amount of time needed to fix a sample for IHC without over-fixing? My training/journal searches have led me to believe that the longer a sample is in formalin, the stronger the protiens are cross-linked, and the more difficult it is to break those bonds with antigen retreival and get your antibody to the target. I generally tell the scientists in my department to fix overnight (most samples vary from about the size of a pea to a lima bean), and then to transfer to 70% Ethanol for me to process for paraffin. The total time in fixative tends to be around 24 hours. Morphology has always been great, and my IHC's work well. This question is mainly for my information, since I have used the same tissue prep protocol for a few years now, I want to make sure I am keeping up with current opinions. Thanks! -Anne This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From browning <@t> HHSC.CA Wed Feb 16 14:25:32 2005 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] HIER and EIER Message-ID: <3AADFB88753AD31189C100902786B91C0E278709@hch_nt_exchange.hhsc.ca> With the limited antibodies that we have to do both EIER and HIER, we perform the enzyme first with good success. -----Original Message----- From: bliven.laura@marshfieldclinic.org [mailto:bliven.laura@marshfieldclinic.org] Sent: Wednesday, February 16, 2005 2:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HIER and EIER When used in combination, is it better to use HIER (Heat Induced Epitope Retrieval) first or EIER (Enzyme Induced Epitope Retrieval) first. Or does it depend upon the antigen of interest? Any references? Thanks, Laura Bliven _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From RCHIOVETTI <@t> aol.com Wed Feb 16 14:44:55 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Antibody Purification for Quantum Dots experiment Message-ID: In a message dated 2/16/2005 11:45:13 AM US Mountain Standard Time, skouko@po-box.mcgill.ca writes: > How do I purify my antibody > without losing half the content (as would happen if I used dialysis). Any > ideas? > Hi Sophia, You could probably get a fairly good yield by using affinity purification with either a Protein G or Protein A - Sepharose column. The exact protocol would depend on the origin of the antibody, IgA / IgM, etc. Protein G seems to be used more frequently than Protein A. For background, see for example: Good luck! Cheers, Bob Chiovetti From Rcartun <@t> harthosp.org Wed Feb 16 15:00:24 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] cd5 and cd23 on B5 fixed tissue Message-ID: How many labs still use B5? We should work towards eliminating it from our environment. 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 >>> Histology SLU 02/16/05 02:23PM >>> Hello All: Is anyone doing these two antibodies on B5 fixed tissue with good results? If you are, and would be willing to share your secrets, any info would be appreciated. Both of the vendors and a couple of others have not used these antibodies with B5 fixation as part of their QC procedures and can not help. Thanks, in advance. Susan --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jhabecke <@t> seattlecca.org Wed Feb 16 15:06:02 2005 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Antibodies to Thy1.1 in FFPE mouse tissue Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAF10@wala01.seattlecca.org> Hello all, I am working with an investigator who would like to detect Thy1.1 (CD90.1) positive cells which were infused into a mouse expressing Thy1.2 (CD90.2). They are doing this retrospectively and already have FFPE tissue (so I can't use frozen tissue). I tried the OX-7 clone from BD with no antigen retrieval, pH 6 tris/steam, EDTA/steam, and proteinase K using the ARK kit for detection on spleens from Thy1.1 mice..... But I didn't get any signal. Has anyone had good results with a Thy1.1 antibody on FFPE mouse tissue? Please let me know! Thanks, Julie Julie Randolph-Habecker, Ph.D. Experimental Histopathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N, G1-300 PO Box 19023 Seattle, WA 98109-1024 Tel: (206) 288-1187 FAX: (206) 288-1345 jhabecke@fhcrc.org This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From jstaruk <@t> masshistology.com Wed Feb 16 15:35:04 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Question on blocking serum In-Reply-To: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAF10@wala01.seattlecca.org> Message-ID: <20050216213507.BYKF25879.out010.verizon.net@FrontOffice> All, I always block non-specific antibodies with a 20% solution of normal serum of the same species the secondary antibody was produced in prior to the initial primary antibody step. What would I use as a blocking solution if the primary antibody is already biotin labeled? A 20% solution of normal serum of the same species the primary antibody was raised in? Anyone planning on invoicing me for their answer need not respond to this question. Thanks in advance Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com From nmuvarak <@t> facstaff.wisc.edu Wed Feb 16 16:14:16 2005 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL E MUVARAK) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Pressure fixation with PFA Message-ID: <83928d838fdf.838fdf83928d@wiscmail.wisc.edu> I was wondering if anyone knows how long is enough to pressure-fix rat brain vessels with PFA? Thanks. Nidal E Muvarak Associate Research Specialist Vascular Tissue Biomechanics Laboratory Department of Biomedical Engineering University of Wisconsin-Madison 1550 Engineering Dr.; Rm. 2158 Madison, WI 53706-1609 Lab: (608) 265-8921; Office: (608) 265-4205; Home: (608) 249-2611; Cell: (608) 332-6068 http://vtb.bme.wisc.edu From AnthonyH <@t> chw.edu.au Wed Feb 16 16:36:32 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Question on blocking serum Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E1B2@simba.kids> Jim, I would use either the same sera that the primary was raised in or whatever sera from another species that I had on hand. I hope that any non-specific binding of the primary AB would be prevented by this blocking sera incubation. 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: Jim Staruk [mailto:jstaruk@masshistology.com] Sent: Thursday, 17 February 2005 8:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Question on blocking serum All, I always block non-specific antibodies with a 20% solution of normal serum of the same species the secondary antibody was produced in prior to the initial primary antibody step. What would I use as a blocking solution if the primary antibody is already biotin labeled? A 20% solution of normal serum of the same species the primary antibody was raised in? Anyone planning on invoicing me for their answer need not respond to this question. Thanks in advance Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From bradyjfulton <@t> yahoo.com Wed Feb 16 17:22:48 2005 From: bradyjfulton <@t> yahoo.com (brady fulton) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Help -- I'm Looking for Carrie Kyle-Byrne Message-ID: <20050216232248.20089.qmail@web53604.mail.yahoo.com> I am hoping to find Carrie Kyle-Byrne. She was formerly with Genentech, then (I think) the California Animal Health and Food Safety Labratory (at U.C. Davis), then Eyelixis, and then Lab Vision. I am interested in some work that she did when she was with Genentech, and would like to get in contact with her to discuss it. She used to post quite a few messages here. Someone told me that she might have moved out of the country, but they didn't know any more than that. Does anyone know how I can reach her or what her current contact info or email address is? Thanks very much. Brady Fulton Chicago, IL bradyjfulton@yahoo.com (312) 377-3246 --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From scoop <@t> mail.nih.gov Wed Feb 16 18:02:33 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] re mouse perfusion set up Message-ID: Many thanks to all the people who gave me suggestions for a more convenient mouse perfusion set up. It sounds like most people make a stage from a block of paraffin and use that until it gets yucky and then make another one. Good, convenient, cheap idea! I'm planning to try it. Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From katri <@t> cogeco.ca Wed Feb 16 20:22:38 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Formalin Fixation times for IHC samples References: <4212150A.9040600@bms.com> Message-ID: <004301c51497$8d44f110$6a9a9618@Katri> Anne, I think you are doing well with your protocol. Although I don't think "overfixation" is as a big problem any more with the advent of different HIER protocols with various buffers being used. Under fixation (less than 24 hours) can be a bigger problem with certain antibodies like Her-2, ER and PR to mention few. This concept of over fixation is a remnant from the time prior to HIER, when we all had a problem trying to demonstrate many antigens with weaker retrievals (proteolytic enzymes). The heat over 60C was actually banned from any immunohistochemistry protocols 20 years ago. My two cents worth... Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Anne C Lewin" To: Sent: Tuesday, February 15, 2005 10:28 AM Subject: [Histonet] Formalin Fixation times for IHC samples > Could I get a general concensus from Histoland about the amount of time > needed to fix a sample for IHC without over-fixing? My training/journal > searches have led me to believe that the longer a sample is in > formalin, the stronger the protiens are cross-linked, and the more > difficult it is to break those bonds with antigen retreival and get your > antibody to the target. I generally tell the scientists in my > department to fix overnight (most samples vary from about the size of a > pea to a lima bean), and then to transfer to 70% Ethanol for me to > process for paraffin. The total time in fixative tends to be around 24 > hours. Morphology has always been great, and my IHC's work well. This > question is mainly for my information, since I have used the same tissue > prep protocol for a few years now, I want to make sure I am keeping up > with current opinions. > Thanks! > -Anne > -------------------------------------------------------------------------------- > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From scoop <@t> mail.nih.gov Wed Feb 16 20:43:58 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] mouse skin biopsies Message-ID: Dear Histonetters, I need to do a skin biopsy on a mouse. I've never done any skin stuff before. Could someone share a protocol with me on how to do this? (even info on human skin biopsies would be helpful). Are skin biopsies usually formalin fixed and paraffin embedded? If anyone has a protocol or a reference I could look up it would be greatly appreciated. Ignorantly yours, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From CrochiereSteve <@t> aol.com Wed Feb 16 21:16:47 2005 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Gunk in stainer drain line Message-ID: Does anyone have a problem with gunk clogging up the drain line on their autostainer. I have a Varistain Gemini, and the line has clogged up with blue goo twice in the past 4 years. What could be the cause and how would I prevent it, short of periodically taking the thing apart and reaming out the hose? Pouring bleach down the drain was suggested, but didn't keep the problem from recurring. Any suggestions? Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 From Kemlo.Rogerson <@t> elht.nhs.uk Thu Feb 17 02:21:44 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Formalin Fixation times for IHC samples[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EFC7@bhrv-nt-11.bhrv.nwest.nhs.uk> Not very good at ICC but I always believed formalin fixation could be washed out; in other words if you over fix the situation is not irreparable. -----Original Message----- From: Anne C Lewin [mailto:anne.lewin@bms.com] Sent: 15 February 2005 15:28 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Formalin Fixation times for IHC samples[Scanned] Could I get a general concensus from Histoland about the amount of time needed to fix a sample for IHC without over-fixing? My training/journal searches have led me to believe that the longer a sample is in formalin, the stronger the protiens are cross-linked, and the more difficult it is to break those bonds with antigen retreival and get your antibody to the target. I generally tell the scientists in my department to fix overnight (most samples vary from about the size of a pea to a lima bean), and then to transfer to 70% Ethanol for me to process for paraffin. The total time in fixative tends to be around 24 hours. Morphology has always been great, and my IHC's work well. This question is mainly for my information, since I have used the same tissue prep protocol for a few years now, I want to make sure I am keeping up with current opinions. Thanks! -Anne From Kemlo.Rogerson <@t> elht.nhs.uk Thu Feb 17 02:23:48 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Gunk in stainer drain line[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EFC9@bhrv-nt-11.bhrv.nwest.nhs.uk> Do you stain Non-Gynae? Specifically breast FNAC's? -----Original Message----- From: CrochiereSteve@aol.com [mailto:CrochiereSteve@aol.com] Sent: 17 February 2005 03:17 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Gunk in stainer drain line[Scanned] Does anyone have a problem with gunk clogging up the drain line on their autostainer. I have a Varistain Gemini, and the line has clogged up with blue goo twice in the past 4 years. What could be the cause and how would I prevent it, short of periodically taking the thing apart and reaming out the hose? Pouring bleach down the drain was suggested, but didn't keep the problem from recurring. Any suggestions? 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 lpwenk <@t> sbcglobal.net Thu Feb 17 04:40:00 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] CAP and proficiency testing References: Message-ID: <007301c514dd$0b84aee0$ac30d445@domainnotset.invalid> Hmm - I thought Proficiency assessment had to be through an OUTSIDE agency, such as CAP or a CAP-alternate approved provider, which assesses how the LAB did. Isn't an INTERNAL evaluation actually a competency assessment, assessing how each PERSON does? As for the general requirements for a proficiency assessment, go to the CAP web page, to the checklists, and click on Laboratory General, Dec. 2004. http://www.cap.org/apps/docs/laboratory_accreditation/checklists/checklistft p.html For more information, including Frequently Asked Questions, go to the CAP page http://www.cap.org/apps/cap.portal?_nfpb=true&_pageLabel=proficiency_testing _page As far as I know, HistoQIP is the only CAP accepted proficiency assessment for histotechs. http://www.cap.org/apps/docs/proficiency_testing/histoqip.html Over 500 labs in the US participate, and twice a year they have to submit slides of H&E/special stains/IHC to be assessed on fixation, processing, embedding, sectioning, staining and coverslipping. Each cycle it is different tissues (gathered from recent surgical cases or controls) and different stains. These slides are assessed by a panel of histotechs and pathologists. Each lab gets a report of how they did, and how they compare with the rest of the participating labs. Along with the report is an explanation of the stains just assessed (how they work, common errors) along with Kodachromes (new - they will be color plates instead of 35 mm slides). And I understand that there will be multiple test questions in the future, so these reports can be used by the lab for training and competency assessment. No, I'm not on the panel, but I think HistoQIP is a great idea. CAP and NSH have worked together on this. Our lab participates, and I personally recommend it for every histology lab. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Angela Bitting" To: ; Sent: Tuesday, February 15, 2005 11:28 AM Subject: Re: [Histonet] CAP and proficiency testing > Our proficiency testing is very simple. We named six general functions > and gave an excellent, good or poor rating. Method was direct > observation. The tester and techs signed and dated it. Repeat it > annually. > > 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 > > >>> 02/15/05 10:45AM >>> > Does anyone know what the requirements are for proficiency testing for > CAP accredited private histology laboratories? > Thanks, > Ron Martin > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Thu Feb 17 05:12:43 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Gunk in stainer drain line Message-ID: In a word, yes. It's disgusting, and presumably severely compromises the efficacy of the wash, inadequate at the best of times. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: CrochiereSteve@aol.com [mailto:CrochiereSteve@aol.com] Sent: 17 February 2005 03:17 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Gunk in stainer drain line Does anyone have a problem with gunk clogging up the drain line on their autostainer. I have a Varistain Gemini, and the line has clogged up with blue goo twice in the past 4 years. What could be the cause and how would I prevent it, short of periodically taking the thing apart and reaming out the hose? Pouring bleach down the drain was suggested, but didn't keep the problem from recurring. Any suggestions? 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 BMolinari <@t> heart.thi.tmc.edu Thu Feb 17 06:20:05 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] mouse skin biopsies Message-ID: Sharon, I have done a few, they were for monitoring the hair growth, so I had only a small tag of skin. They were fixed in 10% NBF and I ran them in a same day run. Seemed to work fine for us. 70% 30min 80% 30min 95% 30min 95% 30min 100% 30min 100% 30min xylene 45min xylene 45min paraffin 30min paraffin 30min paraffin 30min paraffin 30min Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave Houston,TX 77090 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Cooperman Sent: Wednesday, February 16, 2005 8:44 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] mouse skin biopsies Dear Histonetters, I need to do a skin biopsy on a mouse. I've never done any skin stuff before. Could someone share a protocol with me on how to do this? (even info on human skin biopsies would be helpful). Are skin biopsies usually formalin fixed and paraffin embedded? If anyone has a protocol or a reference I could look up it would be greatly appreciated. Ignorantly yours, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfockler <@t> mail1.vcu.edu Thu Feb 17 06:32:49 2005 From: cfockler <@t> mail1.vcu.edu (cfockler@mail1.vcu.edu) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Grossing Tech Message-ID: <200502171232.HAA22089@despina.vcu.edu> Just a quick question.. . what is the average pay increase to go from a standard histotech, to a grossing tech? (with a B.S.) And, how long on average should it take to see it in your job description and your paycheck? Thanks! C. Fockler From japoteete <@t> saintfrancis.com Thu Feb 17 07:34:58 2005 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Gunk in stainer drain line Message-ID: The "blue goo" in the stainers in our laboratory was found to be a variety of slime mold. I can't remember its' scientific name, but we had to use bleach to get rid of it, and the bleach did not prevent it from recurring. Pouring bleach down the drain became part of our cleaning-maintenance procedure. Good luck! Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com -----Original Message----- From: CrochiereSteve@aol.com [mailto:CrochiereSteve@aol.com] Sent: Wednesday, February 16, 2005 9:17 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Gunk in stainer drain line Does anyone have a problem with gunk clogging up the drain line on their autostainer. I have a Varistain Gemini, and the line has clogged up with blue goo twice in the past 4 years. What could be the cause and how would I prevent it, short of periodically taking the thing apart and reaming out the hose? Pouring bleach down the drain was suggested, but didn't keep the problem from recurring. Any suggestions? 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 ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From Kemlo.Rogerson <@t> elht.nhs.uk Thu Feb 17 08:16:49 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Gunk in stainer drain line[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EFD0@bhrv-nt-11.bhrv.nwest.nhs.uk> When we had these problems we asked the Company about how to solve it. They asked if we stained breast FNAC's and we did. They suggested it was the fat from said aspirates; I had always thought it was fungus. We adopted a protocol of bleach and boiling water and that cured it, but gassed the BMS's. We had a similar problem in London too, but that was really due to the sewerage they laughingly call 'drinking water'. The solution was to connect the stainer to piped 'clean' water; but then no-one drinks the water in London they all use chillers. Doesn't explain why the beer 'London pride' is so good, or does it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Poteete, Jacquie A. [mailto:japoteete@saintfrancis.com] Sent: 17 February 2005 13:35 To: CrochiereSteve@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line[Scanned] The "blue goo" in the stainers in our laboratory was found to be a variety of slime mold. I can't remember its' scientific name, but we had to use bleach to get rid of it, and the bleach did not prevent it from recurring. Pouring bleach down the drain became part of our cleaning-maintenance procedure. Good luck! Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com -----Original Message----- From: CrochiereSteve@aol.com [mailto:CrochiereSteve@aol.com] Sent: Wednesday, February 16, 2005 9:17 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Gunk in stainer drain line Does anyone have a problem with gunk clogging up the drain line on their autostainer. I have a Varistain Gemini, and the line has clogged up with blue goo twice in the past 4 years. What could be the cause and how would I prevent it, short of periodically taking the thing apart and reaming out the hose? Pouring bleach down the drain was suggested, but didn't keep the problem from recurring. Any suggestions? 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 ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Thu Feb 17 08:20:38 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Gunk in stainer drain line Message-ID: I can't resist this - what a name! http://botit.botany.wisc.edu/toms_fungi/june99.html Tom Volk's Fungus of the Month for June 1999 This month's fungus is Fuligo septica, the dog vomit slime mold. Well worth a little read. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Poteete, Jacquie A. [mailto:japoteete@saintfrancis.com] Sent: 17 February 2005 13:35 To: 'CrochiereSteve@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line The "blue goo" in the stainers in our laboratory was found to be a variety of slime mold. I can't remember its' scientific name, but we had to use bleach to get rid of it, and the bleach did not prevent it from recurring. Pouring bleach down the drain became part of our cleaning-maintenance procedure. Good luck! Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com -----Original Message----- From: CrochiereSteve@aol.com [mailto:CrochiereSteve@aol.com] Sent: Wednesday, February 16, 2005 9:17 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Gunk in stainer drain line Does anyone have a problem with gunk clogging up the drain line on their autostainer. I have a Varistain Gemini, and the line has clogged up with blue goo twice in the past 4 years. What could be the cause and how would I prevent it, short of periodically taking the thing apart and reaming out the hose? Pouring bleach down the drain was suggested, but didn't keep the problem from recurring. Any suggestions? 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 ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ajennings <@t> unmc.edu Thu Feb 17 08:50:16 2005 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] mouse skin biopsies In-Reply-To: Message-ID: I had the pleasure a few years back to work on the preliminary studies for UpJohn. We had a mouse line that lost its hair and they took advantage of these guys to see if they could regrow hair with the application of product. Depending on what your final needs are this may be of some help. We were looking mostly for morphological changes in hair growth. We fixed our skin punches in modified Carnoy's (95%ETOH/5%AcA) followed by 3X15 min changes of 100% ETOH 2X15min Xylenes 2X30 min paraffin with vacuum. Not sure if the reference has the actual procedure but the above was routine in the lab. Knapp, L.W. and Dawson, W.D. 1991. Morphological Analysis of Hair in the hr-2 Mutant Deer Mouse (Peromyscus maniculatus). J. Heredity 82:431. From pmarcum <@t> polysciences.com Thu Feb 17 09:18:09 2005 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Gunk in stainer drain line In-Reply-To: Message-ID: <000301c51503$e3bd3400$7f00a8c0@PMARCUM2K> If it is a slime mold forming would a PM using hot water and soap with anti bacterial agent help or just using a true anti-bacterial (not Bleach) once a week? Just a thought. Thanks, Pam Marcum? > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Poteete, > Jacquie A. > Sent: Thursday, February 17, 2005 8:35 AM > To: 'CrochiereSteve@aol.com'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Re: Gunk in stainer drain line > > > The "blue goo" in the stainers in our laboratory was found to be a variety > of slime mold. I can't remember its' scientific name, but we had to use > bleach to get rid of it, and the bleach did not prevent it from recurring. > Pouring bleach down the drain became part of our cleaning-maintenance > procedure. Good luck! > > Jacquie Poteete MT(ASCP)QIHC > Lead Technologist, IHC Laboratory > Saint Francis Hospital, Tulsa, OK > japoteete@saintfrancis.com > > -----Original Message----- > From: CrochiereSteve@aol.com [mailto:CrochiereSteve@aol.com] > Sent: Wednesday, February 16, 2005 9:17 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Gunk in stainer drain line > > > Does anyone have a problem with gunk clogging up the drain line on their > autostainer. I have a Varistain Gemini, and the line has clogged up with > blue goo > twice in the past 4 years. What could be the cause and how would I prevent > it, > short of periodically taking the thing apart and reaming out the hose? > Pouring bleach down the drain was suggested, but didn't keep the problem > from > recurring. > Any suggestions? > > 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 > > > > > ********* Email Confidentiality Statement ********* > Visit http://www.saintfrancis.com/emailconf.asp > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From GAshton <@t> PICR.man.ac.uk Thu Feb 17 09:38:33 2005 From: GAshton <@t> PICR.man.ac.uk (Garry Ashton) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] fibroblasts Message-ID: Hi Histonetters, I've been asked if I know of any ICC markers to stain components of extracellular matrix, in particular fibroblasts (in mouse). I've looked in the archives with no real success. Could anybody give me any advice. many thanks Garry PICR UK -------------------------------------------------------- This email is confidential and intended solely for the use of the person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute for Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From Terry.Marshall <@t> rothgen.nhs.uk Thu Feb 17 09:38:20 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Gunk in stainer drain line[Scanned] Message-ID: Kemlo, You believe that (about the breast fat)? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 February 2005 14:17 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line[Scanned] When we had these problems we asked the Company about how to solve it. They asked if we stained breast FNAC's and we did. They suggested it was the fat from said aspirates; I had always thought it was fungus. We adopted a protocol of bleach and boiling water and that cured it, but gassed the BMS's. We had a similar problem in London too, but that was really due to the sewerage they laughingly call 'drinking water'. The solution was to connect the stainer to piped 'clean' water; but then no-one drinks the water in London they all use chillers. Doesn't explain why the beer 'London pride' is so good, or does it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Poteete, Jacquie A. [mailto:japoteete@saintfrancis.com] Sent: 17 February 2005 13:35 To: CrochiereSteve@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line[Scanned] The "blue goo" in the stainers in our laboratory was found to be a variety of slime mold. I can't remember its' scientific name, but we had to use bleach to get rid of it, and the bleach did not prevent it from recurring. Pouring bleach down the drain became part of our cleaning-maintenance procedure. Good luck! Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com -----Original Message----- From: CrochiereSteve@aol.com [mailto:CrochiereSteve@aol.com] Sent: Wednesday, February 16, 2005 9:17 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Gunk in stainer drain line Does anyone have a problem with gunk clogging up the drain line on their autostainer. I have a Varistain Gemini, and the line has clogged up with blue goo twice in the past 4 years. What could be the cause and how would I prevent it, short of periodically taking the thing apart and reaming out the hose? Pouring bleach down the drain was suggested, but didn't keep the problem from recurring. Any suggestions? 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 ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@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 celebrej <@t> HHSC.CA Thu Feb 17 10:04:22 2005 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Grossing Tech Message-ID: <3AADFB88753AD31189C100902786B91C14E6DB69@hch_nt_exchange.hhsc.ca> I don't know about other hospitals, but for the last 12 years all our techs have been grossing without seeing any increase in our paycheques. Grossing has now been classified as 'other duties as assigned'. Great to hear that other sites get compensated for grossing, maybe in the next ten years we will be too? Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca -----Original Message----- From: cfockler@mail1.vcu.edu [mailto:cfockler@mail1.vcu.edu] Sent: Thursday, February 17, 2005 7:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Grossing Tech Just a quick question.. . what is the average pay increase to go from a standard histotech, to a grossing tech? (with a B.S.) And, how long on average should it take to see it in your job description and your paycheck? Thanks! C. Fockler _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From Charles.Embrey <@t> carle.com Thu Feb 17 10:15:50 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Grossing Tech Message-ID: First, my job description would have to be finalized and pay raise set before I opened my first specimen jar. Once you have started the duties your employer has lost his incentive to get this done. As far as pay raise is concerned it would really depend on the extent of your grossing and time involved. Personally I think the raise negotiations should start at $5.00 for small biopsies and increase with the level of specimen difficulty. The biggest problem I have found is that people tend to underestimate their worth. The only other option for grossing is a pathologist or pathologists' assistant and you would still be a steal with a $5.00 raise. If you enjoy grossing you should apply to one of the PA schools. It is only an additional two years of school, one of which is actual grossing at clinical sites. I teach second year PA students from Rosalind Franklin University in Chicago and my two students that graduate in April have already signed great contracts. Good Luck Charles Embrey PA(ASCP) Urbana, IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cfockler@mail1.vcu.edu Sent: Thursday, February 17, 2005 6:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Grossing Tech Just a quick question.. . what is the average pay increase to go from a standard histotech, to a grossing tech? (with a B.S.) And, how long on average should it take to see it in your job description and your paycheck? Thanks! C. Fockler _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kelly.mcqueeney <@t> bms.com Thu Feb 17 10:52:36 2005 From: kelly.mcqueeney <@t> bms.com (Kelly D Mcqueeney) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Frozen storage of tissue and sections, what is the correct temperature? Message-ID: <4214CBD4.7060209@bms.com> I know that frozen tissue storage is a common topic, but I have a hard time finding the right answer throughout the histonet archives. We are currently coating brain in embedding medium and snap freezing brains in isopentane followed by immediate storage at -80C. Then, we bring the brain up to -20C a few hours before sectioning. After sectioning, we dry the slides at RT for several hours and store at -80C until using the tissue for ligand binding, IHC or insitu. This exact protocol has been moderately successful, so far so good. But, some of the staff store frozen brains at -20C. They feel that keeping the tissue at cryostat temperature reduces the amount of freeze artifact. I was always taught to freeze fresh frozen brain at a much lower temperature. None of this has been tested in our lab. Any suggestions for long-term storage of whole brains after collection? Is -80C the correct temperature? Will ice crystals spread at -20C? Will the receptor affinity decrease or protein degrade more rapidly at -20C? Any feedback would be greatly appreciated. Thanks again, Kelly From Kemlo.Rogerson <@t> elht.nhs.uk Thu Feb 17 11:21:52 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Gunk in stainer drain line[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EFD2@bhrv-nt-11.bhrv.nwest.nhs.uk> I'm afraid I don't. What does interest me and was my point, is where do the fungal spores come from? The air or the water? I think it is probably the water, hence the London connection. If you want to reduce the Gunk then deal with the source, the water, not the outcome, the Gunk. Depending of the severity of the problem a 'sterile' source for the water may be the solution (forgive the pun). -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 February 2005 15:38 To: Kemlo Rogerson; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line[Scanned] Kemlo, You believe that (about the breast fat)? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 February 2005 14:17 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line[Scanned] When we had these problems we asked the Company about how to solve it. They asked if we stained breast FNAC's and we did. They suggested it was the fat from said aspirates; I had always thought it was fungus. We adopted a protocol of bleach and boiling water and that cured it, but gassed the BMS's. We had a similar problem in London too, but that was really due to the sewerage they laughingly call 'drinking water'. The solution was to connect the stainer to piped 'clean' water; but then no-one drinks the water in London they all use chillers. Doesn't explain why the beer 'London pride' is so good, or does it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Poteete, Jacquie A. [mailto:japoteete@saintfrancis.com] Sent: 17 February 2005 13:35 To: CrochiereSteve@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line[Scanned] The "blue goo" in the stainers in our laboratory was found to be a variety of slime mold. I can't remember its' scientific name, but we had to use bleach to get rid of it, and the bleach did not prevent it from recurring. Pouring bleach down the drain became part of our cleaning-maintenance procedure. Good luck! Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com -----Original Message----- From: CrochiereSteve@aol.com [mailto:CrochiereSteve@aol.com] Sent: Wednesday, February 16, 2005 9:17 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Gunk in stainer drain line Does anyone have a problem with gunk clogging up the drain line on their autostainer. I have a Varistain Gemini, and the line has clogged up with blue goo twice in the past 4 years. What could be the cause and how would I prevent it, short of periodically taking the thing apart and reaming out the hose? Pouring bleach down the drain was suggested, but didn't keep the problem from recurring. Any suggestions? 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 ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@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 cgin <@t> pen.eiu.edu Thu Feb 17 11:31:05 2005 From: cgin <@t> pen.eiu.edu (ikenwosu) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: service contract Message-ID: <4214d4d9.123.1d23.13454@pen.eiu.edu> Hi, I just wanted to thank everyone who gave me leads as where to find a good service contractor. The gesture was very helpful. Ike Thedore Nwosu Grad. Student Dept. of Biological Sciences Eastern Illinois University 600 Linclon Ave Charleston, IL 61938 From Rcartun <@t> harthosp.org Thu Feb 17 12:23:34 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: B5 fixed tissue Message-ID: Dear Melanie & Histonet: I work with 5 hematopathologists who I have a great deal of respect for and I can tell you that our service is "First Class" all the way. We eliminated B5 about 1.5 years ago and, yes, they were very concerned about the effect it would have on morphology. We tried several of the B5 substitutes, but eventually we ended up fixing all bone marrows in formalin. Their "only" complaint is that the CD15 (Leu-M1) immunoperoxidase stain is weaker on formalin-fixed tissue than with B5. I can also tell you that immunoreactivity for a wide range of proteins is stronger and more reproducible with formalin than with B5. I see no reason to continue using B5. Hopefully, it will be eliminated from all laboratories in the near future. 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 >>> "Trivett, Melanie" 02/16/05 05:51PM >>> we do for our bone marrow trephines. I would love to eliminate it for handling problems, disposal issues and IHC consistency. Our haematologist will not hear of it. I would love to hear any good arguments that you have in addition! He still thinks there is nothing as good as B5 for the morphology of the trephines. Melanie -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Thursday, 17 February 2005 8:00 AM To: histonet@lists.utsouthwestern.edu; sluhisto@yahoo.com Subject: Re: [Histonet] cd5 and cd23 on B5 fixed tissue How many labs still use B5? We should work towards eliminating it from our environment. 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 >>> Histology SLU 02/16/05 02:23PM >>> Hello All: Is anyone doing these two antibodies on B5 fixed tissue with good results? If you are, and would be willing to share your secrets, any info would be appreciated. Both of the vendors and a couple of others have not used these antibodies with B5 fixation as part of their QC procedures and can not help. Thanks, in advance. Susan --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@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 brian.dias <@t> mail.utexas.edu Thu Feb 17 12:44:50 2005 From: brian.dias <@t> mail.utexas.edu (Brian Dias) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Problems with secondary antibodies Message-ID: <5.2.0.9.2.20050217123836.03030d78@mail.utexas.edu> hi everyone, i'm trying to get a double label ICC to work using a Rbt polyclonal (against Glucocorticoid receptor) and a Ms monoclonal (against Tyrosine hydroxylase) primary combination. both of these work in single label runs using Biotinylated secondaries. however, when i switch to double fluorescent runs i run into the following problems: 1. my anti-Rbt FITC doesn't light up any cells at all (in single as well as double studies)(my anti-Ms Tx Red works just fine and i see some nice staining in both my single and double studies) 2. i thought this might be secondary problem, but when i use another Ployclonal Rbt (towards Tyrosine hydroxylase) and anti-Rbt FITC, i see some nice staining comparable to when i use the mouse monoclonal. any suggestions would be welcome. thanks a ton b ____________________________________________________________________________ Brian George Dias Graduate Student (Crews lab) Office Phone #: 512-475-6738 The University of Texas at Austin E-mail: brian.dias@mail.utexas.edu Institute for Neuroscience Website: https://webspace.utexas.edu/bgd85/b.htm Patterson 56 1 University Station Stop C0930 Austin, TX 78712 ____________________________________________________________________________ From dcrippen <@t> buckinstitute.org Thu Feb 17 13:03:08 2005 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Sudan Black B Message-ID: Hi all, I'm hoping to find some help with the protocol for this lipofuscin stain. It instructs to "filter through filter paper and then filter again through a frittered glass filter of medium porosity with suction". Can anyone define "frittered glass filter" for me?? I have my guesses, but I prefer to consult the experts first. Many thanks in advance!! Danielle Crippen From csawrenk <@t> bccancer.bc.ca Thu Feb 17 13:09:42 2005 From: csawrenk <@t> bccancer.bc.ca (Sawrenko, Christina) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] p63/SMHC myosin cocktail for breast Message-ID: <57AD92D1EDD1854BAAC02DD7DDAF0E3601700558@srvex10.phsabc.ehcnet.ca> Good morning all, Does anyone use a cocktail of p63 and smooth muscle heavy chain (SMHC) myosin for breast? One of our pathologists is interested in trying it out. Thanks as always for any help you can provide. Chris Sawrenko Histopathology BC Cancer Agency - VCC Vancouver BC Canada From TillRenee <@t> uams.edu Thu Feb 17 13:22:57 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Oil Red O Message-ID: Hello. We have been doing some Oil Red O stains on cryosections of aorta and have been having problems with the hematoxylin leaching out during mounting. We have tried several different aqueous mounting medias and none have corrected the problem. Could it be the tissue or maybe the type of hematoxylin? Another lab has been doing the stain on liver and hasn't had any problems. Thanks, Renee' Till Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From llewllew <@t> shaw.ca Thu Feb 17 13:32:30 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Sudan Black B References: Message-ID: <000601c51527$6c1b3f40$58034246@yourlk4rlmsu> They are also called "sintered glass filters". They look like a large filter funnel with a rough glass plate permanently bonded just above the constriction. The rough glass plate is made of glass fibres or particles partially melted together to give the desired pore size. We used to use them for filtering our diluent for white cell counts on early model Coulter cell counters (in 1962 when I was a student in hematology). They were used because they could be put under vacuum without distorting and large volumes of the fluid could be filtered fast and efficiently. Bryan Llewellyn ----- Original Message ----- From: "Danielle Crippen" To: Sent: Thursday, February 17, 2005 11:03 AM Subject: [Histonet] Sudan Black B Hi all, I'm hoping to find some help with the protocol for this lipofuscin stain. It instructs to "filter through filter paper and then filter again through a frittered glass filter of medium porosity with suction". Can anyone define "frittered glass filter" for me?? I have my guesses, but I prefer to consult the experts first. Many thanks in advance!! Danielle Crippen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From schnegelsberg <@t> xgene.com Thu Feb 17 12:00:55 2005 From: schnegelsberg <@t> xgene.com (Birthe Schnegelsberg) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] sterile 5mm dermal punch supplier In-Reply-To: <000601c51527$6c1b3f40$58034246@yourlk4rlmsu> Message-ID: Hi. I am looking for sterile 5 mm dermal punches to retrieve tissue grown in 96 well inserts. Can anybody recommend a supplier for this? Thank, Birthe From gu.lang <@t> gmx.at Thu Feb 17 14:12:59 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] formaldehyd concentration and IHC Message-ID: In addition to the fixation-time-discussion a question about the concentration. What formaldehyd concentration do you prefer to get the optimal IHC- result? 4% or higher?; or does it really matter? Gudrun Lang From jkiernan <@t> uwo.ca Thu Feb 17 14:22:19 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Oil Red O References: Message-ID: <4214FCFB.D60F9AF1@uwo.ca> Extraction of haemalum must mean that the aqueous medium is acidic. The acidity might be there initially (it's a good thing for some stains). It may come from atmospheric CO2 (distilled water usually has pH=5 for this reason), or it may be due to aluminium and sulphate ions coming from the stained section. (All haemalum solutions contain a large excess of aluminium sulphate over & above the amount that complexes with haematein; this is necessary for the stain to work properly.) Suggested remedy: After counterstaining, blueing and washing, rinse the sections in a slightly alkaline buffer. (Phosphate, pH 7.4 should be OK.) Then shake off excess buffer and apply the aqueous mountant and coverslip. -- ------------------------------- 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 _______________________________ "Till, Renee" wrote: > > Hello. We have been doing some Oil Red O stains on cryosections of aorta > and have been having problems with the hematoxylin leaching out during > mounting. We have tried several different aqueous mounting medias and > none have corrected the problem. Could it be the tissue or maybe the > type of hematoxylin? Another lab has been doing the stain on liver and > hasn't had any problems. From sluhisto <@t> yahoo.com Thu Feb 17 14:26:46 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] DAB disposal Message-ID: <20050217202646.72601.qmail@web51004.mail.yahoo.com> Hello All: How is everyone disposing of their waste DAB from their automated IHC stainers? Is there a solution to neutralize DAB? Has anyone encountered defending their method of disposition to a CAP inspector? Thanks for any and all replies, in advance. Susan --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From lindas <@t> awesomenet.net Thu Feb 17 14:34:12 2005 From: lindas <@t> awesomenet.net (Linda Davis) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Cell blocks Message-ID: <005b01c51530$0a49cf30$e70ac942@D6JLZ851> Greetings all, Our pathologists have started complaining that our cell blocks are not adequate. I would like to know the method the majority of you prefer to obtain decent cell blocks. Thanks. Linda Davis, H.T. (ASCP), B.S. Rio Grande Regional Hospital McAllen, TX. -------------- next part -------------- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.300 / Virus Database: 265.8.8 - Release Date: 2/14/2005 From liz <@t> premierlab.com Thu Feb 17 14:51:06 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] sterile 5mm dermal punch supplier In-Reply-To: Message-ID: <000401c51532$66ab2f10$76d48a80@AMY> Mercedes Medical (800) 331-2716 or VWR should have them. They are made by Miltex (Dermal Biospy Punch). We use the 8 mm punches for our tissue constructs, and I have seen 6 mm punches, I'm sure they have 5mm punches. 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 Birthe Schnegelsberg Sent: Thursday, February 17, 2005 11:01 AM To: Histonet Subject: [Histonet] sterile 5mm dermal punch supplier Hi. I am looking for sterile 5 mm dermal punches to retrieve tissue grown in 96 well inserts. Can anybody recommend a supplier for this? Thank, Birthe _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ja.mitchell <@t> hosp.wisc.edu Thu Feb 17 14:57:39 2005 From: ja.mitchell <@t> hosp.wisc.edu (Mitchell Jean A.) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] sterile 5mm dermal punch supplier Message-ID: <583D3E9A1E843445BD54E461D1A2F6F31108FAF1@uwhis-xchng2.hosp.wisc.edu> Premier Products Co., 1-888-670-6100 www.premusa.com/medical/dermatology.asp is where I order my sterile dermal punches from. Jean Mitchell University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Birthe Schnegelsberg Sent: Thursday, February 17, 2005 12:01 PM To: Histonet Subject: [Histonet] sterile 5mm dermal punch supplier Hi. I am looking for sterile 5 mm dermal punches to retrieve tissue grown in 96 well inserts. Can anybody recommend a supplier for this? Thank, Birthe _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Feb 17 14:33:28 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] antigen retrieval question Message-ID: <200502172033.j1HKXPFF023793@chip.viawest.net> I have heard that it is not a good idea to do HIER one day and do the IHC the next? Is this so? I heard that the unmasking could actually remask if left sitting a long time before doing the staining. I have a protocol that requires a 3 hr. water bath incubation for AR and it would be more convienent if I could do that one day and do the rest of the procedure the next. Patsy From tissuearray <@t> hotmail.com Thu Feb 17 17:10:55 2005 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] sterile 5mm dermal punch supplier In-Reply-To: Message-ID: Hey Birthe, The link below is for tissue arrays but the information might be helpful for ordering th dermal punch needles. I don't know if they have 5mm but you could look. Go to the middle of the page there are addresses and such on the dermal needles. [1]http://www.arrayworkshop.com/Manual.html > >Hi. >I am looking for sterile 5 mm dermal punches to retrieve tissue grown in 96 >well inserts. >Can anybody recommend a supplier for this? >Thank, Birthe > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. http://www.arrayworkshop.com/Manual.html From jstaruk <@t> masshistology.com Thu Feb 17 17:23:52 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] antigen retrieval question In-Reply-To: <200502172033.j1HKXPFF023793@chip.viawest.net> Message-ID: <20050217232351.IDIO27788.out007.verizon.net@FrontOffice> We tried that once and the control slide (along with the research slides) were completely negative. Not sure if it was a fluke, but we never tried it again! Now everything is done on the same day. Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Thursday, February 17, 2005 3:33 PM To: ihcrg@neo.agsci.colostate.edu Cc: histonet@pathology.swmed.edu Subject: [Histonet] antigen retrieval question I have heard that it is not a good idea to do HIER one day and do the IHC the next? Is this so? I heard that the unmasking could actually remask if left sitting a long time before doing the staining. I have a protocol that requires a 3 hr. water bath incubation for AR and it would be more convienent if I could do that one day and do the rest of the procedure the next. Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MichelM9 <@t> chw.edu.au Thu Feb 17 19:39:22 2005 From: MichelM9 <@t> chw.edu.au (Michelle McDonald) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Histomorphometry image analysis system Message-ID: <1CF2E2E5BB36D5119E7A0008C791F374153F1518@simba.kids> Dear all, A colleague of mine has recently lost his decade old image analysis system which he used to perform histomorphometry on bone sections. He is now in the market for a new analysis system. I was wondering if anyone out there has any recommendations for systems he should consider. The work performed is specifically on bone sections so any advice from people with experience in bone histomorphometry (MAR,BFR,BV/TV etc)would be greatly appreciated. Thank you in advance Michelle Michelle McDonald B.MedSci PhD Student, Research Assistant Orthopaedic Research Dept. The Children's Hospital Westmead. Westmead NSW 2145 Australia Ph. +612 9845 3087 Fax. +612 9845 3078 ********************************************************************** 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 rockbeki <@t> ufl.edu Thu Feb 17 22:59:26 2005 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Aqueous mountant Message-ID: <806632105.1108702766744.JavaMail.osg@osgjas04.cns.ufl.edu> I've been using Crystalmount with AEC and I have had good results with it. It sets hard and has a refractive index close to glass, so technically you don't have to put a coverslip on it, but I find that it improves the resolution to postmount with permount and coverslip, after the crystalmount has dried. -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From deborah.mills <@t> syngenta.com Fri Feb 18 02:21:43 2005 From: deborah.mills <@t> syngenta.com (deborah.mills@syngenta.com) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Cresyl Violet counterstain Message-ID: <7D28784DD2EBD511AC4900508B6219C1056D2FC6@ukapmbx01.ukct.zeneca.com> I am attempting to determine dopaminergic cell loss in the SNpc of mouse brains using a tyrosine hydroxylase antibody. The sections have to be counterstained with cresyl violet to enable me to count CV+ / TH- neurons in this region. However, I am having trouble achieving a good enough contrast between my TH staining (DAB visualisation) and the CV stain to allow me to count the CV neurons. The CV doesn't appear to be staining up the neurons very well. Is there anything I can do to improve the intensity of my staining and be able to pick out the neurons more easily? I have tried increasing the length of time the sections are in CV solution for but all this seems to do is increase the background staining without increasing neuronal staining. I have also tried altering the differentiation steps to no avail, From Kemlo.Rogerson <@t> elht.nhs.uk Fri Feb 18 02:32:50 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Re: Gunk in stainer drain line[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EFD4@bhrv-nt-11.bhrv.nwest.nhs.uk> PS. I keep fish in a pond outside. In Spring you get a sticky algae/ gunk that lasts for a few weeks. It's caused by lack of denitrifying bacteria in the filter (they die over Winter), protein in the water (fish poo and fish food) and increased light levels and a increasing water temperature (over 10 degrees C). To eradicate you put compacted barley bales into the water which decompose to produce an organic acid that kills the gunk. You can also buy Propriety water treatments from the Fish Shop but they only give temporary relief; once you get balance with a good culture of bacteria in your filter and sufficient plant growth to remove the nitrates that accumulate from the breakdown of ammonia by those bacteria, the problem resolves spontaneously. Does that help? -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 February 2005 15:38 To: Kemlo Rogerson; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line[Scanned] Kemlo, You believe that (about the breast fat)? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 February 2005 14:17 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line[Scanned] When we had these problems we asked the Company about how to solve it. They asked if we stained breast FNAC's and we did. They suggested it was the fat from said aspirates; I had always thought it was fungus. We adopted a protocol of bleach and boiling water and that cured it, but gassed the BMS's. We had a similar problem in London too, but that was really due to the sewerage they laughingly call 'drinking water'. The solution was to connect the stainer to piped 'clean' water; but then no-one drinks the water in London they all use chillers. Doesn't explain why the beer 'London pride' is so good, or does it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Poteete, Jacquie A. [mailto:japoteete@saintfrancis.com] Sent: 17 February 2005 13:35 To: CrochiereSteve@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line[Scanned] The "blue goo" in the stainers in our laboratory was found to be a variety of slime mold. I can't remember its' scientific name, but we had to use bleach to get rid of it, and the bleach did not prevent it from recurring. Pouring bleach down the drain became part of our cleaning-maintenance procedure. Good luck! Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com -----Original Message----- From: CrochiereSteve@aol.com [mailto:CrochiereSteve@aol.com] Sent: Wednesday, February 16, 2005 9:17 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Gunk in stainer drain line Does anyone have a problem with gunk clogging up the drain line on their autostainer. I have a Varistain Gemini, and the line has clogged up with blue goo twice in the past 4 years. What could be the cause and how would I prevent it, short of periodically taking the thing apart and reaming out the hose? Pouring bleach down the drain was suggested, but didn't keep the problem from recurring. Any suggestions? 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 ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@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 deborah.mills <@t> syngenta.com Fri Feb 18 02:42:47 2005 From: deborah.mills <@t> syngenta.com (deborah.mills@syngenta.com) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] Marking of mouse brain tissue Message-ID: <7D28784DD2EBD511AC4900508B6219C1056D2FC7@ukapmbx01.ukct.zeneca.com> I am sectioning Para formaldehyde fixed, sucrose cryoprotected mouse brain tissue on the microtome to then perform free floating tyrosine hydroxylase IHC with a cresyl violet counterstain to count CV+/TH- neurons in the SNpc region. Before sectioning I need to mark the tissue so I can load them all onto slides in the same orientation. I have tried puncturing the tissue with various sizes of needle and have found it difficult to get a hole large enough to be visible to the eye yet small enough to avoid the SNpc and avoid the sections breaking apart during the staining process. Is there a dye I could puncture with that wouldn't spread to the rest of the tissue or interfere with my IHC? Any other suggestions? Many thanks in advance. From c.m.vanderloos <@t> amc.uva.nl Fri Feb 18 03:35:24 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] RE: Frozen storage of tissue and sections, what is the correct temperature? Message-ID: <128dfb71289170.1289170128dfb7@amc.uva.nl> Dear Kelly, We always store our (unfixed) cryosections at -80C. IHC staining is successful up to years of storage in this way. In the past we used to store at -20C and this resulted into a loss of specific epitopes like CD25, CD4, CD3 in just a matter of weeks. Don't do this! If you have dried your specimens completely (as you did for a few hours at room temperature) there is no chance that ice-crystals influence the tissue morphology, simply because there is no water left! ----- Original Message ----- From Kelly D Mcqueeney Date Thu, 17 Feb 2005 11:52:36 -0500 To histonet@lists.utsouthwestern.edu Subject [Histonet] Frozen storage of tissue and sections, what is the correct temperature? I know that frozen tissue storage is a common topic, but I have a hard time finding the right answer throughout the histonet archives. We are currently coating brain in embedding medium and snap freezing brains in isopentane followed by immediate storage at -80C. Then, we bring the brain up to -20C a few hours before sectioning. After sectioning, we dry the slides at RT for several hours and store at -80C until using the tissue for ligand binding, IHC or insitu. This exact protocol has been moderately successful, so far so good. But, some of the staff store frozen brains at -20C. They feel that keeping the tissue at cryostat temperature reduces the amount of freeze artifact. I was always taught to freeze fresh frozen brain at a much lower temperature. None of this has been tested in our lab. Any suggestions for long-term storage of whole brains after collection? Is -80C the correct ! temperatu From marytedo <@t> hotmail.com Fri Feb 18 04:04:56 2005 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] formaldehyd concentration and IHC In-Reply-To: Message-ID: If I am not in a mistake, I think you can use Formol Buffer (p.H 7.4-7.6) to IHC. I guess... [i.p.emcrook.gif] MARIA T. DOMINGUEZ ANATOMIC PATHOLOGY SERV. RIO GRANDE'S REG.HOSPITAL RIO GRANDE, TIERRA DEL FUEGO, ARGENTINA. >From: "Gudrun Lang" >Reply-To: gu.lang@gmx.at >To: "Histonetliste (Histonetliste)" >Subject: [Histonet] formaldehyd concentration and IHC >Date: Thu, 17 Feb 2005 21:12:59 +0100 > >In addition to the fixation-time-discussion a question about the >concentration. What formaldehyd concentration do you prefer to get the >optimal IHC- result? > >4% or higher?; or does it really matter? > > > >Gudrun Lang > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar [1]MSN Toolbar Get it now! References 1. http://g.msn.com/8HMBEN/2752??PS=47575 From bhewlett <@t> cogeco.ca Fri Feb 18 06:41:18 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] formaldehyd concentration and IHC References: <20050217202055.B2BD322FA@fep5.cogeco.net> Message-ID: <001401c515b7$24b690b0$6500a8c0@mainbox> Gudrun, A minimum fixation time of 24 hours, in the commonly used concentration of 4% w/v formaldehyde in phosphate buffer pH 7.2-7.4, has worked well in my hands for over 40 years of IHC, using hundreds of different antibodies. Bryan ----- Original Message ----- From: "Gudrun Lang" To: "Histonetliste (Histonetliste)" Sent: Thursday, February 17, 2005 3:12 PM Subject: [Histonet] formaldehyd concentration and IHC > In addition to the fixation-time-discussion a question about the > concentration. What formaldehyd concentration do you prefer to get the > optimal IHC- result? > > 4% or higher?; or does it really matter? > > > > Gudrun Lang > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From kmerriam2003 <@t> yahoo.com Fri Feb 18 06:42:15 2005 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] antigen retrieval question In-Reply-To: <20050217232351.IDIO27788.out007.verizon.net@FrontOffice> Message-ID: <20050218124216.16307.qmail@web52508.mail.yahoo.com> Patsy, I have done this many times before, and have never had any issues with re-masking. Why would the sites be re-masked if they are not subjected to formalin again? If I am staining a big batch of slides (anything over 100 slides at a time), I do the depar and retrieval on the first day and all of the antibody stuff on the second day. Maybe it just depends on the protein/antibody? This has peaked my interest. I am curious to see if anyone else has encountered this problem. Kim Merriam Novartis Cambridge, MA Jim Staruk wrote: We tried that once and the control slide (along with the research slides) were completely negative. Not sure if it was a fluke, but we never tried it again! Now everything is done on the same day. Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Thursday, February 17, 2005 3:33 PM To: ihcrg@neo.agsci.colostate.edu Cc: histonet@pathology.swmed.edu Subject: [Histonet] antigen retrieval question I have heard that it is not a good idea to do HIER one day and do the IHC the next? Is this so? I heard that the unmasking could actually remask if left sitting a long time before doing the staining. I have a protocol that requires a 3 hr. water bath incubation for AR and it would be more convienent if I could do that one day and do the rest of the procedure the next. 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 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From JQB7 <@t> CDC.GOV Fri Feb 18 06:33:30 2005 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] re-embedding blocks Message-ID: Hi everybody: Just curious as to what oven temperature is generally used for melting down paraffin blocks for re-embedding. Is there a temperature that is generally considered too high either for the paraffin or for the tissue? Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 From TMcNemar <@t> lmhealth.org Fri Feb 18 07:48:49 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] DAB disposal Message-ID: <6CD94D97ED7D924BA5C2B588FA95282139678A@nt_exchange.lmhealth.org> We have ours hauled away. We drain directly into a 20 gallon carboy. They have to come and get it about every 3-4 months. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Histology SLU [mailto:sluhisto@yahoo.com] Sent: Thursday, February 17, 2005 3:27 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] DAB disposal Hello All: How is everyone disposing of their waste DAB from their automated IHC stainers? Is there a solution to neutralize DAB? Has anyone encountered defending their method of disposition to a CAP inspector? Thanks for any and all replies, in advance. Susan --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kelly.mcqueeney <@t> bms.com Fri Feb 18 07:47:11 2005 From: kelly.mcqueeney <@t> bms.com (Kelly D Mcqueeney) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] antigen retrieval question In-Reply-To: <20050217232351.IDIO27788.out007.verizon.net@FrontOffice> References: <20050217232351.IDIO27788.out007.verizon.net@FrontOffice> Message-ID: <4215F1DF.10101@bms.com> I have never seen a problem when using antibodies that require an overnight incubation. Jim Staruk wrote: >We tried that once and the control slide (along with the research slides) >were completely negative. Not sure if it was a fluke, but we never tried it >again! Now everything is done on the same day. > >Jim > >______________________ > Jim Staruk >Mass Histology Service >www.masshistology.com > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg >Sent: Thursday, February 17, 2005 3:33 PM >To: ihcrg@neo.agsci.colostate.edu >Cc: histonet@pathology.swmed.edu >Subject: [Histonet] antigen retrieval question > >I have heard that it is not a good idea to do HIER one day and do the IHC >the next? Is this so? I heard that the unmasking could actually remask if >left sitting a long time before doing the staining. I have a protocol that >requires a 3 hr. water bath incubation for AR and it would be more >convienent if I could do that one day and do the rest of the procedure the >next. >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 TMcNemar <@t> lmhealth.org Fri Feb 18 07:56:29 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:38 2005 Subject: [Histonet] re-embedding blocks Message-ID: <6CD94D97ED7D924BA5C2B588FA95282139678B@nt_exchange.lmhealth.org> We just use the embedding station. Set them on the hot plate or set them into the molten paraffin. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Bartlett, Jeanine [mailto:JQB7@CDC.GOV] Sent: Friday, February 18, 2005 7:34 AM To: Histonet Subject: [Histonet] re-embedding blocks Hi everybody: Just curious as to what oven temperature is generally used for melting down paraffin blocks for re-embedding. Is there a temperature that is generally considered too high either for the paraffin or for the tissue? Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Fri Feb 18 12:00:30 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Marking of mouse brain tissue In-Reply-To: <7D28784DD2EBD511AC4900508B6219C1056D2FC7@ukapmbx01.ukct.zeneca.com> References: <7D28784DD2EBD511AC4900508B6219C1056D2FC7@ukapmbx01.ukct.zeneca.com> Message-ID: <42162D3E.8080209@umdnj.edu> Hi Deborah: Hmmm ... sounds like an MPTP/neuroprotection experiment. :-) What I do is notch the cortex (I cut a V out of the cortex with a razor blade) on the (animal's) right side. This works for me but if you cut far into the brain stem the cortex may separate during staining and then you have to match sections, not fun. You could also use a 25 gauge hypo needle to punch a hole on the right side in the region of the superior colliculus, or far enough dorsal to avoid the SNpc. I have found that examining free-floating sections against a black background allows one to see a great deal of detail. Perhaps a bit of india ink in the hypo might work? The clinical lab people on this list use india ink or a commercially available indelible marking pen to mark tissue so they can orient it for sectioning, that sounds like a good thing to try. I'm sure someone will provide the name of that device. "Sharpie" ink might work but it is soluble in alcohols and xylene. Have fun counting all those cells! Geoff deborah.mills@syngenta.com wrote: >I am sectioning Para formaldehyde fixed, sucrose cryoprotected mouse brain >tissue on the microtome to then perform free floating tyrosine hydroxylase >IHC with a cresyl violet counterstain to count CV+/TH- neurons in the SNpc >region. Before sectioning I need to mark the tissue so I can load them all >onto slides in the same orientation. I have tried puncturing the tissue >with various sizes of needle and have found it difficult to get a hole large >enough to be visible to the eye yet small enough to avoid the SNpc and avoid >the sections breaking apart during the staining process. Is there a dye I >could puncture with that wouldn't spread to the rest of the tissue or >interfere with my IHC? Any other suggestions? > >Many thanks in advance. > >_______________________________________________ >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 GDawson <@t> dynacaremilwaukee.com Fri Feb 18 08:17:41 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] antigen retrieval question Message-ID: Patsy, In my experience, staining quality decreases as the time between HIER and staining increases. I believe the epitopes you retrieve with heat do not stay open indefinitely and will, in fact, begin to slowly close very shortly after HIER. So, for the best staining, I would recommend no wait time at all between retrieval and staining. Do you have a tech that comes in earlier than you? Perhaps they could begin the 3 hr. water bath incubation for you before you get in so you don't have to wait around. I have done this myself and it works well if the early tech is available. Good Luck, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Thursday, February 17, 2005 2:33 PM To: ihcrg@neo.agsci.colostate.edu Cc: histonet@pathology.swmed.edu Subject: [Histonet] antigen retrieval question I have heard that it is not a good idea to do HIER one day and do the IHC the next? Is this so? I heard that the unmasking could actually remask if left sitting a long time before doing the staining. I have a protocol that requires a 3 hr. water bath incubation for AR and it would be more convienent if I could do that one day and do the rest of the procedure the next. Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jstaruk <@t> masshistology.com Fri Feb 18 08:39:56 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] antigen retrieval question In-Reply-To: <20050218124216.16307.qmail@web52508.mail.yahoo.com> Message-ID: <0IC400L583EDOR30@vms040.mailsrvcs.net> If I remember correctly, mine was an anti-collagen I or II that did not work after sitting in buffer overnight. I also know that one of my part-timers tried this at the hospital he works at and they also had disastrous results on day #2. ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Merriam Sent: Friday, February 18, 2005 7:42 AM To: Jim Staruk; 'Patsy Ruegg'; ihcrg@neo.agsci.colostate.edu Cc: histonet@pathology.swmed.edu Subject: RE: [Histonet] antigen retrieval question Patsy, I have done this many times before, and have never had any issues with re-masking. Why would the sites be re-masked if they are not subjected to formalin again? If I am staining a big batch of slides (anything over 100 slides at a time), I do the depar and retrieval on the first day and all of the antibody stuff on the second day. Maybe it just depends on the protein/antibody? This has peaked my interest. I am curious to see if anyone else has encountered this problem. Kim Merriam Novartis Cambridge, MA From bhewlett <@t> cogeco.ca Fri Feb 18 09:48:33 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] antigen retrieval question References: Message-ID: <003101c515d1$4db5b7b0$6500a8c0@mainbox> Patsy, I have often placed sections in hypotonic TBS pH7.6, after the cool down period, left them for a couple of hours at RT and also overnight, even over the weekend at 4C, without deleterious effects on any of the markers I have used. I have heard of people having loss of staining following such delays however, I have never seen it. In fact, prolonged (O/N to 48 hour) soaking in hypotonic TBS was a very common way of unmasking antigens for IHC prior to the days of HIER! In any case, should such loss of staining occur surely the positive tissue control would reveal it. I think that it's worth a test prior to committing your entire set of slides. Regards, Bryan P.S. did you get the pictures OK? ----- Original Message ----- From: "Dawson, Glen" Cc: Sent: Friday, February 18, 2005 9:17 AM Subject: RE: [Histonet] antigen retrieval question > Patsy, > > In my experience, staining quality decreases as the time between HIER and > staining increases. I believe the epitopes you retrieve with heat do not > stay open indefinitely and will, in fact, begin to slowly close very shortly > after HIER. So, for the best staining, I would recommend no wait time at > all between retrieval and staining. > > Do you have a tech that comes in earlier than you? Perhaps they could begin > the 3 hr. water bath incubation for you before you get in so you don't have > to wait around. I have done this myself and it works well if the early tech > is available. > > Good Luck, > > Glen Dawson BS, HT & QIHC (ASCP) > IHC Manager > Milwaukee, WI > > -----Original Message----- > From: Patsy Ruegg [mailto:pruegg@ihctech.net] > Sent: Thursday, February 17, 2005 2:33 PM > To: ihcrg@neo.agsci.colostate.edu > Cc: histonet@pathology.swmed.edu > Subject: [Histonet] antigen retrieval question > > > I have heard that it is not a good idea to do HIER one day and do the IHC > the next? Is this so? I heard that the unmasking could actually remask if > left sitting a long time before doing the staining. I have a protocol that > requires a 3 hr. water bath incubation for AR and it would be more > convienent if I could do that one day and do the rest of the procedure the > next. > 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 vbaker60 <@t> yahoo.com Fri Feb 18 09:50:58 2005 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] antigen retrieval question In-Reply-To: <0IC400L583EDOR30@vms040.mailsrvcs.net> Message-ID: <20050218155058.17447.qmail@web50109.mail.yahoo.com> Patsy, I'm sort of coming in on the middle of this so I hope I don't make a major gaff here. When doing the long term HIER take your protocol up to the primary antibody. Apply the primary and cover the slides with strips of parafilm. Then incubate the slides overnight at 4ºC in a sealed humid chamber. Next day you can complete the balance of your protocol. I've done with with a variety of ab's on a variety of species. Hope this helps. Vikki Baker Mt. Sinai School of Medicine --- Jim Staruk wrote: > If I remember correctly, mine was an anti-collagen I > or II that did not work > after sitting in buffer overnight. I also know that > one of my part-timers > tried this at the hospital he works at and they also > had disastrous results > on day #2. > > ______________________ > Jim Staruk > Mass Histology Service > www.masshistology.com > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Kim Merriam > Sent: Friday, February 18, 2005 7:42 AM > To: Jim Staruk; 'Patsy Ruegg'; > ihcrg@neo.agsci.colostate.edu > Cc: histonet@pathology.swmed.edu > Subject: RE: [Histonet] antigen retrieval question > > Patsy, > > I have done this many times before, and have never > had any issues with > re-masking. Why would the sites be re-masked if > they are not subjected to > formalin again? If I am staining a big batch of > slides (anything over 100 > slides at a time), I do the depar and retrieval on > the first day and all of > the antibody stuff on the second day. Maybe it just > depends on the > protein/antibody? > > This has peaked my interest. I am curious to see if > anyone else has > encountered this problem. > > Kim Merriam > Novartis > Cambridge, MA > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. http://promotions.yahoo.com/new_mail From JQB7 <@t> CDC.GOV Fri Feb 18 10:16:10 2005 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] re-embedding blocks Message-ID: Thanks. I am usually dealing with large volumes of outside blocks embedded in who knows what kind of paraffin. -----Original Message----- From: Tom McNemar [mailto:TMcNemar@lmhealth.org] Sent: Friday, February 18, 2005 8:56 AM To: Bartlett, Jeanine; Histonet Subject: RE: [Histonet] re-embedding blocks We just use the embedding station. Set them on the hot plate or set them into the molten paraffin. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Bartlett, Jeanine [mailto:JQB7@CDC.GOV] Sent: Friday, February 18, 2005 7:34 AM To: Histonet Subject: [Histonet] re-embedding blocks Hi everybody: Just curious as to what oven temperature is generally used for melting down paraffin blocks for re-embedding. Is there a temperature that is generally considered too high either for the paraffin or for the tissue? Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Feb 18 10:58:22 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] re-embedding blocks In-Reply-To: <6CD94D97ED7D924BA5C2B588FA95282139678B@nt_exchange.lmhealth.org> Message-ID: <200502181658.j1IGwIFF010221@chip.viawest.net> I put mine in the paraffin bath on the embedding station in a mold to catch the tissue when it melts. It is set at 58-60 dc. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Friday, February 18, 2005 6:56 AM To: 'Bartlett, Jeanine'; Histonet Subject: RE: [Histonet] re-embedding blocks We just use the embedding station. Set them on the hot plate or set them into the molten paraffin. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Bartlett, Jeanine [mailto:JQB7@CDC.GOV] Sent: Friday, February 18, 2005 7:34 AM To: Histonet Subject: [Histonet] re-embedding blocks Hi everybody: Just curious as to what oven temperature is generally used for melting down paraffin blocks for re-embedding. Is there a temperature that is generally considered too high either for the paraffin or for the tissue? Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From am458 <@t> cornell.edu Fri Feb 18 11:17:30 2005 From: am458 <@t> cornell.edu (Amanda MacFarlane) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] IHC on sections from beta-gal stained tissues Message-ID: <2723.128.253.96.73.1108747050.squirrel@128.253.96.73> I currently am characterizing some transgenic mice by performing beta-gal staining on whole tissues. The procedure includes a step for gluteraldehyde fixation before staining. I freeze the tissues on isopentane and cryosection to determine the localization of the beta-gal stain. I was wondering if it's also possible to perform IHC on these sections and if so are there (general) protocols out there. Will antigen retrievel be necessary, etc.? Thanks in advance. Amanda Amanda MacFarlane Division of Nutritional Sciences Cornell University From Sue.Kapoor <@t> uhsi.org Fri Feb 18 11:47:53 2005 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] eosin and microwave tissue processors Message-ID: <61E9F2400F53D5119CFC00508B44E33B02E9E067@khmcexch.uhsi.org> Hi all, Does anyone using a microwave tissue processor use Eosin in their alcohol? Or know of a reason why it shouldn't be used? We put eosin in our alcohol on our Tissue-Tek and have grown accustomed to our bxs being a nice pink. We've just gotten our first Energy Beam microwave tissue processor and have found it hard to embed "naked" bxs. Thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From kelly.mcqueeney <@t> bms.com Fri Feb 18 13:54:16 2005 From: kelly.mcqueeney <@t> bms.com (Kelly D Mcqueeney) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Dry paraformaldehyde vapor Message-ID: <421647E8.9050604@bms.com> In order to avoid tritium contamination of screens, I have been fixing my slides with paraformaldehyde vapor. PF powder is placed in the bottom of a dessicator with slides. Air is removed from the dessicator and the air-tight chamber exposes the tissue to paraformaldehyde overnight. Has anyone used this method? I am having a hard time understanding why the powder alone is sufficient for fixation. Liquid fixative is not an option because the receptor/lingand complex is washed off. The protocol is vaguely outlined in Biotechiques 26:432-434; Liberatore, et. al. 1999. Thanks for your input, Kelly From lsb <@t> jax.org Fri Feb 18 14:06:29 2005 From: lsb <@t> jax.org (Lesley S. Bechtold) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] pathology databases Message-ID: <6.0.3.0.0.20050218150428.026980d0@aretha.jax.org> Hi Everyone, How are you managing your pathology data? Do you use an electronic database of some sort? Did you buy it or make it? Can you attach results from other labs like blood chemistry and pictures (X-rays, digital images of slides) ? I'd be especially interested in anyone who's managing data for animal (non-human) tissues. Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 From djamesnz <@t> orcon.net.nz Fri Feb 18 14:27:09 2005 From: djamesnz <@t> orcon.net.nz (Darren James) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Bone saw and bone protocols[Scanned] In-Reply-To: <6AB79F0BA7C4914BA8DD8F5E0C21B9752E2928@AHEXMAIL01.xalderhey.com> Message-ID: <200502182027.j1IKRJv9002940@dbmail-mx3.orcon.net.nz> Hi David, For a cheap bone saw I have heard of labs using a tile saw. These have diamond blades and are often water cooled. They are available at many hardware stores. Hope this helps Darren -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Muskett David Sent: 16 February 2005 21:10 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone saw and bone protocols[Scanned] Dear All I am currently reviewing our protocols for the processing of bone. Would anybody be willing to share protocols and processing schedules? I am also interested in buying a bone saw, preferably one which allows thin sections to be cut easily. Can anybody recommend me a firm or a saw? 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Feb 18 14:46:02 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Tungsten carbide knives Message-ID: <200502182045.j1IKjvFF023979@chip.viawest.net> Does anyone have a D profile tungsten carbide knife they could sell me? Patsy From mcauliff <@t> umdnj.edu Fri Feb 18 18:03:42 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Dry paraformaldehyde vapor In-Reply-To: <421647E8.9050604@bms.com> References: <421647E8.9050604@bms.com> Message-ID: <4216825E.2050301@umdnj.edu> Hot paraformaldehyd vapors were used to induce fluorescence of aminergic neurotransmitters in freeze-dried tissues. This method dates from the early 1960's. Osmium vapors have been used by protozoologists to fix drops of protozoa on coverslips for .... long before I became a biologist. One does not have to get one's nose too close to a bottle of paraformaldehyde to realize that the vapors are very noxious. I think vapor fixation has been used to preserve secretions of the respiratory system but I can't come up with a reference off the top of my head. Geoff Kelly D Mcqueeney wrote: > In order to avoid tritium contamination of screens, I have been fixing > my slides with paraformaldehyde vapor. PF powder is placed in the > bottom of a dessicator with slides. Air is removed from the dessicator > and the air-tight chamber exposes the tissue to paraformaldehyde > overnight. Has anyone used this method? I am having a hard time > understanding why the powder alone is sufficient for fixation. Liquid > fixative is not an option because the receptor/lingand complex is > washed off. The protocol is vaguely outlined in Biotechiques > 26:432-434; Liberatore, et. al. 1999. > > Thanks for your input, > Kelly > > _______________________________________________ > 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 bhewlett <@t> cogeco.ca Fri Feb 18 15:33:18 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Dry paraformaldehyde vapor References: <421647E8.9050604@bms.com> Message-ID: <001601c51601$7678be10$6500a8c0@mainbox> Kelly, Vapour fixation, mainly with formaldehyde and osmium fixatives is a practice of long standing (1944?) in histology and cytology. It has also been applied to freeze dried tissues. Probably the most famous application was by Falck (1962) who described vapour fixation at elevated temperatures with formaldehyde and the subsequent conversion of catecholamines and 5-HT to fluorescent condensation products, i.e formaldehyde-induced fluorescence. Paraformaldehyde is a solid polymerized form of formaldehyde (polyoxymethylene). Under reduced pressure, and more readily with elevated temperature, it gases off as formaldehyde. Check 'Histochemistry: theoretical and applied' by the late A.G. Everson Pearse. The father of histochemistry. Bryan ----- Original Message ----- From: "Kelly D Mcqueeney" To: "Histonet" Sent: Friday, February 18, 2005 2:54 PM Subject: [Histonet] Dry paraformaldehyde vapor > In order to avoid tritium contamination of screens, I have been fixing > my slides with paraformaldehyde vapor. PF powder is placed in the bottom > of a dessicator with slides. Air is removed from the dessicator and the > air-tight chamber exposes the tissue to paraformaldehyde overnight. Has > anyone used this method? I am having a hard time understanding why the > powder alone is sufficient for fixation. Liquid fixative is not an > option because the receptor/lingand complex is washed off. The protocol > is vaguely outlined in Biotechiques 26:432-434; Liberatore, et. al. 1999. > > Thanks for your input, > Kelly > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From histophilhuff <@t> yahoo.com Fri Feb 18 16:10:09 2005 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] sterile 5mm dermal punch supplier In-Reply-To: Message-ID: <20050218221009.34552.qmail@web30809.mail.mud.yahoo.com> We get ours from Delasco (800 831 6273) http://www.delasco.com/pcat/1/Sharps/Fray%5FDisposable/dlmif016/ They have from 1.5 mm to 8 mm. Phil Thom Jensen wrote: Hey Birthe, The link below is for tissue arrays but the information might be helpful for ordering th dermal punch needles. I don't know if they have 5mm but you could look. Go to the middle of the page there are addresses and such on the dermal needles. [1]http://www.arrayworkshop.com/Manual.html > >Hi. >I am looking for sterile 5 mm dermal punches to retrieve tissue grown in 96 >well inserts. >Can anybody recommend a supplier for this? >Thank, Birthe > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. http://www.arrayworkshop.com/Manual.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? The all-new My Yahoo! – What will yours do? From northma <@t> ohsu.edu Fri Feb 18 17:45:04 2005 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Oil Red O Message-ID: I cover initially with Crystal Mount and let it dry at room temp. After drying, it can be dipped into xylene and coverslipped with resinous medium. Mary North OHSU Neuromuscular Lab Portland, OR >>> "Till, Renee" 02/17/05 11:22 AM >>> Hello. We have been doing some Oil Red O stains on cryosections of aorta and have been having problems with the hematoxylin leaching out during mounting. We have tried several different aqueous mounting medias and none have corrected the problem. Could it be the tissue or maybe the type of hematoxylin? Another lab has been doing the stain on liver and hasn't had any problems. Thanks, Renee' Till 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 SUSANLMCCOY <@t> aol.com Fri Feb 18 18:34:03 2005 From: SUSANLMCCOY <@t> aol.com (SUSANLMCCOY@aol.com) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Job Oppurtunity (Las Vegas, Nevada) Message-ID: <7B1E371D.635B627A.3C887B41@aol.com> F/T Histology Position Sunrise Hospital and Children's Hospital Medical Center 3186 Maryland Parkway Las Vegas, Nevada 89109 For Details Contact: Mary Ann Haynes 702 836-3971 (M-F) From djamesnz <@t> orcon.net.nz Sat Feb 19 04:18:54 2005 From: djamesnz <@t> orcon.net.nz (Darren James) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] DAB disposal In-Reply-To: <20050217202646.72601.qmail@web51004.mail.yahoo.com> Message-ID: <200502191019.j1JAJ7U0002663@dbmail-mx3.orcon.net.nz> Hi, I always thought addition of bleach neutralises DAB. Please correct me if I am wrong. Regards Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: 18 February 2005 09:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] DAB disposal Hello All: How is everyone disposing of their waste DAB from their automated IHC stainers? Is there a solution to neutralize DAB? Has anyone encountered defending their method of disposition to a CAP inspector? Thanks for any and all replies, in advance. Susan --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From djamesnz <@t> orcon.net.nz Sat Feb 19 04:32:43 2005 From: djamesnz <@t> orcon.net.nz (Darren James) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Re: Gunk in stainer drain line In-Reply-To: <000301c51503$e3bd3400$7f00a8c0@PMARCUM2K> Message-ID: <200502191032.j1JAWmFC007840@dbmail-mx3.orcon.net.nz> I agree. I have experienced this also and a weekly dose of bleach or an antibacterial agent seemed to work (hot water is good too) Regards Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pamela Marcum Sent: 18 February 2005 04:18 To: Poteete, Jacquie A.; CrochiereSteve@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Gunk in stainer drain line If it is a slime mold forming would a PM using hot water and soap with anti bacterial agent help or just using a true anti-bacterial (not Bleach) once a week? Just a thought. Thanks, Pam Marcum? > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Poteete, > Jacquie A. > Sent: Thursday, February 17, 2005 8:35 AM > To: 'CrochiereSteve@aol.com'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Re: Gunk in stainer drain line > > > The "blue goo" in the stainers in our laboratory was found to be a variety > of slime mold. I can't remember its' scientific name, but we had to use > bleach to get rid of it, and the bleach did not prevent it from recurring. > Pouring bleach down the drain became part of our cleaning-maintenance > procedure. Good luck! > > Jacquie Poteete MT(ASCP)QIHC > Lead Technologist, IHC Laboratory > Saint Francis Hospital, Tulsa, OK > japoteete@saintfrancis.com > > -----Original Message----- > From: CrochiereSteve@aol.com [mailto:CrochiereSteve@aol.com] > Sent: Wednesday, February 16, 2005 9:17 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Gunk in stainer drain line > > > Does anyone have a problem with gunk clogging up the drain line on their > autostainer. I have a Varistain Gemini, and the line has clogged up with > blue goo > twice in the past 4 years. What could be the cause and how would I prevent > it, > short of periodically taking the thing apart and reaming out the hose? > Pouring bleach down the drain was suggested, but didn't keep the problem > from > recurring. > Any suggestions? > > 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 > > > > > ********* Email Confidentiality Statement ********* > Visit http://www.saintfrancis.com/emailconf.asp > > _______________________________________________ > Histonet mailing list > Histonet@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 skouko <@t> po-box.mcgill.ca Sat Feb 19 12:27:18 2005 From: skouko <@t> po-box.mcgill.ca (skouko@po-box.mcgill.ca) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] DAB Disposal Message-ID: <1108837638.42178506bd194@www.webmail.mcgill.ca> You are right. We do the same thing in our lab to neutralize it. We pour all the DAB solution in a beaker full of bleach and rinse the flasks and all "DAB-tainted glassware" with bleach as well. Cheers! Sophia Koukoui Behavioral Neuroscience Chaudhuri Vision Laboratory Hi, I always thought addition of bleach neutralises DAB. Please correct me if I am wrong. Regards Darren James From Histopatty <@t> aol.com Sat Feb 19 15:15:00 2005 From: Histopatty <@t> aol.com (Histopatty@aol.com) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Plant histology Message-ID: <193.39d577e1.2f490654@aol.com> Dose any one on the list have any knowledge or experience in plant histology. Specifically I am trying to demonstrate mitosis in onion roots. The slides I have access to are faded and the details are not crisp enough for photography. Any help on processing protocols and staining techniques would be appreciated. Patty.Eneff@hcahealthcare.com or Histopatty@aol.com From bhewlett <@t> cogeco.ca Sat Feb 19 16:36:09 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Plant histology References: <193.39d577e1.2f490654@aol.com> Message-ID: <099801c516d3$697d5860$6500a8c0@mainbox> Patty, I can help, but are you trying to restore the faded stains or wishing to make new preparations? Bryan ----- Original Message ----- From: To: Sent: Saturday, February 19, 2005 4:15 PM Subject: [Histonet] Plant histology > Dose any one on the list have any knowledge or experience in plant histology. > Specifically I am trying to demonstrate mitosis in onion roots. The slides > I have access to are faded and the details are not crisp enough for > photography. Any help on processing protocols and staining techniques would be > appreciated. > Patty.Eneff@hcahealthcare.com > or Histopatty@aol.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Krat18 <@t> aol.com Sun Feb 20 08:05:51 2005 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] DAB disposal Message-ID: <1e8.355697ad.2f49f33f@aol.com> _DAB-Out_ (http://www.trianglebiomedical.com/products/dab_out.html) _http://www.trianglebiomedical.com/assets/msds/DabOut.pdf_ (http://www.trianglebiomedical.com/assets/msds/DabOut.pdf) _BBC Biochemical : Products_ (http://www.bbcus.com/products.html?pc=28&pid=435) Hi, Susan, Triangle Biomedical Products and BBC Biochemical both have a disposal system for DAB, probably both the same system, so you'd have to compare prices, etc. One of these sites (Triangle) has the MSDS info, in case you'd like to see that. Hope this helps. Karen_Raterman@ssmhc.com From maxim_71 <@t> mail.ru Sun Feb 20 11:48:09 2005 From: maxim_71 <@t> mail.ru (Maxim) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] re-embedding blocks Message-ID: <1102172984.20050220204809@mail.ru> Dear Jeanine! I'm melt down the paraffin blocks for 60 deg 20 min. Then carry specimen in the xylene (60 deg), 2 changes 30 min each. This procedure is complite removed paraffin without serious changes of tissue structures. Maxim Peshkov, Department of biopsy and cytological research Pathological and anatomical bureau Taganrog Russia, 347942 mailto:maxim_71@mail.ru From sbindu20002000 <@t> yahoo.co.in Sun Feb 20 21:23:43 2005 From: sbindu20002000 <@t> yahoo.co.in (s bindu) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] DAB disposal Message-ID: <20050221032343.60323.qmail@web8508.mail.in.yahoo.com> Hi, In our lab, we are using bleaching powder as a neutralising agent for DAB. Regards Bindu.S Research Assistant, Histopathology Lab SCTIMST,TVM. sbindu20002000@yahoo.co.in Yahoo! India Matrimony: Find your life partneronline. From jhofecker <@t> yahoo.com Mon Feb 21 07:27:47 2005 From: jhofecker <@t> yahoo.com (Jennifer Hofecker) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] RE: eosin and microwave processing Message-ID: <20050221132747.2375.qmail@web21005.mail.yahoo.com> Hi Sue, I have used microwave processing in the past and we did put eosin in our isopropyl alcohols. It never seemed to cause any problems except pink paraffin sometimes! Definitely better than "naked" biopsies:) Good Luck, Jennifer Hofecker Hi all, Does anyone using a microwave tissue processor use Eosin in their alcohol? Or know of a reason why it shouldn't be used? We put eosin in our alcohol on our Tissue-Tek and have grown accustomed to our bxs being a nice pink. We've just gotten our first Energy Beam microwave tissue processor and have found it hard to embed "naked" bxs. Thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 __________________________________ Do you Yahoo!? All your favorites on one personal page – Try My Yahoo! http://my.yahoo.com From MDiCarlo <@t> KaleidaHealth.Org Mon Feb 21 07:35:29 2005 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] DAB disposal Message-ID: Darren 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 Darren James Sent: Saturday, February 19, 2005 05:19 To: 'Histology SLU'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] DAB disposal Hi, I always thought addition of bleach neutralises DAB. Please correct me if I am wrong. Regards Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: 18 February 2005 09:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] DAB disposal Hello All: How is everyone disposing of their waste DAB from their automated IHC stainers? Is there a solution to neutralize DAB? Has anyone encountered defending their method of disposition to a CAP inspector? Thanks for any and all replies, in advance. Susan --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email 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 MDiCarlo <@t> KaleidaHealth.Org Mon Feb 21 07:41:16 2005 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] DAB disposal Message-ID: Darren, When I worked in Pathology we neutralized the DAB waste container by adding 12-15 ml. of 30% Hydrogen Peroxide and letting it sit for 24 hours and then drain the liquid down the drain and put the jell0-like material into a jug for waste. 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 Darren James Sent: Saturday, February 19, 2005 05:19 To: 'Histology SLU'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] DAB disposal Hi, I always thought addition of bleach neutralises DAB. Please correct me if I am wrong. Regards Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: 18 February 2005 09:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] DAB disposal Hello All: How is everyone disposing of their waste DAB from their automated IHC stainers? Is there a solution to neutralize DAB? Has anyone encountered defending their method of disposition to a CAP inspector? Thanks for any and all replies, in advance. Susan --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email 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 browning <@t> HHSC.CA Mon Feb 21 09:02:54 2005 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] DAB disposal Message-ID: <3AADFB88753AD31189C100902786B91C0E27872B@hch_nt_exchange.hhsc.ca> We used to add excess bleach to the DAB waste, and once the solution was clear, threw it down the sink with lots of running tap water. However, I understand that even tho the DAB itself may have been neutralized, the new product from the bleaching reaction was toxic itself, but never tested to prove that. So, being proactive, we now collect all DAB waste and have it removed as chemical disposal. The volume of DAB we produce daily is too large to treat with permanganate or hydrogen peroxide/peroxidase. -----Original Message----- From: Histology SLU [mailto:sluhisto@yahoo.com] Sent: Thursday, February 17, 2005 3:27 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] DAB disposal Hello All: How is everyone disposing of their waste DAB from their automated IHC stainers? Is there a solution to neutralize DAB? Has anyone encountered defending their method of disposition to a CAP inspector? Thanks for any and all replies, in advance. Susan --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From herme013 <@t> umn.edu Mon Feb 21 09:14:30 2005 From: herme013 <@t> umn.edu (Yves Heremans) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] (no subject) Message-ID: <48E993A9-841B-11D9-A11C-000393BA7938@umn.edu> Dear All, I was wondering whether 4% formalin used for fixation before RNA in situ hybridization has to made up RNAse-free or does the fact that it is a fixative renders it de facto RNAse-free ? Yves From POWELL_SA <@t> Mercer.edu Mon Feb 21 10:05:36 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] GSH meeting Message-ID: Hi All, Posting this again and this one has the registration form. Sorry for the omission in the last post. If the program below does not print out for you, email me and I will send an attachment of the program in Word to you personally. Shirley Powell Once again in the year 2005 the Georgia Society of Cytology and the Georgia Society for Histotechnology have joined to present a combined annual symposium. This year we are not only repeating the joint venture but we are also returning to a previous location ? the Hyatt Regency Suites Perimeter Northwest ? Atlanta. The Hyatt features luxuriously decorated rooms, indoor swimming pool and health club as well as an excellent on site restaurant. The hotel offers complimentary shuttle service within a 5 mile radius and covers a wide variety of restaurant and evening entertainment opportunities. A block of rooms is reserved until February 18, 2005. Mention the Georgia Society of Histotechnology to receive the single/double rate of $84.00, triple/quad rate of $94.00 and Regency Suite rate of $375.00. Call 770-956-1234 or fax 770-916-1120 to make your reservations. Joint Sessions of histology and cytology will be held featuring current topics of common interest. Registration for your respective program gives you access to the other society?s program at no additional charge. A special treat this year will be the dinner theater with a scientific flavor. Many scientific companies will be present to exhibit laboratory products and services along with the most current products and technologies for both cytology and histology. Following the Dinner Theater an opportunity of mix and mingle with venders and colleagues in the exhibit area will be available. Drawings for completed Vender Bingo as well as other drawings and prizes will be held at this time. You must be present to win. For more information on the area/events visit www.accessatlanta.com. Room Reservations: The host hotel is the Hyatt Regency Suites Perimeter Northwest ? Atlanta. Room reservations should be made directly with the Hyatt by calling 770-956-1234 or faxing 770-916-1120. When making your reservation, identify yourself as an attendee of the Georgia Society of Histotechnology/Cytology meeting. The following rates are guaranteed until Feb. 18, 2005 Single/double occupancy $ 84.00 Triple/quad occupancy $ 94.00 Regency Suite $375.00 Check-in time 3:00 p.m. Check-out time 12:00 p.m. Directions: From I-285 take I-75 North. Take Exit 260 Windy Hill Road; go east ? miles. The Hyatt Regency suites will be on the left, before the Powers Ferry intersection. Program Histology Cytology Saturday, March 5, 2005 7:30 ? 8:30 a.m. Registration 8:00 ? 9:00 a.m. Registration 8:30 a.m. ? 12:00 p.m. 9:00 ? 10:00 a.m. Lymphoma Panel Workups Atypical Glandular Cells of Undetermined Significance Laura Hughs Dr. Stephen Lau, Grady Health System Sponsored by Cell Marque Corp. 10:00 ?10:30 a.m. 9:30 ? 10:00 a.m. Break ? View Exhibits Break ? View Exhibits 10:30 ? 11:30 Lung Fine Needle Aspiration Dr. Roger Lane, Southeastern Pathology 11:30 ? 12:00 Pap Smear Litigation Update Dr. Roger Lan 12:00 ? 1:30 p.m. Lunch ? View Exhibits 12:00 ? 1:30 p.m. Lunch ? View Exhibits Joint Cytology/Histology Sessions 1:30 - 2:30 p.m. Obesity: What?s Happening in Our Society? Annie B. Carr, MS, RD, CDC 2:30 ? 3:00 p.m. Break ? View Exhibits 3:00 ? 4:30 p.m. Case Studies in Forensic Pathology Geoffry Smith M.D. Fulton Cty Med Examiner 4:30 ? 5:45 p.m Nipple Duct Lavage; Cytology and Histology Joanne Piratzky, M.D. 5:45 p.m. GSH General Membership Meeting 5:45 p.m. GSC General Membership Meeting 6:30 ? 10:30 p.m. Dinner Theater/Exhibits/Cash Bar Sunday, March 6, 2005 7:30-8:00 a.m. Registration 8:00 ? 9:00 a.m. Registration 8:00 ? 10:30 a.m. 9:00- 10:00 a.m. Microwave: The Whole Enchilada Urocyt Donna Willis HT/HTL(ASCP) Brian Hancock, Cytyc Corp 10:30 ? 11:00 a.m. 10:00 ? 10:30 a.m. Break Break 11:00 a.m. ? 1:00 p.m. 10:30 ? 11:30 a.m. The Fun Side of Presenting a Workshop FNA, Preparation and Fundamentals Linda Jenkins HT(ASCP) Dr. Julie Baird, Grady Health System 1:30 ? GSH BOD meeting Symposium Registration: To register, complete the Registration form using with either the Histology or Cytology portion only. Registrations postmarked after February 21, 2005 must include a late fee of $10.00. Histology: Full symposium registration fee includes Saturday and Sunday symposium sessions (including the ability to attend Cytology lectures at no additional charge), Saturday lunch break, Saturday Dinner Theater and Exhibit viewing and all morning and afternoon breaks. One day only fees are available for either Saturday or Sunday and include only activities and sessions scheduled for that day. Additional Saturday Dinner Theater tickets are $30.00 and MUST be ordered in advance. Registrations postmarked after February 21, 2005 must include a late fee of $10.00. Your annual GSH dues may be paid with your registration, entitling you to the member discount. Annual dues are $10.00 for the calendar year 2005. Make check payable to: Georgia Society for Histotechnology. Cytology: Full symposium registration fee includes Saturday and Sunday symposium sessions (including the ability to attend Histology lectures at no additional charge), Saturday lunch break, Saturday Dinner theater and Exhibit viewing and all morning and afternoon breaks and GSC annual dues. Additional Saturday Dinner Theater tickets are $30.00 and MUST be ordered in advance. Registrations postmarked after February 21, 2005 must include a late fee of $10.00. Make check payable to: Georgia Society of Cytology. Program Cancellation Policies: Refunds can only be issued for cancellation requests received by February 21, 2005. NO refunds will be made after that date for any reason. Attendance by a substitute person is encouraged in lieu of cancellation. Please return registration form and fees (for both cytology and histology) to: Connie Wavrin GSC/GSH Symposium Chairperson 2667 Meadow Court Chamblee, GA 30341 We are unable to accept credit card registrations. Additional information is available from: Histology Cytology Venders Connie Wavrin Shirley VanDuzer Ruby Huggins-Rodriquez w. ph. 678-443-2332 w. ph. 404-321-6111 ext 4081 h. ph. 770-513-3290 fax: 678-443-2339 fax: 404-235-3007 email: Rodriquezr@bellsouth.net email: cwavrin@labmd.org email: Shirley.vanduzer@med.va.gov GSH NEWS UPDATE The good news continues! Dues were free in 2003; half price in 2004 and due to good financial health dues will remain half price in 2005. That?s just $10.00! Name: _________________________________________________________________ Home Mailing Address: __________________________________________________ City, State, Zip Code: _____________________________________________________ Organization/Employer: ____________________________________________________ Work Mailing Address: ____________________________________________________ City, State, Zip Code: ______________________________________________________ W.Ph: ( )__________________H.Ph.: ( )_______________ Fax: ( )___________ e-mail:__________________________________________________________________ Dues must be current and paid in full to be eligible for member discount. Include with symposium registration or send to: Shirley Powell 156 Oakridge Ave Macon, GA 31204 Registration Form Complete either Cytology or Histology portion and fill name, address, etc. Please see Symposium Registration for additional information and instructions. Cytology Full Symposium Fee $ 80.00 Saturday Attendance only $ 60.00 Sunday Attendance only $ 30.00 Registration Fee $ 10.00 Late Fee (Postmark after Feb. 21, 2005) $ 10.00 $ 10.00 Additional Saturday Dinner Theater $ 30.00 Total Histology Full Symposium Fee Member $ 80.00 Non-member $100.00 Saturday Attendance only Member $ 60.00 Non-member $ 75.00 Sunday Attendance only Member $ 30.00 Non-member $ 50.00 Registration Fee $ 25.00 $ 25.00 Late Fee (Postmark after Feb. 21, 2005) $ 10.00 GSH Dues (1/2 price in 2005 or $10) $ 10.00 Additional Saturday Dinner Theater $ 30.00 Total Name: __________________________________________________________________ Home Mailing Address: ____________________________________________________ City, State, Zip Code: ______________________________________________________ Organization/Employer: ____________________________________________________ Work Mailing Address: ____________________________________________________ City, State, Zip Code: ______________________________________________________ W.Ph: ( )_______________H.Ph.: ( )_____________Fax: ( )_________ email:___________________________________________________________________ Return completed form and money to: Connie Wavrin, Symposium Chair 2667 Meadow Court Chamblee, GA 30341 From jkiernan <@t> uwo.ca Mon Feb 21 10:15:05 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] DAB disposal References: <20050221032343.60323.qmail@web8508.mail.in.yahoo.com> Message-ID: <421A0909.74BF79AF@uwo.ca> A Google search brings up, at the top of the list, a note by H. Horn in Biotech Histochem. 2002 Jul;77(4):229. http://taylorandfrancis.metapress.com/app/home/contribution.asp?wasp=1d8cff1kwr7qwg58al1y&referrer=parent&backto=issue,8,10;journal,11,19;linkingpublicationresults,1:105250,1 The second item (of 54) is a detailed protocol using permanganate (from the Histonet archives, no less), by K. Bowden, at: http://www.histosearch.com/histonet/May03A/Re.DABremovalA.html A few years ago there was much Histonet discussion of this subject. Then (and now, too, it seems) nearly everyone oxidized surplus DAB with bleach. Some evidence (or ?suspicion) was cited, saying that the bleach + oxidized DAB mixture might be mutagenic to bacteria. After oxidation of DAB by a solution containing potassium permanganate and sulphuric acid, the resulting goo did not threaten bacteria with mutation. The inference, I think, is that if you drink the stuff, but not enough to kill you, it won't give you cancer. Bleach and potassium permanganate are both good disinfectants, killing bacteria when quite dilute. One does wonder if testing such substances for mutagenicity is meaningful. Be that as it may, acidified permanganate is the politically correct way to get rid of DAB. It gets nearly a page in a book: Lunn G & Sansone EB (1990) Destruction of Hazardous Chemicals in the Laboratory. New York: Wiley, 271 pages. [I've not checked to see if there is a more recent edition; it seems likely.] John Kiernan London, Canada. ------------------- s bindu wrote: > > Hi, > > In our lab, we are using bleaching powder as a neutralising agent for DAB. > > Regards > Bindu.S > Research Assistant, > Histopathology Lab > SCTIMST,TVM. > sbindu20002000@yahoo.co.in > > > Yahoo! India Matrimony: Find your life partneronline. > _______________________________________________ From jkiernan <@t> uwo.ca Mon Feb 21 10:15:42 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] re-embedding blocks References: Message-ID: <421A092E.16E4AB8@uwo.ca> I don't know about "generally". When I do this (which isn't often) I use the same temperature as for infiltration, which in my lab is about 60C for a 58C wax. It works. Why do otherwise? With regard to "a temperature that is generally considered too high either for the paraffin or for the tissue", you can expect a variety of replies because this is controversial. A popular notion is that too-hot makes the tissue unduly hard to cut. I don't agree, but that's another story, and there's plenty of HISTOMYTHOLOGY in this area. John Kiernan Dept of anatomy & Cell Biology University of Western Ontario London, Canada. _______________________________________________ Bartlett, Jeanine" wrote: > > Hi everybody: > > Just curious as to what oven temperature is generally used for melting > down paraffin blocks for re-embedding. Is there a temperature that is > generally considered too high either for the paraffin or for the tissue? > > Thanks! > Jeanine Bartlett, HT(ASCP) > Centers for Disease Control and Prevention > Infectious Disease Pathology Activity > 1600 Clifton Road, MS/G-32 > Atlanta, GA 30333 _______________________________________ From Matthew_Frank <@t> URMC.Rochester.edu Mon Feb 21 10:23:44 2005 From: Matthew_Frank <@t> URMC.Rochester.edu (Frank, Matthew) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Plant histology Message-ID: I use root tip sections fixed in CRAF and stained with Flemming's Triple stain (Safranin O, Crystal Violet and Orange G). Xylene, Ethanol changes to hydrate, water rinse, Safranin O 1% aqueous for 1.5 hours, 15-25 secondes with 1% Iodine and 1% KI in 80 % ethanol, 5 minutes in 1%aq crsytal violet, 30 seconds of same iodine solution, rinsed with 100% ethanol, 0.25 % solution of clove oil for about 5 minutes (be careful not to take out all of the crystal violet), rinse with xylene, clear and mount. -----Original Message----- From: Histopatty@aol.com [mailto:Histopatty@aol.com] Sent: Saturday, February 19, 2005 4:15 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Plant histology Dose any one on the list have any knowledge or experience in plant histology. Specifically I am trying to demonstrate mitosis in onion roots. The slides I have access to are faded and the details are not crisp enough for photography. Any help on processing protocols and staining techniques would be appreciated. Patty.Eneff@hcahealthcare.com or Histopatty@aol.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ja.mitchell <@t> hosp.wisc.edu Mon Feb 21 11:11:50 2005 From: ja.mitchell <@t> hosp.wisc.edu (Mitchell Jean A.) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Slide Folders Message-ID: <583D3E9A1E843445BD54E461D1A2F6F30F83F529@uwhis-xchng2.hosp.wisc.edu> I am looking for particular slide folders that hold 20 slides. I know that there are varieties available in heavy duty cardboard & in plastic. What I am hoping to find are the plastic slide folders in different colors. Anyone know of a possible source for colored slide folders that hold 20 slides? Thanks, Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI From ccrowder <@t> mail.vetmed.lsu.edu Mon Feb 21 11:20:47 2005 From: ccrowder <@t> mail.vetmed.lsu.edu (Cheryl Crowder) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Vibratome info Message-ID: Hi all - One research lab is doing confocal microscopy. They were given a virtatome and are trying (without success) to get 50 um sections. If any of you know the intricasies of the virbatome and are willing to correspond or talk to the tech that is trying to work with the virbatome, would you contact me and I will pass the information on. I know that she has the e-mail address and phone number for Virbratome, but would also like to have contact with working techs. Thank you in advance, Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 From David.Edmondson <@t> christie-tr.nwest.nhs.uk Mon Feb 21 11:23:35 2005 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Slide Folders Message-ID: We have some plasic folders from Surgipath out on the corridor shelf. For a week or two we used them for incubations but that was so many years ago. I have no idea whether they are still in the catalogue or what colours they did have. Dave Histology Christie Hosp Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mitchell Jean A. Sent: 21 February 2005 17:12 To: Histonet Subject: [Histonet] Slide Folders I am looking for particular slide folders that hold 20 slides. I know that there are varieties available in heavy duty cardboard & in plastic. What I am hoping to find are the plastic slide folders in different colors. Anyone know of a possible source for colored slide folders that hold 20 slides? Thanks, Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ******************************************************************************************************************* This e-mail and any files transmitted with it are confidential and solely for the use of the intended recipient. If you have received this e-mail in error you should not disseminate, distribute or copy it. Please notify the sender immediately and delete this e-mail from your system. ******************************************************************************************************************* From JEllin <@t> yumaregional.org Mon Feb 21 11:29:38 2005 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Grossing Tech Message-ID: Charles you mentioned that this needs to be done before you start, how can a person go about asking for this once he is in this situation. and were is the PA schools that are specific to Pathologist Assistants?? Jesus Ellin Yuma Regional Medical Center >>> "Charles.Embrey" 02/17/05 09:15AM >>> First, my job description would have to be finalized and pay raise set before I opened my first specimen jar. Once you have started the duties your employer has lost his incentive to get this done. As far as pay raise is concerned it would really depend on the extent of your grossing and time involved. Personally I think the raise negotiations should start at $5.00 for small biopsies and increase with the level of specimen difficulty. The biggest problem I have found is that people tend to underestimate their worth. The only other option for grossing is a pathologist or pathologists' assistant and you would still be a steal with a $5.00 raise. If you enjoy grossing you should apply to one of the PA schools. It is only an additional two years of school, one of which is actual grossing at clinical sites. I teach second year PA students from Rosalind Franklin University in Chicago and my two students that graduate in April have already signed great contracts. Good Luck Charles Embrey PA(ASCP) Urbana, IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cfockler@mail1.vcu.edu Sent: Thursday, February 17, 2005 6:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Grossing Tech Just a quick question.. . what is the average pay increase to go from a standard histotech, to a grossing tech? (with a B.S.) And, how long on average should it take to see it in your job description and your paycheck? Thanks! C. Fockler _______________________________________________ Histonet mailing list Histonet@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 Charles.Embrey <@t> carle.com Mon Feb 21 11:49:26 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Grossing Tech Message-ID: Once you have already started you will probably have to appeal to the Pathologists' sense of fair play. Here are the training sites: Training Programs for Pathologists' Assistants The National Accrediting Agency for Clinical Laboratory Sciences (NAACLS) board of directors voted in 1995 to accept the Essentials of Accredited Training Programs for the Pathologists' Assistant, establishing a process of accreditation for pathologists' assistant training programs. Adoption of NAACLS Essentials ensures a uniform standard of excellence for the training of pathologists assistants. The following training programs have been accredited by NAACLS and provide academic and practical training leading to either a master's degree or a bachelor's degree. Duke University - MHS, 2 years Claudia Brady,MHS, Program Director Department of Pathology PO Box 3712 Durham, NC 27710 919.684.2159 Tara Mann - student liaison 535 Crossview Drive Durham, NC 27703 mann0025@mc.duke.edu Quinnipiac University - MHS, 2 years Scott Farber, Graduate Admissions Director 275 Mount Carmel Ave Hamden, CT 06518 203.582.8795 graduate@quinnipiac.edu ------ Leo Kelly, MHS, Clinical Coordinator (contact for employment opportunities) 203.932.5711 x4758 Ray Schneider - student liaison Norwalk Hospital Dept. of Pathology 24 Stevens St. Norwalk, CT 06856 (203)-852-2657 raymond.schneider@norwalkhealth.org University of Maryland - MS, 2 years Raymond Jones, PhD, Program Director Department of Pathology 22 S Greene St Baltimore, MD 21201 410.328.1221 rjones@som.umaryland.edu Katie Flickinger - student liaison Dept. of Pathology 600 N. Wolfe St. Pathology B 107A Baltimore, MD 21287-6667 (443)-287-6134 kflicki1@jhmi.edu Wayne State University - BS, 2 years Peter Frade, PhD, Program Director Department of Mortuary Sciences 5439 Woodward Ave Detroit, MI 48202 313.577.2050 ab8123@wayne.edu Leena Budev and Anne Hildebrandt - student liasions Anatomic Pathologists' Assistant Program Wayne State University Dept. of Mortuary Science 5439 Woodward Ave. Detroit, MI 48202 (313)-966-0575 lbudev@dmc.org and arh1214@aol.com Ohio State University - MS, 2 years Charles Hitchcock, MD, PhD, Program Director Gretchen Staschiak, Pathology Education Coordinator N-308A Doan Hall 410 W Tenth Ave Columbus, Ohio 43210 614.293.3055 staschiak.2@osu.edu Sandra Banky - student liaison OSU Medical Center 417 East Doan Hall 410 West 10th Avenue Columbus, OH 43210 (614)-293-4875 banky-1@medctr.osu.edu Rosalind Franklin University of Medicine and Science - MS, 2 years John Vitale, MHS, Program Director & Clinical Coordinator Pathologists' Assistant Department 3333 Green Bay Road North Chicago, IL 60064 847.578.8638 john.vitale@rosalindfranklin.edu Trina A.Sherlitz, PT, MS, Clinical Coordinator trina.sherlitz@rosalindfranklin.edu Karen Skish - student liaison (630) 926-9561 karenskish@comcast.net Please Note: Graduates of non-NAACLS accredited training programs are eligible for membership; however, at least one year of work experience is required to become eligible for the AAPA Fellowship Exam. If you have any problems or questions concerning accreditation, please contact: Daniel Tice Staff Program Coordinator dtice@naacls.org or Dr. Olive Kimball Executive Director kimball@naacls.org Further information about NAACLS (National Accrediting Agency for Clinical Laboratory Sciences) may be obtained from www.naacls.org or naaclsinfo@naacls.org. -----Original Message----- From: Jesus Ellin [mailto:JEllin@yumaregional.org] Sent: Monday, February 21, 2005 11:30 AM To: Charles.Embrey; cfockler@mail1.vcu.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Grossing Tech Charles you mentioned that this needs to be done before you start, how can a person go about asking for this once he is in this situation. and were is the PA schools that are specific to Pathologist Assistants?? Jesus Ellin Yuma Regional Medical Center >>> "Charles.Embrey" 02/17/05 09:15AM >>> First, my job description would have to be finalized and pay raise set before I opened my first specimen jar. Once you have started the duties your employer has lost his incentive to get this done. As far as pay raise is concerned it would really depend on the extent of your grossing and time involved. Personally I think the raise negotiations should start at $5.00 for small biopsies and increase with the level of specimen difficulty. The biggest problem I have found is that people tend to underestimate their worth. The only other option for grossing is a pathologist or pathologists' assistant and you would still be a steal with a $5.00 raise. If you enjoy grossing you should apply to one of the PA schools. It is only an additional two years of school, one of which is actual grossing at clinical sites. I teach second year PA students from Rosalind Franklin University in Chicago and my two students that graduate in April have already signed great contracts. Good Luck Charles Embrey PA(ASCP) Urbana, IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cfockler@mail1.vcu.edu Sent: Thursday, February 17, 2005 6:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Grossing Tech Just a quick question.. . what is the average pay increase to go from a standard histotech, to a grossing tech? (with a B.S.) And, how long on average should it take to see it in your job description and your paycheck? Thanks! C. Fockler _______________________________________________ Histonet mailing list Histonet@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 DavidH <@t> marketlabinc.com Mon Feb 21 12:35:18 2005 From: DavidH <@t> marketlabinc.com (David Haagsma) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Slide Folders Message-ID: <19E3602A16438E48B51A4250CA04B5F6099753@exchange.marketlab.com> Market Lab has them in Blue, Green, Grey, White and Yellow. Dave Haagsma MarketLab Inc. I am looking for particular slide folders that hold 20 slides. I know that there are varieties available in heavy duty cardboard & in plastic. What I am hoping to find are the plastic slide folders in different colors. Anyone know of a possible source for colored slide folders that hold 20 slides? Thanks, Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI From pathrm35 <@t> adelphia.net Mon Feb 21 14:39:09 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] alcohol and xylene recycling units Message-ID: <8305682.1109018349658.JavaMail.root@web8.mail.adelphia.net> I was wondering what recycling units some of you were using along with their pros and cons. You can contact me directly if you wish. Sales reps are also welcome to contact me if they wish. Thanks, Ron Martin From strande <@t> umdnj.edu Mon Feb 21 15:01:22 2005 From: strande <@t> umdnj.edu (Louise Strande) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Negative Rabbit Contrl Message-ID: <003901c51858$7f9950f0$c165db82@chsmail.root.cooperhealth.edu> Does anyone have a source for a good normal rabbit IgG to use for negative control immunohistochemical applications? I have human tissue or human cytospins and occassionally my primary antibody is grown in rabbit. In order to run a non immunized rabbit negative control, I have purchased normal rabbit IgG from DAKO, Zymed, Santa Cruz, Lab Visions. Every one I try gives positive staining on the negative slide. I have done the appropriate tests and the staining is not coming from streptaviden, or the link anibody, but from the rabbit IgG itself. I do not have this problem with Mouse negative control IgG or goat negative control IgG. Any suggestions, other than not buying any antibodies rasied in rabbit? Louise Strande, MS Cooper University Hospital Department of Surgery Division of Surgical Research Phone : (856) 757-7827 From Jackie.O'Connor <@t> abbott.com Mon Feb 21 15:12:46 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Negative Rabbit Contrl Message-ID: Try Biocare.net They have a pretty good line. Jackie O'Connor Assistant Scientist Cancer Research GPRD-Abbott Laboratories Louise Strande Sent by: histonet-bounces@lists.utsouthwestern.edu 02/21/2005 03:01 PM To: Histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Negative Rabbit Contrl Does anyone have a source for a good normal rabbit IgG to use for negative control immunohistochemical applications? I have human tissue or human cytospins and occassionally my primary antibody is grown in rabbit. In order to run a non immunized rabbit negative control, I have purchased normal rabbit IgG from DAKO, Zymed, Santa Cruz, Lab Visions. Every one I try gives positive staining on the negative slide. I have done the appropriate tests and the staining is not coming from streptaviden, or the link anibody, but from the rabbit IgG itself. I do not have this problem with Mouse negative control IgG or goat negative control IgG. Any suggestions, other than not buying any antibodies rasied in rabbit? Louise Strande, MS Cooper University Hospital Department of Surgery Division of Surgical Research Phone : (856) 757-7827 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Mon Feb 21 16:48:26 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] (no subject) Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E1C0@simba.kids> I would assume that the formalin would inactivate the RNAses. At least from literature experience, mRNA has been regularly demonstrated in FFPE tissues. 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: Yves Heremans [mailto:herme013@umn.edu] Sent: Tuesday, 22 February 2005 2:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Dear All, I was wondering whether 4% formalin used for fixation before RNA in situ hybridization has to made up RNAse-free or does the fact that it is a fixative renders it de facto RNAse-free ? Yves _______________________________________________ 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 dsuer <@t> comcast.net Mon Feb 21 17:22:21 2005 From: dsuer <@t> comcast.net (dsuer@comcast.net) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] her-2 nuclear staining Message-ID: <022120052322.12388.421A6D2C000EC4BD0000306422070016419D0A9A9C0B@comcast.net> Occasionally we have a her-2 with irregular nuclear staining.It seems to be happening on breast needle cores. Could it be over or under fixation? If anyone has any information I would greatly appreciate it. D. Suermann HT(ASCP) From liz <@t> premierlab.com Mon Feb 21 18:01:15 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] MMP's Message-ID: <000001c51871$a071a2a0$76d48a80@AMY> Does anyone know what MMP's can be stained for in formic acid decalcifed rat joints. Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From tahseen <@t> brain.net.pk Mon Feb 21 10:02:09 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] special microtome for Left hander. Message-ID: <005401c5182e$b399f1e0$972bfea9@m7c0y4> Hi Everyone, I was wondering if anyone has any idea or uses the special microtome for Left handed. Thanks in advance Regards Muhammad Tahseen Laboratory Supervisor SKMCH & RC Lahore Pakistan. From JQB7 <@t> CDC.GOV Mon Feb 21 18:18:02 2005 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] special microtome for Left hander. Message-ID: The Microm Ergostar is an ambidextrous machine and it works great! Jeanine Bartlett CDC/Atlanta -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Muhammad Tahseen Sent: Mon 2/21/2005 11:02 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] special microtome for Left hander. Hi Everyone, I was wondering if anyone has any idea or uses the special microtome for Left handed. Thanks in advance Regards Muhammad Tahseen Laboratory Supervisor SKMCH & RC Lahore Pakistan. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JQB7 <@t> CDC.GOV Mon Feb 21 18:20:26 2005 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] re-embedding blocks Message-ID: We often get large volumes of blocks from other countries that must be reembedded. I melt them in a tray in our paraffin oven at 60C but some others in the lab feel that it should be done at a higher temperature to speed up the process. -----Original Message----- From: John Kiernan [mailto:jkiernan@uwo.ca] Sent: Mon 2/21/2005 11:15 AM To: Bartlett, Jeanine Cc: Histonet Subject: Re: [Histonet] re-embedding blocks I don't know about "generally". When I do this (which isn't often) I use the same temperature as for infiltration, which in my lab is about 60C for a 58C wax. It works. Why do otherwise? With regard to "a temperature that is generally considered too high either for the paraffin or for the tissue", you can expect a variety of replies because this is controversial. A popular notion is that too-hot makes the tissue unduly hard to cut. I don't agree, but that's another story, and there's plenty of HISTOMYTHOLOGY in this area. John Kiernan Dept of anatomy & Cell Biology University of Western Ontario London, Canada. _______________________________________________ Bartlett, Jeanine" wrote: > > Hi everybody: > > Just curious as to what oven temperature is generally used for melting > down paraffin blocks for re-embedding. Is there a temperature that is > generally considered too high either for the paraffin or for the tissue? > > Thanks! > Jeanine Bartlett, HT(ASCP) > Centers for Disease Control and Prevention > Infectious Disease Pathology Activity > 1600 Clifton Road, MS/G-32 > Atlanta, GA 30333 _______________________________________ From stamptrain <@t> yahoo.com Mon Feb 21 19:12:24 2005 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] special microtome for Left hander. In-Reply-To: <005401c5182e$b399f1e0$972bfea9@m7c0y4> Message-ID: <20050222011224.40151.qmail@web31106.mail.mud.yahoo.com> Am not aware of them, altho I use the Microm. Many years ago I learned that being left handed in the lab ( and I _am_ left handed--very dominantly so) could be a handicap, so I have learned to use my right hand--to accomodate those who cannot adapt to a left handed world! As a result I am competently ambidextrous for those lab procedures that require a "right handed" approach. Makes me think that maybe us left handers are far more adaptable than those who always work from the right! Roger Moretz Dept of Toxicology Boehringer Ingelheim Pharmaceuticals --- Muhammad Tahseen wrote: > Hi Everyone, > > I was wondering if anyone has any idea or uses the > special microtome for Left handed. > > Thanks in advance > Regards > Muhammad Tahseen > Laboratory Supervisor > SKMCH & RC > Lahore Pakistan. > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. http://promotions.yahoo.com/new_mail From stamptrain <@t> yahoo.com Mon Feb 21 19:19:03 2005 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Vibratome info In-Reply-To: Message-ID: <20050222011903.42061.qmail@web31106.mail.mud.yahoo.com> Could you describe more clearly the problems you are having? I have successfully used a vibratome for years (and sporadically at that), always getting 50um sections of the samples I am sectioning. Have done everything from cns to liver and kidney and nearly everything else in between. Successfully mounting your samples is the first goal. Second, I have always preferred a #22 scalpel blade to a razor blade--using only the straight portion of the blade of course. Third, maintaining a cool temperature in the bath (with ice or the chiller if the vibratome is so equipped) is essential. Finally, don't try to do mammoth samples--altho I have done up to 25x25mm samples without a problem. If you have done all of these, please provide more details and I'll see if I can help. Roger Moretz, Ph.D Dept of Toxicology Boehringer Ingelheim Pharmaceuticals --- Cheryl Crowder wrote: > Hi all - One research lab is doing confocal > microscopy. They were given a > virtatome and are trying (without success) to get 50 > um sections. If any of > you know the intricasies of the virbatome and are > willing to correspond or > talk to the tech that is trying to work with the > virbatome, would you > contact me and I will pass the information on. I > know that she has the > e-mail address and phone number for Virbratome, but > would also like to have > contact with working techs. Thank you in advance, > Cheryl > > > Cheryl Crowder, BA, HTL(ASCP) > Chief Technologist > Anatomic Pathology > Department of Pathobiological Sciences > School of Veterinary Medicine > Louisiana State University > Skip Bertman Drive > Baton Rouge, LA 70803 > > 225-578-9734 > FAX: 225-578-9720 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Do you Yahoo!? Yahoo! Mail - now with 250MB free storage. Learn more. http://info.mail.yahoo.com/mail_250 From robinmax <@t> rogers.com Mon Feb 21 19:28:19 2005 From: robinmax <@t> rogers.com (Robin Maxwell-Nunn) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] xylene free mounting media Message-ID: <001d01c5187d$cab0a500$6501a8c0@NEWCOMPUTER> Hi We have a tech in our lab who is sensitive to xylene (exhibits headaches, puffy eyes and itchy skin). We are switching to Formula 83 by CBG Biotech for frozen sections but still need a mountant that is both compatible with this and xylene free. Does anyone have any suggestions? thanks, Robin Maxwell-Nunn, MLT St Mary's General Hospital, Kitchener, Ontario, Canada. From ajennings <@t> unmc.edu Mon Feb 21 19:33:54 2005 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] special microtome for Left hander. Message-ID: I always thought they were all made for left handers, my right hand is turning a balanced wheel, controlling speed, the left hand is the one I use to guide the ribbon and smooth out the section. I believe that takes far more dexterity and a steady hand. From Kemlo.Rogerson <@t> elht.nhs.uk Tue Feb 22 02:02:43 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] re-embedding blocks[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EFEB@bhrv-nt-11.bhrv.nwest.nhs.uk> If you keep boiling an egg, does it get harder the longer you boil it? Is a fried egg harder than a boiled egg? I don't know cos I'm not allowed to eat them. -----Original Message----- From: Bartlett, Jeanine [mailto:JQB7@CDC.GOV] Sent: 22 February 2005 00:20 To: jkiernan@uwo.ca Cc: Histonet Subject: RE: [Histonet] re-embedding blocks[Scanned] We often get large volumes of blocks from other countries that must be reembedded. I melt them in a tray in our paraffin oven at 60C but some others in the lab feel that it should be done at a higher temperature to speed up the process. -----Original Message----- From: John Kiernan [mailto:jkiernan@uwo.ca] Sent: Mon 2/21/2005 11:15 AM To: Bartlett, Jeanine Cc: Histonet Subject: Re: [Histonet] re-embedding blocks I don't know about "generally". When I do this (which isn't often) I use the same temperature as for infiltration, which in my lab is about 60C for a 58C wax. It works. Why do otherwise? With regard to "a temperature that is generally considered too high either for the paraffin or for the tissue", you can expect a variety of replies because this is controversial. A popular notion is that too-hot makes the tissue unduly hard to cut. I don't agree, but that's another story, and there's plenty of HISTOMYTHOLOGY in this area. John Kiernan Dept of anatomy & Cell Biology University of Western Ontario London, Canada. _______________________________________________ Bartlett, Jeanine" wrote: > > Hi everybody: > > Just curious as to what oven temperature is generally used for melting > down paraffin blocks for re-embedding. Is there a temperature that is > generally considered too high either for the paraffin or for the tissue? > > Thanks! > Jeanine Bartlett, HT(ASCP) > Centers for Disease Control and Prevention > Infectious Disease Pathology Activity > 1600 Clifton Road, MS/G-32 > Atlanta, GA 30333 _______________________________________ From ian.montgomery <@t> bio.gla.ac.uk Tue Feb 22 05:01:54 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Mouse footpads. Message-ID: <6.2.1.2.2.20050222105711.03039dd0@udcf.gla.ac.uk> Just about to start a study on mouse footpads, routine, IHC etc. My intention is to fix with formaldehyde then decalcify using EDTA but as mice as such wee devils are there any hints and tips I should be aware of? Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From ian.montgomery <@t> bio.gla.ac.uk Tue Feb 22 05:05:19 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Left-handed microtome. Message-ID: <6.2.1.2.2.20050222110203.03039b30@udcf.gla.ac.uk> As a left hander I've always found a microtome to be ideal. You turn the handle with your right hand then manipulate the sections with your left, the dominant hand. What could be better? Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From kelly.mcqueeney <@t> bms.com Tue Feb 22 07:52:05 2005 From: kelly.mcqueeney <@t> bms.com (Kelly D Mcqueeney) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Negative Rabbit Contrl In-Reply-To: <003901c51858$7f9950f0$c165db82@chsmail.root.cooperhealth.edu> References: <003901c51858$7f9950f0$c165db82@chsmail.root.cooperhealth.edu> Message-ID: <421B3905.10501@bms.com> Louis, I have seen the same problem with rabbit IgG. I always use the rabbit IgG from Vector or Jackson as a control. Once you receive the IgG, immediately aliquot (5ul if possible) and store at -20 or -80C. Once you thaw and use an aliquot, immediately throw it away. Do not leave at 4C or re-freeze. I only get negative results with a fresh, frozen aliquot used once. Good luck, Kelly Louise Strande wrote: >Does anyone have a source for a good normal rabbit IgG to use for negative control immunohistochemical applications? I have human tissue or human cytospins and occassionally my primary antibody is grown in rabbit. In order to run a non immunized rabbit negative control, I have purchased normal rabbit IgG from DAKO, Zymed, Santa Cruz, Lab Visions. Every one I try gives positive staining on the negative slide. I have done the appropriate tests and the staining is not coming from streptaviden, or the link anibody, but from the rabbit IgG itself. I do not have this problem with Mouse negative control IgG or goat negative control IgG. Any suggestions, other than not buying any antibodies rasied in rabbit? > >Louise Strande, MS >Cooper University Hospital >Department of Surgery >Division of Surgical Research >Phone : (856) 757-7827 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From JNocito <@t> Pathreflab.com Tue Feb 22 08:32:08 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Grossing Tech In-Reply-To: Message-ID: Chuck, is any reason given as to why all the programs are up north? Is it a supply and demand issue? Joe Nocito -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Charles.Embrey Sent: Monday, February 21, 2005 11:49 AM To: Jesus Ellin; cfockler@mail1.vcu.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Grossing Tech Once you have already started you will probably have to appeal to the Pathologists' sense of fair play. Here are the training sites: Training Programs for Pathologists' Assistants The National Accrediting Agency for Clinical Laboratory Sciences (NAACLS) board of directors voted in 1995 to accept the Essentials of Accredited Training Programs for the Pathologists' Assistant, establishing a process of accreditation for pathologists' assistant training programs. Adoption of NAACLS Essentials ensures a uniform standard of excellence for the training of pathologists assistants. The following training programs have been accredited by NAACLS and provide academic and practical training leading to either a master's degree or a bachelor's degree. Duke University - MHS, 2 years Claudia Brady,MHS, Program Director Department of Pathology PO Box 3712 Durham, NC 27710 919.684.2159 Tara Mann - student liaison 535 Crossview Drive Durham, NC 27703 mann0025@mc.duke.edu Quinnipiac University - MHS, 2 years Scott Farber, Graduate Admissions Director 275 Mount Carmel Ave Hamden, CT 06518 203.582.8795 graduate@quinnipiac.edu ------ Leo Kelly, MHS, Clinical Coordinator (contact for employment opportunities) 203.932.5711 x4758 Ray Schneider - student liaison Norwalk Hospital Dept. of Pathology 24 Stevens St. Norwalk, CT 06856 (203)-852-2657 raymond.schneider@norwalkhealth.org University of Maryland - MS, 2 years Raymond Jones, PhD, Program Director Department of Pathology 22 S Greene St Baltimore, MD 21201 410.328.1221 rjones@som.umaryland.edu Katie Flickinger - student liaison Dept. of Pathology 600 N. Wolfe St. Pathology B 107A Baltimore, MD 21287-6667 (443)-287-6134 kflicki1@jhmi.edu Wayne State University - BS, 2 years Peter Frade, PhD, Program Director Department of Mortuary Sciences 5439 Woodward Ave Detroit, MI 48202 313.577.2050 ab8123@wayne.edu Leena Budev and Anne Hildebrandt - student liasions Anatomic Pathologists' Assistant Program Wayne State University Dept. of Mortuary Science 5439 Woodward Ave. Detroit, MI 48202 (313)-966-0575 lbudev@dmc.org and arh1214@aol.com Ohio State University - MS, 2 years Charles Hitchcock, MD, PhD, Program Director Gretchen Staschiak, Pathology Education Coordinator N-308A Doan Hall 410 W Tenth Ave Columbus, Ohio 43210 614.293.3055 staschiak.2@osu.edu Sandra Banky - student liaison OSU Medical Center 417 East Doan Hall 410 West 10th Avenue Columbus, OH 43210 (614)-293-4875 banky-1@medctr.osu.edu Rosalind Franklin University of Medicine and Science - MS, 2 years John Vitale, MHS, Program Director & Clinical Coordinator Pathologists' Assistant Department 3333 Green Bay Road North Chicago, IL 60064 847.578.8638 john.vitale@rosalindfranklin.edu Trina A.Sherlitz, PT, MS, Clinical Coordinator trina.sherlitz@rosalindfranklin.edu Karen Skish - student liaison (630) 926-9561 karenskish@comcast.net Please Note: Graduates of non-NAACLS accredited training programs are eligible for membership; however, at least one year of work experience is required to become eligible for the AAPA Fellowship Exam. If you have any problems or questions concerning accreditation, please contact: Daniel Tice Staff Program Coordinator dtice@naacls.org or Dr. Olive Kimball Executive Director kimball@naacls.org Further information about NAACLS (National Accrediting Agency for Clinical Laboratory Sciences) may be obtained from www.naacls.org or naaclsinfo@naacls.org. -----Original Message----- From: Jesus Ellin [mailto:JEllin@yumaregional.org] Sent: Monday, February 21, 2005 11:30 AM To: Charles.Embrey; cfockler@mail1.vcu.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Grossing Tech Charles you mentioned that this needs to be done before you start, how can a person go about asking for this once he is in this situation. and were is the PA schools that are specific to Pathologist Assistants?? Jesus Ellin Yuma Regional Medical Center >>> "Charles.Embrey" 02/17/05 09:15AM >>> First, my job description would have to be finalized and pay raise set before I opened my first specimen jar. Once you have started the duties your employer has lost his incentive to get this done. As far as pay raise is concerned it would really depend on the extent of your grossing and time involved. Personally I think the raise negotiations should start at $5.00 for small biopsies and increase with the level of specimen difficulty. The biggest problem I have found is that people tend to underestimate their worth. The only other option for grossing is a pathologist or pathologists' assistant and you would still be a steal with a $5.00 raise. If you enjoy grossing you should apply to one of the PA schools. It is only an additional two years of school, one of which is actual grossing at clinical sites. I teach second year PA students from Rosalind Franklin University in Chicago and my two students that graduate in April have already signed great contracts. Good Luck Charles Embrey PA(ASCP) Urbana, IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cfockler@mail1.vcu.edu Sent: Thursday, February 17, 2005 6:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Grossing Tech Just a quick question.. . what is the average pay increase to go from a standard histotech, to a grossing tech? (with a B.S.) And, how long on average should it take to see it in your job description and your paycheck? Thanks! C. Fockler _______________________________________________ Histonet mailing list Histonet@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 vazquezr <@t> ohsu.edu Tue Feb 22 09:05:02 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] re-embedding blocks Message-ID: Jeanine, Check your paraffin package and it should tell you the melting point of your paraffin. Robyn >>> "Bartlett, Jeanine" 02/18/05 4:33 AM >>> Hi everybody: Just curious as to what oven temperature is generally used for melting down paraffin blocks for re-embedding. Is there a temperature that is generally considered too high either for the paraffin or for the tissue? Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Tue Feb 22 09:16:25 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] CPT coding for IHC Message-ID: Are any of you using the new CPT code (88360) for IHC/semiquantitative analysis? If so, why is 88360 used on the technical side as well (there is no difference in the amount of technical work performed)? The only difference I see is on the professional side with the interpretation. Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From JQB7 <@t> CDC.GOV Tue Feb 22 09:16:05 2005 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] re-embedding blocks Message-ID: I was just wondering how far above the melting point you could safely go. -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: Tuesday, February 22, 2005 10:05 AM To: Bartlett, Jeanine; histonet@pathology.swmed.edu Subject: Re: [Histonet] re-embedding blocks Jeanine, Check your paraffin package and it should tell you the melting point of your paraffin. Robyn >>> "Bartlett, Jeanine" 02/18/05 4:33 AM >>> Hi everybody: Just curious as to what oven temperature is generally used for melting down paraffin blocks for re-embedding. Is there a temperature that is generally considered too high either for the paraffin or for the tissue? Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.Eyres <@t> sanofi-aventis.com Tue Feb 22 09:22:37 2005 From: Stephen.Eyres <@t> sanofi-aventis.com (Stephen.Eyres@sanofi-aventis.com) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Sakura TDE 30 Decalcifier System Message-ID: Hi, Does anyone have any experience of using this system? If not, what do folks use to speed up their decalcifying system? I am particularly interested in people working with rodent samples/ Thanks 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 Pawel.Pasierbek <@t> imp.univie.ac.at Tue Feb 22 09:26:24 2005 From: Pawel.Pasierbek <@t> imp.univie.ac.at (Pawel,Pasierbek) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] collagen gell autofluorescence Message-ID: <1E6148FED8920445AC79AEB51579E5238069C1@EXCHSRV.imp.univie.ac.at> Dear All, One of the users of our microscopy facility is thrying to image cells grown in the collagen gel (DB). After immunostaings she gets a lot of autofluorescence (as I believe) comming from the gel itself - it is visible when imaged with all 3 lasers 488nm, 543nm, 633nm. Do you have any hint how to get around this problem? All the best from Vienna, Pawel -------------------------------------------------------------------------- Pawel Pasierbek, PhD Biooptics IMP (Research Institute of Molecular Pathology) Dr. Bohr-Gasse 7 1030 Vienna Austria Telephone: +43 (1) 79730591 Fax: +43 (1) 7987153 http://www.imp.univie.ac.at From Charles.Embrey <@t> carle.com Tue Feb 22 09:42:14 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Grossing Tech Message-ID: Hell Joe, you're in San Antonio. Almost anywhere in the U.S. is "up north" to you. The most southern program is at Duke and all the rest are either in New England or near the great lakes. There is nothing west of the Mississippi and I do think that it is because of the population centers. Ohio State and Rosalind Franklin are the newest schools and I am sure more will open because of the increasing demand for PA's. Chuck -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Tuesday, February 22, 2005 8:32 AM To: Charles.Embrey; Jesus Ellin; cfockler@mail1.vcu.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Grossing Tech Chuck, is any reason given as to why all the programs are up north? Is it a supply and demand issue? Joe Nocito -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Charles.Embrey Sent: Monday, February 21, 2005 11:49 AM To: Jesus Ellin; cfockler@mail1.vcu.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Grossing Tech Once you have already started you will probably have to appeal to the Pathologists' sense of fair play. Here are the training sites: Training Programs for Pathologists' Assistants The National Accrediting Agency for Clinical Laboratory Sciences (NAACLS) board of directors voted in 1995 to accept the Essentials of Accredited Training Programs for the Pathologists' Assistant, establishing a process of accreditation for pathologists' assistant training programs. Adoption of NAACLS Essentials ensures a uniform standard of excellence for the training of pathologists assistants. The following training programs have been accredited by NAACLS and provide academic and practical training leading to either a master's degree or a bachelor's degree. Duke University - MHS, 2 years Claudia Brady,MHS, Program Director Department of Pathology PO Box 3712 Durham, NC 27710 919.684.2159 Tara Mann - student liaison 535 Crossview Drive Durham, NC 27703 mann0025@mc.duke.edu Quinnipiac University - MHS, 2 years Scott Farber, Graduate Admissions Director 275 Mount Carmel Ave Hamden, CT 06518 203.582.8795 graduate@quinnipiac.edu ------ Leo Kelly, MHS, Clinical Coordinator (contact for employment opportunities) 203.932.5711 x4758 Ray Schneider - student liaison Norwalk Hospital Dept. of Pathology 24 Stevens St. Norwalk, CT 06856 (203)-852-2657 raymond.schneider@norwalkhealth.org University of Maryland - MS, 2 years Raymond Jones, PhD, Program Director Department of Pathology 22 S Greene St Baltimore, MD 21201 410.328.1221 rjones@som.umaryland.edu Katie Flickinger - student liaison Dept. of Pathology 600 N. Wolfe St. Pathology B 107A Baltimore, MD 21287-6667 (443)-287-6134 kflicki1@jhmi.edu Wayne State University - BS, 2 years Peter Frade, PhD, Program Director Department of Mortuary Sciences 5439 Woodward Ave Detroit, MI 48202 313.577.2050 ab8123@wayne.edu Leena Budev and Anne Hildebrandt - student liasions Anatomic Pathologists' Assistant Program Wayne State University Dept. of Mortuary Science 5439 Woodward Ave. Detroit, MI 48202 (313)-966-0575 lbudev@dmc.org and arh1214@aol.com Ohio State University - MS, 2 years Charles Hitchcock, MD, PhD, Program Director Gretchen Staschiak, Pathology Education Coordinator N-308A Doan Hall 410 W Tenth Ave Columbus, Ohio 43210 614.293.3055 staschiak.2@osu.edu Sandra Banky - student liaison OSU Medical Center 417 East Doan Hall 410 West 10th Avenue Columbus, OH 43210 (614)-293-4875 banky-1@medctr.osu.edu Rosalind Franklin University of Medicine and Science - MS, 2 years John Vitale, MHS, Program Director & Clinical Coordinator Pathologists' Assistant Department 3333 Green Bay Road North Chicago, IL 60064 847.578.8638 john.vitale@rosalindfranklin.edu Trina A.Sherlitz, PT, MS, Clinical Coordinator trina.sherlitz@rosalindfranklin.edu Karen Skish - student liaison (630) 926-9561 karenskish@comcast.net Please Note: Graduates of non-NAACLS accredited training programs are eligible for membership; however, at least one year of work experience is required to become eligible for the AAPA Fellowship Exam. If you have any problems or questions concerning accreditation, please contact: Daniel Tice Staff Program Coordinator dtice@naacls.org or Dr. Olive Kimball Executive Director kimball@naacls.org Further information about NAACLS (National Accrediting Agency for Clinical Laboratory Sciences) may be obtained from www.naacls.org or naaclsinfo@naacls.org. -----Original Message----- From: Jesus Ellin [mailto:JEllin@yumaregional.org] Sent: Monday, February 21, 2005 11:30 AM To: Charles.Embrey; cfockler@mail1.vcu.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Grossing Tech Charles you mentioned that this needs to be done before you start, how can a person go about asking for this once he is in this situation. and were is the PA schools that are specific to Pathologist Assistants?? Jesus Ellin Yuma Regional Medical Center >>> "Charles.Embrey" 02/17/05 09:15AM >>> First, my job description would have to be finalized and pay raise set before I opened my first specimen jar. Once you have started the duties your employer has lost his incentive to get this done. As far as pay raise is concerned it would really depend on the extent of your grossing and time involved. Personally I think the raise negotiations should start at $5.00 for small biopsies and increase with the level of specimen difficulty. The biggest problem I have found is that people tend to underestimate their worth. The only other option for grossing is a pathologist or pathologists' assistant and you would still be a steal with a $5.00 raise. If you enjoy grossing you should apply to one of the PA schools. It is only an additional two years of school, one of which is actual grossing at clinical sites. I teach second year PA students from Rosalind Franklin University in Chicago and my two students that graduate in April have already signed great contracts. Good Luck Charles Embrey PA(ASCP) Urbana, IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cfockler@mail1.vcu.edu Sent: Thursday, February 17, 2005 6:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Grossing Tech Just a quick question.. . what is the average pay increase to go from a standard histotech, to a grossing tech? (with a B.S.) And, how long on average should it take to see it in your job description and your paycheck? Thanks! C. Fockler _______________________________________________ Histonet mailing list Histonet@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 Shawnster73 <@t> aol.com Tue Feb 22 09:44:21 2005 From: Shawnster73 <@t> aol.com (Shawnster73@aol.com) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Block Message-ID: <334C1E06.792809AF.3F5C7C89@aol.com> I am currently trying to stain mouse xenograft tissue using a p53 antibody. I have worked up this antibody on human breast cancer using the Ventana AB block and amplifier and have gotten beautiful results. I am now looking for a way to block and amplify the tissue using reagents that do not contain mouse. Any suggestions would be greatly appreciated. Michelle HT(ASCP) From JWEEMS <@t> sjha.org Tue Feb 22 09:55:09 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Grossing Tech Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507BD1@sjhaexc02.sjha.org> There is also a program at the University of Maryland. j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Charles.Embrey Sent: Tuesday, February 22, 2005 10:42 AM To: Joe Nocito; Jesus Ellin; cfockler@mail1.vcu.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Grossing Tech Hell Joe, you're in San Antonio. Almost anywhere in the U.S. is "up north" to you. The most southern program is at Duke and all the rest are either in New England or near the great lakes. There is nothing west of the Mississippi and I do think that it is because of the population centers. Ohio State and Rosalind Franklin are the newest schools and I am sure more will open because of the increasing demand for PA's. Chuck -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Tuesday, February 22, 2005 8:32 AM To: Charles.Embrey; Jesus Ellin; cfockler@mail1.vcu.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Grossing Tech Chuck, is any reason given as to why all the programs are up north? Is it a supply and demand issue? Joe Nocito -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Charles.Embrey Sent: Monday, February 21, 2005 11:49 AM To: Jesus Ellin; cfockler@mail1.vcu.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Grossing Tech Once you have already started you will probably have to appeal to the Pathologists' sense of fair play. Here are the training sites: Training Programs for Pathologists' Assistants The National Accrediting Agency for Clinical Laboratory Sciences (NAACLS) board of directors voted in 1995 to accept the Essentials of Accredited Training Programs for the Pathologists' Assistant, establishing a process of accreditation for pathologists' assistant training programs. Adoption of NAACLS Essentials ensures a uniform standard of excellence for the training of pathologists assistants. The following training programs have been accredited by NAACLS and provide academic and practical training leading to either a master's degree or a bachelor's degree. Duke University - MHS, 2 years Claudia Brady,MHS, Program Director Department of Pathology PO Box 3712 Durham, NC 27710 919.684.2159 Tara Mann - student liaison 535 Crossview Drive Durham, NC 27703 mann0025@mc.duke.edu Quinnipiac University - MHS, 2 years Scott Farber, Graduate Admissions Director 275 Mount Carmel Ave Hamden, CT 06518 203.582.8795 graduate@quinnipiac.edu ------ Leo Kelly, MHS, Clinical Coordinator (contact for employment opportunities) 203.932.5711 x4758 Ray Schneider - student liaison Norwalk Hospital Dept. of Pathology 24 Stevens St. Norwalk, CT 06856 (203)-852-2657 raymond.schneider@norwalkhealth.org University of Maryland - MS, 2 years Raymond Jones, PhD, Program Director Department of Pathology 22 S Greene St Baltimore, MD 21201 410.328.1221 rjones@som.umaryland.edu Katie Flickinger - student liaison Dept. of Pathology 600 N. Wolfe St. Pathology B 107A Baltimore, MD 21287-6667 (443)-287-6134 kflicki1@jhmi.edu Wayne State University - BS, 2 years Peter Frade, PhD, Program Director Department of Mortuary Sciences 5439 Woodward Ave Detroit, MI 48202 313.577.2050 ab8123@wayne.edu Leena Budev and Anne Hildebrandt - student liasions Anatomic Pathologists' Assistant Program Wayne State University Dept. of Mortuary Science 5439 Woodward Ave. Detroit, MI 48202 (313)-966-0575 lbudev@dmc.org and arh1214@aol.com Ohio State University - MS, 2 years Charles Hitchcock, MD, PhD, Program Director Gretchen Staschiak, Pathology Education Coordinator N-308A Doan Hall 410 W Tenth Ave Columbus, Ohio 43210 614.293.3055 staschiak.2@osu.edu Sandra Banky - student liaison OSU Medical Center 417 East Doan Hall 410 West 10th Avenue Columbus, OH 43210 (614)-293-4875 banky-1@medctr.osu.edu Rosalind Franklin University of Medicine and Science - MS, 2 years John Vitale, MHS, Program Director & Clinical Coordinator Pathologists' Assistant Department 3333 Green Bay Road North Chicago, IL 60064 847.578.8638 john.vitale@rosalindfranklin.edu Trina A.Sherlitz, PT, MS, Clinical Coordinator trina.sherlitz@rosalindfranklin.edu Karen Skish - student liaison (630) 926-9561 karenskish@comcast.net Please Note: Graduates of non-NAACLS accredited training programs are eligible for membership; however, at least one year of work experience is required to become eligible for the AAPA Fellowship Exam. If you have any problems or questions concerning accreditation, please contact: Daniel Tice Staff Program Coordinator dtice@naacls.org or Dr. Olive Kimball Executive Director kimball@naacls.org Further information about NAACLS (National Accrediting Agency for Clinical Laboratory Sciences) may be obtained from www.naacls.org or naaclsinfo@naacls.org. -----Original Message----- From: Jesus Ellin [mailto:JEllin@yumaregional.org] Sent: Monday, February 21, 2005 11:30 AM To: Charles.Embrey; cfockler@mail1.vcu.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Grossing Tech Charles you mentioned that this needs to be done before you start, how can a person go about asking for this once he is in this situation. and were is the PA schools that are specific to Pathologist Assistants?? Jesus Ellin Yuma Regional Medical Center >>> "Charles.Embrey" 02/17/05 09:15AM >>> First, my job description would have to be finalized and pay raise set before I opened my first specimen jar. Once you have started the duties your employer has lost his incentive to get this done. As far as pay raise is concerned it would really depend on the extent of your grossing and time involved. Personally I think the raise negotiations should start at $5.00 for small biopsies and increase with the level of specimen difficulty. The biggest problem I have found is that people tend to underestimate their worth. The only other option for grossing is a pathologist or pathologists' assistant and you would still be a steal with a $5.00 raise. If you enjoy grossing you should apply to one of the PA schools. It is only an additional two years of school, one of which is actual grossing at clinical sites. I teach second year PA students from Rosalind Franklin University in Chicago and my two students that graduate in April have already signed great contracts. Good Luck Charles Embrey PA(ASCP) Urbana, IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cfockler@mail1.vcu.edu Sent: Thursday, February 17, 2005 6:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Grossing Tech Just a quick question.. . what is the average pay increase to go from a standard histotech, to a grossing tech? (with a B.S.) And, how long on average should it take to see it in your job description and your paycheck? Thanks! C. Fockler _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From timothy.m.coskran <@t> pfizer.com Tue Feb 22 10:19:35 2005 From: timothy.m.coskran <@t> pfizer.com (Coskran, Timothy M) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Negative Rabbit Control Message-ID: <599089EF29304C44AF1D84B1C223AC6313EB2E@groamrexm03.amer.pfizer.com> For those using negative Rabbit IgG, how do you determine the dilution of your negative. Are you matching the IgG concentration of your negative rabbit IgG to that of the primary antibody at its working dilution? What do you do when a vendor does not know the IgG concentration of a rabbit polyclonal primary antibody? We try to match the isotype and Ig concentration for negative controls when using monoclonals, but this seems to be a difficult task for rabbit polyclonal antibodies for which you don't have a starting Ig concentration. Tim Coskran Pfizer LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From Lizbeth_Kelly <@t> hgsi.com Tue Feb 22 11:53:09 2005 From: Lizbeth_Kelly <@t> hgsi.com (Lizbeth_Kelly@hgsi.com) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] MSH Seminar - Saturday March 5, 2005 Message-ID: This is posted as an invitation to join the Maryland Society of Histotechnologists by attending a seminar on (Saturday) March 5, 2005 in Glen Burnie, Maryland. This seminar is sponsored by Dakocytomation on "How to Select the Best Immunostaining System for your Laboratory". Incorporated will be detection kits for high sensitivity, double labeling and animal tissue, description in chemistry, the complexity and sensitivity of each and dilutions. Level will be basic to intermediate. Continental breakfast and lunch will be provided by Dakocytomation. Contact hours will be 2.0 (awarded by the National Society of Histotechnology). For Pre-registration please contact Terri DeCarli at 410-787-4546. Lizbeth Kelly, HT(ASCP), Q-IHC Maryland Society of Histotechnologists Board Member 240-314-4400 x 2325 From katri <@t> cogeco.ca Tue Feb 22 12:02:37 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] her-2 nuclear staining References: <022120052322.12388.421A6D2C000EC4BD0000306422070016419D0A9A9C0B@comcast.net> Message-ID: <004a01c51908$b19175a0$6a9a9618@Katri> The cases that you find nuclear staining, how long have they been fixed? My understanding of doing Her-2 is that the tissue has to be fixed minimum 24 hours in 10% NBF (regardless of the size) to avoid false +/- staining. "Underfixation" is generally bigger problem than "overfixation" and I have wittnessed false nuclear staining with some antibodies. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: To: Cc: Sent: Monday, February 21, 2005 6:22 PM Subject: [Histonet] her-2 nuclear staining > Occasionally we have a her-2 with irregular nuclear staining.It seems to > be happening on breast needle cores. Could it be over or under fixation? > If anyone has any information I would greatly appreciate it. D. > Suermann HT(ASCP) > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From traphp <@t> parknicollet.com Tue Feb 22 12:05:18 2005 From: traphp <@t> parknicollet.com (Traphagan, Patricia) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Help Message-ID: We are looking for a formic acid decalcifier for our bone marrows. We have been told that this type of decalcifier works best on bone marrows when a Kappa and Lambda IP stain is requested. We have tried the BBC formic acid decalcifier, but found that the length of time it needed to be in the decal was long (up to 4 fours) and the core was still calcified. Are there any suggest on what could work? Thanks PRIVACY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain business confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If this e-mail was not intended for you, please notify the sender by reply e-mail that you received this in error. Destroy all copies of the original message and attachments. From SCheasty <@t> memorialcare.org Tue Feb 22 12:55:41 2005 From: SCheasty <@t> memorialcare.org (Sandy Cheasty) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Mircowave GMS for Pneumocystis Message-ID: <59FE4C55358C6A42B84BAEC6092CEDBC05ACF5A7@sbnt7> Can someone please recommend a source of reagents and procedure for a GMS to demonstrate pneumocystis? Thanks, Sandy ______________________________________________________________________________ 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. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm From Rachael_Emerson <@t> URMC.Rochester.edu Tue Feb 22 13:12:56 2005 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] alk phos, again Message-ID: Other there other methods to block endogenous alkaline phosphatase? I am using mouse embryos, frozen sectioned. I've tried: levamisole at varying concentrations: still background heated PBS at varying times: too strong, blocked my signal 15% acetic acid at 15": too strong, blocked my signal 5% acetic acid at 15": still background usually I did a combo of levamisole and another treatment. I appreciate any thoughts....... Thanks! Rachael From JWEEMS <@t> sjha.org Tue Feb 22 13:12:31 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Shandon Varistainer 24-3 Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507BE1@sjhaexc02.sjha.org> Does anyone have any parts for this instrument? Thanks, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From GDawson <@t> dynacaremilwaukee.com Tue Feb 22 13:15:48 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] CPT coding for IHC Message-ID: I think it is to avoid confusion. If a pathology report lists 88360 X 6 but we are billing 88342 X 6, problems and confusion may arise. Although the reimbursement is the same, coding everything as 88342's for the technical portion may not be the easy way out. My in-house IHC is billed directly from the pathology report so I don't have the option of coding them myself, so the pathologist determines which are 88360's. I do, however, bill my outside clients myself and I use the 88360 coding where appropriate so that the technical billing matches their reports. Just a thought, Glen Dawson -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, February 22, 2005 9:16 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CPT coding for IHC Are any of you using the new CPT code (88360) for IHC/semiquantitative analysis? If so, why is 88360 used on the technical side as well (there is no difference in the amount of technical work performed)? The only difference I see is on the professional side with the interpretation. Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barb.Richmond <@t> mckennan.org Tue Feb 22 13:19:58 2005 From: Barb.Richmond <@t> mckennan.org (Barb Richmond) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] stain for calcium oxalate on paraffin Message-ID: <9953A44C9B142646860F379C8EBC891109EFFB18@msexchange.phs-sfalls.amck.net> We are looking for a stain that we can use on a paraffin embedded renal biopsy to determine oxalate crystals. Does anyone know of any?? ____________________ 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 akbitting <@t> geisinger.edu Tue Feb 22 13:22:44 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] JCAHO vs CAP Message-ID: I am working with a lab that dropped their CAP accreditation. They're up for JCAHO inspection soon and I'm trying to help them get ready. I understand that JCAHO will defer to CAP. I've been getting them ready by following CAPs checklist, but is there something I might miss for a JCAHO inspection? Anyone have experience with the differences? 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 joeamateur <@t> hotmail.com Tue Feb 22 14:27:53 2005 From: joeamateur <@t> hotmail.com (Jack England) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] need manual for AO 855 cryostat Message-ID: Aloha Histonetters, Is there any chance anyone out there can email/fax/sell me a copy of the manual for an AO 855 cryostat? My lab just bought one (without a manual) and want me to make it dance. Right now, I'm at the poke-it-with-a-stick stage, since I have no experience whatsoever with cryosectioning. Can any of you kind people help? --Aloha, Jack England Tissue Genesis, Inc. http://www.tissuegenesis.com _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From hborgeri <@t> wfubmc.edu Tue Feb 22 14:18:06 2005 From: hborgeri <@t> wfubmc.edu (Hermina Borgerink) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] stain for calcium oxalate on paraffin Message-ID: <9AEEF1FB6254224AA355ED285F8491650999CB50@EXCHVS2.medctr.ad.wfubmc.edu> Pizzolato's stain is generally used for the demonstration of calcium oxalate crystals in paraffin. The complete technique is in Theory and Practice of Histotechnology by Sheehan and Hrapchak, p. 228. 1. Hydrate slides 2. Mix equal volumes of 30% hydrogen peroxide and 5% silver nitrate. 3. Flood slides with the mixture and expose to a 60-watt tungsten-filament electric lamp or a 25-watt fluorescent bulb. The light should be approximately 15 cm above the sections. Exposure varies from 15 to 30 minutes. If excessive bubbling is observed re place the regent mixture with a fresh one. 4. Rinse thoroughly in distilled or deionized water. 5. Counterstain with 0.1% safranin in 1% acetic acid for 2-3 minutes. 6. Rinse in distilled or deionized water, dehydrate, clear and coverslip. Calcium oxalate - black Nuclei - red Note: Calcium fluoride and calcium and barium sulfates do not react by this method. Hermina Hermina M. Borgerink, BA, HTL(ASCP)QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax. (336) 716-1515 e-mail hborgeri@wfubmc.edu "Treat a man as he appears to be, and you make him worse. But treat a man as if he already were what he potentially could be, and you make him what he should be." (Goethe) This electronic message, including any attachments, is confidential and proprietary and is solely for the intended recipient. If you are not the intended recipient, this message was sent to you in error and you are hereby advised that any review, disclosure, copying, distribution or use of this message, or any of the information included therein, is unauthorized and strictly prohibited. If you have received this electronic transmission in error, please immediately notify the sender by reply and permanently delete all copies of this message and its attachments. Thank you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Tuesday, February 22, 2005 2:20 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] stain for calcium oxalate on paraffin We are looking for a stain that we can use on a paraffin embedded renal biopsy to determine oxalate crystals. Does anyone know of any?? ____________________ 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 Kathy.Paton <@t> waitematadhb.govt.nz Tue Feb 22 16:23:43 2005 From: Kathy.Paton <@t> waitematadhb.govt.nz (Kathy Paton (WDHB)) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Left-handed microtome. Message-ID: I couldn't agree more. I have always thought that the microtome was invented by a true blue lefty. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ian Montgomery Sent: Wednesday, 23 February 2005 00:05 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Left-handed microtome. As a left hander I've always found a microtome to be ideal. You turn the handle with your right hand then manipulate the sections with your left, the dominant hand. What could be better? Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jstaruk <@t> masshistology.com Tue Feb 22 17:23:45 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Left-handed microtome. In-Reply-To: Message-ID: <0ICC0099V6AUF8B1@vms040.mailsrvcs.net> I remember an older histologist I met in Florida who had the microtome facing away from him as he cut ribbons! Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathy Paton (WDHB) Sent: Tuesday, February 22, 2005 5:24 PM To: Ian Montgomery; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Left-handed microtome. I couldn't agree more. I have always thought that the microtome was invented by a true blue lefty. (snip) From histology.bc <@t> shaw.ca Tue Feb 22 19:02:20 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] stain for calcium oxalate on paraffin In-Reply-To: <9953A44C9B142646860F379C8EBC891109EFFB18@msexchange.phs-sfalls.amck.net> References: <9953A44C9B142646860F379C8EBC891109EFFB18@msexchange.phs-sfalls.amck.net> Message-ID: <421BD61C.6090300@shaw.ca> The quickest and easiest approach is to view your existing H&E sections using a polarized light microscope. Calcium oxalate crystals are strongly birefringest and are impossible to miss. Would I be correct to assume you have sections of kidney from an ethylene glycol (antifreeze) poisoning? The "stains" suggested in some texts are essentially modifications of von Kossa's silver substitution method for the calcium salts usually associated with bone or chronic inflammatory reactions. They work, but are not specific for calcium oxalate as they will also react with any other insoluble calcium salts that happen to be present. Give the polarizer a try, I think you will be happy with the results. Best of luck, Paul Bradbury Kamloops, BC, Canada From jkiernan <@t> uwo.ca Tue Feb 22 22:32:27 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] stain for calcium oxalate on paraffin References: <9953A44C9B142646860F379C8EBC891109EFFB18@msexchange.phs-sfalls.amck.net> <421BD61C.6090300@shaw.ca> Message-ID: <421C075B.1D83B3C5@uwo.ca> Paul's advice is very good. I've never seen an oxalosis kidney, but I have looked at sections of bits of plants; they often contain layers of cells that are almost filled with star-like growths of calcium oxalate. Each limb of a star is bright with crossed polars, and the birefringence changes as you rotate the stage. The principal von Kossa variant for Ca oxalate is that of Pizzolato (1964). Sections are treated with 1% silver nitrate in 15% (Yes! 15%) hydrogen peroxide in bright light. This must be done after removing calcium carbonate and phosphate deposits (12% acetic acid, 15 minutes at 22C; a warm room temp). If the acid treatment is omitted, water-insoluble carbonates and phosphates also yield black deposits of silver. Pure calcium oxalate does not give a dark deposit with the usual von Kossa method. Refs. Pizzolato P (1964) J Histochem Cytochem 12: 333 Pearse AGE (1985) Histochemistry Theoretical and Applied, 4th edn, Vol 2. Chapter 20. Plant histology books, especially by Berlyn and by Ruzin. John Kiernan Dept of anatomy & Cell Biology University of Western Ontario London, Canada. ---------------------------------- Paul Bradbury wrote: > > The quickest and easiest approach is to view your existing H&E sections > using a polarized light microscope. Calcium oxalate crystals are > strongly birefringest and are impossible to miss. Would I be correct to > assume you have sections of kidney from an ethylene glycol (antifreeze) > poisoning? > > The "stains" suggested in some texts are essentially modifications of > von Kossa's silver substitution method for the calcium salts usually > associated with bone or chronic inflammatory reactions. They work, but > are not specific for calcium oxalate as they will also react with any > other insoluble calcium salts that happen to be present. > > Give the polarizer a try, I think you will be happy with the results. > > Best of luck, > > Paul Bradbury > Kamloops, BC, > Canada > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From swaram <@t> myrealbox.com Wed Feb 23 00:03:10 2005 From: swaram <@t> myrealbox.com (Swaram) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Telomerase Ab for Paraffin tissue In-Reply-To: <9953A44C9B142646860F379C8EBC891109EFFB18@msexchange.phs-sfalls.amck.net> References: <9953A44C9B142646860F379C8EBC891109EFFB18@msexchange.phs-sfalls.amck.net> Message-ID: <421C1C9E.8050705@myrealbox.com> Hi Histonetters, I am working on Telomerase (hTERT) IHC on paraffin fixed tissue. I just wanted to know if there are problems with Novocastra's NCL-hTERT antibody. I heard that it may be non-specific for Telomerase. Does anyone have experience using fluorescent IHC with hTERT on paraffin or fixed cells. I would also be very happy if someone can send me a picture of how it looks. I find that it is localized in the nucleolus with some cells also showing a nucleoplasmic localization in addition to the nucleolus. Is this correct..? Thanks Swaram -- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.300 / Virus Database: 266.3.0 - Release Date: 2/21/2005 From pex0220 <@t> yahoo.com.cn Wed Feb 23 05:00:34 2005 From: pex0220 <@t> yahoo.com.cn (=?gb2312?q?=CC=EC=20=D0=C1?=) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] hello Message-ID: <20050223110034.30928.qmail@web15508.mail.cnb.yahoo.com> hello,Histonetter, I am really nice to send a Email to you! I am having some difficulties in my experiments. At present, I do double-immunofluorescence in bone tissue, but I find that it is not easy,including embedding, cutting, staining and microscopy. If someone gains some experience about it, I hope that he or she can give me some helps( for example, providing a protocol about double immunofluorescence, or give some advice about it). If ok, I will very happy! I am looking forward for you. Thanks very much! Qian --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From Don.Birgerson <@t> leica-microsystems.com Wed Feb 23 08:13:32 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] need manual for AO 855 cryostat Message-ID: Hi Jack, I can get you a copy of the operating manual. American Optical (AO), Reichert, and Cambridge are some of the companies that make up Leica-Microsystems. The AO 855 is the outer cabinet for the 975C Cryocut cryostat. The 855 cabinet +840 microtome = AO 975C Cryostat. Give me a call on the 800 number. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 7:00 to 4:00 CT "Jack England" To: Histonet@lists.utsouthwestern.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] need manual for AO 855 cryostat western.edu 02/22/2005 02:27 PM Aloha Histonetters, Is there any chance anyone out there can email/fax/sell me a copy of the manual for an AO 855 cryostat? My lab just bought one (without a manual) and want me to make it dance. Right now, I'm at the poke-it-with-a-stick stage, since I have no experience whatsoever with cryosectioning. Can any of you kind people help? --Aloha, Jack England Tissue Genesis, Inc. http://www.tissuegenesis.com _________________________________________________________________ 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 ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From froyer <@t> bitstream.net Wed Feb 23 08:43:48 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:39 2005 Subject: [Histonet] Left-handed microtome. In-Reply-To: <0ICC0099V6AUF8B1@vms040.mailsrvcs.net> References: <0ICC0099V6AUF8B1@vms040.mailsrvcs.net> Message-ID: <421C96A4.4050903@bitstream.net> Jim Staruk wrote: >I remember an older histologist I met in Florida who had the microtome >facing away from him as he cut ribbons! > >Jim > >______________________ > Jim Staruk >Mass Histology Service >www.masshistology.com > Poor fellow... didn't know whether he was coming or going. Obviously due to all those years of unbridled exposure to xylene & formalin (or is that formaldehyde? ...I can never remember). ;-) ~ Ford Ford M. Royer, MT(ASCP) Midwest Science Biocenter, Inc. Minneapolis, MN 800-745-4869 From Rosemary_Dunn <@t> ssmhc.com Wed Feb 23 09:47:09 2005 From: Rosemary_Dunn <@t> ssmhc.com (Rosemary_Dunn@ssmhc.com) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Motorized cryostat Message-ID: I would like feedback from anyone using the CRYOTOME SME THERMAL ELECTRON motorized cryostat. We are looking to purchase one and I would appreciate hearing from someone currently using it. Thank you, Rose From funderwood <@t> mcohio.org Wed Feb 23 10:01:53 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:40 2005 Subject: [BULK] - Re: [Histonet] Left-handed microtome. Message-ID: Or, perhaps the pathologist wanted sections from the opposite side of the block. >>> Ford Royer 02/23/05 09:43AM >>> Jim Staruk wrote: >I remember an older histologist I met in Florida who had the microtome >facing away from him as he cut ribbons! > >Jim > >______________________ > Jim Staruk >Mass Histology Service >www.masshistology.com > Poor fellow... didn't know whether he was coming or going. Obviously due to all those years of unbridled exposure to xylene & formalin (or is that formaldehyde? ...I can never remember). ;-) ~ Ford Ford M. Royer, MT(ASCP) Midwest Science Biocenter, Inc. Minneapolis, MN 800-745-4869 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Wed Feb 23 10:36:25 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Polka dots again Message-ID: I must have tempted fate mentioning pink disease about one week ago. We are in the second day of the worst outbreak I have seen by a country mile. Virtually every section is affected. Yuck. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk From RBARNHART <@t> summithealth.org Wed Feb 23 12:17:29 2005 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Polka dots again Message-ID: Maybe it is like a virus and we passed it on to you. Sorry :( As long as our water bath temperature doesn't get to high the polka dots stay away. >>> "Marshall Terry Dr,Consultant Histopathologist" 2/23/2005 11:36:25 AM >>> I must have tempted fate mentioning pink disease about one week ago. We are in the second day of the worst outbreak I have seen by a country mile. Virtually every section is affected. Yuck. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RossS <@t> BaylorHealth.edu Wed Feb 23 12:31:25 2005 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:24:40 2005 Subject: [BULK] - Re: [Histonet] Left-handed microtome. Message-ID: <74A8B7A904152141BD5AA2761F5D22340E4806@BHDAEXCH11.bhcs.pvt> I had a resident come in looking at at a block contemplating ordering a special stain. She looked and looked at the top of the block where tissue is placed when grossing. Finally she said. "I need to order a special on this block, but there isn't any more tissue." The tech took the block out of her hand and turned it over. "OOOHHH, there it is." Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Wednesday, February 23, 2005 10:02 AM To: froyer@bitstream.net; histonet@lists.utsouthwestern.edu Subject: [BULK] - Re: [Histonet] Left-handed microtome. Or, perhaps the pathologist wanted sections from the opposite side of the block. >>> Ford Royer 02/23/05 09:43AM >>> Jim Staruk wrote: >I remember an older histologist I met in Florida who had the microtome >facing away from him as he cut ribbons! > >Jim > >______________________ > Jim Staruk >Mass Histology Service >www.masshistology.com > Poor fellow... didn't know whether he was coming or going. Obviously due to all those years of unbridled exposure to xylene & formalin (or is that formaldehyde? ...I can never remember). ;-) ~ Ford Ford M. Royer, MT(ASCP) Midwest Science Biocenter, Inc. Minneapolis, MN 800-745-4869 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From pathrm35 <@t> adelphia.net Wed Feb 23 12:38:47 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Fisher rep Message-ID: <1452316.1109183927552.JavaMail.root@web5.mail.adelphia.net> Does anyone know who the Fisher rep for southeast Florida is? From petepath <@t> yahoo.com Wed Feb 23 12:43:09 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] oxalate crystals Message-ID: <20050223184309.76376.qmail@web30403.mail.mud.yahoo.com> Calcium oxylate crystals are a common finding in tissues which most experienced pathologists will recognize on H & E. As Paul Bradbury suggests they will polarize brightly. On H & E the have a slight browinsh cast and will show radial striations when polarized. If you are looking at ethylene glycol ( antifreeze) poisoning they will be very numerous in tubular lumina. If you want a control they are very common in normal and diseased thyroid, in inceasing frequency with age. More than half of patients over 60 will have many of them. Try polarizing a piece of thyroid if you want to see them. Stephen Peters M.D. Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 Attending Pathologist Hackensack University Medical Center 201 996 4836 From Kemlo.Rogerson <@t> elht.nhs.uk Wed Feb 23 12:52:17 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Polka dots again[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F006@bhrv-nt-11.bhrv.nwest.nhs.uk> I never knew how you knew my middle name was Richard!! Amazing....... -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 23 February 2005 16:36 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Polka dots again[Scanned] I must have tempted fate mentioning pink disease about one week ago. We are in the second day of the worst outbreak I have seen by a country mile. Virtually every section is affected. Yuck. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RossS <@t> BaylorHealth.edu Wed Feb 23 12:57:58 2005 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Histogel cutting tips Message-ID: <74A8B7A904152141BD5AA2761F5D22340E4807@BHDAEXCH11.bhcs.pvt> We are using histogel to keep some thin liver biopsies from breaking apart during processing/embedding. When the techs go to cut the slides, the tissue pops out of the block on a pretty regular basis. Has anyone else had this problem? Anyone have any tips or solutions? Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From tsanger <@t> biology2.wustl.edu Wed Feb 23 13:04:06 2005 From: tsanger <@t> biology2.wustl.edu (Thomas Sanger) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] plant embedding and sectioning tips Message-ID: <8F939618D1D01941B8EC772D5813FEF309C06B@biology2.bioserver.wustl.edu> Dear all, A colleague of mine wants to take 2-5 micrometer thick sections of soft plant roots but I have no experience working with these tissues. Can anyone direct me how to do this? The proper embedding material? Described protocols? Tricks of the trade? Thanks, Thom Sanger tsanger@biology2.wustl.edu From TJJ <@t> Stowers-Institute.org Wed Feb 23 13:26:24 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Automated immunostainer for research applications Message-ID: Hi all (and apologies to those on both lists), For those of you doing research histology and immunohistochemistry, which automated immunostainer do you use? Do you do any overnight incubations with it? For those of you who have automated stainers and formerly did overnight (4C) incubations, what modifications did you make to your staining protocol to achieve the same result? Also, do you always use the substrate on instrument, or do you do that via microscope? Some immunostainers require minimum amounts of primary antibody (~500 microliters), even if staining only one slide. Some times we don't have that much antibody to use. Are there any tips to get around this with the exception of manually pipetting as if doing a titration run? Thanks for your input! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From SCheasty <@t> memorialcare.org Wed Feb 23 13:27:33 2005 From: SCheasty <@t> memorialcare.org (Sandy Cheasty) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Mircowave GMS for Pneumocystis Additional Info Message-ID: <59FE4C55358C6A42B84BAEC6092CEDBC05ACF5AC@sbnt7> We are having trouble using our non-microwave GMS method with the microwave. For fungus it is fine, but for the longer pneumocystis time we get too much back ground precipitate. I'm looking for a clean microwave GMS pneumocystis procedure. If you have a good method in your lab, please include the reagent vendor... Thanks, Sandy ______________________________________________________________________________ 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. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm From liz <@t> premierlab.com Wed Feb 23 13:28:15 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] alcohol fixation and confocal microscopy Message-ID: <000001c519dd$d5247410$76d48a80@AMY> Hello everyone Some one had told me that alcohol fixation will not work for confocal microscopy. Is there any truth to this? I know that formaldehyde and paraformaldehye fixatives lead to autofluroescence, so why would alcohol fixation cause problems. 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 tpmorken <@t> labvision.com Wed Feb 23 14:22:39 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: pathologists in the lab (shivers down the spine...) was "left handed microtome" Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CA47@usca0082k08.labvision.apogent.com> "OOOHHH, there it is." Oh, the days in the lab... My favorite is the pathologist who came in late in the evening, opened the (already running) tissue processor and then spent two hours grossing, occasionally lobbing a cassette into the processor. Next day he (and all the other pathologists) was mad that his blocks were were two hours late - but soon he was hiding under his desk after he explained what he had done and all the other pathologists turned on him. Luckily, only his blocks were wrecked - he had lobbed them in while the processor was late in 100%, and did not fully dehydrate. It is truly amazing how ignorant most pathology residents are about basic histology procedures. Our lab director was enlightened and he made all the path residents spend 5 full days in the histo lab following the tissue through the entire process from accession to H&E, then specials, IHC, EM etc. They were chaffing by the second day, but by the 5th they were very, very appreciative of all the work that went on to get their slides and stains done. It made life a lot easier later because they actually understand what was going on - and knew it was better to ask a tech what to do rather than just blindly (arrogantly?) do what ever they felt like doing. Tim Morken I had a resident come in looking at at a block contemplating ordering a special stain. She looked and looked at the top of the block where tissue is placed when grossing. Finally she said. "I need to order a special on this block, but there isn't any more tissue." The tech took the block out of her hand and turned it over. "OOOHHH, there it is." Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Wednesday, February 23, 2005 10:02 AM To: froyer@bitstream.net; histonet@lists.utsouthwestern.edu Subject: [BULK] - Re: [Histonet] Left-handed microtome. Or, perhaps the pathologist wanted sections from the opposite side of the block. >>> Ford Royer 02/23/05 09:43AM >>> Jim Staruk wrote: >I remember an older histologist I met in Florida who had the microtome >facing away from him as he cut ribbons! > >Jim > >______________________ > Jim Staruk >Mass Histology Service >www.masshistology.com > Poor fellow... didn't know whether he was coming or going. Obviously due to all those years of unbridled exposure to xylene & formalin (or is that formaldehyde? ...I can never remember). ;-) ~ Ford Ford M. Royer, MT(ASCP) Midwest Science Biocenter, Inc. Minneapolis, MN 800-745-4869 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JosefaNava <@t> texashealth.org Wed Feb 23 15:31:23 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] "Premiere" charge slides Message-ID: <2C515C1049EAF5459EFD8C9B929078A4194474@phdex03.txhealth.org> Hello there, Has anyone been using this brand of charged slides "PREMIERE" to do their immuno stains using Ventana immuno machines. I will appreciate any feedback you can give me regarding this brand. Thank you. Josie The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From la.sebree <@t> hosp.wisc.edu Wed Feb 23 16:00:01 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] "Premiere" charge slides Message-ID: We just (today) received a sample of these to try so we also would like to hear how they fair on VMS instruments. 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 Nava, Josefa Sent: Wednesday, February 23, 2005 3:31 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] "Premiere" charge slides Hello there, Has anyone been using this brand of charged slides "PREMIERE" to do their immuno stains using Ventana immuno machines. I will appreciate any feedback you can give me regarding this brand. Thank you. Josie The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From petepath <@t> yahoo.com Wed Feb 23 16:18:15 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] pathologists in the lab Message-ID: <20050223221815.63753.qmail@web30403.mail.mud.yahoo.com> I could not agree with you more. A generous histotech taught me how to cut and stain as a resident and I ended up cutting a handful of my autopsies. It taught me a great deal about the problems a histotech faces and insite into the artifacts I was producing my self. This also helped me to become a better frozen sectionist. Never take for granted that patholgists understand the details in histology. Some do, many don't. It can get you into trouble. And be care trusting them to load the processor. I would be happy if all my residents could trim a block to a unifom 3 mm thickness! Stephen Peters M.D. Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 Attending Pathologist Hackensack University Medical Center 201 996 4836 From kgrobert <@t> rci.rutgers.edu Wed Feb 23 16:34:38 2005 From: kgrobert <@t> rci.rutgers.edu (kgrobert@rci.rutgers.edu) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: pathologists in the lab (shivers down the spine...) was "left handed microtome" In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328CA47@usca0082k08.labvision.apogen t.com> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328CA47@usca0082k08.labvision.apogent.com> Message-ID: <15483.128.6.148.191.1109198078.squirrel@webmail.rci.rutgers.edu> We're just a small research and teaching lab, but I have taught all sorts of people about histology-from high-schoolers on up to full professors, and every time my boss teaches his Toxicologic Pathology graduate course, I get to teach the lab portion of it, where the students learn almost the same thing-from animal necropsy all the way to staining. So why isn't Histology part of Medical School/Pathology training? Kathleen Principal Lab Technician Neurotoxicology Labs Rutgers University > "OOOHHH, there it is." > > Oh, the days in the lab... My favorite is the pathologist who came in late > in the evening, opened the (already running) tissue processor and then > spent > two hours grossing, occasionally lobbing a cassette into the processor. > Next > day he (and all the other pathologists) was mad that his blocks were were > two hours late - but soon he was hiding under his desk after he explained > what he had done and all the other pathologists turned on him. Luckily, > only > his blocks were wrecked - he had lobbed them in while the processor was > late > in 100%, and did not fully dehydrate. > > It is truly amazing how ignorant most pathology residents are about basic > histology procedures. Our lab director was enlightened and he made all the > path residents spend 5 full days in the histo lab following the tissue > through the entire process from accession to H&E, then specials, IHC, EM > etc. They were chaffing by the second day, but by the 5th they were very, > very appreciative of all the work that went on to get their slides and > stains done. It made life a lot easier later because they actually > understand what was going on - and knew it was better to ask a tech what > to > do rather than just blindly (arrogantly?) do what ever they felt like > doing. > > Tim Morken > > > > > I had a resident come in looking at at a block contemplating ordering a > special stain. She looked and looked at the top of the block where tissue > is placed when grossing. > > Finally she said. "I need to order a special on this block, but there > isn't > any more tissue." > > The tech took the block out of her hand and turned it over. > > > "OOOHHH, there it is." > > Ross M Stapf > Histopathology Manager > Baylor University Medical Center > 3500 Gaston Ave. > Dallas, TX 75246 > 214-820-2465 > > 214-820-4110 fax > RossS@baylorhealth.edu > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred > Underwood > Sent: Wednesday, February 23, 2005 10:02 AM > To: froyer@bitstream.net; histonet@lists.utsouthwestern.edu > Subject: [BULK] - Re: [Histonet] Left-handed microtome. > > > Or, perhaps the pathologist wanted sections from the opposite side of the > block. > >>>> Ford Royer 02/23/05 09:43AM >>> > Jim Staruk wrote: > >>I remember an older histologist I met in Florida who had the > microtome >>facing away from him as he cut ribbons! >> >>Jim >> >>______________________ >> Jim Staruk >>Mass Histology Service >>www.masshistology.com >> > Poor fellow... didn't know whether he was coming or going. Obviously due > to > all those years of unbridled exposure to xylene & > > formalin (or is that formaldehyde? ...I can never remember). > > ;-) ~ Ford > Ford M. Royer, MT(ASCP) > Midwest Science Biocenter, Inc. > Minneapolis, MN > 800-745-4869 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > This e-mail, facsimile, or letter and any files or attachments transmitted > with it contains information that is confidential and privileged. This > information is intended only for the use of the individual(s) and > entity(ies) to whom it is addressed. If you are the intended recipient, > further disclosures are prohibited without proper authorization. If you > are > not the intended recipient, any disclosure, copying, printing, or use of > this information is strictly prohibited and possibly a violation of > federal > or state law and regulations. If you have received this information in > error, please notify Baylor Health Care System immediately at > 1-866-402-1661 > or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its > subsidiaries, and affiliates hereby claim all applicable privileges > related > to this information. > > _______________________________________________ > Histonet mailing list > Histonet@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 TheBestTime23 <@t> aol.com Wed Feb 23 16:42:03 2005 From: TheBestTime23 <@t> aol.com (TheBestTime23@aol.com) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] richard-allen type 9 paraffin Message-ID: Hello everyone, The lab that I work for recently decided to switch to Richard-Allen type 9 paraffin. I was wondering if anyone has experience with this paraffin and could give some advice on techniques used in cutting it. Opinions of this product would also be greatly appreciated. Thanks in advance, Megan From madbaza <@t> powerup.com.au Wed Feb 23 16:42:15 2005 From: madbaza <@t> powerup.com.au (Barry Madigan) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Formalin Fixation times for IHC samples In-Reply-To: <004301c51497$8d44f110$6a9a9618@Katri> Message-ID: <20050223224218.HQYY20373.smta07.mail.ozemail.net@WORKSTATION1> I find the major impediment to good quality Immunohistochemistry is turn around times, which has been a buzz word here in Australia for a number of years. The result of this management tool has led to poorly fixed and processed tissue samples that are unable to withstand the harsh treatment of heat retrieval. Thanks to Murphy's Law, we usually find that the most vital block for diagnosis is always the one that is poorly preserved. We receive material for Immunohistochemistry from a large number of hospitals in Queensland (Australia). Unfortunately there is no uniformity in the way specimens are fixed or processed by these referring laboratories. Both lymph nodes and breast tissue are the major specimens affected by under fixation. With lymph nodes the peripheral staining is excellent but the morphology of the centre is compromised. The central structures of these specimens have not been strengthened by fixation and as a result the morphology is damaged by heat retrieval. With breast specimens there is an increase risk of tissue lifting from the slide. Try to convince Pathologists that fixation is the problem is just about impossible. We place control material, which has received at least 24 hours fixation on the same slide. The control material is always perfect and the Pathologist always concurs but questions why the test morphology is damaged. It's great to get pathology reports out quickly but at what diagnostic cost. How many other laboratories experience this problem? Barry Madigan Brisbane, Queensland, Australia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Katri Tuomala Sent: Thursday, 17 February 2005 12:23 PM To: Anne C Lewin; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Formalin Fixation times for IHC samples Anne, I think you are doing well with your protocol. Although I don't think "overfixation" is as a big problem any more with the advent of different HIER protocols with various buffers being used. Under fixation (less than 24 hours) can be a bigger problem with certain antibodies like Her-2, ER and PR to mention few. This concept of over fixation is a remnant from the time prior to HIER, when we all had a problem trying to demonstrate many antigens with weaker retrievals (proteolytic enzymes). The heat over 60C was actually banned from any immunohistochemistry protocols 20 years ago. My two cents worth... Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Anne C Lewin" To: Sent: Tuesday, February 15, 2005 10:28 AM Subject: [Histonet] Formalin Fixation times for IHC samples > Could I get a general concensus from Histoland about the amount of time > needed to fix a sample for IHC without over-fixing? My training/journal > searches have led me to believe that the longer a sample is in > formalin, the stronger the protiens are cross-linked, and the more > difficult it is to break those bonds with antigen retreival and get your > antibody to the target. I generally tell the scientists in my > department to fix overnight (most samples vary from about the size of a > pea to a lima bean), and then to transfer to 70% Ethanol for me to > process for paraffin. The total time in fixative tends to be around 24 > hours. Morphology has always been great, and my IHC's work well. This > question is mainly for my information, since I have used the same tissue > prep protocol for a few years now, I want to make sure I am keeping up > with current opinions. > Thanks! > -Anne > ---------------------------------------------------------------------------- ---- > _______________________________________________ > Histonet mailing list > Histonet@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 Wed Feb 23 16:58:54 2005 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Cell Block Systems Message-ID: <5CB39BCA5724F349BCB748675C6CA1A202C75663@SJMEMXMB02.stjude.sjcrh.local> Hello Everyone, Does anyone know of a cell block system that can capture a small amount of cells for a FFPE block? We currently have the HistoGel system and it works great when plenty of cells are present however it does not work well with minute cells. Thanks, 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 From mcauliff <@t> umdnj.edu Wed Feb 23 20:04:03 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: pathologists in the lab (shivers down thespine...) In-Reply-To: <15483.128.6.148.191.1109198078.squirrel@webmail.rci.rutgers.edu> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328CA47@usca0082k08.labvision.apogent.com> <15483.128.6.148.191.1109198078.squirrel@webmail.rci.rutgers.edu> Message-ID: <421D3613.9060003@umdnj.edu> Two reasons for not training medical students: 1. Not enough time in the curriculum. 2. Most medical students won't be pathologists, so why train them to process tissue and stain slides? Those who train in Pathology should know what is involved in tissue preparation and know enough not to demonstrate their ignorance. Geoff kgrobert@rci.rutgers.edu wrote: >We're just a small research and teaching lab, but I have taught all sorts >of people about histology-from high-schoolers on up to full professors, >and every time my boss teaches his Toxicologic Pathology graduate course, >I get to teach the lab portion of it, where the students learn almost the >same thing-from animal necropsy all the way to staining. > >So why isn't Histology part of Medical School/Pathology training? > >Kathleen >Principal Lab Technician >Neurotoxicology Labs >Rutgers University > > > > > > >>"OOOHHH, there it is." >> >>Oh, the days in the lab... My favorite is the pathologist who came in late >>in the evening, opened the (already running) tissue processor and then >>spent >>two hours grossing, occasionally lobbing a cassette into the processor. >>Next >>day he (and all the other pathologists) was mad that his blocks were were >>two hours late - but soon he was hiding under his desk after he explained >>what he had done and all the other pathologists turned on him. Luckily, >>only >>his blocks were wrecked - he had lobbed them in while the processor was >>late >>in 100%, and did not fully dehydrate. >> >>It is truly amazing how ignorant most pathology residents are about basic >>histology procedures. Our lab director was enlightened and he made all the >>path residents spend 5 full days in the histo lab following the tissue >>through the entire process from accession to H&E, then specials, IHC, EM >>etc. They were chaffing by the second day, but by the 5th they were very, >>very appreciative of all the work that went on to get their slides and >>stains done. It made life a lot easier later because they actually >>understand what was going on - and knew it was better to ask a tech what >>to >>do rather than just blindly (arrogantly?) do what ever they felt like >>doing. >> >>Tim Morken >> >> >> >> >>I had a resident come in looking at at a block contemplating ordering a >>special stain. She looked and looked at the top of the block where tissue >>is placed when grossing. >> >>Finally she said. "I need to order a special on this block, but there >>isn't >>any more tissue." >> >>The tech took the block out of her hand and turned it over. >> >> >>"OOOHHH, there it is." >> >>Ross M Stapf >>Histopathology Manager >>Baylor University Medical Center >>3500 Gaston Ave. >>Dallas, TX 75246 >>214-820-2465 >> >>214-820-4110 fax >>RossS@baylorhealth.edu >> >> >> >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred >>Underwood >>Sent: Wednesday, February 23, 2005 10:02 AM >>To: froyer@bitstream.net; histonet@lists.utsouthwestern.edu >>Subject: [BULK] - Re: [Histonet] Left-handed microtome. >> >> >>Or, perhaps the pathologist wanted sections from the opposite side of the >>block. >> >> >> >>>>>Ford Royer 02/23/05 09:43AM >>> >>>>> >>>>> >>Jim Staruk wrote: >> >> >> >>>I remember an older histologist I met in Florida who had the >>> >>> >>microtome >> >> >>>facing away from him as he cut ribbons! >>> >>>Jim >>> >>>______________________ >>> Jim Staruk >>>Mass Histology Service >>>www.masshistology.com >>> >>> >>> >>Poor fellow... didn't know whether he was coming or going. Obviously due >>to >>all those years of unbridled exposure to xylene & >> >>formalin (or is that formaldehyde? ...I can never remember). >> >>;-) ~ Ford >>Ford M. Royer, MT(ASCP) >>Midwest Science Biocenter, Inc. >>Minneapolis, MN >>800-745-4869 >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >> >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >>This e-mail, facsimile, or letter and any files or attachments transmitted >>with it contains information that is confidential and privileged. This >>information is intended only for the use of the individual(s) and >>entity(ies) to whom it is addressed. If you are the intended recipient, >>further disclosures are prohibited without proper authorization. If you >>are >>not the intended recipient, any disclosure, copying, printing, or use of >>this information is strictly prohibited and possibly a violation of >>federal >>or state law and regulations. If you have received this information in >>error, please notify Baylor Health Care System immediately at >>1-866-402-1661 >>or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its >>subsidiaries, and affiliates hereby claim all applicable privileges >>related >>to this information. >> >>_______________________________________________ >>Histonet mailing list >>Histonet@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 > > > > -- -- ********************************************** 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 bills <@t> icpmr.wsahs.nsw.gov.au Wed Feb 23 17:08:30 2005 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Formalin Fixation times for IHC samples In-Reply-To: <20050223224218.HQYY20373.smta07.mail.ozemail.net@wsahs.nsw.gov.au> Message-ID: <000201c519fc$97e09ce0$0ecd080a@wsahs.nsw.gov.au> Hi Barry, I couldn't agree more. We are a major centre for NSW Breast Screen Western and The Breast Cancer Institute and the surgeons perform their operations on Wednesdays and Thursdays then have a meeting Monday and Tuesday and expect the results including Immunohistochemistry at these meetings. Fixation for most specimens is minimal at best and the results show poorly on Immunohistochemistry. As you said incomplete fixation means sections looking tatty or falling off. The positive controls, usually collected after the specimen has fixed for at least another 24 hours, always have great morphology and staining. We find it is very much surgeon driven as they want results for meetings for discussion about course of treatment and for discusion with patients. Many do not understand the importance of good fixation to the diagnostic process. Automation (Tissue Processors, Staining Machines, Automated coverslippers and automated Immunohistochemistry) helps a little with TAT but these are only some facets of the total process which begins with fixation. Microwave fixation helps but is not the answer to all the problems. Until someone discovers a revolutionary new fixation method I will still stick to "good old formaldehyde". I have tried several other formaldehyde substitutes over many years but the pathologists still prefer the known artefacts produced by formaldehyde. Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Barry Madigan Sent: Thursday, 24 February 2005 9:42 AM To: Histonet@lists.utsouthwestern.edu Cc: 'Katri Tuomala' Subject: RE: [Histonet] Formalin Fixation times for IHC samples I find the major impediment to good quality Immunohistochemistry is turn around times, which has been a buzz word here in Australia for a number of years. The result of this management tool has led to poorly fixed and processed tissue samples that are unable to withstand the harsh treatment of heat retrieval. Thanks to Murphy's Law, we usually find that the most vital block for diagnosis is always the one that is poorly preserved. We receive material for Immunohistochemistry from a large number of hospitals in Queensland (Australia). Unfortunately there is no uniformity in the way specimens are fixed or processed by these referring laboratories. Both lymph nodes and breast tissue are the major specimens affected by under fixation. With lymph nodes the peripheral staining is excellent but the morphology of the centre is compromised. The central structures of these specimens have not been strengthened by fixation and as a result the morphology is damaged by heat retrieval. With breast specimens there is an increase risk of tissue lifting from the slide. Try to convince Pathologists that fixation is the problem is just about impossible. We place control material, which has received at least 24 hours fixation on the same slide. The control material is always perfect and the Pathologist always concurs but questions why the test morphology is damaged. It's great to get pathology reports out quickly but at what diagnostic cost. How many other laboratories experience this problem? Barry Madigan Brisbane, Queensland, Australia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Katri Tuomala Sent: Thursday, 17 February 2005 12:23 PM To: Anne C Lewin; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Formalin Fixation times for IHC samples Anne, I think you are doing well with your protocol. Although I don't think "overfixation" is as a big problem any more with the advent of different HIER protocols with various buffers being used. Under fixation (less than 24 hours) can be a bigger problem with certain antibodies like Her-2, ER and PR to mention few. This concept of over fixation is a remnant from the time prior to HIER, when we all had a problem trying to demonstrate many antigens with weaker retrievals (proteolytic enzymes). The heat over 60C was actually banned from any immunohistochemistry protocols 20 years ago. My two cents worth... Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Anne C Lewin" To: Sent: Tuesday, February 15, 2005 10:28 AM Subject: [Histonet] Formalin Fixation times for IHC samples > Could I get a general concensus from Histoland about the amount of time > needed to fix a sample for IHC without over-fixing? My training/journal > searches have led me to believe that the longer a sample is in > formalin, the stronger the protiens are cross-linked, and the more > difficult it is to break those bonds with antigen retreival and get your > antibody to the target. I generally tell the scientists in my > department to fix overnight (most samples vary from about the size of a > pea to a lima bean), and then to transfer to 70% Ethanol for me to > process for paraffin. The total time in fixative tends to be around 24 > hours. Morphology has always been great, and my IHC's work well. This > question is mainly for my information, since I have used the same tissue > prep protocol for a few years now, I want to make sure I am keeping up > with current opinions. > Thanks! > -Anne > ---------------------------------------------------------------------------- ---- > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This 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 tpmorken <@t> labvision.com Wed Feb 23 17:33:32 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Formalin Fixation times for IHC samples Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CA4D@usca0082k08.labvision.apogent.com> I thought Dr Leong converted all you in Oz to microwave fixation! Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bill Sinai Sent: Wednesday, February 23, 2005 3:09 PM To: 'Barry Madigan'; histonet (E-mail) Subject: RE: [Histonet] Formalin Fixation times for IHC samples Hi Barry, I couldn't agree more. We are a major centre for NSW Breast Screen Western and The Breast Cancer Institute and the surgeons perform their operations on Wednesdays and Thursdays then have a meeting Monday and Tuesday and expect the results including Immunohistochemistry at these meetings. Fixation for most specimens is minimal at best and the results show poorly on Immunohistochemistry. As you said incomplete fixation means sections looking tatty or falling off. The positive controls, usually collected after the specimen has fixed for at least another 24 hours, always have great morphology and staining. We find it is very much surgeon driven as they want results for meetings for discussion about course of treatment and for discusion with patients. Many do not understand the importance of good fixation to the diagnostic process. Automation (Tissue Processors, Staining Machines, Automated coverslippers and automated Immunohistochemistry) helps a little with TAT but these are only some facets of the total process which begins with fixation. Microwave fixation helps but is not the answer to all the problems. Until someone discovers a revolutionary new fixation method I will still stick to "good old formaldehyde". I have tried several other formaldehyde substitutes over many years but the pathologists still prefer the known artefacts produced by formaldehyde. Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Barry Madigan Sent: Thursday, 24 February 2005 9:42 AM To: Histonet@lists.utsouthwestern.edu Cc: 'Katri Tuomala' Subject: RE: [Histonet] Formalin Fixation times for IHC samples I find the major impediment to good quality Immunohistochemistry is turn around times, which has been a buzz word here in Australia for a number of years. The result of this management tool has led to poorly fixed and processed tissue samples that are unable to withstand the harsh treatment of heat retrieval. Thanks to Murphy's Law, we usually find that the most vital block for diagnosis is always the one that is poorly preserved. We receive material for Immunohistochemistry from a large number of hospitals in Queensland (Australia). Unfortunately there is no uniformity in the way specimens are fixed or processed by these referring laboratories. Both lymph nodes and breast tissue are the major specimens affected by under fixation. With lymph nodes the peripheral staining is excellent but the morphology of the centre is compromised. The central structures of these specimens have not been strengthened by fixation and as a result the morphology is damaged by heat retrieval. With breast specimens there is an increase risk of tissue lifting from the slide. Try to convince Pathologists that fixation is the problem is just about impossible. We place control material, which has received at least 24 hours fixation on the same slide. The control material is always perfect and the Pathologist always concurs but questions why the test morphology is damaged. It's great to get pathology reports out quickly but at what diagnostic cost. How many other laboratories experience this problem? Barry Madigan Brisbane, Queensland, Australia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Katri Tuomala Sent: Thursday, 17 February 2005 12:23 PM To: Anne C Lewin; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Formalin Fixation times for IHC samples Anne, I think you are doing well with your protocol. Although I don't think "overfixation" is as a big problem any more with the advent of different HIER protocols with various buffers being used. Under fixation (less than 24 hours) can be a bigger problem with certain antibodies like Her-2, ER and PR to mention few. This concept of over fixation is a remnant from the time prior to HIER, when we all had a problem trying to demonstrate many antigens with weaker retrievals (proteolytic enzymes). The heat over 60C was actually banned from any immunohistochemistry protocols 20 years ago. My two cents worth... Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Anne C Lewin" To: Sent: Tuesday, February 15, 2005 10:28 AM Subject: [Histonet] Formalin Fixation times for IHC samples > Could I get a general concensus from Histoland about the amount of > time needed to fix a sample for IHC without over-fixing? My > training/journal searches have led me to believe that the longer a > sample is in formalin, the stronger the protiens are cross-linked, and > the more difficult it is to break those bonds with antigen retreival > and get your antibody to the target. I generally tell the scientists > in my department to fix overnight (most samples vary from about the > size of a pea to a lima bean), and then to transfer to 70% Ethanol for > me to process for paraffin. The total time in fixative tends to be > around 24 hours. Morphology has always been great, and my IHC's work > well. This question is mainly for my information, since I have used > the same tissue prep protocol for a few years now, I want to make sure > I am keeping up with current opinions. Thanks! > -Anne > ---------------------------------------------------------------------------- ---- > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Wed Feb 23 17:32:38 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] richard-allen type 9 paraffin In-Reply-To: Message-ID: <000201c519ff$f6145650$76d48a80@AMY> We use this paraffin in both the processor and for embedding. It works well for bone sections. I cuts just fine and we don't have a problem with it. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of TheBestTime23@aol.com Sent: Wednesday, February 23, 2005 3:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] richard-allen type 9 paraffin Hello everyone, The lab that I work for recently decided to switch to Richard-Allen type 9 paraffin. I was wondering if anyone has experience with this paraffin and could give some advice on techniques used in cutting it. Opinions of this product would also be greatly appreciated. Thanks in advance, Megan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bhewlett <@t> cogeco.ca Wed Feb 23 17:37:59 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Formalin Fixation times for IHC samples References: <20050223224218.HQYY20373.smta07.mail.ozemail.net@WORKSTATION1> Message-ID: <002a01c51a00$b69b2430$6500a8c0@mainbox> Barry, You asked >>How many other laboratories experience this problem? Answer, The majority of histopathology labs in Canada and the US! For over 25 years, some of us have been on a crusade to correct this. The result, When shown the undeniable evidence, as you have done, the IHC technologists get it immediately but few others choose to listen. And I thought that we were in the era of 'evidence-based' medicine!!! Bryan Hewlett Quality Management Program-Laboratory Services Ontario, Canada. ----- Original Message ----- From: "Barry Madigan" To: Cc: "'Katri Tuomala'" Sent: Wednesday, February 23, 2005 5:42 PM Subject: RE: [Histonet] Formalin Fixation times for IHC samples > I find the major impediment to good quality Immunohistochemistry is turn > around times, which has been a buzz word here in Australia for a number of > years. The result of this management tool has led to poorly fixed and > processed tissue samples that are unable to withstand the harsh treatment of > heat retrieval. > Thanks to Murphy's Law, we usually find that the most vital block for > diagnosis is always the one that is poorly preserved. > > We receive material for Immunohistochemistry from a large number of > hospitals in Queensland (Australia). Unfortunately there is no uniformity in > the way specimens are fixed or processed by these referring laboratories. > Both lymph nodes and breast tissue are the major specimens affected by under > fixation. > With lymph nodes the peripheral staining is excellent but the morphology of > the centre is compromised. > The central structures of these specimens have not been strengthened by > fixation and as a result the morphology is damaged by heat retrieval. > With breast specimens there is an increase risk of tissue lifting from the > slide. > > Try to convince Pathologists that fixation is the problem is just about > impossible. > We place control material, which has received at least 24 hours fixation on > the same slide. The control material is always perfect and the Pathologist > always concurs but questions why the test morphology is damaged. > > It's great to get pathology reports out quickly but at what diagnostic cost. > How many other laboratories experience this problem? > > Barry Madigan > Brisbane, Queensland, Australia > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Katri > Tuomala > Sent: Thursday, 17 February 2005 12:23 PM > To: Anne C Lewin; Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Formalin Fixation times for IHC samples > > Anne, > I think you are doing well with your protocol. Although I don't think > "overfixation" is as a big problem any more with the advent of different > HIER protocols with various buffers being used. Under fixation (less than 24 > > hours) can be a bigger problem with certain antibodies like Her-2, ER and PR > > to mention few. This concept of over fixation is a remnant from the time > prior to HIER, when we all had a problem trying to demonstrate many antigens > > with weaker retrievals (proteolytic enzymes). The heat over 60C was actually > > banned from any immunohistochemistry protocols 20 years ago. > My two cents worth... > > Katri > > Katri Tuomala > Hamilton, Ontario, Canada > > > ----- Original Message ----- > From: "Anne C Lewin" > To: > Sent: Tuesday, February 15, 2005 10:28 AM > Subject: [Histonet] Formalin Fixation times for IHC samples > > > > Could I get a general concensus from Histoland about the amount of time > > needed to fix a sample for IHC without over-fixing? My training/journal > > searches have led me to believe that the longer a sample is in > > formalin, the stronger the protiens are cross-linked, and the more > > difficult it is to break those bonds with antigen retreival and get your > > antibody to the target. I generally tell the scientists in my > > department to fix overnight (most samples vary from about the size of a > > pea to a lima bean), and then to transfer to 70% Ethanol for me to > > process for paraffin. The total time in fixative tends to be around 24 > > hours. Morphology has always been great, and my IHC's work well. This > > question is mainly for my information, since I have used the same tissue > > prep protocol for a few years now, I want to make sure I am keeping up > > with current opinions. > > Thanks! > > -Anne > > > > > -------------------------------------------------------------------------- -- > ---- > > > > _______________________________________________ > > Histonet mailing list > > Histonet@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 jryan <@t> sleh.com Wed Feb 23 17:54:05 2005 From: jryan <@t> sleh.com (John Ryan) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] I'm sorry I can not reply immediately because I am out of the office, returning on Monday 2/28/2005. Message-ID: I'm sorry I can not reply immediately because I am out of the office, returning on Monday 2/28/2005. +++++CONFIDENTIALITY NOTICE+++++ The information in this e-mail may be confidential and/or privileged. If you are not the intended recipient or an authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments, if any, or the information contained herein is prohibited. If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail from your computer system. Thank you. From portera203 <@t> yahoo.com Wed Feb 23 19:22:44 2005 From: portera203 <@t> yahoo.com (Amy S. Porter, HT(ASCP) QIHC) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Automated immunostainer for research applications Message-ID: <20050224012244.53485.qmail@web40905.mail.yahoo.com> Teri - We have a Dako Autostainer in our lab which is strictly research no service and find it very flexible and user friendly. We still do all of our overnight incubations at 4C with the exception of Dystrophin antibodies which we do at room temperature. We do all of our substrate incubations on the instrument - using standardized system protocols. The Dako is nice due to the fact it allows you to dispense at three different areas on the slide using a minimum amount of 100 microliters in each position on the slide. Hope this helps out. "Johnson, Teri" wrote:Hi all (and apologies to those on both lists), For those of you doing research histology and immunohistochemistry, which automated immunostainer do you use? Do you do any overnight incubations with it? For those of you who have automated stainers and formerly did overnight (4C) incubations, what modifications did you make to your staining protocol to achieve the same result? Also, do you always use the substrate on instrument, or do you do that via microscope? Some immunostainers require minimum amounts of primary antibody (~500 microliters), even if staining only one slide. Some times we don't have that much antibody to use. Are there any tips to get around this with the exception of manually pipetting as if doing a titration run? Thanks for your input! 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 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!? Yahoo! Mail - Find what you need with new enhanced search. Learn more. From djohnson14 <@t> hotmail.com Wed Feb 23 19:22:04 2005 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Silane coated sldies Message-ID: I have a question in the same vein as Josefa's question. Why do Silane coated slides conflict with and/or cause problems on Ventana's stainer? >From: "Nava, Josefa" >To: "'histonet@pathology.swmed.edu'" >Subject: [Histonet] "Premiere" charge slides >Date: Wed, 23 Feb 2005 15:31:23 -0600 > >Hello there, > >Has anyone been using this brand of charged slides "PREMIERE" to do their >immuno stains using Ventana immuno machines. I will appreciate any >feedback you can give me regarding this brand. Thank you. > > > >Josie > > > >The information contained in this message and any attachments is intended >only for the use of the individual or entity to which it is addressed, and >may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from >disclosure under applicable law. If you are not the intended recipient, >you are prohibited from copying, distributing, or using the information. >Please contact the sender immediately by return e-mail and delete the >original message from your system. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cbass <@t> bidmc.harvard.edu Wed Feb 23 19:33:36 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] HER-2 and survivin antibodies Message-ID: <1A48364E-8604-11D9-BFF6-000A95EC3B78@bidmc.harvard.edu> Hello, Could anyone recommend a HER-2 and survivin antibody? I am looking for something that could be used for either IHC or western blotting. I will be using primarily breast cancer cell lines such as MDA-MB-453 among others. Any recommendations would be welcomed. Also, if anyone has a very simple protocol for IHC with these cell lines I would greatly appreciate it. Thanks, Caroline Bass From dharclerode <@t> macropore.com Wed Feb 23 19:49:05 2005 From: dharclerode <@t> macropore.com (Donna Harclerode) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: Automated immunostainer for research applications (Johnson, Teri) Message-ID: 8. Automated immunostainer for research applications (Johnson, Teri) Message: 8 Date: Wed, 23 Feb 2005 13:26:24 -0600 Hi all (and apologies to those on both lists), For those of you doing research histology and immunohistochemistry, which automated immunostainer do you use? Do you do any overnight incubations with it? For those of you who have automated stainers and formerly did overnight (4C) incubations, what modifications did you make to your staining protocol to achieve the same result? Also, do you always use the substrate on instrument, or do you do that via microscope? Some immunostainers require minimum amounts of primary antibody (~500 microliters), even if staining only one slide. Some times we don't have that much antibody to use. Are there any tips to get around this with the exception of manually pipetting as if doing a titration run? Thanks for your input! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org Hi Terri This is a bit more info than what I wrote on the other list I use a "semi automated" system, the Shandon Sequenza. I do research staining and have not found any (to date) systems that would allow me the flexibility to incubate at 4 oC overnight or some of the other things I wanted to do. I run anywhere from 10-90+ slides at a time with LSAB and DAB and it is no problem. The Sequenza system has a coverplate on one slide that keeps 80ul on the slide. The carriers make their own humidity chamber with 10 slides in each carrier and I think they are great. I load the slides (after H2O2 blocking for paraffin), block with the Dako endogenous peroxidase block for frozens, rinse in buffer and then primary etc. (I do not do a separate protein block, just make up all the abs in Dako antibody diluent and that has worked for many years for me) Once the slides are loaded I just add the reagents (100ul) or buffer to the top. The only tricky part was learning to load the slides on the coverplate. After the slides are loaded I always fill the reservoir with PBS to check for misloaded slides. If the slides are loaded correctly the buffer drains slowly about 3-5 minutes. If it drains too fast I know I did not get the slide in correctly. I reuse the coverplates (they say of course use only once) but as long as I only rinse them in DI water they last about a month. If you use soap they are ruined. I use them for frozen, cytospin and paraffin sections (paraffin usually are the overnight ones. The other really nice thing is you can put tissue on most of the slide and 100ul is plenty of antibody for each slide (I always use 100 ul of antibody to be sure all the buffers are removed and replaced by reagents). I do not DAB in the chambers, I remove the slides for that so I can monitor the stain. I have done fluorescent labeling in the chambers and that has worked well. Anyhow the web info which is http://www.thermo.com/com/cda/product/detail/1,1055,1000005630785,00.htm l I did not buy the holder for the racks, but just the slide racks and coverplates. Good luck with whatever you get and if you have any questions I forgot to address, just let me know Donna Harclerode, HT(ASCP)HTL, QIHC Immunohistochemist MacroPore Biosurgery, Inc 6740 Top Gun St. San Diego, CA 92121 858-458-0900 ext 322 dharclerode@macropore.com From c.m.vanderloos <@t> amc.uva.nl Thu Feb 24 02:44:19 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: alk phos, again Message-ID: <160775516075cf.16075cf1607755@amc.uva.nl> Hi Rachael, You wrote that you consider the observed "background" as endogenous alk. phos. activity. Please realize that real endogenous alk. phos. activity shows up specifically in vessels for example rather than an overall non-specific background staining. The fact that high concentrations of acetic acid did kill your background doesn't say anything as it also killed your specific signal. The Dako Dual Endogenous Enzyme Block reagent (S2003) blocks both endogenous peroxidase and alk. phosphatase activities. However, as I wrote on Histonet before it may affect your epitopes as well. There is another solution, a quite mysterious one I should admit but it works good. It's both a fixative and blocking reagent for endogenous peroxidase and alk. phosphatase activities. Prepare 4% sodium nitrite and 4% pararosaniline solutions and mix 1:1 for exactly 1 minute under the fume hood. Than, mix 2 ml of diazotized pararosaniline with 200 ml distilled water and store at 4C in the dark. Fix your cryosections for exactly 2 min in cold fixative and wash with TBS or PBS. Ref: Schrijver et al. JHC 48:95-103, 2000. Again, after using this fixative/blocking reagent you have to check whether or not your epitopes are affected or not. Hope this helps, 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 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From "Emerson, Rachael" Date Tue, 22 Feb 2005 14:12:56 -0500 To "'histonet@lists.utsouthwestern.edu'" Subject [Histonet] alk phos, again Other there other methods to block endogenous alkaline phosphatase? I am using mouse embryos, frozen sectioned. I've tried: levamisole at varying concentrations: still background heated PBS at varying times: too strong, blocked my signal 15% acetic acid at 15": too strong, blocked my signal 5% acetic acid at 15": still background usually I did a combo of levamisole and another treatment. I appreciate any thoughts....... Thanks! Rachael References 1. mailto:c.m.vanderloos@amc.uva.nl From Myri37 <@t> aol.com Thu Feb 24 03:25:13 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] (no subject) Message-ID: <03AF12B0.13D2F211.0005167B@aol.com> Hi dear histonetters, i am searching a technique and special stains, for staining very small collagen fibers, on metallic biomaterial. Do you have an experience with this ? Thahk you very much in advance for any help. myriam Natural Implant From Myri37 <@t> aol.com Thu Feb 24 03:26:07 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] collagen fibers Message-ID: <06AB602F.19F40B24.0005167B@aol.com> Hi dear histonetters, i am searching a technique and special stains, for staining very small collagen fibers, on metallic biomaterial. Do you have an experience with this ? Thahk you very much in advance for any help. myriam Natural Implant From jstaruk <@t> masshistology.com Thu Feb 24 07:46:21 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] collagen fibers In-Reply-To: <06AB602F.19F40B24.0005167B@aol.com> Message-ID: <0ICF00HJN50MP080@vms044.mailsrvcs.net> My favorite is Sirius Red using a polarized light. Just did a bunch of livers yesterday and they came out great. Nice, delicate, single collagen fibers glowed brilliantly. Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Myri37@aol.com Sent: Thursday, February 24, 2005 4:26 AM To: histonet@pathology.swmed.edu Subject: [Histonet] collagen fibers Hi dear histonetters, i am searching a technique and special stains, for staining very small collagen fibers, on metallic biomaterial. Do you have an experience with this ? Thahk you very much in advance for any help. myriam Natural Implant _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ajennings <@t> unmc.edu Thu Feb 24 08:06:00 2005 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Formalin Fixation times for IHC samples In-Reply-To: <20050223224218.HQYY20373.smta07.mail.ozemail.net@WORKSTATION1> Message-ID: Great timing on this topic I have been looking for a review article that actually puts in print the data collected on over fixation or under fixation in samples that will be used for IHC. To include the eventual breakdown of formalin and need for 10-20 times volume for proper fixation. Is there such a paper with all of information that histologist reverberate and others ignore? From brett_connolly <@t> merck.com Thu Feb 24 08:19:46 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Formalin Fixation times for IHC samples Message-ID: Take a look at the J. Histotechnology 24:3, Sept 2001....special issue on Fixation. It includes a couple of articles dealing with fixation and IHC. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of ajennings@unmc.edu Sent: Thursday, February 24, 2005 9:06 AM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Formalin Fixation times for IHC samples Great timing on this topic I have been looking for a review article that actually puts in print the data collected on over fixation or under fixation in samples that will be used for IHC. To include the eventual breakdown of formalin and need for 10-20 times volume for proper fixation. Is there such a paper with all of information that histologist reverberate and others ignore? _______________________________________________ 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 pruegg <@t> ihctech.net Thu Feb 24 08:33:45 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: [IHCRG] Automated immunostainer for research applications In-Reply-To: Message-ID: <200502241434.j1OEXwAG038716@pro12.abac.com> In my experience drying even after DAB is not good at least in some cases the tissue does not far well and the morphology can be adversely affected. Patsy -----Original Message----- From: ihcrg-bounces@neo.agsci.colostate.edu [mailto:ihcrg-bounces@neo.agsci.colostate.edu] On Behalf Of Ni, Ruoyu Sent: Thursday, February 24, 2005 7:26 AM To: 'Patsy Ruegg'; leharkins@yahoo.com; TJJ@stowers-institute.org; ihcrg@neo.agsci.colostate.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [IHCRG] Automated immunostainer for research applications I think, at this point(after DAB and washing), it does not matter if the slides are dry or not. Ruoyu Ni, MLT Molecular Histology Mount Sinai Hospital Toronto, M5G1N6 CANADA -----Original Message----- From: Patsy Ruegg [mailto:pruegg@msn.com] Sent: February 24, 2005 9:12 AM To: leharkins@yahoo.com; TJJ@stowers-institute.org; ihcrg@neo.agsci.colostate.edu Cc: histonet@lists.utsouthwestern.edu Subject: Re: [IHCRG] Automated immunostainer for research applications I use the DAKO autostainer overnight somewhat different from Loui. I set the run to go with usually a 1-2 hr. incubation of primary and complete run, my last wash is dih20 so after the run finishes the autostainer will apply dih20 once every hour until I end the program the next day, this way I do not waste buffer as after the run finishes dih20 can be applied to keep the sections from drying out. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 cell 720-281-5406 wk email pruegg@ihctech.net hm email pruegg@msn.com web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. ----Original Message Follows---- From: lualhati harkins To: "Johnson, Teri" , ihcrg@neo.agsci.colostate.edu CC: Histonet Subject: Re: [IHCRG] Automated immunostainer for research applications Date: Wed, 23 Feb 2005 18:12:23 -0800 (PST) Hi Teri, In response to your first question on overnight incubation( automated immunostainers) 1) Overnight incubation is achieved on Biogenex I6000 , with Optimax and also with DAKO Autostainer through setting up the program on delayed run. The deparaffinized slides are loaded into the machine and set on continious buffer run to prevent slides from drying up. Antibody incubation time- i optimize my antibody dilution to allow 1-2 hours of incubation at room temperature.This allow me to use very low dilution of primary antibody which answer your other question: 2)" what if you have only 500 microliter of antibody available to use" I was able to use 200microliter ( Biogenex i 6000or Optimax) or less than 200 in some instances with Dako Autostainer. If you do not have any time as well as enough antibody for optimization: a ) find supporting data such as : dilution used for Immunofluorescence- ( frozen)- By previous experience i found out that if immunofluorescence working concentration (frozen) is 1:500 then i will use 1:50 as dilution for paraffin sections b) if i still do not have enough antibody to come up with this dilution, then i will use 1:100 and run an overnight incubation in the refrigerator. c) western blot data is useful also: if westernblot working concentration is 1:500 , then start a working dilution at 1:50 for paraffin sections, or if using frozen then use 1:250 ( this will really need some optimization For overnight incubation, start with 1:100 for paraffin sections, frozen sections could possibly work at 1:500 overnight, but not a sure thing; I have better luck for sure signal to stay around 1:300.( these are just guidelines) This dilution gets complicated depending on how much protein concentration , and the type of protein and also where the localizationis expected : nuclear, cytoplasmic/membrane. Please contact me directly for detailed and in depth discussion of other data determining dilution. To prevent using too much buffer, i also adjusted my secondary antibody and HRP label incubation time for 45 minutes each. With respect to chromogen use, for research purposes, i prefer to develop chromogen and observe signals through te microscope. For diagnostic runs, i optimize the chromogen development to run in the machine for puposes of other personnell using the machine when i am on vacation Finally to give an example of overnight/delayed run in the machine: Start machine at 6Pm Delay-(4hrs) -start at 10PM Antibody Incubation- done at 12AM Secondary/label done at 1:30 AM Buffer rinse until morning- You can adjust the delay for however long you want- I hope the information will be useful. I hope i was able to help you Sincerely Loui "Johnson, Teri" wrote: Hi all (and apologies to those on both lists), For those of you doing research histology and immunohistochemistry, which automated immunostainer do you use? Do you do any overnight incubations with it? For those of you who have automated stainers and formerly did overnight (4C) incubations, what modifications did you make to your staining protocol to achieve the same result? Also, do you always use the substrate on instrument, or do you do that via microscope? Some immunostainers require minimum amounts of primary antibody (~500 microliters), even if staining only one slide. Some times we don't have that much antibody to use. Are there any tips to get around this with the exception of manually pipetting as if doing a titration run? Thanks for your input! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ IHCRG mailing list IHCRG@neo.agsci.colostate.edu http://neo.agsci.colostate.edu/mailman/listinfo/ihcrg **************************************************************** Please note: Protocol and technical information provided by the undersigned are developed on site with conditions limited to fixation of tissues,reagents,control tissues and execution of protocol which is characteristic only of our laboratory and therefore we do not bear neither responsibility nor liability for results outside of our laboratory. It is our courteous gesture to share information that we deemed might be useful to laboratorians and technologists. ****************************************************************** Loui Harkins , M.S. QIHC Biologist, Immunohistochemistry/Path.& lab. Medicine DEpt. Of Veterans Affairs,700 South, 19th Street Birmingham, Alabama 35233 (Research Consultant) Dept .of Surgery, Neurosurgery Division University of Alabama in Birmingham 205-933-8101 ext.6719; fax 205-558-4817 or 558-7034 _______________________________________________ IHCRG mailing list IHCRG@neo.agsci.colostate.edu http://neo.agsci.colostate.edu/mailman/listinfo/ihcrg _______________________________________________ IHCRG mailing list IHCRG@neo.agsci.colostate.edu http://neo.agsci.colostate.edu/mailman/listinfo/ihcrg _______________________________________________ IHCRG mailing list IHCRG@neo.agsci.colostate.edu http://neo.agsci.colostate.edu/mailman/listinfo/ihcrg From pruegg <@t> ihctech.net Thu Feb 24 08:48:37 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: [IHCRG] Automated immunostainer for research applications In-Reply-To: <20050224021224.2943.qmail@web50504.mail.yahoo.com> Message-ID: <200502241449.j1OEmrKG049372@pro12.abac.com> As for antibody amount. On the DAKO depending on the size of the tissue I use 100 microliters to 200 microliters, I always use a pap pen to isolate the reagent over the section, I put all my sections at the bottom of the slide as the reagent tends to draw up towards the label, I just draw one line straight across the slide just above the section at the bottom to keep the reagent down at the bottom over the section, I deparaffinize, rehydrate to 95% alcohol and then let the sections dry, I then draw the pap pen line while the section is dry before rinsing with dih20 and then buffer, some pap pens these days even survive HIER (BioCare's does), I have even noticed that this one is still there after passing thru alcohols and xylene to coverslip, others I have used will not survive the solvents. As Loui, if I am developing a protocol, I check the DAB development microscopically until I get an optimized time, I then program the machine for that time for that protocol. I am an advocate for longer incubation times with more dilute antibodies, I feel you get cleaner staining and more efficient use of the antibodies. I also dilute detections and use them for longer periods in this same way. I know in clinical practice everybody is in a hurry but I think you pay in cost of reagents and signal/noise by using antibodies too concentrated for short times. Patsy -----Original Message----- From: ihcrg-bounces@neo.agsci.colostate.edu [mailto:ihcrg-bounces@neo.agsci.colostate.edu] On Behalf Of lualhati harkins Sent: Wednesday, February 23, 2005 7:12 PM To: Johnson, Teri; ihcrg@neo.agsci.colostate.edu Cc: Histonet Subject: Re: [IHCRG] Automated immunostainer for research applications Hi Teri, In response to your first question on overnight incubation( automated immunostainers) 1) Overnight incubation is achieved on Biogenex I6000 , with Optimax and also with DAKO Autostainer through setting up the program on delayed run. The deparaffinized slides are loaded into the machine and set on continious buffer run to prevent slides from drying up. Antibody incubation time- i optimize my antibody dilution to allow 1-2 hours of incubation at room temperature.This allow me to use very low dilution of primary antibody which answer your other question: 2)" what if you have only 500 microliter of antibody available to use" I was able to use 200microliter ( Biogenex i 6000or Optimax) or less than 200 in some instances with Dako Autostainer. If you do not have any time as well as enough antibody for optimization: a ) find supporting data such as : dilution used for Immunofluorescence- ( frozen)- By previous experience i found out that if immunofluorescence working concentration (frozen) is 1:500 then i will use 1:50 as dilution for paraffin sections b) if i still do not have enough antibody to come up with this dilution, then i will use 1:100 and run an overnight incubation in the refrigerator. c) western blot data is useful also: if westernblot working concentration is 1:500 , then start a working dilution at 1:50 for paraffin sections, or if using frozen then use 1:250 ( this will really need some optimization For overnight incubation, start with 1:100 for paraffin sections, frozen sections could possibly work at 1:500 overnight, but not a sure thing; I have better luck for sure signal to stay around 1:300.( these are just guidelines) This dilution gets complicated depending on how much protein concentration , and the type of protein and also where the localizationis expected : nuclear, cytoplasmic/membrane. Please contact me directly for detailed and in depth discussion of other data determining dilution. To prevent using too much buffer, i also adjusted my secondary antibody and HRP label incubation time for 45 minutes each. With respect to chromogen use, for research purposes, i prefer to develop chromogen and observe signals through te microscope. For diagnostic runs, i optimize the chromogen development to run in the machine for puposes of other personnell using the machine when i am on vacation Finally to give an example of overnight/delayed run in the machine: Start machine at 6Pm Delay-(4hrs) -start at 10PM Antibody Incubation- done at 12AM Secondary/label done at 1:30 AM Buffer rinse until morning- You can adjust the delay for however long you want- I hope the information will be useful. I hope i was able to help you Sincerely Loui "Johnson, Teri" wrote: Hi all (and apologies to those on both lists), For those of you doing research histology and immunohistochemistry, which automated immunostainer do you use? Do you do any overnight incubations with it? For those of you who have automated stainers and formerly did overnight (4C) incubations, what modifications did you make to your staining protocol to achieve the same result? Also, do you always use the substrate on instrument, or do you do that via microscope? Some immunostainers require minimum amounts of primary antibody (~500 microliters), even if staining only one slide. Some times we don't have that much antibody to use. Are there any tips to get around this with the exception of manually pipetting as if doing a titration run? Thanks for your input! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ IHCRG mailing list IHCRG@neo.agsci.colostate.edu http://neo.agsci.colostate.edu/mailman/listinfo/ihcrg **************************************************************** Please note: Protocol and technical information provided by the undersigned are developed on site with conditions limited to fixation of tissues,reagents,control tissues and execution of protocol which is characteristic only of our laboratory and therefore we do not bear neither responsibility nor liability for results outside of our laboratory. It is our courteous gesture to share information that we deemed might be useful to laboratorians and technologists. ****************************************************************** Loui Harkins , M.S. QIHC Biologist, Immunohistochemistry/Path.& lab. Medicine DEpt. Of Veterans Affairs,700 South, 19th Street Birmingham, Alabama 35233 (Research Consultant) Dept .of Surgery, Neurosurgery Division University of Alabama in Birmingham 205-933-8101 ext.6719; fax 205-558-4817 or 558-7034 _______________________________________________ IHCRG mailing list IHCRG@neo.agsci.colostate.edu http://neo.agsci.colostate.edu/mailman/listinfo/ihcrg From ajennings <@t> unmc.edu Thu Feb 24 08:51:24 2005 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Formalin Fixation times for IHC samples In-Reply-To: Message-ID: Thanks Brett I knew I didn't make my preaching up all by myself. I pulled the Fixation issue and it is a good resource to support my requests on tissue handling. From mucram11 <@t> comcast.net Thu Feb 24 09:05:18 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Formalin Fixation times for IHC samples Message-ID: <022420051505.2169.421DED2D000B5F43000008792207003201CECE030E9D0C9A03@comcast.net> I agree with Brett about the article however, a great deal of what we do in histology is empirical or just accepted as the way things are always done. Over the past decade as IHC has become so important and time has been shortened for many laboratories the arguments about fixation are increasing. The time tissue is in fixative should include the not only processor time but the time from grossing through processor fixation. If the sample comes and is grossed early (mid morning) and placed in a holding container of fixative it may have different for better or worse staining than later arriving specimens. The tissue that comes in at 2PM and gets grossed in at 3PM and placed in the holding container for 30 minutes to an hour and then on the processor will often appear very different or "under fixed" in comparison. We don't look at those differences in time of fixation as often as we should and we are not always allowed to make up the time required in fixative. If the specimen goes in late enough the fixation may be completed more by alcohols during dehydration than 10% NBF and this can effect staining as well. Pam Marcum -------------- Original message -------------- > Take a look at the J. Histotechnology 24:3, Sept 2001....special issue on > Fixation. It includes a couple of articles dealing with fixation and IHC. > > Brett > > Brett M. Connolly, Ph.D. > Merck & Co., Inc. > MRL, Imaging Research > WP26A-3000 > PO Box 4 > West Point, PA 19486 > PH 215-652-2501 > fax. 215-652-2075 > e-mail. brett_connolly@merck.com > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > ajennings@unmc.edu > Sent: Thursday, February 24, 2005 9:06 AM > To: Histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Formalin Fixation times for IHC samples > > > > > > > Great timing on this topic > > I have been looking for a review article that actually puts in print the > data collected on over fixation or under fixation in samples that will be > used for IHC. To include the eventual breakdown of formalin and need for > 10-20 times volume for proper fixation. Is there such a paper with all of > information that histologist reverberate and others ignore? > > > _______________________________________________ > 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. > ------------------------------------------------------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pex0220 <@t> yahoo.com.cn Thu Feb 24 09:25:12 2005 From: pex0220 <@t> yahoo.com.cn (=?gb2312?q?=CC=EC=20=D0=C1?=) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] double immunofluorescence in bone tissue Message-ID: <20050224152512.28562.qmail@web15505.mail.cnb.yahoo.com> Hi, Dear histonetter, I have had some difficulties in do double immunofluorescence in bone tissues. Firstly, I find that it is not easy to cut the paraffin blocks of bone and muscle. Secondly, I do double immunofluorescence by general protocol, but my results are not good. in the section, part tissues were stained, but part tissues were not stained.I do not know why this results appear. I think that double immunofluorescence in bone and muscle tissues is different from that in other tissues. I check some articles, but I can not find a good article, so if you gain some experience about it, I hope you can give me a protocol or some advice. thank you very much! Guofeng --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From TillRenee <@t> uams.edu Thu Feb 24 09:48:05 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] cryosections Message-ID: Hello. Our research group just purchased our first cryostat. Until now, anytime cryosections were needed we had to send out our tissues to another lab. Is there a good book or other source for general information about cryosectioning? I am the only histotech in the group, but have had virtually no experience with cryosections. I know the other general techs that also do some histology are going to come to me with all their questions. We basically need to know anything and everything. Due to the volume of tissues, sectioning won't be performed right as the tissues are removed from the animal. How do you store them? Do you fix, and when? What type of slides are best? How do you store the slides after cryosectioning? Are there specific considerations to take into account as far as fixing, or anything else depending on the tissue or what stains will be performed on it? For example, I know already that there will be some heart and aorta sectioned for fat stains, some pig gi, possibly mammary gland, uterus, and mouse lung with b-gal. Thanks in advance for any help or suggestions you can give me. Renee' From hhawkins <@t> UTMB.EDU Thu Feb 24 10:50:35 2005 From: hhawkins <@t> UTMB.EDU (Hawkins, Hal K.) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Formalin Fixation times for IHC samples Message-ID: <8D6F233E2A5D574B929F3944F3316FD01A5902@EXCH2K3.utmb.edu> One factor that has not been mentioned yet with regard to this discussion of fixation time is the effect of agitation. Those of us who do autopsies know well that there is often poor penetration of fixative between samples resting in a static container. Agitation during fixation can reduce this problem and reduce the time necessary for fixation by maintaining a nearly constant concentration of fixative at the surface of each piece of tissue. Hal Hawkins, UT Medical Branch, Galveston, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mucram11@comcast.net Sent: Thursday, February 24, 2005 9:05 AM To: Connolly, Brett M; 'ajennings@unmc.edu'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Formalin Fixation times for IHC samples I agree with Brett about the article however, a great deal of what we do in histology is empirical or just accepted as the way things are always done. Over the past decade as IHC has become so important and time has been shortened for many laboratories the arguments about fixation are increasing. The time tissue is in fixative should include the not only processor time but the time from grossing through processor fixation. If the sample comes and is grossed early (mid morning) and placed in a holding container of fixative it may have different for better or worse staining than later arriving specimens. The tissue that comes in at 2PM and gets grossed in at 3PM and placed in the holding container for 30 minutes to an hour and then on the processor will often appear very different or "under fixed" in comparison. We don't look at those differences in time of fixation as often as we should and we are not always allowed to make up the time required in fixative. If the specimen goes in late enough the fixation may be completed more by alcohols during dehydration than 10% NBF and this can effect staining as well. Pam Marcum -------------- Original message -------------- > Take a look at the J. Histotechnology 24:3, Sept 2001....special issue on > Fixation. It includes a couple of articles dealing with fixation and IHC. > > Brett > > Brett M. Connolly, Ph.D. > Merck & Co., Inc. > MRL, Imaging Research > WP26A-3000 > PO Box 4 > West Point, PA 19486 > PH 215-652-2501 > fax. 215-652-2075 > e-mail. brett_connolly@merck.com > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > ajennings@unmc.edu > Sent: Thursday, February 24, 2005 9:06 AM > To: Histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Formalin Fixation times for IHC samples > > > > > > > Great timing on this topic > > I have been looking for a review article that actually puts in print the > data collected on over fixation or under fixation in samples that will be > used for IHC. To include the eventual breakdown of formalin and need for > 10-20 times volume for proper fixation. Is there such a paper with all of > information that histologist reverberate and others ignore? > > > _______________________________________________ > 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. > ------------------------------------------------------------------------ ------ > > _______________________________________________ > Histonet mailing list > Histonet@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 kelly.mcqueeney <@t> bms.com Thu Feb 24 10:54:03 2005 From: kelly.mcqueeney <@t> bms.com (Kelly D Mcqueeney) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] cryosections In-Reply-To: References: Message-ID: <421E06AB.80807@bms.com> Renee, You should insist on an in-depth training from the vendor. Ask for a 4 hour training from their cryostat expert, not some sales person. They should offer it to you. The technique is standard but varies from user to user. You'll have to find your way. I am going to give you our standard protocol for brain IHC. 1) Remove fresh tissue, dip in mounting medium (we use Shandon embedding medium, or you can skip this step) and immediately freeze in ice-cold isopentane (keep on dry ice for 30 minutes prior to freezing). For rat brain, we freeze for 20 seconds. Remove from isopentane and place tissue in a conical tube (some people use molds). Leave the tube on dry ice and immediately store at -70C. 2) Remove tissue from -70C and place in -20C freezer for 1 hour to equilibrate. 3) Drop embedding medium on chuck, add tissue and freeze base with cytocool. Cover tissue with embedding medium (we dip and freeze with cytocool). 4) Place chuck in cryostat and section tissue. I use probe on plus slides from Fisher or Superfrost from VWR. 5) Once sections are mounted on slides, dry slides completely for 1 hour RT and store at -80C until ready to use. 6) Remove slides from freezer and thaw at RT for 15 minutes. Place slides in fix and stain or perform IHC. There are many different methods for sectioning/storing, etc. Times, tissue, temperature and protocols will vary from lab to lab or person to person. Have fun! Kelly Till, Renee wrote: >Hello. Our research group just purchased our first cryostat. Until now, >anytime cryosections were needed we had to send out our tissues to >another lab. Is there a good book or other source for general >information about cryosectioning? I am the only histotech in the group, >but have had virtually no experience with cryosections. I know the other >general techs that also do some histology are going to come to me with >all their questions. We basically need to know anything and everything. >Due to the volume of tissues, sectioning won't be performed right as the >tissues are removed from the animal. How do you store them? Do you fix, >and when? What type of slides are best? How do you store the slides >after cryosectioning? Are there specific considerations to take into >account as far as fixing, or anything else depending on the tissue or >what stains will be performed on it? For example, I know already that >there will be some heart and aorta sectioned for fat stains, some pig >gi, possibly mammary gland, uterus, and mouse lung with b-gal. > >Thanks in advance for any help or suggestions you can give me. > > > >Renee' > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From petepath <@t> yahoo.com Thu Feb 24 11:08:11 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] cryosections Message-ID: <20050224170811.73986.qmail@web30405.mail.mud.yahoo.com> Renee, I have a tutorial on frozen section technique on my web site. http:pathologyinnovations.com/frozen_section_technique.htm I do not know of any books that contain this information but it is everything I have learned the hard way (trial and error) over the years. It is based on experience in surgical pathology but I believe many of these concepts will apply to a variety of applications.I hope you find it useful. If anyone cares to add anything to this or criticize the content I wecome any comments and new information and will credit them on the web site. Stephen Stephen Peters M.D. Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com Senior Attending Pathologist Hackensack University Medical Center 201 996 4836 From froyer <@t> bitstream.net Thu Feb 24 11:40:26 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: pathologists in the lab (shivers down thespine...) In-Reply-To: <421D3613.9060003@umdnj.edu> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328CA47@usca0082k08.labvision.apogent.com> <15483.128.6.148.191.1109198078.squirrel@webmail.rci.rutgers.edu> <421D3613.9060003@umdnj.edu> Message-ID: <421E118A.7050007@bitstream.net> I agree that there is precious little time to include anatomical and/or clinical pathology practical training in the medical school curriculum. And in-depth training in any lab procedure really isn't necessary. But exposure to the 'World of the Lab' can be beneficial no matter how superficial, and not necessarily during undergraduate medical school or during internships. Where it worked at one place, that I am aware, was to include exposure to the laboratory during residency programs. At the last lab I worked in, the hospital had a fairly large Family Practice Residency Program. With the help and input of our pathologists and MT/HT staff, the Medical Director of the Residency Program included a one-week rotation of all the third year residents through the lab. (they would be scheduled two at a time over a period of a year so we were not overwhelmed with too many residents at one time.) It also helped that the Medical Director was very supportive of the lab and believed that this knowledge was essential. The residents spent one day in each department (Chemistry, Hematology/Microbiology, Blood Bank/Coagulation, Histology/Cytology, and Pathology) following a Med Tech or Histotech around. They saw for themselves how the lab worked on a practical daily basis and how crazy our world could be at times. They walked a mile in our shoes. Bonds were formed that lasted for years after they graduated. They came to appreciate what we do, why we do it (i.e. why we insist on proper specimen collection/handling, etc.), and why it may "take so long" to get results back. Barriers were torn down and communication lines were opened up. For the first time, I was working in a hospital where the medical staff would actually call the lab to consult directly with a MT or HT. Us "lab rats" were no longer looked upon on as those mysterious creatures that dwelled unseen in the bowels of the hospital, but as actual important members of the medical team... there to save lives and stamp out disease, just like the doctors and nurses. An interesting side note.... while I was involved in this program there were three F.P. residents, over a period of years, that went on to specialize in Pathology. They enjoyed the "week in the lab" experience so much that it led them to a vocation that they had not thought of originally . I would recommend looking into this if you are in a facility that has residency programs. One 5-day week out of a four-year residency is not too much to ask. ~ Ford Ford M. Royer, MT(ASCP) Midwest Science & Biocenter, Inc. Minneapolis, MN Geoff McAuliffe wrote: > Two reasons for not training medical students: > 1. Not enough time in the curriculum. > 2. Most medical students won't be pathologists, so why train them to > process tissue and stain slides? Those who train in Pathology should > know what is involved in tissue preparation and know enough not to > demonstrate their ignorance. > > Geoff > > kgrobert@rci.rutgers.edu wrote: > >> We're just a small research and teaching lab, but I have taught all >> sorts >> of people about histology-from high-schoolers on up to full professors, >> and every time my boss teaches his Toxicologic Pathology graduate >> course, >> I get to teach the lab portion of it, where the students learn almost >> the >> same thing-from animal necropsy all the way to staining. >> >> So why isn't Histology part of Medical School/Pathology training? >> >> Kathleen >> Principal Lab Technician >> Neurotoxicology Labs >> Rutgers University >> >> >> >> >> >> >>> "OOOHHH, there it is." >>> >>> Oh, the days in the lab... My favorite is the pathologist who came >>> in late >>> in the evening, opened the (already running) tissue processor and then >>> spent >>> two hours grossing, occasionally lobbing a cassette into the processor. >>> Next >>> day he (and all the other pathologists) was mad that his blocks were >>> were >>> two hours late - but soon he was hiding under his desk after he >>> explained >>> what he had done and all the other pathologists turned on him. Luckily, >>> only >>> his blocks were wrecked - he had lobbed them in while the processor was >>> late >>> in 100%, and did not fully dehydrate. >>> >>> It is truly amazing how ignorant most pathology residents are about >>> basic >>> histology procedures. Our lab director was enlightened and he made >>> all the >>> path residents spend 5 full days in the histo lab following the tissue >>> through the entire process from accession to H&E, then specials, >>> IHC, EM >>> etc. They were chaffing by the second day, but by the 5th they were >>> very, >>> very appreciative of all the work that went on to get their slides and >>> stains done. It made life a lot easier later because they actually >>> understand what was going on - and knew it was better to ask a tech >>> what >>> to >>> do rather than just blindly (arrogantly?) do what ever they felt like >>> doing. >>> >>> Tim Morken >>> >>> >>> >>> >>> I had a resident come in looking at at a block contemplating ordering a >>> special stain. She looked and looked at the top of the block where >>> tissue >>> is placed when grossing. >>> >>> Finally she said. "I need to order a special on this block, but there >>> isn't >>> any more tissue." >>> >>> The tech took the block out of her hand and turned it over. >>> >>> >>> "OOOHHH, there it is." >>> >>> Ross M Stapf >>> Histopathology Manager >>> Baylor University Medical Center >>> 3500 Gaston Ave. >>> Dallas, TX 75246 >>> 214-820-2465 >>> >>> 214-820-4110 fax >>> RossS@baylorhealth.edu >>> >>> >>> >>> -----Original Message----- >>> From: histonet-bounces@lists.utsouthwestern.edu >>> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred >>> Underwood >>> Sent: Wednesday, February 23, 2005 10:02 AM >>> To: froyer@bitstream.net; histonet@lists.utsouthwestern.edu >>> Subject: [BULK] - Re: [Histonet] Left-handed microtome. >>> >>> >>> Or, perhaps the pathologist wanted sections from the opposite side >>> of the >>> block. >>> >>> >>> >>>>>> Ford Royer 02/23/05 09:43AM >>> >>>>>> >>>>> >>> Jim Staruk wrote: >>> >>> >>> >>>> I remember an older histologist I met in Florida who had the >>>> >>> >>> microtome >>> >>> >>>> facing away from him as he cut ribbons! >>>> >>>> Jim >>>> >>>> ______________________ >>>> Jim Staruk >>>> Mass Histology Service >>>> www.masshistology.com >>>> >>>> >>> >>> Poor fellow... didn't know whether he was coming or going. Obviously >>> due >>> to >>> all those years of unbridled exposure to xylene & >>> >>> formalin (or is that formaldehyde? ...I can never remember). >>> >>> ;-) ~ Ford >>> Ford M. Royer, MT(ASCP) >>> Midwest Science Biocenter, Inc. >>> Minneapolis, MN >>> 800-745-4869 >>> >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >>> This e-mail, facsimile, or letter and any files or attachments >>> transmitted >>> with it contains information that is confidential and privileged. This >>> information is intended only for the use of the individual(s) and >>> entity(ies) to whom it is addressed. If you are the intended recipient, >>> further disclosures are prohibited without proper authorization. If you >>> are >>> not the intended recipient, any disclosure, copying, printing, or >>> use of >>> this information is strictly prohibited and possibly a violation of >>> federal >>> or state law and regulations. If you have received this information in >>> error, please notify Baylor Health Care System immediately at >>> 1-866-402-1661 >>> or via e-mail at privacy@baylorhealth.edu. Baylor Health Care >>> System, its >>> subsidiaries, and affiliates hereby claim all applicable privileges >>> related >>> to this information. >>> >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@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 mhorne <@t> upei.ca Thu Feb 24 11:06:16 2005 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] lectins Message-ID: <421DDF57.27008.414902B@localhost> Hello Everyone , Has anyone out there conjugated gold colloid to lectins? Some gold conjugated lectins I can buy but others I may have to make. Thanks , Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada From jfish <@t> gladstone.ucsf.edu Thu Feb 24 12:21:04 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: pathologists in the lab (shivers down thespine...) In-Reply-To: <421E118A.7050007@bitstream.net> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328CA47@usca0082k08.labvision.apogent.co m> <15483.128.6.148.191.1109198078.squirrel@webmail.rci.rutgers.edu> <421D3613.9060003@umdnj.edu> <421E118A.7050007@bitstream.net> Message-ID: Hello, just my two cents worth, I agree that if there were a way the residents should spend at least a short period of time in a lab so that they can see several different staining procedures from beginning to end, from the actually biopsy procedure to the staining procedure. They should see some slides of properly and improperly handled samples, just to see what a small mistake can lead to further down the line. On a more personal note: There have been times when I have been the patient lying on the examination table, that I have wondered if the doctor is handling my biopsy correctly. I wonder if she knows what that small piece of tissue was about to go through, either freezing or fixation and paraffin embedding, and whether or not she knows that the way she handles it now can change dramatically the outcome of whatever tests she orders. I know she has an idea of how precious it is, but does she know how one small moment can change the results, such as letting the tissue sit on the table to dry slightly, or smashing it, or not placing it in enough fixative, correct fixative, etc. If she had spent a week in a lab, she would know more about the steps that the tissue goes through, and the precise decisions histologists (or other lab personnel) make concerning the handling of the tissue once it is in their hands. One more side note: My husband had his "dreaded" but necessary Colonoscopy last fall and had a biopsy of one polyp taken. We waited on pins and needles (no pun intended) for the results for two weeks. I could not believe it when they could not provide test results on my husband's biopsy because the tissue was "too destroyed to do the ordered tests". He actually had to go through another round of tests and exams because, most likely, the doctor probably didn't handle the original sample correctly. Notice I don't blame the lab personnel! Take care, Jo Dee Fish ********************************************************************** ********** Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 734-5766 Fax: (415) 355-0230 E-mail: jfish@gladstone.ucsf.edu Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158 From pwebster <@t> hei.org Thu Feb 24 12:23:17 2005 From: pwebster <@t> hei.org (Paul Webster) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] lectins In-Reply-To: <421DDF57.27008.414902B@localhost> Message-ID: Jeurgen Roth and John Lucocq did lots of studies conjugating lectins to colloidal gold particles about 20 years ago. I think they finally concluded that it was more efficient to use sequential labeling protocols with secondary labeling reagents for detecting lectins. That is, you react the lectin with the section, bind an anti-lectin antibody to the lectin and then visualize with a gold-conjugated secondary antibody (or protein a gold). One method they described used fetuin-gold to detect lectin bound to the section (Light and electron microscopic demonstration of sialic acid residues with the lectin from Limax flavus: a cytochemical affinity technique with the use of fetuin-gold complexes. J Histochem Cytochem. 1984 Nov;32(11):1167-76.). Paul Webster. Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057-1922 Phone: 213 273 8026 Fax: 213 13 739 E-mail: pwebster@hei.org On 2/24/05 6:06 AM, "Margaret Horne" wrote: > Hello Everyone , > Has anyone out there conjugated gold colloid to lectins? > Some gold conjugated lectins I can buy but others I may have to > make. > > Thanks , > Margaret > > > > > Margaret Horne , > Histology Teaching Assistant, > Dept. of B.SC., > Atlantic Veterinary College, U.P.E.I., > 550 University Ave., Charlottetown, > P.E.I., C1A 4P3 > Canada > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From plucas <@t> biopath.org Thu Feb 24 12:30:44 2005 From: plucas <@t> biopath.org (Paula Lucas) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] CAP Immunohistochemistry Tissue Microarray Program Message-ID: <004b01c51a9e$f4397b90$7c01a8c0@biopath.org> My administrator asked me to research this clinical trial, so of course I turn to the experts. Here is what the advertisement says: This is a program that stains and interprets tissue arrays for HER2, CD117, ER, CD20 and EGFR. Comparing results to our peers. It validates our testing strategy and demonstrates eligibility for participation in NCI funded clinical trials for HER2. Has anyone heard of this program and if so, could you tell me anything and everything about it? Thanks in advance. Paula Bio-Path Medical Group From RossS <@t> BaylorHealth.edu Thu Feb 24 12:41:55 2005 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: pathologists in the lab (shivers down thespine...) Message-ID: <74A8B7A904152141BD5AA2761F5D22340E480B@BHDAEXCH11.bhcs.pvt> Our new residents after a few weeks on their first Gross Room rotation spend a day in histology to see the blocks they grossed get embedded and cut. They get to see first hand the results of their work. It helps tremendously. This was not always done consistently here. Hence the story I shared earlier. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From info <@t> instrumedics.com Thu Feb 24 13:38:35 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] cryosections References: <421E06AB.80807@bms.com> Message-ID: <01aa01c51aa8$73a15cf0$6401a8c0@INSTRUMEDICS22> Frozen sectioning can be difficult and requires a skillful hand. It is often hard to produce a flat, unfolded, fully intact section using the conventional method of retrieving a section on a room temperature slide. The result of melting the unfixed section to mount it on the warm slide degrades the morphology. With the CryoJane Tape-Transfer process the section is captured on a COLD tape as it is being cut . The section is flat, uncompressed, intact and as thin a 2 microns. The section is transferred to a COLD slide. The section is still frozen and the morphology preserved A choice of fixation is then made depending on the staining protocol to follow. This method makes it possible to cut hard tissue or fatty tissue which is often very problematic. The CryoJane process is easy to learn and makes preparing difficult frozen sections easy. For the details of the CryoJane process please visit our web site www.instrumedics.com Bernice Instrumedics ----- Original Message ----- From: "Kelly D Mcqueeney" To: "Till, Renee" Cc: Sent: Thursday, February 24, 2005 11:54 AM Subject: Re: [Histonet] cryosections > Renee, > > You should insist on an in-depth training from the vendor. Ask for a 4 > hour training from their cryostat expert, not some sales person. They > should offer it to you. The technique is standard but varies from user to > user. You'll have to find your way. I am going to give you our standard > protocol for brain IHC. > > 1) Remove fresh tissue, dip in mounting medium (we use Shandon embedding > medium, or you can skip this step) and immediately freeze in ice-cold > isopentane (keep on dry ice for 30 minutes prior to freezing). For rat > brain, we freeze for 20 seconds. Remove from isopentane and place tissue > in a conical tube (some people use molds). Leave the tube on dry ice and > immediately store at -70C. > > 2) Remove tissue from -70C and place in -20C freezer for 1 hour to > equilibrate. > > 3) Drop embedding medium on chuck, add tissue and freeze base with > cytocool. Cover tissue with embedding medium (we dip and freeze with > cytocool). > > 4) Place chuck in cryostat and section tissue. I use probe on plus slides > from Fisher or Superfrost from VWR. > > 5) Once sections are mounted on slides, dry slides completely for 1 hour > RT and store at -80C until ready to use. > > 6) Remove slides from freezer and thaw at RT for 15 minutes. Place slides > in fix and stain or perform IHC. > > There are many different methods for sectioning/storing, etc. Times, > tissue, temperature and protocols will vary from lab to lab or person to > person. > > Have fun! > Kelly > > Till, Renee wrote: > >>Hello. Our research group just purchased our first cryostat. Until now, >>anytime cryosections were needed we had to send out our tissues to >>another lab. Is there a good book or other source for general >>information about cryosectioning? I am the only histotech in the group, >>but have had virtually no experience with cryosections. I know the other >>general techs that also do some histology are going to come to me with >>all their questions. We basically need to know anything and everything. >>Due to the volume of tissues, sectioning won't be performed right as the >>tissues are removed from the animal. How do you store them? Do you fix, >>and when? What type of slides are best? How do you store the slides >>after cryosectioning? Are there specific considerations to take into >>account as far as fixing, or anything else depending on the tissue or >>what stains will be performed on it? For example, I know already that >>there will be some heart and aorta sectioned for fat stains, some pig >>gi, possibly mammary gland, uterus, and mouse lung with b-gal. >>Thanks in advance for any help or suggestions you can give me. >> >> >>Renee' >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > From petepath <@t> yahoo.com Thu Feb 24 13:57:23 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Re: cryosections Message-ID: <20050224195723.9044.qmail@web30410.mail.mud.yahoo.com> Sorry I left the // out of the address for the frozen section tutorial. Fere is the complete link http://pathologyinnovations.com/frozen_section_technique.htm Stephen Peters M.D. Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com Senior Attending Pathologist Hackensack University Medical Center 201 996 4836 From maria <@t> ski.org Thu Feb 24 14:14:30 2005 From: maria <@t> ski.org (Maria Mejia) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Re: cryosections References: <20050224195723.9044.qmail@web30410.mail.mud.yahoo.com> Message-ID: <421E35A6.7010104@ski.org> Dr. Peters, please let us (histonet) know when you have completed & have added some original techniques for sur. path dissections to your website. Maria Bartola Mejia Neurohistologist Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Stephen Peters M.D. wrote: >Sorry I left the // out of the address for the frozen section tutorial. Fere is the complete link >http://pathologyinnovations.com/frozen_section_technique.htm > > > >Stephen Peters M.D. >Pathology Innovations, LLC >410 Old Mill Lane, >Wyckoff, NJ 07481 >201 847 7600 >www.pathologyinnovations.com > >Senior Attending Pathologist >Hackensack University Medical Center >201 996 4836 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Daydawning <@t> wideopenwest.com Thu Feb 24 15:38:18 2005 From: Daydawning <@t> wideopenwest.com (Dawn Truscott) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] (no subject) Message-ID: <001301c51ab9$280deb80$6401a8c0@wowway.com> Don't dis "just the sales person" A good percentage of our sales force are histotechs!! Dawn M. Truscott, HT(ASCP) Richard Allan Scientific Renee, You should insist on an in-depth training from the vendor. Ask for a 4 hour training from their cryostat expert, not some sales person. They should offer it to you. The technique is standard but varies from user to user. You'll have to find your way. I am going to give you our standard protocol for brain IHC. Dawn M. Truscott Recycling New Year's resolutions since 1987. From kelly.mcqueeney <@t> bms.com Thu Feb 24 16:14:38 2005 From: kelly.mcqueeney <@t> bms.com (Kelly D Mcqueeney) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] (no subject) In-Reply-To: <001301c51ab9$280deb80$6401a8c0@wowway.com> References: <001301c51ab9$280deb80$6401a8c0@wowway.com> Message-ID: <421E51CE.1080702@bms.com> I just had a salesperson try to train us on a new cryostat from Richard Allan and it was the first time she used it. We had to put it together and teach ourselves, it was a disaster. My point for Renee, make sure they know how to use the machine. Dawn Truscott wrote: >Don't dis "just the sales person" A good percentage of our sales force are histotechs!! > >Dawn M. Truscott, HT(ASCP) >Richard Allan Scientific > > > > > >Renee, > >You should insist on an in-depth training from the vendor. Ask for a 4 >hour training from their cryostat expert, not some sales person. They >should offer it to you. The technique is standard but varies from user >to user. You'll have to find your way. I am going to give you our >standard protocol for brain IHC. >Dawn M. Truscott > >Recycling New Year's resolutions since 1987. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From TIGERGIL <@t> aol.com Fri Feb 25 00:29:30 2005 From: TIGERGIL <@t> aol.com (TIGERGIL@aol.com) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] stent techniques anyone??? Message-ID: <81.222d9ee0.2f501fca@aol.com> ok...here is one where i need some guidance.....post mortem manangement of coronary artery stents....those little metal mesh things...should i push the clot out and section it?? gil corrigan msmdphd tigergil@aol.com best practise anyone??? ps we are starting a computer section in the aafs ...in case you known some forensic nurds...send them to tigergil@aol.com From ssq5977 <@t> yahoo.com.cn Fri Feb 25 01:25:53 2005 From: ssq5977 <@t> yahoo.com.cn (=?gb2312?q?=CA=E7=C7=ED=20=C9=F2?=) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Is there any DVD about how to make frozen section? Message-ID: <20050225072553.28001.qmail@web15604.mail.cnb.yahoo.com> Hi,everybody, I am a layman in IHC.I heard from one of my friend that some lab would provide DVD on how to make frosen sections and so on.Would anyone give me some hints? Thanks a lot. Shuqiong Shen Jinling hospital Nanjing China --------------------------------- Do You Yahoo!? 150ÍòÇúMP3·è¿ñËÑ£¬´øÄú´³ÈëÒôÀÖµîÌà ÃÀÅ®Ã÷ÐÇÓ¦Óо¡ÓУ¬ËѱéÃÀͼ¡¢ÑÞͼºÍ¿áͼ 1G¾ÍÊÇ1000Õ×£¬ÑÅ»¢µçÓÊ×ÔÖúÀ©ÈÝ£¡ From jean.gillson <@t> elht.nhs.uk Fri Feb 25 05:02:57 2005 From: jean.gillson <@t> elht.nhs.uk (Jean Gillson) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] 34BE12 on prostate[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05AC0348@bhrv-nt-11.bhrv.nwest.nhs.uk> Dear all fellow histologist, We seem to have a recurrent problem with 34BE12 IHC staining on both prostate chippings and prostatectomy specimens. Some basal cells stain whilst others that should stain are negative. Our procedure is Trypsin for 12 minute at pH 7.8 and 37 degrees C. We use a Dako autostainer machine and our antibody is from Dako. Previous spec sheets state Trypsinisation but our most recent one states heat retrieval. So as a test we did both retrieval methods. Oddly enough, microwaved in Dako retrieval solution was slightly better in prostatectomy specimens than trypsin (still uneven staining) and worst in chipping specimens. Can anyone offer any advice? What antibody panels are people doing for prostate cases? Help desperately needed!! Jean Gillson BMS2 Histology Department Blackburn Royal Infirmary Blackburn UK From petepath <@t> yahoo.com Fri Feb 25 06:41:54 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] stent techniques anyone??? Message-ID: <20050225124154.22991.qmail@web30407.mail.mud.yahoo.com> I had do do this on a research project on pig carotids where the goal was to see the reaction to the stent which was buried in an organizing fibrous tissue responce. I took a strong sissor and cut the vessel transversely into 2-3mm rings. Using a forceps I was able to pull the now cut wires making up the stent out of the tissue leaving the tissue pretty much intact. It came out quite well. PS don't use your favorite sissor! Stephen Peters M.D. Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com Senior Attending Pathologist Hackensack University Medical Center 201 996 4836 From TMcNemar <@t> lmhealth.org Fri Feb 25 06:45:03 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] CAP Immunohistochemistry Tissue Microarray Program Message-ID: <6CD94D97ED7D924BA5C2B588FA95282139678D@nt_exchange.lmhealth.org> Not much to tell, it's prety much like any other survey. They send you 1 slide with multiple tissue samples. You stain it, the pathologists reads it and you send it back. We did it one year. It's the same four antibodies every year so I haven't bought it again. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Paula Lucas [mailto:plucas@biopath.org] Sent: Thursday, February 24, 2005 1:31 PM To: Histonet (E-mail) Subject: [Histonet] CAP Immunohistochemistry Tissue Microarray Program My administrator asked me to research this clinical trial, so of course I turn to the experts. Here is what the advertisement says: This is a program that stains and interprets tissue arrays for HER2, CD117, ER, CD20 and EGFR. Comparing results to our peers. It validates our testing strategy and demonstrates eligibility for participation in NCI funded clinical trials for HER2. Has anyone heard of this program and if so, could you tell me anything and everything about it? Thanks in advance. Paula Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Fri Feb 25 08:16:00 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] xylene free mounting media Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3C96@exchsrv01.barrynet.barry.edu> If one sticks to stains that are fast in absolute alcohol, one can mount from absolute alcohol in Euparal (gum sandarac), which uses eucalyptol as a solvent. Hematoxylin fades slowly in Euparal, but is said to keep virtually forever in Euparal Vert. 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 Robin Maxwell-Nunn Sent: Monday, February 21, 2005 8:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] xylene free mounting media Hi We have a tech in our lab who is sensitive to xylene (exhibits headaches, puffy eyes and itchy skin). We are switching to Formula 83 by CBG Biotech for frozen sections but still need a mountant that is both compatible with this and xylene free. Does anyone have any suggestions? thanks, Robin Maxwell-Nunn, MLT St Mary's General Hospital, Kitchener, Ontario, Canada. _______________________________________________ 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 katri <@t> cogeco.ca Fri Feb 25 10:38:08 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] 34BE12 on prostate[Scanned] References: <1030B679AD69D6119C3F00080210DD9D05AC0348@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: <001601c51b58$63715190$6a9a9618@Katri> Hi Jean, Yes, clone 34 BE12 can be a problematic one. Over the years we have struggled same as you, but seem to have pretty good handle on it now. Our antibody is same as yours and we find, that more intense HIER is required. We are using citrate buffer (EDTA or others may be better, you'll have to try) in Decloaker. Our citrate buffer is home made and we add Tween 20 into it. Our dilution of the primary antibody now is 1:30. I hope this helps... Katri Katri Tuomala Hamilton,Ontario, Canada ----- Original Message ----- From: "Jean Gillson" To: Sent: Friday, February 25, 2005 6:02 AM Subject: [Histonet] 34BE12 on prostate[Scanned] > Dear all fellow histologist, > > We seem to have a recurrent problem with 34BE12 IHC staining on both > prostate chippings and prostatectomy specimens. Some basal cells stain > whilst others that should stain are negative. Our procedure is Trypsin > for > 12 minute at pH 7.8 and 37 degrees C. We use a Dako autostainer machine > and > our antibody is from Dako. Previous spec sheets state Trypsinisation but > our most recent one states heat retrieval. So as a test we did both > retrieval methods. Oddly enough, microwaved in Dako retrieval solution > was > slightly better in prostatectomy specimens than trypsin (still uneven > staining) and worst in chipping specimens. Can anyone offer any advice? > What antibody panels are people doing for prostate cases? Help > desperately > needed!! > > Jean Gillson BMS2 > > Histology Department > > Blackburn Royal Infirmary > > Blackburn > > UK > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.J.Scholz <@t> osfhealthcare.org Fri Feb 25 11:01:23 2005 From: Stephen.J.Scholz <@t> osfhealthcare.org (Scholz, Stephen J.) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] JACHO Compliance Message-ID: <7F1312711CA7474A89B3DF8BA0BA54D0F5F190@pmc-rfd-mx01.intranet.osfnet.org> Hello all; We are about to have a JACHO inspection and a problem was brought to my attention. It involves using two identifiers on all laboratory specimens. The below paragraph is from JCAHO FAQ web site for laboratory compliance with the National Patient Safety Goals. This particular statement seems to answer the question about use of only one identifying number on a pathology or any lab specimen. Please let me know your interpretation and how your laboratories comply to it. Currently we have only the case number (hand written) on the blocks and only the case number (hand written) on the slides until after staining at which point there are printed labels put on them. Take note that this FAQ was updated in January, 2005. FAQ-We use a label for the initial patient sample that contains only one unique identifier, a number, to identify laboratory specimens. This unique identifier can be traced to multiple patient identifiers. Does this meet the intent of the goal? ANS-No. The intent is to use two separate identifiers on the specimen label. Confidence in an accurate identification improves as the number of identifiers increases, depending upon their uniqueness. A single identifier can be more easily misread, resulting in avoidable errors. Bar coding that includes two or more person-specific identifiers (not room number) will comply with this requirement. (New 1/18/05) What are suggestions to my problem. Stephen J. Scholz HT(ASCP) Histology Coordinator OSF St. Anthony Medical Center Rockford IL Phone: 815-395-5410 Fax: 815-395-5364 From Kristopher.Kalleberg <@t> unilever.com Fri Feb 25 12:34:23 2005 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] magnesium detection Message-ID: Does anyone know of a way to detect magnesium or calcium, through IHC or fluorescence, in skin? What about quinalizarin? THanks. Kris L Kalleberg Histologist Unilever R&D 45 River Rd. Edgewater, NJ 07020 201 840 2472 From dswarts <@t> pitt.edu Fri Feb 25 12:35:48 2005 From: dswarts <@t> pitt.edu (J. Douglas Swarts) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] rescue sections for staining Message-ID: <421F7004.5010509@pitt.edu> Hello, We are trying to stain some sections cut about ten years ago. They are rat cranial bases including the middle ear which is the area we are interested in. When we stain the sections with H&E we get no hematoxylin staining, ie. no nuclei at all, but the eosin gives a nice uniform pink color. The specimens were decalcified in 10% formic acid which was neutralized with sodium sulfate according to the histologists notes. The sections stained at the time show very little hematoxylin staining. So the questions are: Are these sections overdecalcified? If so is there any technique I can use to rescue them? Did the neutralization with sodium sulfate create the problem? If so can I do anything to reverse the effect? Finally, if one or the other of these situations are true and cannot be remedied is there another staining protocol that will give me a "reasonable" level of morphologic detail? Thanks for your help. Doug From ddeibler <@t> centexpathlab.com Fri Feb 25 12:47:43 2005 From: ddeibler <@t> centexpathlab.com (David Deibler) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Equipment Message-ID: <000801c51b6a$7e380110$2f01010a@tamtron.ctpl.dns> We are looking for an old Shandon portable coverslipping hood. Our lab needs to buy on for IHC. If anyone has a used one let me know , we would love to purchase it. David Deibler ddeibler@centexpathlab.com From la.sebree <@t> hosp.wisc.edu Fri Feb 25 13:16:22 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] 35BH11 Message-ID: Hi all, I'm wondering if there is a reference lab out there (preferably offering "stain only") that does 35BH11 stain. I'm assuming that's a clone but I don't have any other name for it. 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 lcheung <@t> cvmed.stanford.edu Fri Feb 25 13:14:52 2005 From: lcheung <@t> cvmed.stanford.edu (Cheung, Lauren) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] paraffin embedding of mouse tails Message-ID: <8022E9EE6D5ED511A39200A0C9DED0E902FF358E@cvexchange> Hi everyone, I work with mouse tail sections, each approximately 4-5 mm in thickness. I have been attempting to embed them in paraffin for sectioning however sectioning them hasn't been very successful. I can't obtain a ribbon and the sections seem to break out of the paraffin and break apart. I currently fix the tails in 4% Paraformaldehyde overnight and then place them in EDTA for 7-10 days for decalcification. Afterwards, I dehydrate them through a series of graded alcohol steps, clear them in xylenes, and then let them sit in paraffin. I am wondering if anyone has a good protocol including times for each step which I could test out on my samples. Any advice would be greatly appreciated. Thanks so much, Lauren Lauren Cheung Stanford University Rockson Lab Lab: 650-723-5354 Email: Lcheung@cvmed.stanford.edu From jlinda <@t> ces.clemson.edu Fri Feb 25 13:48:30 2005 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] RE: stent techniques anyone??? Message-ID: <5.2.1.1.2.20050225140622.02910608@mailhost.ces.clemson.edu> You stated: "ok...here is one where i need some guidance.....post mortem manangement of coronary artery stents....those little metal mesh things...should i push the clot out and section it?? gil corrigan msmdphd tigergil@aol.com best practise anyone??? ps we are starting a computer section in the aafs ...in case you known some forensic nurds...send them to tigergil@aol.com" Gil, Removing the stent or clot can remove a lot of valuable information regarding biocompatibility. A better choice would be to embed the section in plastic (methyl methacrylate, glycol methacrylate, etc.) and then section on an automated microtome with tungsten carbide blade or slab section with a diamond coated blade and grind the sample to the desired thinness. Of course, this requires specialized equipment and technical skill that many routine clinical labs may not have. You might want to consider sending these out for contract work. The private contract labs that process stents on a routine basis have some really good procedures BUT cannot (unfortunately) share with others as they are bound by non-disclosure agreements. If you are familiar with plastics, you may want to give it a try! Good Luck, Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo/histo.htm+ From carolb <@t> mail.phys.mcw.edu Fri Feb 25 14:21:06 2005 From: carolb <@t> mail.phys.mcw.edu (Carol Bobrowitz) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] GFP Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A9640@thor.phys.mcw.edu> Has anyone worked with GFP tissue? I have found GFP is surviving frozen sections, acetone and 100% alcohol. I am wondering about FFPE tissue. I'm thinking the heat during processing would destroy the GFP. Does formalin fixation effect the GFP? Is it possible to fix the tissue in an alcoholic fixative and manually hand process at room temperature and infiltrate into plastic? Would the plastic medium destroy the GFP? What about celloidin embedded tissue? This is not my choice Any information would be helpful. I thank everyone on the histonet that has in the past and future helped me. I really appreciate your help. Thanks, Carol Ann Bobrowitz Histology Laboratory Department of Physiology - Room 541 Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, Wisconsin 53226 414-456-8179 FAX 414-456-6546 cbobrowi@mcw.edu From straussj <@t> upstate.edu Fri Feb 25 14:39:58 2005 From: straussj <@t> upstate.edu ( Judy Strauss) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] CXCR4 & CXCL12 antibodies Message-ID: Can anyone recommend antibodies to CXCR4 (fusin) and CXCL12 that are suitable for immunohistochemistry in formalin fixed paraffin embedded rat tissue? Thanks, Judith Strauss SUNY Upstate Medical University Department of Orthopedic Surgery IHP room 3118 505 Irving Avenue Syracuse, NY 13120 phone: (315) 464-9960 fax: (315) 464-6638 From TheBestTime23 <@t> aol.com Fri Feb 25 15:43:50 2005 From: TheBestTime23 <@t> aol.com (TheBestTime23@aol.com) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] richard-allen type 9 paraffin Message-ID: <92.2169ca98.2f50f616@aol.com> Thanks for your reply. How long have you been using this paraffin? What temperatures are you using in your water baths and processors? What do you use to keep your blocks chilled (if anything)? I'm having a little difficulty with this new paraffin. I thought it was something that I needed time getting used to, but it has been almost 2 months of using it and I am still having problems (and so are the 3 other people I work with). Problems include compression, fragile ribbons and wrinkles. I am now taking an excessive amount of time and care cutting my blocks only to discover minute folds that could not be seen on the waterbath. We cut our routine sections at 4 microns and our temperatures are where they should be for what the company recommends (59 in processor and holding tanks and waterbath between 35-40). I just thought someone that had worked with this paraffin for a while could give me some tips. Otherwise I might have to start wearing a wig : ) Thank you! Megan From lindas <@t> awesomenet.net Fri Feb 25 16:00:17 2005 From: lindas <@t> awesomenet.net (Linda Davis) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Re:35BH11 Message-ID: <006001c51b85$642517c0$de0ac942@D6JLZ851> Linda, Esoterix will stain 35BH11 if you send them your block. They will send the block and slides back to you for your pathologist to interpert the stain. Linda Davis Rio Grande Regional Hospital McAllen, TX. (956) 632-6402 Fax (956) 632-6641 -------------- next part -------------- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.300 / Virus Database: 266.3.0 - Release Date: 2/21/2005 From lindas <@t> awesomenet.net Fri Feb 25 16:22:53 2005 From: lindas <@t> awesomenet.net (Linda Davis) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Fw: Re:35BH11 Message-ID: <004001c51b88$8c565ee0$6d0ac942@D6JLZ851> Linda, Esoterix will stain 35BH11 if you send them your block. They will send the block and slides back to you for your pathologist to interpert the stain. Linda Davis Rio Grande Regional Hospital McAllen, TX. (956) 632-6402 Fax (956) 632-6641 -------------- next part -------------- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.300 / Virus Database: 266.3.0 - Release Date: 2/21/2005 From cfockler <@t> mail1.vcu.edu Fri Feb 25 16:55:56 2005 From: cfockler <@t> mail1.vcu.edu (cfockler@mail1.vcu.edu) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Histotech Marketability Message-ID: <200502252255.RAA22846@arrakis.vcu.edu> Hello histonetters. . .just a quick question. What is the marketability (pay scale ect.) of a tech with a BS in biology/chemistry, 2 years of experience with proven competency in all aspects, grosses, and has IHC and IF experience? I am looking to get feedback from all over the country. Any information would be greatly appreciated! Thanks. Candyce Fockler Histotechnician Anatomic Pathology From mhirano <@t> ku.edu Fri Feb 25 17:48:01 2005 From: mhirano <@t> ku.edu (HIRANO Makoto) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Square "sample stage" Message-ID: I am a beginner in frozen sectioning. Now I have a silly question. What is the formal name of a metal, square "sample stage" or "chunk" for cryostat? I mean the 'partner' of a cryomold. I have visited some web sites and consulted some catalogues, but I cannot find out the name. I have absolutely to order some. Thanks a lot in advance. Makoto HIRANO, MD Department of Molecular Biosciences University of Kansas E-mail: mhirano@ku.edu From hlukey <@t> msn.com Fri Feb 25 21:45:10 2005 From: hlukey <@t> msn.com (Hugh Luk) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] richard-allen type 9 paraffin Message-ID: Megan, Just my 2 cents. If any of this helps, great. I've found this is one of the hardest, polymer-rich paraffins available. I find it similar to Surgipath's standard embedding paraffin and Cardinal's (old S/P) Ameraffin. It sections and ribbons well. Currently, however, I am using Ameraffin (also made by Richard Allan), which I find very interchangable with type 9. We modified our tissue processing (a little more time in wax-multiple changes totalling around 2 hours at 58 to 62C- and maybe more alcohol), deparaffinization (more time in solvent), and two, different microtome blades. Sakura's Accu-Edge HIGH profile blades are honed perfectly to cut ANYTHING at 4 microns with this wax. The "Slight compression" is balanced with "Ease of ribboning." We sometimes use a wider angle on the blade holder to increase the rake angle. For really thin sections, "Sharper" blades (like Duraedge, Leica's, or Richard Allan's blades) are required. For example, on delicate biopsies and lymph nodes, 1 or 2 microns is no problem! I also like the way the paraffin holds its shape on the water bath (~38-44C). I learned histology using Paraplast X-tra. I've tried Paraplast, Paraplast plus (with DMSO), Fisher Tissue Prep and Tissue Prep2 embedding medias. When we had a new supervisor at QMC, she changed the paraffin to a harder wax. We complained about it because it felt different, but I came to really like it. It became my standard where ever I work (thanks Jackie O'). If you are experimenting, you will need to do full switch-overs, every instrument using wax should be changed. Mixing paraffins of different grades is cheating the experiment. My conclusion is they are all pretty good. The paraffin is only for support; and with a little tinkering, I like to think I can use any one of them at any time. Hugh Luk, HTL (ASCP) Former histopathology supervisor CLH hluk@crch.hawaii.edu Cancer Research Center of Hawaii 1236 Lauhala Street, Room 316 Honolulu, HI 96813 Tel: (808) 440-5238 Fax: (808) 586-2982 >From: TheBestTime23@aol.com >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] richard-allen type 9 paraffin >Date: Wed, 23 Feb 2005 17:42:03 EST > >Hello everyone, > >The lab that I work for recently decided to switch to Richard-Allen type 9 >paraffin. I was wondering if anyone has experience with this paraffin and >could give some advice on techniques used in cutting it. Opinions of this >product would also be greatly appreciated. > >Thanks in advance, >Megan > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LuckG <@t> empirehealth.org Sat Feb 26 01:53:17 2005 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Histotech Marketability Message-ID: Candyce, Request that the HR dept of your current employer do a job code market survey for your geographic area. After they tell you your salary is equivalent to current market conditions ask them how long it would take to fill a vacancy in your area. When they say "We don't know" ask them to try to fill a position (real or not). Then inform them what a "histo'temp" would cost ($57-60 hr.) to fill in while ???? There is a good deal of upward pressure on histotech salaries (well overdue) and given a suitable fit now is the time to look towards the future for new opportunities An individual's concept of their marketability may differ substantially from their employer's but this is limited only by their current work environment. I know, blah, blah, blah. We have a full-time, M-F, day shift, histotech position open. This is a seasoned dept looking towards the future to lay the foundation of a new staff to take over (+/-5 yrs) as we retire. We have a current opportunity for someone either interested in the eventual department manager's role or as a traditional bench histotech. Click on the web links that follow to get a snapshot of our institution and of Spokane: www.deaconessmedicalcenter.org & www.visitspokane.com. Remember, "making a living" is not the same thing as "making a life". Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: cfockler@mail1.vcu.edu [mailto:cfockler@mail1.vcu.edu] Sent: Friday, February 25, 2005 2:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histotech Marketability Hello histonetters. . .just a quick question. What is the marketability (pay scale ect.) of a tech with a BS in biology/chemistry, 2 years of experience with proven competency in all aspects, grosses, and has IHC and IF experience? I am looking to get feedback from all over the country. Any information would be greatly appreciated! Thanks. Candyce Fockler Histotechnician Anatomic Pathology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfockler <@t> mail1.vcu.edu Sat Feb 26 08:17:23 2005 From: cfockler <@t> mail1.vcu.edu (cfockler@mail1.vcu.edu) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] HT vs. HLT In-Reply-To: <20050226140426.7869.qmail@web81003.mail.yahoo.com> Message-ID: <200502261417.JAA24219@despina.vcu.edu> What exactly is the difference between and HT and an HLT. ASCP makes this distinction with regards to their wage and salary survey. Is one certified and the other not? Any information would be greatly appreciated! Thanks. From tdemuth <@t> tgen.org Sat Feb 26 09:52:48 2005 From: tdemuth <@t> tgen.org (Tim Demuth) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Collagen-I gel, dye and autofluorescence Message-ID: Hi there, we are currently trying to cut FFPE sections of multicellular tumor-spheroids grown in collagen-I gel (bovine). There are two major problems: 1) Since the spheroid is so small (diameter ~200-500?m) and very pale it is extremely hard to identify in the fixed and embedded collagen block. We would like to use a dye to visualize the spheroid prior to embedding, so we have some guidance as to where and when to expect the spheroid so we don't have to cut through the whole block. Eosin works fine for that purpose, however it's autofluorescence interferes with fluorescence IHC. Is there any dye that doesn't show autof.? 2) We also experienced the spheroid to pop out of the gel when sectioning resulting in an empty spot on the slide. Any suggestions on how to improve fixation (currently 10% neutral buffered PFA for 24hrs) technique? Thanks for the input, it's greatly appreciated! Tim Translational Genomics Research Institute, TGEN Phoenix AZ From marjoh3 <@t> telus.net Sat Feb 26 13:45:13 2005 From: marjoh3 <@t> telus.net (Marilyn Johnson) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Toxoplasmosis in Sheep by Immunohistochemistry Message-ID: <001001c51c3b$b040eba0$6401a8c0@VALUED20606295> Hi Histonetters in Veterinary Labs, We are initiating a surveillance project involving toxoplasmosis in sheep. Liver sections will be submitted for H&E and Immunohistochemistry staining. I am currently using the Dako LSAB2 detection system for IHC staining. Would any veterinary labs. provide me with information for purchasing the source of primary antibody, including company information (catalog nos., cost, dilutions, methods, etc.)? Thanks in advance to any replies. Greatly appreciated! Marilyn Johnson Food Safety Division Alberta Agriculture Edmonton, Alberta, Canada. From Shawnster73 <@t> aol.com Sat Feb 26 16:15:34 2005 From: Shawnster73 <@t> aol.com (Shawnster73@aol.com) Date: Fri Sep 16 15:24:40 2005 Subject: [Histonet] Block Message-ID: My antibody is a rabbit polyclonal, but the tissue is mouse xenograft tissue of human prostate cancer. In a message dated 2/23/2005 9:06:59 AM Eastern Standard Time, anh2006@med.cornell.edu writes: Is your antibody a mouse monoclonal? More info is needed before I can help. >I am currently trying to stain mouse xenograft tissue using a p53 >antibody. I have worked up this antibody on human breast cancer >using the Ventana AB block and amplifier and have gotten beautiful >results. I am now looking for a way to block and amplify the tissue >using reagents that do not contain mouse. Any suggestions would be >greatly appreciated. > >Michelle HT(ASCP) > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- From billingconsultants <@t> yahoo.com Sat Feb 26 23:05:42 2005 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Histology Training Opportunities in KY? Message-ID: <20050227050542.10159.qmail@web54205.mail.yahoo.com> Hi everyone, Would any of you know of any training opportunities in Kentucky for someone interested in becoming a histology technician? I have a friend in Lexington that has a BS in biology and is really interested in breaking into the field. She would love to find a place - even volunteer her time if necessary for the chance to learn. Any suggestions you may have would be greatly appreciated. Louri Roberts-Caldwell 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. magnesium detection (Kristopher Kalleberg) 2. rescue sections for staining (J. Douglas Swarts) 3. Equipment (David Deibler) 4. 35BH11 (Sebree Linda A.) 5. paraffin embedding of mouse tails (Cheung, Lauren) 6. RE: stent techniques anyone??? (Linda Jenkins) 7. GFP (Carol Bobrowitz) 8. CXCR4 & CXCL12 antibodies ( Judy Strauss) 9. Re: richard-allen type 9 paraffin (TheBestTime23@aol.com) 10. Re:35BH11 (Linda Davis) 11. Fw: Re:35BH11 (Linda Davis) 12. Histotech Marketability (cfockler@mail1.vcu.edu) 13. Square "sample stage" (HIRANO Makoto) 14. RE: richard-allen type 9 paraffin (Hugh Luk) 15. RE: Histotech Marketability (Luck, Greg D.) 16. HT vs. HLT (cfockler@mail1.vcu.edu) 17. Collagen-I gel, dye and autofluorescence (Tim Demuth) ---------------------------------------------------------------------- Message: 1 Date: Fri, 25 Feb 2005 13:34:23 -0500 From: "Kristopher Kalleberg" Subject: [Histonet] magnesium detection To: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: TEXT/PLAIN; CHARSET=US-ASCII Does anyone know of a way to detect magnesium or calcium, through IHC or fluorescence, in skin? What about quinalizarin? THanks. Kris L Kalleberg Histologist Unilever R&D 45 River Rd. Edgewater, NJ 07020 201 840 2472 ------------------------------ Message: 2 Date: Fri, 25 Feb 2005 13:35:48 -0500 From: "J. Douglas Swarts" Subject: [Histonet] rescue sections for staining To: histonet@lists.utsouthwestern.edu Message-ID: <421F7004.5010509@pitt.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hello, We are trying to stain some sections cut about ten years ago. They are rat cranial bases including the middle ear which is the area we are interested in. When we stain the sections with H&E we get no hematoxylin staining, ie. no nuclei at all, but the eosin gives a nice uniform pink color. The specimens were decalcified in 10% formic acid which was neutralized with sodium sulfate according to the histologists notes. The sections stained at the time show very little hematoxylin staining. So the questions are: Are these sections overdecalcified? If so is there any technique I can use to rescue them? Did the neutralization with sodium sulfate create the problem? If so can I do anything to reverse the effect? Finally, if one or the other of these situations are true and cannot be remedied is there another staining protocol that will give me a "reasonable" level of morphologic detail? Thanks for your help. Doug ------------------------------ Message: 3 Date: Fri, 25 Feb 2005 12:47:43 -0600 From: "David Deibler" Subject: [Histonet] Equipment To: Message-ID: <000801c51b6a$7e380110$2f01010a@tamtron.ctpl.dns> Content-Type: text/plain; charset="iso-8859-1" We are looking for an old Shandon portable coverslipping hood. Our lab needs to buy on for IHC. If anyone has a used one let me know , we would love to purchase it. David Deibler ddeibler@centexpathlab.com ------------------------------ Message: 4 Date: Fri, 25 Feb 2005 13:16:22 -0600 From: "Sebree Linda A." Subject: [Histonet] 35BH11 To: "Histonet \(E-mail\)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi all, I'm wondering if there is a reference lab out there (preferably offering "stain only") that does 35BH11 stain. I'm assuming that's a clone but I don't have any other name for it. 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 ------------------------------ Message: 5 Date: Fri, 25 Feb 2005 11:14:52 -0800 From: "Cheung, Lauren" Subject: [Histonet] paraffin embedding of mouse tails To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <8022E9EE6D5ED511A39200A0C9DED0E902FF358E@cvexchange> Content-Type: text/plain; charset="iso-8859-1" Hi everyone, I work with mouse tail sections, each approximately 4-5 mm in thickness. I have been attempting to embed them in paraffin for sectioning however sectioning them hasn't been very successful. I can't obtain a ribbon and the sections seem to break out of the paraffin and break apart. I currently fix the tails in 4% Paraformaldehyde overnight and then place them in EDTA for 7-10 days for decalcification. Afterwards, I dehydrate them through a series of graded alcohol steps, clear them in xylenes, and then let them sit in paraffin. I am wondering if anyone has a good protocol including times for each step which I could test out on my samples. Any advice would be greatly appreciated. Thanks so much, Lauren Lauren Cheung Stanford University Rockson Lab Lab: 650-723-5354 Email: Lcheung@cvmed.stanford.edu ------------------------------ Message: 6 Date: Fri, 25 Feb 2005 14:48:30 -0500 From: Linda Jenkins Subject: [Histonet] RE: stent techniques anyone??? To: histonet@lists.utsouthwestern.edu Message-ID: <5.2.1.1.2.20050225140622.02910608@mailhost.ces.clemson.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed You stated: "ok...here is one where i need some guidance.....post mortem manangement of coronary artery stents....those little metal mesh things...should i push the clot out and section it?? gil corrigan msmdphd tigergil@aol.com best practise anyone??? ps we are starting a computer section in the aafs ...in case you known some forensic nurds...send them to tigergil@aol.com" Gil, Removing the stent or clot can remove a lot of valuable information regarding biocompatibility. A better choice would be to embed the section in plastic (methyl methacrylate, glycol methacrylate, etc.) and then section on an automated microtome with tungsten carbide blade or slab section with a diamond coated blade and grind the sample to the desired thinness. Of course, this requires specialized equipment and technical skill that many routine clinical labs may not have. You might want to consider sending these out for contract work. The private contract labs that process stents on a routine basis have some really good procedures BUT cannot (unfortunately) share with others as they are bound by non-disclosure agreements. If you are familiar with plastics, you may want to give it a try! Good Luck, Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo/histo.htm+ ------------------------------ Message: 7 Date: Fri, 25 Feb 2005 14:21:06 -0600 From: Carol Bobrowitz Subject: [Histonet] GFP To: "'Histonet (E-mail)" Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A9640@thor.phys.mcw.edu> Content-Type: text/plain; charset="iso-8859-1" Has anyone worked with GFP tissue? I have found GFP is surviving frozen sections, acetone and 100% alcohol. I am wondering about FFPE tissue. I'm thinking the heat during processing would destroy the GFP. Does formalin fixation effect the GFP? Is it possible to fix the tissue in an alcoholic fixative and manually hand process at room temperature and infiltrate into plastic? Would the plastic medium destroy the GFP? What about celloidin embedded tissue? This is not my choice Any information would be helpful. I thank everyone on the histonet that has in the past and future helped me. I really appreciate your help. Thanks, Carol Ann Bobrowitz Histology Laboratory Department of Physiology - Room 541 Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, Wisconsin 53226 414-456-8179 FAX 414-456-6546 cbobrowi@mcw.edu ------------------------------ Message: 8 Date: Fri, 25 Feb 2005 15:39:58 -0500 From: " Judy Strauss" Subject: [Histonet] CXCR4 & CXCL12 antibodies To: Message-ID: Content-Type: text/plain; charset=US-ASCII Can anyone recommend antibodies to CXCR4 (fusin) and CXCL12 that are suitable for immunohistochemistry in formalin fixed paraffin embedded rat tissue? Thanks, Judith Strauss SUNY Upstate Medical University Department of Orthopedic Surgery IHP room 3118 505 Irving Avenue Syracuse, NY 13120 phone: (315) 464-9960 fax: (315) 464-6638 ------------------------------ Message: 9 Date: Fri, 25 Feb 2005 16:43:50 EST From: TheBestTime23@aol.com Subject: Re: [Histonet] richard-allen type 9 paraffin To: liz@premierlab.com Cc: histonet@lists.utsouthwestern.edu Message-ID: <92.2169ca98.2f50f616@aol.com> Content-Type: text/plain; charset="US-ASCII" Thanks for your reply. How long have you been using this paraffin? What temperatures are you using in your water baths and processors? What do you use to keep your blocks chilled (if anything)? I'm having a little difficulty with this new paraffin. I thought it was something that I needed time getting used to, but it has been almost 2 months of using it and I am still having problems (and so are the 3 other people I work with). Problems include compression, fragile ribbons and wrinkles. I am now taking an excessive amount of time and care cutting my blocks only to discover minute folds that could not be seen on the waterbath. We cut our routine sections at 4 microns and our temperatures are where they should be for what the company recommends (59 in processor and holding tanks and waterbath between 35-40). I just thought someone that had worked with this paraffin for a while could give me some tips. Otherwise I might have to start wearing a wig : ) Thank you! Megan ------------------------------ Message: 10 Date: Fri, 25 Feb 2005 16:00:17 -0600 From: "Linda Davis" Subject: [Histonet] Re:35BH11 To: Message-ID: <006001c51b85$642517c0$de0ac942@D6JLZ851> Content-Type: text/plain; charset="iso-8859-1" Linda, Esoterix will stain 35BH11 if you send them your block. They will send the block and slides back to you for your pathologist to interpert the stain. Linda Davis Rio Grande Regional Hospital McAllen, TX. (956) 632-6402 Fax (956) 632-6641 -------------- next part -------------- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.300 / Virus Database: 266.3.0 - Release Date: 2/21/2005 ------------------------------ Message: 11 Date: Fri, 25 Feb 2005 16:22:53 -0600 From: "Linda Davis" Subject: [Histonet] Fw: Re:35BH11 To: Message-ID: <004001c51b88$8c565ee0$6d0ac942@D6JLZ851> Content-Type: text/plain; charset="iso-8859-1" Linda, Esoterix will stain 35BH11 if you send them your block. They will send the block and slides back to you for your pathologist to interpert the stain. Linda Davis Rio Grande Regional Hospital McAllen, TX. (956) 632-6402 Fax (956) 632-6641 -------------- next part -------------- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.300 / Virus Database: 266.3.0 - Release Date: 2/21/2005 ------------------------------ Message: 12 Date: Fri, 25 Feb 2005 17:55:56 -0500 From: Subject: [Histonet] Histotech Marketability To: histonet@lists.utsouthwestern.edu Message-ID: <200502252255.RAA22846@arrakis.vcu.edu> Content-Type: text/plain; charset=iso-8859-1 Hello histonetters. . .just a quick question. What is the marketability (pay scale ect.) of a tech with a BS in biology/chemistry, 2 years of experience with proven competency in all aspects, grosses, and has IHC and IF experience? I am looking to get feedback from all over the country. Any information would be greatly appreciated! Thanks. Candyce Fockler Histotechnician Anatomic Pathology ------------------------------ Message: 13 Date: Fri, 25 Feb 2005 17:48:01 -0600 From: "HIRANO Makoto" Subject: [Histonet] Square "sample stage" To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" I am a beginner in frozen sectioning. Now I have a silly question. What is the formal name of a metal, square "sample stage" or "chunk" for cryostat? I mean the 'partner' of a cryomold. I have visited some web sites and consulted some catalogues, but I cannot find out the name. I have absolutely to order some. Thanks a lot in advance. Makoto HIRANO, MD Department of Molecular Biosciences University of Kansas E-mail: mhirano@ku.edu ------------------------------ Message: 14 Date: Fri, 25 Feb 2005 17:45:10 -1000 From: "Hugh Luk" Subject: RE: [Histonet] richard-allen type 9 paraffin To: TheBestTime23@aol.com Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Megan, Just my 2 cents. If any of this helps, great. I've found this is one of the hardest, polymer-rich paraffins available. I find it similar to Surgipath's standard embedding paraffin and Cardinal's (old S/P) Ameraffin. It sections and ribbons well. Currently, however, I am using Ameraffin (also made by Richard Allan), which I find very interchangable with type 9. We modified our tissue processing (a little more time in wax-multiple changes totalling around 2 hours at 58 to 62C- and maybe more alcohol), deparaffinization (more time in solvent), and two, different microtome blades. Sakura's Accu-Edge HIGH profile blades are honed perfectly to cut ANYTHING at 4 microns with this wax. The "Slight compression" is balanced with "Ease of ribboning." We sometimes use a wider angle on the blade holder to increase the rake angle. For really thin sections, "Sharper" blades (like Duraedge, Leica's, or Richard Allan's blades) are required. For example, on delicate biopsies and lymph nodes, 1 or 2 microns is no problem! I also like the way the paraffin holds its shape on the water bath (~38-44C). I learned histology using Paraplast X-tra. I've tried Paraplast, Paraplast plus (with DMSO), Fisher Tissue Prep and Tissue Prep2 embedding medias. When we had a new supervisor at QMC, she changed the paraffin to a harder wax. We complained about it because it felt different, but I came to really like it. It became my standard where ever I work (thanks Jackie O'). If you are experimenting, you will need to do full switch-overs, every instrument using wax should be changed. Mixing paraffins of different grades is cheating the experiment. My conclusion is they are all pretty good. The paraffin is only for support; and with a little tinkering, I like to think I can use any one of them at any time. Hugh Luk, HTL (ASCP) Former histopathology supervisor CLH hluk@crch.hawaii.edu Cancer Research Center of Hawaii 1236 Lauhala Street, Room 316 Honolulu, HI 96813 Tel: (808) 440-5238 Fax: (808) 586-2982 >From: TheBestTime23@aol.com >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] richard-allen type 9 paraffin >Date: Wed, 23 Feb 2005 17:42:03 EST > >Hello everyone, > >The lab that I work for recently decided to switch to Richard-Allen type 9 >paraffin. I was wondering if anyone has experience with this paraffin and >could give some advice on techniques used in cutting it. Opinions of this >product would also be greatly appreciated. > >Thanks in advance, >Megan > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Fri, 25 Feb 2005 23:53:17 -0800 From: "Luck, Greg D." Subject: RE: [Histonet] Histotech Marketability To: "'cfockler@mail1.vcu.edu'" , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Candyce, Request that the HR dept of your current employer do a job code market survey for your geographic area. After they tell you your salary is equivalent to current market conditions ask them how long it would take to fill a vacancy in your area. When they say "We don't know" ask them to try to fill a position (real or not). Then inform them what a "histo'temp" would cost ($57-60 hr.) to fill in while ???? There is a good deal of upward pressure on histotech salaries (well overdue) and given a suitable fit now is the time to look towards the future for new opportunities An individual's concept of their marketability may differ substantially from their employer's but this is limited only by their current work environment. I know, blah, blah, blah. We have a full-time, M-F, day shift, histotech position open. This is a seasoned dept looking towards the future to lay the foundation of a new staff to take over (+/-5 yrs) as we retire. We have a current opportunity for someone either interested in the eventual department manager's role or as a traditional bench histotech. Click on the web links that follow to get a snapshot of our institution and of Spokane: www.deaconessmedicalcenter.org & www.visitspokane.com. Remember, "making a living" is not the same thing as "making a life". Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: cfockler@mail1.vcu.edu [mailto:cfockler@mail1.vcu.edu] Sent: Friday, February 25, 2005 2:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histotech Marketability Hello histonetters. . .just a quick question. What is the marketability (pay scale ect.) of a tech with a BS in biology/chemistry, 2 years of experience with proven competency in all aspects, grosses, and has IHC and IF experience? I am looking to get feedback from all over the country. Any information would be greatly appreciated! Thanks. Candyce Fockler Histotechnician Anatomic Pathology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Sat, 26 Feb 2005 09:17:23 -0500 From: Subject: [Histonet] HT vs. HLT To: histonet@lists.utsouthwestern.edu Message-ID: <200502261417.JAA24219@despina.vcu.edu> Content-Type: text/plain; charset=iso-8859-1 What exactly is the difference between and HT and an HLT. ASCP makes this distinction with regards to their wage and salary survey. Is one certified and the other not? Any information would be greatly appreciated! Thanks. ------------------------------ Message: 17 Date: Sat, 26 Feb 2005 08:52:48 -0700 From: Tim Demuth Subject: [Histonet] Collagen-I gel, dye and autofluorescence To: Message-ID: Content-Type: text/plain; charset="ISO-8859-1" Hi there, we are currently trying to cut FFPE sections of multicellular tumor-spheroids grown in collagen-I gel (bovine). There are two major problems: 1) Since the spheroid is so small (diameter ~200-500µm) and very pale it is extremely hard to identify in the fixed and embedded collagen block. We would like to use a dye to visualize the spheroid prior to embedding, so we have some guidance as to where and when to expect the spheroid so we don't === message truncated === --------------------------------- Do you Yahoo!? Yahoo! Sports - Sign up for Fantasy Baseball. From FrozenInk <@t> aol.com Sun Feb 27 07:44:12 2005 From: FrozenInk <@t> aol.com (FrozenInk@aol.com) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Looking for C4d info Message-ID: Is there a C4d polyclonal antibody out there that is FDA approved? We perform a lot of these,but can not bill for them. Any info would help Gary From pathrm35 <@t> adelphia.net Sun Feb 27 08:59:35 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Florida histotechs grossing derm specimens Message-ID: <13166132.1109516375976.JavaMail.root@web1.mail.adelphia.net> Does anyone know what the reqiurements are in the state of Florida for histotechs grossing tissue? In particular, a private dermpath laboratory with small skin biopsies. What certifications, state licenses and/or college degree are required? Thanks, Ron Martin From jstaruk <@t> masshistology.com Sun Feb 27 10:32:46 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Florida histotechs grossing derm specimens In-Reply-To: <13166132.1109516375976.JavaMail.root@web1.mail.adelphia.net> Message-ID: <3rr3lu$j6sgt5@mxip08a.cluster1.charter.net> Ron, I can't answer your particular question, but it does raise another: Should the "grosser" carry professional liability insurance? If a histotech (or anyone else) working in a private lab loses a specimen for any reason, he will more than likely be sued by someone. Believe me, it happens! I would address this issue and get something in writing before I ever touched a specimen. Jim ____________________ Jim Staruk www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pathrm35@adelphia.net Sent: Sunday, February 27, 2005 10:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Florida histotechs grossing derm specimens Does anyone know what the reqiurements are in the state of Florida for histotechs grossing tissue? In particular, a private dermpath laboratory with small skin biopsies. What certifications, state licenses and/or college degree are required? Thanks, Ron Martin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Sun Feb 27 17:15:21 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Histology Training Opportunities in KY? References: <20050227050542.10159.qmail@web54205.mail.yahoo.com> Message-ID: <010f01c51d22$3695aec0$65d0d445@domainnotset.invalid> Maybe she could print a notice in the Kentucky newsletter. http://www.nsh.org/pubs/statenewsltrs.html Or maybe put up a notice at the state histology symposium March 18-19, 2005 Kentucky Society for Histotechnology Louisville, KY Contact: Renee Matherly, 502-852-5587 Email: rmath0516@aol.com Just a couple of ideas, in case you get no other responses. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Billing Consultants, LLC" To: Sent: Sunday, February 27, 2005 12:05 AM Subject: [Histonet] Histology Training Opportunities in KY? > Hi everyone, > > Would any of you know of any training opportunities in Kentucky for someone interested in becoming a histology technician? I have a friend in Lexington that has a BS in biology and is really interested in breaking into the field. She would love to find a place - even volunteer her time if necessary for the chance to learn. > > Any suggestions you may have would be greatly appreciated. > > Louri Roberts-Caldwell > > 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. magnesium detection (Kristopher Kalleberg) > 2. rescue sections for staining (J. Douglas Swarts) > 3. Equipment (David Deibler) > 4. 35BH11 (Sebree Linda A.) > 5. paraffin embedding of mouse tails (Cheung, Lauren) > 6. RE: stent techniques anyone??? (Linda Jenkins) > 7. GFP (Carol Bobrowitz) > 8. CXCR4 & CXCL12 antibodies ( Judy Strauss) > 9. Re: richard-allen type 9 paraffin (TheBestTime23@aol.com) > 10. Re:35BH11 (Linda Davis) > 11. Fw: Re:35BH11 (Linda Davis) > 12. Histotech Marketability (cfockler@mail1.vcu.edu) > 13. Square "sample stage" (HIRANO Makoto) > 14. RE: richard-allen type 9 paraffin (Hugh Luk) > 15. RE: Histotech Marketability (Luck, Greg D.) > 16. HT vs. HLT (cfockler@mail1.vcu.edu) > 17. Collagen-I gel, dye and autofluorescence (Tim Demuth) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 25 Feb 2005 13:34:23 -0500 > From: "Kristopher Kalleberg" > Subject: [Histonet] magnesium detection > To: "histonet@lists.utsouthwestern.edu" > > Message-ID: > > Content-Type: TEXT/PLAIN; CHARSET=US-ASCII > > Does anyone know of a way to detect magnesium or calcium, through IHC or > fluorescence, in skin? What about quinalizarin? THanks. > > Kris L Kalleberg > Histologist > Unilever R&D > 45 River Rd. > Edgewater, NJ 07020 > 201 840 2472 > > > > > ------------------------------ > > Message: 2 > Date: Fri, 25 Feb 2005 13:35:48 -0500 > From: "J. Douglas Swarts" > Subject: [Histonet] rescue sections for staining > To: histonet@lists.utsouthwestern.edu > Message-ID: <421F7004.5010509@pitt.edu> > Content-Type: text/plain; charset=ISO-8859-1; format=flowed > > Hello, > We are trying to stain some sections cut about ten years ago. They are > rat cranial bases including the middle ear which is the area we are > interested in. When we stain the sections with H&E we get no > hematoxylin staining, ie. no nuclei at all, but the eosin gives a nice > uniform pink color. > > The specimens were decalcified in 10% formic acid which was neutralized > with sodium sulfate according to the histologists notes. The sections > stained at the time show very little hematoxylin staining. So the > questions are: > > Are these sections overdecalcified? If so is there any technique I can > use to rescue them? > > Did the neutralization with sodium sulfate create the problem? If so > can I do anything to reverse the effect? > > Finally, if one or the other of these situations are true and cannot be > remedied is there another staining protocol that will give me a > "reasonable" level of morphologic detail? > > Thanks for your help. > > Doug > > > > > ------------------------------ > > Message: 3 > Date: Fri, 25 Feb 2005 12:47:43 -0600 > From: "David Deibler" > > Subject: [Histonet] Equipment > To: > Message-ID: <000801c51b6a$7e380110$2f01010a@tamtron.ctpl.dns> > Content-Type: text/plain; charset="iso-8859-1" > > We are looking for an old Shandon portable coverslipping > hood. Our lab needs to buy on for IHC. If anyone has a > used one let me know , we would love to purchase it. > > David Deibler > ddeibler@centexpathlab.com > > ------------------------------ > > Message: 4 > Date: Fri, 25 Feb 2005 13:16:22 -0600 > From: "Sebree Linda A." > Subject: [Histonet] 35BH11 > To: "Histonet \(E-mail\)" > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > Hi all, > > I'm wondering if there is a reference lab out there (preferably offering "stain only") that does 35BH11 stain. I'm assuming that's a clone but I don't have any other name for it. > > 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 > > > > > > ------------------------------ > > Message: 5 > Date: Fri, 25 Feb 2005 11:14:52 -0800 > From: "Cheung, Lauren" > Subject: [Histonet] paraffin embedding of mouse tails > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: <8022E9EE6D5ED511A39200A0C9DED0E902FF358E@cvexchange> > Content-Type: text/plain; charset="iso-8859-1" > > Hi everyone, > > I work with mouse tail sections, each approximately 4-5 mm in thickness. I > have been attempting to embed them in paraffin for sectioning however > sectioning them hasn't been very successful. I can't obtain a ribbon and the > sections seem to break out of the paraffin and break apart. I currently fix > the tails in 4% Paraformaldehyde overnight and then place them in EDTA for > 7-10 days for decalcification. Afterwards, I dehydrate them through a series > of graded alcohol steps, clear them in xylenes, and then let them sit in > paraffin. I am wondering if anyone has a good protocol including times for > each step which I could test out on my samples. Any advice would be greatly > appreciated. > > Thanks so much, > Lauren > > > Lauren Cheung > Stanford University > Rockson Lab > Lab: 650-723-5354 > Email: Lcheung@cvmed.stanford.edu > > > > ------------------------------ > > Message: 6 > Date: Fri, 25 Feb 2005 14:48:30 -0500 > From: Linda Jenkins > Subject: [Histonet] RE: stent techniques anyone??? > To: histonet@lists.utsouthwestern.edu > Message-ID: > <5.2.1.1.2.20050225140622.02910608@mailhost.ces.clemson.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > > You stated: > "ok...here is one where i need some guidance.....post mortem manangement of > coronary artery stents....those little metal mesh things...should i push the > clot out and section it?? > gil corrigan msmdphd tigergil@aol.com best practise anyone??? > ps we are starting a computer section in the aafs ...in case you known some > forensic nurds...send them to tigergil@aol.com" > > Gil, > Removing the stent or clot can remove a lot of valuable > information regarding biocompatibility. A better choice would be to embed > the section in plastic (methyl methacrylate, glycol methacrylate, etc.) and > then section on an automated microtome with tungsten carbide blade or slab > section with a diamond coated blade and grind the sample to the desired > thinness. Of course, this requires specialized equipment and technical > skill that many routine clinical labs may not have. You might want to > consider sending these out for contract work. > The private contract labs that process stents on a routine basis > have some really good procedures BUT cannot (unfortunately) share with > others as they are bound by non-disclosure agreements. > If you are familiar with plastics, you may want to give it a try! > Good Luck, > Linda > > Linda Jenkins, HT > Clemson University > Dept. of Bioengineering > Clemson, SC 29634-0905 > 864.656.5553 > http://www.ces.clemson.edu/bio/research/histo/histo.htm+ > > > > > ------------------------------ > > Message: 7 > Date: Fri, 25 Feb 2005 14:21:06 -0600 > From: Carol Bobrowitz > Subject: [Histonet] GFP > To: "'Histonet (E-mail)" > Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A9640@thor.phys.mcw.edu> > Content-Type: text/plain; charset="iso-8859-1" > > Has anyone worked with GFP tissue? > > I have found GFP is surviving frozen sections, acetone and 100% alcohol. > > I am wondering about FFPE tissue. I'm thinking the heat during processing > would destroy the GFP. > > Does formalin fixation effect the GFP? > > Is it possible to fix the tissue in an alcoholic fixative and manually hand > process at room temperature and infiltrate into plastic? Would the plastic > medium destroy the GFP? > > What about celloidin embedded tissue? This is not my choice > > Any information would be helpful. > > I thank everyone on the histonet that has in the past and future helped me. > I really appreciate your help. > > Thanks, > > Carol Ann Bobrowitz > Histology Laboratory > Department of Physiology - Room 541 > Medical College of Wisconsin > 8701 Watertown Plank Road > Milwaukee, Wisconsin 53226 > 414-456-8179 > FAX 414-456-6546 > cbobrowi@mcw.edu > > > > > ------------------------------ > > Message: 8 > Date: Fri, 25 Feb 2005 15:39:58 -0500 > From: " Judy Strauss" > Subject: [Histonet] CXCR4 & CXCL12 antibodies > To: > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > Can anyone recommend antibodies to CXCR4 (fusin) and CXCL12 that are > suitable for immunohistochemistry in formalin fixed paraffin embedded rat > tissue? > > Thanks, > > > Judith Strauss > SUNY Upstate Medical University > Department of Orthopedic Surgery > IHP room 3118 > 505 Irving Avenue > Syracuse, NY 13120 > > phone: (315) 464-9960 > fax: (315) 464-6638 > > > > > > ------------------------------ > > Message: 9 > Date: Fri, 25 Feb 2005 16:43:50 EST > From: TheBestTime23@aol.com > Subject: Re: [Histonet] richard-allen type 9 paraffin > To: liz@premierlab.com > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <92.2169ca98.2f50f616@aol.com> > Content-Type: text/plain; charset="US-ASCII" > > Thanks for your reply. How long have you been using this paraffin? What > temperatures are you using in your water baths and processors? What do you use > to keep your blocks chilled (if anything)? > > I'm having a little difficulty with this new paraffin. I thought it was > something that I needed time getting used to, but it has been almost 2 months of > using it and I am still having problems (and so are the 3 other people I > work with). Problems include compression, fragile ribbons and wrinkles. I am > now taking an excessive amount of time and care cutting my blocks only to > discover minute folds that could not be seen on the waterbath. We cut our > routine sections at 4 microns and our temperatures are where they should be for > what the company recommends (59 in processor and holding tanks and waterbath > between 35-40). I just thought someone that had worked with this paraffin for a > while could give me some tips. Otherwise I might have to start wearing a > wig : ) > > Thank you! > Megan > > > ------------------------------ > > Message: 10 > Date: Fri, 25 Feb 2005 16:00:17 -0600 > From: "Linda Davis" > > Subject: [Histonet] Re:35BH11 > To: > Message-ID: <006001c51b85$642517c0$de0ac942@D6JLZ851> > Content-Type: text/plain; charset="iso-8859-1" > > Linda, > > Esoterix will stain 35BH11 if you send them your block. They will send the block and slides back to you for your pathologist to interpert the stain. > > Linda Davis > Rio Grande Regional Hospital > McAllen, TX. > (956) 632-6402 > Fax (956) 632-6641 > -------------- next part -------------- > No virus found in this outgoing message. > Checked by AVG Anti-Virus. > Version: 7.0.300 / Virus Database: 266.3.0 - Release Date: 2/21/2005 > > ------------------------------ > > Message: 11 > Date: Fri, 25 Feb 2005 16:22:53 -0600 > From: "Linda Davis" > > Subject: [Histonet] Fw: Re:35BH11 > To: > Message-ID: <004001c51b88$8c565ee0$6d0ac942@D6JLZ851> > Content-Type: text/plain; charset="iso-8859-1" > > > Linda, > > Esoterix will stain 35BH11 if you send them your block. They will send the block and slides back to you for your pathologist to interpert the stain. > > Linda Davis > Rio Grande Regional Hospital > McAllen, TX. > (956) 632-6402 > Fax (956) 632-6641 > -------------- next part -------------- > No virus found in this outgoing message. > Checked by AVG Anti-Virus. > Version: 7.0.300 / Virus Database: 266.3.0 - Release Date: 2/21/2005 > > ------------------------------ > > Message: 12 > Date: Fri, 25 Feb 2005 17:55:56 -0500 > From: > Subject: [Histonet] Histotech Marketability > To: histonet@lists.utsouthwestern.edu > Message-ID: <200502252255.RAA22846@arrakis.vcu.edu> > Content-Type: text/plain; charset=iso-8859-1 > > Hello histonetters. . .just a quick question. What is the > marketability (pay scale ect.) of a tech with a BS in > biology/chemistry, 2 years of experience with proven competency in all > aspects, grosses, and has IHC and IF experience? I am looking to get > feedback from all over the country. Any information would be greatly > appreciated! > Thanks. > > Candyce Fockler > Histotechnician > Anatomic Pathology > > > > ------------------------------ > > Message: 13 > Date: Fri, 25 Feb 2005 17:48:01 -0600 > From: "HIRANO Makoto" > Subject: [Histonet] Square "sample stage" > To: > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > > I am a beginner in frozen sectioning. > Now I have a silly question. > > What is the formal name of a metal, square "sample stage" or "chunk" for > cryostat? > I mean the 'partner' of a cryomold. > I have visited some web sites and consulted some catalogues, but I cannot > find out the name. > I have absolutely to order some. > > Thanks a lot in advance. > > Makoto HIRANO, MD > Department of Molecular Biosciences > University of Kansas > E-mail: mhirano@ku.edu > > > > > ------------------------------ > > Message: 14 > Date: Fri, 25 Feb 2005 17:45:10 -1000 > From: "Hugh Luk" > Subject: RE: [Histonet] richard-allen type 9 paraffin > To: TheBestTime23@aol.com > Cc: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; format=flowed > > Megan, > > Just my 2 cents. If any of this helps, great. > > I've found this is one of the hardest, polymer-rich paraffins available. I > find it similar to Surgipath's standard embedding paraffin and Cardinal's > (old S/P) Ameraffin. It sections and ribbons well. Currently, however, I am > using Ameraffin (also made by Richard Allan), which I find very > interchangable with type 9. > > We modified our tissue processing (a little more time in wax-multiple > changes totalling around 2 hours at 58 to 62C- and maybe more alcohol), > deparaffinization (more time in solvent), and two, different microtome > blades. Sakura's Accu-Edge HIGH profile blades are honed perfectly to cut > ANYTHING at 4 microns with this wax. The "Slight compression" is balanced > with "Ease of ribboning." We sometimes use a wider angle on the blade > holder to increase the rake angle. > > For really thin sections, "Sharper" blades (like Duraedge, Leica's, or > Richard Allan's blades) are required. For example, on delicate biopsies and > lymph nodes, 1 or 2 microns is no problem! I also like the way the paraffin > holds its shape on the water bath (~38-44C). > > I learned histology using Paraplast X-tra. I've tried Paraplast, Paraplast > plus (with DMSO), Fisher Tissue Prep and Tissue Prep2 embedding medias. > When we had a new supervisor at QMC, she changed the paraffin to a harder > wax. We complained about it because it felt different, but I came to really > like it. It became my standard where ever I work (thanks Jackie O'). > > If you are experimenting, you will need to do full switch-overs, every > instrument using wax should be changed. Mixing paraffins of different > grades is cheating the experiment. > > My conclusion is they are all pretty good. The paraffin is only for > support; and with a little tinkering, I like to think I can use any one of > them at any time. > > Hugh Luk, HTL (ASCP) > Former histopathology supervisor CLH > hluk@crch.hawaii.edu > Cancer Research Center of Hawaii > 1236 Lauhala Street, Room 316 > Honolulu, HI 96813 > Tel: (808) 440-5238 > Fax: (808) 586-2982 > > > >From: TheBestTime23@aol.com > >To: histonet@lists.utsouthwestern.edu > >Subject: [Histonet] richard-allen type 9 paraffin > >Date: Wed, 23 Feb 2005 17:42:03 EST > > > >Hello everyone, > > > >The lab that I work for recently decided to switch to Richard-Allen type 9 > >paraffin. I was wondering if anyone has experience with this paraffin and > >could give some advice on techniques used in cutting it. Opinions of this > >product would also be greatly appreciated. > > > >Thanks in advance, > >Megan > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ------------------------------ > > Message: 15 > Date: Fri, 25 Feb 2005 23:53:17 -0800 > From: "Luck, Greg D." > Subject: RE: [Histonet] Histotech Marketability > To: "'cfockler@mail1.vcu.edu'" , > histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain > > Candyce, > > Request that the HR dept of your current employer do a job code market > survey for your geographic area. After they tell you your salary is > equivalent to current market conditions ask them how long it would take to > fill a vacancy in your area. When they say "We don't know" ask them to try > to fill a position (real or not). Then inform them what a "histo'temp" > would cost ($57-60 hr.) to fill in while ???? There is a good deal of > upward pressure on histotech salaries (well overdue) and given a suitable > fit now is the time to look towards the future for new opportunities An > individual's concept of their marketability may differ substantially from > their employer's but this is limited only by their current work environment. > I know, blah, blah, blah. We have a full-time, M-F, day shift, histotech > position open. This is a seasoned dept looking towards the future to lay > the foundation of a new staff to take over (+/-5 yrs) as we retire. We have > a current opportunity for someone either interested in the eventual > department manager's role or as a traditional bench histotech. Click on the > web links that follow to get a snapshot of our institution and of Spokane: > www.deaconessmedicalcenter.org & www.visitspokane.com. > Remember, "making a living" is not the same thing as "making a life". > Thanks, Greg > > Greg Luck, BS, HT(ASCP) > Anatomic Pathology Supervisor > Deaconess Medical Center > 800 W. 5th Ave > Spokane, WA 99204 > Phone 509.473.7077 > Fax 509.473.7133 > luckg@empirehealth.org > > > > -----Original Message----- > From: cfockler@mail1.vcu.edu [mailto:cfockler@mail1.vcu.edu] > Sent: Friday, February 25, 2005 2:56 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histotech Marketability > > Hello histonetters. . .just a quick question. What is the marketability > (pay scale ect.) of a tech with a BS in biology/chemistry, 2 years of > experience with proven competency in all aspects, grosses, and has IHC and > IF experience? I am looking to get feedback from all over the country. Any > information would be greatly appreciated! > Thanks. > > Candyce Fockler > Histotechnician > Anatomic Pathology > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 16 > Date: Sat, 26 Feb 2005 09:17:23 -0500 > From: > Subject: [Histonet] HT vs. HLT > To: histonet@lists.utsouthwestern.edu > Message-ID: <200502261417.JAA24219@despina.vcu.edu> > Content-Type: text/plain; charset=iso-8859-1 > > What exactly is the difference between and HT and an HLT. ASCP makes > this distinction with regards to their wage and salary survey. Is one > certified and the other not? Any information would be greatly > appreciated! Thanks. > > > > > > > ------------------------------ > > Message: 17 > Date: Sat, 26 Feb 2005 08:52:48 -0700 > From: Tim Demuth > Subject: [Histonet] Collagen-I gel, dye and autofluorescence > To: > Message-ID: > Content-Type: text/plain; charset="ISO-8859-1" > > Hi there, > > we are currently trying to cut FFPE sections of multicellular > tumor-spheroids grown in collagen-I gel (bovine). There are two major > problems: > 1) Since the spheroid is so small (diameter ~200-500?m) and very pale it is > extremely hard to identify in the fixed and embedded collagen block. We > would like to use a dye to visualize the spheroid prior to embedding, so we > have some guidance as to where and when to expect the spheroid so we don't > > === message truncated === > > --------------------------------- > Do you Yahoo!? > Yahoo! Sports - Sign up for Fantasy Baseball. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Sun Feb 27 17:29:59 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] HT vs. HLT References: <200502261417.JAA24219@despina.vcu.edu> Message-ID: <011801c51d24$41d78a40$65d0d445@domainnotset.invalid> According to the ASCP Board of Registry (BOR) HT and HTL exams: - HT must now be trained through a NAACLS accredited program (high school graduate through associate degree), OR have a minimum of an associate degree with 12 credits bio/chem and 1 year on-the-job training - HTL must have a baccalaureate degree with a minimum of 20 credits bio/chem, and then be EITHER trained through a NAACLS accredited histotech program OR be trained on the job for 1 year. - both HT and HTL are tested on all areas of routine histology - fixation, processing, frozen sectioning, decalcification, embedding, sectioning, theory of staining, H&E, histology special stains, lab math, equipment - HTL (but NOT HT) are also tested on: all areas of immunohistochemistry, more in depth in chemistry principles/theories, glycol methacrylates, enzyme histochemical staining, biochemistry of staining, management theories, education methodologies, federal regulations - HTL are tested on a lot more troubleshooting and problem-solving. The following is the exam content guidelines for HT and HTL: ftp://ftp.ascp.org/Grab/PDFs/BORpdf/ht.pdf On the outline, the "*" denotes HTL only are tested in that area. Those without the "*" mean both HT and HTL are tested, but again, the HTL have the harder questions that are troubleshooting ("what went wrong") and problem-solving ("what needs to be done to correct it"). Of course, your work can set up any criteria and name that they want for the histotech jobs. It may be something that you need to work with your human resources department, to get "standardization" of titles and job responsibilities, in regards to what other labs in you area are using as well as what ASCP BOR uses. Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Saturday, February 26, 2005 9:17 AM Subject: [Histonet] HT vs. HLT > What exactly is the difference between and HT and an HLT. ASCP makes > this distinction with regards to their wage and salary survey. Is one > certified and the other not? Any information would be greatly > appreciated! Thanks. > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Sun Feb 27 17:33:50 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Histotech Marketability References: <200502252255.RAA22846@arrakis.vcu.edu> Message-ID: <011f01c51d24$cb781580$65d0d445@domainnotset.invalid> ASCP Board of Registry (BOR) just put on line their latest salary survey (it's from the Fall 2003 - it takes about 1 year for it to be analyzed) http://www.ascp.org/bor/center/center_research.asp Click on 2003 Wage and Vacancy Survey I'm afraid their is no survey out their that delineates pay scales to exact job responsibility. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Friday, February 25, 2005 5:55 PM Subject: [Histonet] Histotech Marketability > Hello histonetters. . .just a quick question. What is the > marketability (pay scale ect.) of a tech with a BS in > biology/chemistry, 2 years of experience with proven competency in all > aspects, grosses, and has IHC and IF experience? I am looking to get > feedback from all over the country. Any information would be greatly > appreciated! > Thanks. > > Candyce Fockler > Histotechnician > Anatomic Pathology > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From WWmn916 <@t> aol.com Sun Feb 27 23:55:28 2005 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Slide flats Message-ID: <193.3a71f685.2f540c50@aol.com> Hello everyone, This may be an odd question, but can anyone tell me why cardboard slide flats are so expensive? Fisher sells a case of about 72 slide flats for about $1,000.00!!!! That leads me to my next question. Are there any suggestions for getting them someplace cheaper (Ebay?). Does anyone have an accounting system set-up for making sure slide flats get returned to the originating facility? Thanks, Deb King, HT(ASCP) Sacramento, CA From Krat18 <@t> aol.com Mon Feb 28 06:52:04 2005 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Slide flats Message-ID: <6.3fcdf8b3.2f546df4@aol.com> Don't know if this will be helpful, but my Surgipath rep just told me that they will be shortly coming out with plastic slide trays that have folding covers, like the old cardboard ones. Don't know cost. From Krat18 <@t> aol.com Mon Feb 28 07:43:15 2005 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Slide flats Message-ID: Oh, one more thing I wanted to mention. Our lab sends out our slides in plastic slide boxes that hold about 25-30 slides, MUCH less expensive to buy and easier for the couriers to deal with. Karen Raterman St. Mary's Health Center _Karen_Raterman@ssmhc.com_ (mailto:Karen_Raterman@ssmhc.com) _krat18@aol.com_ (mailto:krat18@aol.com) From Don.Birgerson <@t> leica-microsystems.com Mon Feb 28 07:51:24 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Square "sample stage" Message-ID: Hi Hirano, We currently call these "specimen stages". Several synonyms come to mind specimen stages, specimen chucks, object chucks or object stages. Give me a call and I will be happy to help you. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "HIRANO Makoto" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Square "sample stage" western.edu 02/25/2005 05:48 PM I am a beginner in frozen sectioning. Now I have a silly question. What is the formal name of a metal, square "sample stage" or "chunk" for cryostat? I mean the 'partner' of a cryomold. I have visited some web sites and consulted some catalogues, but I cannot find out the name. I have absolutely to order some. Thanks a lot in advance. Makoto HIRANO, MD Department of Molecular Biosciences University of Kansas E-mail: mhirano@ku.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From ecl <@t> itsa.ucsf.edu Mon Feb 28 08:46:50 2005 From: ecl <@t> itsa.ucsf.edu (Ellen Liebenberg) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Re: Mouse tail sectioning In-Reply-To: <200502251801.j1PI1muY025104@itsa.ucsf.edu> References: <200502251801.j1PI1muY025104@itsa.ucsf.edu> Message-ID: <6.0.1.1.2.20050228064227.023fdb90@itsa.ucsf.edu> We routinely section mouse tail using a protocol similar to yours. However, I leave the tissue in 70% alcohol overnight, or longer, before processing. It seems necessary for good dehydration. I also use Clearite instead of xylene. Ellen Liebenebrg At 10:01 AM 2/25/2005, you 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. lectins (Margaret Horne) > 2. Re: RE: pathologists in the lab (shivers down thespine...) > (Jo Dee Fish) > 3. Re: lectins (Paul Webster) > 4. CAP Immunohistochemistry Tissue Microarray Program (Paula Lucas) > 5. RE: RE: pathologists in the lab (shivers down thespine...) > (Stapf, Ross) > 6. Re: cryosections (Instrumedics) > 7. Re: cryosections (Stephen Peters M.D.) > 8. Re: Re: cryosections (Maria Mejia) > 9. (no subject) (Dawn Truscott) > 10. Re: (no subject) (Kelly D Mcqueeney) > 11. stent techniques anyone??? (TIGERGIL@aol.com) > 12. Is there any DVD about how to make frozen section? > (=?gb2312?q?=CA=E7=C7=ED=20=C9=F2?=) > 13. 34BE12 on prostate[Scanned] (Jean Gillson) > 14. stent techniques anyone??? (Stephen Peters M.D.) > 15. RE: CAP Immunohistochemistry Tissue Microarray Program > (Tom McNemar) > 16. RE: xylene free mounting media (Smith, Allen) > 17. Re: 34BE12 on prostate[Scanned] (Katri Tuomala) > 18. JACHO Compliance (Scholz, Stephen J.) > > >---------------------------------------------------------------------- > >Message: 1 >Date: Thu, 24 Feb 2005 14:06:16 ADT >From: "Margaret Horne" >Subject: [Histonet] lectins >To: histonet@lists.utsouthwestern.edu >Message-ID: <421DDF57.27008.414902B@localhost> >Content-Type: text/plain; charset=US-ASCII > >Hello Everyone , > Has anyone out there conjugated gold colloid to lectins? >Some gold conjugated lectins I can buy but others I may have to >make. > > Thanks , > Margaret > > > > >Margaret Horne , >Histology Teaching Assistant, >Dept. of B.SC., >Atlantic Veterinary College, U.P.E.I., >550 University Ave., Charlottetown, >P.E.I., C1A 4P3 >Canada > > > > >------------------------------ > >Message: 2 >Date: Thu, 24 Feb 2005 10:21:04 -0800 >From: Jo Dee Fish >Subject: Re: [Histonet] RE: pathologists in the lab (shivers down > thespine...) >To: Ford Royer >Cc: histonet@lists.utsouthwestern.edu >Message-ID: >Content-Type: text/plain; charset="us-ascii" ; format="flowed" > >Hello, just my two cents worth, > >I agree that if there were a way the residents should spend at least >a short period of time in a lab so that they can see several >different staining procedures from beginning to end, from the >actually biopsy procedure to the staining procedure. They should see >some slides of properly and improperly handled samples, just to see >what a small mistake can lead to further down the line. > >On a more personal note: >There have been times when I have been the patient lying on the >examination table, that I have wondered if the doctor is handling my >biopsy correctly. I wonder if she knows what that small piece of >tissue was about to go through, either freezing or fixation and >paraffin embedding, and whether or not she knows that the way she >handles it now can change dramatically the outcome of whatever tests >she orders. I know she has an idea of how precious it is, but does >she know how one small moment can change the results, such as letting >the tissue sit on the table to dry slightly, or smashing it, or not >placing it in enough fixative, correct fixative, etc. If she had >spent a week in a lab, she would know more about the steps that the >tissue goes through, and the precise decisions histologists (or other >lab personnel) make concerning the handling of the tissue once it is >in their hands. > >One more side note: >My husband had his "dreaded" but necessary Colonoscopy last fall and >had a biopsy of one polyp taken. We waited on pins and needles (no >pun intended) for the results for two weeks. I could not believe it >when they could not provide test results on my husband's biopsy >because the tissue was "too destroyed to do the ordered tests". He >actually had to go through another round of tests and exams because, >most likely, the doctor probably didn't handle the original sample >correctly. Notice I don't blame the lab personnel! > >Take care, >Jo Dee Fish >********************************************************************** >********** > >Jo Dee Fish >Research Technologist III >Gladstone Institute of Cardiovascular Disease > >Telephone: (415) 734-5766 >Fax: (415) 355-0230 >E-mail: jfish@gladstone.ucsf.edu > >Mailing address: >The J. David Gladstone Institutes >1650 Owens Street >San Francisco, CA 94158 > > > >------------------------------ > >Message: 3 >Date: Thu, 24 Feb 2005 10:23:17 -0800 >From: Paul Webster >Subject: Re: [Histonet] lectins >To: Margaret Horne , > >Message-ID: >Content-Type: text/plain; charset="US-ASCII" > >Jeurgen Roth and John Lucocq did lots of studies conjugating lectins to >colloidal gold particles about 20 years ago. I think they finally concluded >that it was more efficient to use sequential labeling protocols with >secondary labeling reagents for detecting lectins. That is, you react the >lectin with the section, bind an anti-lectin antibody to the lectin and then >visualize with a gold-conjugated secondary antibody (or protein a gold). > >One method they described used fetuin-gold to detect lectin bound to the >section (Light and electron microscopic demonstration of sialic acid >residues with the lectin from Limax flavus: a cytochemical affinity >technique with the use of fetuin-gold complexes. J Histochem Cytochem. 1984 >Nov;32(11):1167-76.). > >Paul Webster. > > > > >Paul Webster, Ph.D. >House Ear Institute >2100 West Third Street >Los Angeles >CA 90057-1922 >Phone: 213 273 8026 >Fax: 213 13 739 >E-mail: pwebster@hei.org > > > > > > >On 2/24/05 6:06 AM, "Margaret Horne" wrote: > > > Hello Everyone , > > Has anyone out there conjugated gold colloid to lectins? > > Some gold conjugated lectins I can buy but others I may have to > > make. > > > > Thanks , > > Margaret > > > > > > > > > > Margaret Horne , > > Histology Teaching Assistant, > > Dept. of B.SC., > > Atlantic Veterinary College, U.P.E.I., > > 550 University Ave., Charlottetown, > > P.E.I., C1A 4P3 > > Canada > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > >------------------------------ > >Message: 4 >Date: Thu, 24 Feb 2005 10:30:44 -0800 >From: "Paula Lucas" >Subject: [Histonet] CAP Immunohistochemistry Tissue Microarray Program >To: "Histonet \(E-mail\)" >Message-ID: <004b01c51a9e$f4397b90$7c01a8c0@biopath.org> >Content-Type: text/plain; charset="iso-8859-1" > >My administrator asked me to research this clinical trial, so of course I >turn to the experts. > >Here is what the advertisement says: > >This is a program that stains and interprets tissue arrays for HER2, CD117, >ER, CD20 and EGFR. Comparing results to our peers. > >It validates our testing strategy and demonstrates eligibility for >participation in NCI funded clinical trials for HER2. > >Has anyone heard of this program and if so, could you tell me anything and >everything about it? > >Thanks in advance. > >Paula >Bio-Path Medical Group > > > > >------------------------------ > >Message: 5 >Date: Thu, 24 Feb 2005 12:41:55 -0600 >From: "Stapf, Ross" >Subject: RE: [Histonet] RE: pathologists in the lab (shivers down > thespine...) >To: "Ford Royer" , > >Message-ID: > <74A8B7A904152141BD5AA2761F5D22340E480B@BHDAEXCH11.bhcs.pvt> >Content-Type: text/plain; charset="us-ascii" > > >Our new residents after a few weeks on their first Gross Room rotation >spend a day in histology to see the blocks they grossed get embedded and >cut. They get to see first hand the results of their work. It helps >tremendously. > >This was not always done consistently here. Hence the story I shared >earlier. > >Ross M Stapf >Histopathology Manager >Baylor University Medical Center >3500 Gaston Ave. >Dallas, TX 75246 >214-820-2465 >214-820-4110 fax >RossS@baylorhealth.edu > > > > >This e-mail, facsimile, or letter and any files or attachments transmitted >with it contains information that is confidential and privileged. This >information is intended only for the use of the individual(s) and >entity(ies) to whom it is addressed. If you are the intended recipient, >further disclosures are prohibited without proper authorization. If you >are not the intended recipient, any disclosure, copying, printing, or use >of this information is strictly prohibited and possibly a violation of >federal or state law and regulations. If you have received this >information in error, please notify Baylor Health Care System immediately >at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health >Care System, its subsidiaries, and affiliates hereby claim all applicable >privileges related to this information. > > > >------------------------------ > >Message: 6 >Date: Thu, 24 Feb 2005 14:38:35 -0500 >From: "Instrumedics" >Subject: Re: [Histonet] cryosections >To: "Kelly D Mcqueeney" >Cc: histonet@lists.utsouthwestern.edu >Message-ID: <01aa01c51aa8$73a15cf0$6401a8c0@INSTRUMEDICS22> >Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=response > >Frozen sectioning can be difficult and requires a skillful hand. It is often >hard to produce a flat, unfolded, fully intact section using the >conventional method of retrieving a section on a room temperature slide. >The result of melting the unfixed section to mount it on the warm slide >degrades the morphology. > >With the CryoJane Tape-Transfer process the section is captured on a COLD >tape as it is being cut . The section is flat, uncompressed, intact and as >thin a 2 microns. The section is transferred to a COLD slide. The section >is still frozen and the morphology preserved A choice of fixation is then >made depending on the staining protocol to follow. This method makes it >possible to cut hard tissue or fatty tissue which is often very problematic. > >The CryoJane process is easy to learn and makes preparing difficult frozen >sections easy. > >For the details of the CryoJane process please visit our web site >www.instrumedics.com > >Bernice >Instrumedics > >----- Original Message ----- >From: "Kelly D Mcqueeney" >To: "Till, Renee" >Cc: >Sent: Thursday, February 24, 2005 11:54 AM >Subject: Re: [Histonet] cryosections > > > > Renee, > > > > You should insist on an in-depth training from the vendor. Ask for a 4 > > hour training from their cryostat expert, not some sales person. They > > should offer it to you. The technique is standard but varies from user to > > user. You'll have to find your way. I am going to give you our standard > > protocol for brain IHC. > > > > 1) Remove fresh tissue, dip in mounting medium (we use Shandon embedding > > medium, or you can skip this step) and immediately freeze in ice-cold > > isopentane (keep on dry ice for 30 minutes prior to freezing). For rat > > brain, we freeze for 20 seconds. Remove from isopentane and place tissue > > in a conical tube (some people use molds). Leave the tube on dry ice and > > immediately store at -70C. > > > > 2) Remove tissue from -70C and place in -20C freezer for 1 hour to > > equilibrate. > > > > 3) Drop embedding medium on chuck, add tissue and freeze base with > > cytocool. Cover tissue with embedding medium (we dip and freeze with > > cytocool). > > > > 4) Place chuck in cryostat and section tissue. I use probe on plus slides > > from Fisher or Superfrost from VWR. > > > > 5) Once sections are mounted on slides, dry slides completely for 1 hour > > RT and store at -80C until ready to use. > > > > 6) Remove slides from freezer and thaw at RT for 15 minutes. Place slides > > in fix and stain or perform IHC. > > > > There are many different methods for sectioning/storing, etc. Times, > > tissue, temperature and protocols will vary from lab to lab or person to > > person. > > > > Have fun! > > Kelly > > > > Till, Renee wrote: > > > >>Hello. Our research group just purchased our first cryostat. Until now, > >>anytime cryosections were needed we had to send out our tissues to > >>another lab. Is there a good book or other source for general > >>information about cryosectioning? I am the only histotech in the group, > >>but have had virtually no experience with cryosections. I know the other > >>general techs that also do some histology are going to come to me with > >>all their questions. We basically need to know anything and everything. > >>Due to the volume of tissues, sectioning won't be performed right as the > >>tissues are removed from the animal. How do you store them? Do you fix, > >>and when? What type of slides are best? How do you store the slides > >>after cryosectioning? Are there specific considerations to take into > >>account as far as fixing, or anything else depending on the tissue or > >>what stains will be performed on it? For example, I know already that > >>there will be some heart and aorta sectioned for fat stains, some pig > >>gi, possibly mammary gland, uterus, and mouse lung with b-gal. > >>Thanks in advance for any help or suggestions you can give me. > >> > >> > >>Renee' > >> > >> > >> > >>_______________________________________________ > >>Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >> > > > > > > > > > > > >------------------------------ > >Message: 7 >Date: Thu, 24 Feb 2005 11:57:23 -0800 (PST) >From: "Stephen Peters M.D." >Subject: [Histonet] Re: cryosections >To: Histonet@lists.utsouthwestern.edu >Message-ID: <20050224195723.9044.qmail@web30410.mail.mud.yahoo.com> >Content-Type: text/plain; charset=us-ascii > >Sorry I left the // out of the address for the frozen section tutorial. >Fere is the complete link >http://pathologyinnovations.com/frozen_section_technique.htm > > > >Stephen Peters M.D. >Pathology Innovations, LLC >410 Old Mill Lane, >Wyckoff, NJ 07481 >201 847 7600 >www.pathologyinnovations.com > >Senior Attending Pathologist >Hackensack University Medical Center >201 996 4836 > > > >------------------------------ > >Message: 8 >Date: Thu, 24 Feb 2005 12:14:30 -0800 >From: Maria Mejia >Subject: Re: [Histonet] Re: cryosections >To: "Stephen Peters M.D." >Cc: Histonet@lists.utsouthwestern.edu >Message-ID: <421E35A6.7010104@ski.org> >Content-Type: text/plain; charset=us-ascii; format=flowed > >Dr. Peters, please let us (histonet) know when you have completed & have >added >some original techniques for sur. path dissections to your website. > >Maria Bartola Mejia >Neurohistologist >Smith-Kettlewell Eye Research Institute >San Francisco, CA 94115 > > >Stephen Peters M.D. wrote: > > >Sorry I left the // out of the address for the frozen section tutorial. > Fere is the complete link > >http://pathologyinnovations.com/frozen_section_technique.htm > > > > > > > >Stephen Peters M.D. > >Pathology Innovations, LLC > >410 Old Mill Lane, > >Wyckoff, NJ 07481 > >201 847 7600 > >www.pathologyinnovations.com > > > >Senior Attending Pathologist > >Hackensack University Medical Center > >201 996 4836 > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > >------------------------------ > >Message: 9 >Date: Thu, 24 Feb 2005 16:38:18 -0500 >From: "Dawn Truscott" >Subject: [Histonet] (no subject) >To: >Message-ID: <001301c51ab9$280deb80$6401a8c0@wowway.com> >Content-Type: text/plain; charset="iso-8859-1" > >Don't dis "just the sales person" A good percentage of our sales force >are histotechs!! > >Dawn M. Truscott, HT(ASCP) >Richard Allan Scientific > > > > > >Renee, > >You should insist on an in-depth training from the vendor. Ask for a 4 >hour training from their cryostat expert, not some sales person. They >should offer it to you. The technique is standard but varies from user >to user. You'll have to find your way. I am going to give you our >standard protocol for brain IHC. >Dawn M. Truscott > >Recycling New Year's resolutions since 1987. > >------------------------------ > >Message: 10 >Date: Thu, 24 Feb 2005 17:14:38 -0500 >From: Kelly D Mcqueeney >Subject: Re: [Histonet] (no subject) >To: Dawn Truscott >Cc: histonet@lists.utsouthwestern.edu >Message-ID: <421E51CE.1080702@bms.com> >Content-Type: text/plain; format=flowed; charset=us-ascii > >I just had a salesperson try to train us on a new cryostat from Richard >Allan and it was the first time she used it. We had to put it together >and teach ourselves, it was a disaster. My point for Renee, make sure >they know how to use the machine. > >Dawn Truscott wrote: > > >Don't dis "just the sales person" A good percentage of our sales force > are histotechs!! > > > >Dawn M. Truscott, HT(ASCP) > >Richard Allan Scientific > > > > > > > > > > > >Renee, > > > >You should insist on an in-depth training from the vendor. Ask for a 4 > >hour training from their cryostat expert, not some sales person. They > >should offer it to you. The technique is standard but varies from user > >to user. You'll have to find your way. I am going to give you our > >standard protocol for brain IHC. > >Dawn M. Truscott > > > >Recycling New Year's resolutions since 1987. > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > >------------------------------ > >Message: 11 >Date: Fri, 25 Feb 2005 01:29:30 EST >From: TIGERGIL@aol.com >Subject: [Histonet] stent techniques anyone??? >To: histonet@pathology.swmed.edu >Message-ID: <81.222d9ee0.2f501fca@aol.com> >Content-Type: text/plain; charset="US-ASCII" > >ok...here is one where i need some guidance.....post mortem manangement of >coronary artery stents....those little metal mesh things...should i push the >clot out and section it?? >gil corrigan msmdphd tigergil@aol.com best practise anyone??? >ps we are starting a computer section in the aafs ...in case you known some >forensic nurds...send them to tigergil@aol.com > > >------------------------------ > >Message: 12 >Date: Fri, 25 Feb 2005 15:25:53 +0800 (CST) >From: =?gb2312?q?=CA=E7=C7=ED=20=C9=F2?= >Subject: [Histonet] Is there any DVD about how to make frozen section? >To: histonet@lists.utsouthwestern.edu >Message-ID: <20050225072553.28001.qmail@web15604.mail.cnb.yahoo.com> >Content-Type: text/plain; charset=gb2312 > >Hi,everybody, > I am a layman in IHC.I heard from one of my friend that some lab would > provide DVD on how to make frosen sections and so on.Would anyone give > me some hints? > Thanks a lot. >Shuqiong Shen >Jinling hospital >Nanjing China > > > >--------------------------------- >Do You Yahoo!? >150????MP3???????????????????????? >?????????????????????????????????????? >1G????1000?????????????????????? > >------------------------------ > >Message: 13 >Date: Fri, 25 Feb 2005 11:02:57 -0000 >From: "Jean Gillson" >Subject: [Histonet] 34BE12 on prostate[Scanned] >To: >Message-ID: > ><1030B679AD69D6119C3F00080210DD9D05AC0348@bhrv-nt-11.bhrv.nwest.nhs.uk> > >Content-Type: text/plain; charset="iso-8859-1" > >Dear all fellow histologist, > >We seem to have a recurrent problem with 34BE12 IHC staining on both >prostate chippings and prostatectomy specimens. Some basal cells stain >whilst others that should stain are negative. Our procedure is Trypsin for >12 minute at pH 7.8 and 37 degrees C. We use a Dako autostainer machine and >our antibody is from Dako. Previous spec sheets state Trypsinisation but >our most recent one states heat retrieval. So as a test we did both >retrieval methods. Oddly enough, microwaved in Dako retrieval solution was >slightly better in prostatectomy specimens than trypsin (still uneven >staining) and worst in chipping specimens. Can anyone offer any advice? >What antibody panels are people doing for prostate cases? Help desperately >needed!! > >Jean Gillson BMS2 > >Histology Department > >Blackburn Royal Infirmary > >Blackburn > >UK > > > >------------------------------ > >Message: 14 >Date: Fri, 25 Feb 2005 04:41:54 -0800 (PST) >From: "Stephen Peters M.D." >Subject: [Histonet] stent techniques anyone??? >To: Histonet@lists.utsouthwestern.edu >Message-ID: <20050225124154.22991.qmail@web30407.mail.mud.yahoo.com> >Content-Type: text/plain; charset=us-ascii > >I had do do this on a research project on pig carotids where the goal was >to see the reaction to the stent which was buried in an organizing fibrous >tissue responce. I took a strong sissor and cut the vessel transversely >into 2-3mm rings. Using a forceps I was able to pull the now cut wires >making up the stent out of the tissue leaving the tissue pretty much >intact. It came out quite well. >PS don't use your favorite sissor! > > >Stephen Peters M.D. >Pathology Innovations, LLC >410 Old Mill Lane, >Wyckoff, NJ 07481 >201 847 7600 >www.pathologyinnovations.com > >Senior Attending Pathologist >Hackensack University Medical Center >201 996 4836 > > > >------------------------------ > >Message: 15 >Date: Fri, 25 Feb 2005 07:45:03 -0500 >From: Tom McNemar >Subject: RE: [Histonet] CAP Immunohistochemistry Tissue Microarray > Program >To: 'Paula Lucas' , "Histonet (E-mail)" > >Message-ID: > <6CD94D97ED7D924BA5C2B588FA95282139678D@nt_exchange.lmhealth.org> >Content-Type: text/plain; charset="iso-8859-1" > >Not much to tell, it's prety much like any other survey. They send you 1 >slide with multiple tissue samples. You stain it, the pathologists reads it >and you send it back. > >We did it one year. It's the same four antibodies every year so I haven't >bought it again. > >Tom Mc Nemar HT(ASCP) >Histology Supervisor >Licking Memorial Hospital >Newark, Ohio 43055 > > > >-----Original Message----- >From: Paula Lucas [mailto:plucas@biopath.org] >Sent: Thursday, February 24, 2005 1:31 PM >To: Histonet (E-mail) >Subject: [Histonet] CAP Immunohistochemistry Tissue Microarray Program > > >My administrator asked me to research this clinical trial, so of course I >turn to the experts. > >Here is what the advertisement says: > >This is a program that stains and interprets tissue arrays for HER2, CD117, >ER, CD20 and EGFR. Comparing results to our peers. > >It validates our testing strategy and demonstrates eligibility for >participation in NCI funded clinical trials for HER2. > >Has anyone heard of this program and if so, could you tell me anything and >everything about it? > >Thanks in advance. > >Paula >Bio-Path Medical Group > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >------------------------------ > >Message: 16 >Date: Fri, 25 Feb 2005 09:16:00 -0500 >From: "Smith, Allen" >Subject: RE: [Histonet] xylene free mounting media >To: "Robin Maxwell-Nunn" >Cc: histonet@lists.utsouthwestern.edu >Message-ID: > <4C051EAE581BB646BF53A749A73FBA2D1F3C96@exchsrv01.barrynet.barry.edu> >Content-Type: text/plain; charset="us-ascii" > >If one sticks to stains that are fast in absolute alcohol, one can mount >from absolute alcohol in Euparal (gum sandarac), which uses eucalyptol as a >solvent. Hematoxylin fades slowly in Euparal, but is said to keep virtually >forever in Euparal Vert. > >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 Robin >Maxwell-Nunn >Sent: Monday, February 21, 2005 8:28 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] xylene free mounting media > > >Hi >We have a tech in our lab who is sensitive to xylene (exhibits headaches, >puffy eyes and itchy skin). We are switching to Formula 83 by CBG Biotech >for frozen sections but still need a mountant that is both compatible with >this and xylene free. Does anyone have any suggestions? > >thanks, >Robin Maxwell-Nunn, MLT >St Mary's General Hospital, >Kitchener, Ontario, Canada. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >The information transmitted is intended only for the person or entity to >which it is addressed and may contain confidential, and/or privileged >material. No confidentiality or privilege is waived or lost by any errant >transmission. If you receive this message in error, please immediately >delete it and all copies of it from your system and notify the >sender. E-mail transmission cannot be guaranteed to be secure or >error-free as information could be intercepted, corrupted, lost, >destroyed, arrive late or incomplete, or contain viruses. >Barry University - Miami Shores, FL (http://www.barry.edu) > > > >------------------------------ > >Message: 17 >Date: Fri, 25 Feb 2005 11:38:08 -0500 >From: "Katri Tuomala" >Subject: Re: [Histonet] 34BE12 on prostate[Scanned] >To: "Jean Gillson" , > >Message-ID: <001601c51b58$63715190$6a9a9618@Katri> >Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=original > >Hi Jean, >Yes, clone 34 BE12 can be a problematic one. Over the years we have >struggled same as you, but seem to have pretty good handle on it now. Our >antibody is same as yours and we find, that more intense HIER is required. >We are using citrate buffer (EDTA or others may be better, you'll have to >try) in Decloaker. Our citrate buffer is home made and we add Tween 20 into >it. Our dilution of the primary antibody now is 1:30. >I hope this helps... >Katri > >Katri Tuomala >Hamilton,Ontario, Canada > >----- Original Message ----- >From: "Jean Gillson" >To: >Sent: Friday, February 25, 2005 6:02 AM >Subject: [Histonet] 34BE12 on prostate[Scanned] > > > > Dear all fellow histologist, > > > > We seem to have a recurrent problem with 34BE12 IHC staining on both > > prostate chippings and prostatectomy specimens. Some basal cells stain > > whilst others that should stain are negative. Our procedure is Trypsin > > for > > 12 minute at pH 7.8 and 37 degrees C. We use a Dako autostainer machine > > and > > our antibody is from Dako. Previous spec sheets state Trypsinisation but > > our most recent one states heat retrieval. So as a test we did both > > retrieval methods. Oddly enough, microwaved in Dako retrieval solution > > was > > slightly better in prostatectomy specimens than trypsin (still uneven > > staining) and worst in chipping specimens. Can anyone offer any advice? > > What antibody panels are people doing for prostate cases? Help > > desperately > > needed!! > > > > Jean Gillson BMS2 > > > > Histology Department > > > > Blackburn Royal Infirmary > > > > Blackburn > > > > UK > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >------------------------------ > >Message: 18 >Date: Fri, 25 Feb 2005 11:01:23 -0600 >From: "Scholz, Stephen J." >Subject: [Histonet] JACHO Compliance >To: >Message-ID: > ><7F1312711CA7474A89B3DF8BA0BA54D0F5F190@pmc-rfd-mx01.intranet.osfnet.org> > >Content-Type: text/plain; charset="us-ascii" > >Hello all; > >We are about to have a JACHO inspection and a problem was brought to my >attention. It involves using two identifiers on all laboratory specimens. > >The below paragraph is from JCAHO FAQ web site for laboratory compliance >with the National Patient Safety Goals. This particular statement seems >to answer the question about use of only one identifying number on a >pathology or any lab specimen. Please let me know your interpretation and >how your laboratories comply to it. Currently we have only the case >number (hand written) on the blocks and only the case number (hand >written) on the slides until after staining at which point there are >printed labels put on them. > >Take note that this FAQ was updated in January, 2005. > > >FAQ-We use a label for the initial patient sample that contains only one >unique identifier, a number, to identify laboratory specimens. This >unique identifier can be traced to multiple patient identifiers. Does this >meet the intent of the goal? >ANS-No. The intent is to use two separate identifiers on the specimen >label. Confidence in an accurate identification improves as the number of >identifiers increases, depending upon their uniqueness. A single >identifier can be more easily misread, resulting in avoidable errors. Bar >coding that includes two or more person-specific identifiers (not room >number) will comply with this requirement. (New 1/18/05) > > > >What are suggestions to my problem. > > >Stephen J. Scholz HT(ASCP) >Histology Coordinator >OSF St. Anthony Medical Center >Rockford IL > >Phone: 815-395-5410 >Fax: 815-395-5364 > > >------------------------------ > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >End of Histonet Digest, Vol 15, Issue 39 >**************************************** From colins <@t> thor.uk.com Mon Feb 28 08:49:09 2005 From: colins <@t> thor.uk.com (Colin Smith) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] microtome suitable for beginner-Eurocentric Message-ID: <3E985CD1D7CA89499D1902BC3202AF3CDBCE81@uk-res-srv-03.thorspecialities.com> Dear Histonetters, can anyone recommend a manual rotary microtome suitable for a low through-put lab. I would prefer to source the instrument from the UK or Europe. Many Thanks, Colin Smith In Vitro Toxicologist Thor Specialities (UK) Ltd Wincham Avenue Northwich Cheshire CW9 6GB Tel: +44 1606 818873 Fax: +44 1606 818801 colins@thor.uk.com The information contained in this e-mail may be confidential and may also be legally privileged. It is intended solely for the addressee(s). If you are not the intended recipient then please delete forthwith. Views expressed in this email are views of the individual and are not necessarily the views of this company. ************************************************************************************************** e-mail scanned for Viruses by Thor Specialities (UK) Limited using Gfi Mail Security ************************************************************************************************** From jfish <@t> gladstone.ucsf.edu Mon Feb 28 10:47:53 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Slide flats In-Reply-To: <193.3a71f685.2f540c50@aol.com> References: <193.3a71f685.2f540c50@aol.com> Message-ID: Hi Deb, Try www.tedpella.com. They have them for $378. Their hinged slide boxes are cheap, too. No affiliation, I just like their prices! Take care, Jo Dee At 12:55 AM -0500 2/28/05, WWmn916@aol.com wrote: >Hello everyone, > >This may be an odd question, but can anyone tell me why cardboard slide >flats are so expensive? Fisher sells a case of about 72 slide flats >for about >$1,000.00!!!! > >That leads me to my next question. Are there any suggestions for getting >them someplace cheaper (Ebay?). > >Does anyone have an accounting system set-up for making sure slide flats get >returned to the originating facility? > >Thanks, >Deb King, HT(ASCP) >Sacramento, CA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** ********** Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 734-5766 Fax: (415) 355-0230 E-mail: jfish@gladstone.ucsf.edu Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158 From Heather.A.Harper <@t> pcola.med.navy.mil Mon Feb 28 10:45:30 2005 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] (no subject) Message-ID: <807FE48C5A7CC940B973B58D32E7014318A7648D@nhpens-exch1.pcola.med.navy.mil> Hi everybody, It seems as though from reading the emails that a lot of histology techs are questioning whether they should gross or not. Well even according to the CLIA 88 rules, if you have an Associates Degree or higher you qualify to gross. Sad part is, it doesn't specifically say.."histo techs". Personally, it's a serious task and I can't imagine if there is a problem that the Dr. would take the blame. The histo tech would be the one to go down in flames. According to my pathologist, he received a hospital survey and was shocked to see that there are a lot of techs grossing, not just small specimens but big specimens that have no education. Hospitals just want histo techs to gross so they can have you do 2 jobs for the price of one. I'm not comfortable grossing and I probably will refuse to gross. I think CAP needs to implement a set of regulations for histo techs. We have been too long in the gray area unlike most fields, who have specific regulations to what they can and can not do. Heather A. Harper From jtcoppens <@t> ucdavis.edu Mon Feb 28 11:52:40 2005 From: jtcoppens <@t> ucdavis.edu (john coppens) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] GFP In-Reply-To: <59DF6640CDC0D7119DF000D0B761393C1A9640@thor.phys.mcw.edu> References: <59DF6640CDC0D7119DF000D0B761393C1A9640@thor.phys.mcw.edu> Message-ID: i have been working with some mice that express GFP in the Clara Cells of the lung. the signal is quite strong in the cells that are positive. i have had positive results under the following situations: Frozen Tissue, my experience was similar to yours. Whole Tissue, 4% 2hr paraformaldehyde fixed and unfixed. these were viewed under PBS with immersion objectives. The signal lasts for weeks when the tissue is fixed. I can look at it all day with no noticeable photo-bleaching. I only use the unfixed tissue to verify a that mouse is positive for the protein. Paraffin Embedded, fixed 2 hours in 4% paraformaldehyde, then transferred to 50% EtOH. I would lower the temperature of the paraffin pot to 56% if you normally run it higher. The signal is fair but bleaches easily. If you want it stronger and longer lasting signal you can do IHC with a GFP antibody and fluorescent secondary. GMA Embedded, I believe we followed our standard protocol for GMA. There was a signal but if I remember right, it bleached out very quickly in the 1um sections. Good luck. john >Has anyone worked with GFP tissue? > >I have found GFP is surviving frozen sections, acetone and 100% alcohol. > >I am wondering about FFPE tissue. I'm thinking the heat during processing >would destroy the GFP. > >Does formalin fixation effect the GFP? > >Is it possible to fix the tissue in an alcoholic fixative and manually hand >process at room temperature and infiltrate into plastic? Would the plastic >medium destroy the GFP? > >What about celloidin embedded tissue? This is not my choice > >Any information would be helpful. > >I thank everyone on the histonet that has in the past and future helped me. >I really appreciate your help. > >Thanks, > >Carol Ann Bobrowitz >Histology Laboratory >Department of Physiology - Room 541 >Medical College of Wisconsin >8701 Watertown Plank Road >Milwaukee, Wisconsin 53226 >414-456-8179 >FAX 414-456-6546 >cbobrowi@mcw.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- John Coppens Veterinary Medicine - Anatomy, Physiology and Cell Biology 1 Shields Ave. UC Davis Davis, CA 95616 530-754-8141 jcoppens@ucdavis.edu From mfox <@t> memhosp.com Mon Feb 28 12:33:08 2005 From: mfox <@t> memhosp.com (Matt Fox) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] pH meter question. Message-ID: <422363E4.4053220E@memhosp.com> What type of pH meter would you recommend for a routine histology lab? From adelsyscarol <@t> yahoo.com Mon Feb 28 13:04:59 2005 From: adelsyscarol <@t> yahoo.com (Carol Wilson) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Looking for Hypercenter XP Operators Manual Message-ID: <20050228190459.51611.qmail@web51509.mail.yahoo.com> I'm looking for a Shandon Hypercenter XP tissue processor operators manual from someone who isn't using it anymore. I'm willing to pay for at least postage/shipping, and possibly a small cost. Please contact me via my email address. Thanks Carol Wilson, HT(ASCP) Adelsys, Inc. --------------------------------- Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. From laurie.colbert <@t> huntingtonhospital.com Mon Feb 28 13:07:13 2005 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] CAP question - liquid nitrogen Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5C22F@EXCHANGE1.huntingtonhospital.com> I am asking this question for another lab: Does anyone out there have a procedure on the handling of liquid nitrogen that they would be willing to share? This is in reference to a new question on the CAP checklist. It's on the Lab General checklist, GEN.70525 - "Are adequate policies, procedures, and practices in place for the use of liquid nitrogen?" I would appreciate any help. Thanks! Laurie Colbert Huntington Memorial Hospital Pasadena, CA From ohenry <@t> dfw.net Mon Feb 28 13:08:12 2005 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Please, no replys in Digest form. Message-ID: <000f01c51dc8$e90041a0$3fdd3040@Nationwide.net> P-l-e-a-s-e, when you send/reply to Histonet don't send the complete digest. Please copy/cut/paste, what you need. Some days it is impossible to read, even to scroll thru my digest, because of persons who send a complete digest with their reply/question/answer, etc. I just gave up trying to read the digest that came thru a short while ago. Several people had replied back, EACH sending an older digest , making the current digest impossible to read. Susan Owens-TX ohenry@dfw.net From Jenny-Oblander <@t> omrf.ouhsc.edu Mon Feb 28 13:22:15 2005 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] ASCP Message-ID: I have a question about the ASCP registry, some histotechs and I were talking about paying your ASCP dues every year and one said they have never paid since they were initially registered. I guess my question is do you have to maintain your dues with ASCP to continue to be registered? I always assumed that you need to pay dues every year to be registered but I really don't know. Thanks Jenny J.Oblander, HT (A.S.C.P.) Comparative Medicine Oklahoma Medical Research Foundation MS#32 825 NE 13th St. Okc,Ok 73104 jenny-oblander@omrf.ouhsc.edu 405-271-7083 From dholmes <@t> anatomy.umsmed.edu Mon Feb 28 13:34:23 2005 From: dholmes <@t> anatomy.umsmed.edu (Dianne Holmes) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] "Flying" brains !! Message-ID: HELP! a colleague is cutting rat brains on a cryostat and is having a terrible time with static causing the sections to "jump and fly" around in the cryo-cabinet. Has anyone got a remedy for reducing the static ? From pruegg <@t> ihctech.net Mon Feb 28 13:40:36 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] ASCP In-Reply-To: Message-ID: <200502281940.j1SJeafR026380@chip.viawest.net> HT/HTL certification is for life if registered before 2005. It is not required to pay yearly dues to ASCP (although there are lots of benefits to doing so) to maintain your certification. Starting in 2005 CEU's will be required to maintain certificiation. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jenny Oblander Sent: Monday, February 28, 2005 12:22 PM To: Histonet (E-mail) Subject: [Histonet] ASCP I have a question about the ASCP registry, some histotechs and I were talking about paying your ASCP dues every year and one said they have never paid since they were initially registered. I guess my question is do you have to maintain your dues with ASCP to continue to be registered? I always assumed that you need to pay dues every year to be registered but I really don't know. Thanks Jenny J.Oblander, HT (A.S.C.P.) Comparative Medicine Oklahoma Medical Research Foundation MS#32 825 NE 13th St. Okc,Ok 73104 jenny-oblander@omrf.ouhsc.edu 405-271-7083 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ASelf <@t> gmhsc.com Mon Feb 28 13:45:31 2005 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] PR (+) control tissue Message-ID: <39836CD6DB61654E8F95A35898C92186D70DD9@exchange.gmhpost.com> Hey Netters, Does anyone have any extra PR (+) control tissue that they would be willing to share with me? Thanks in advance, Amy NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. From kelly.mcqueeney <@t> bms.com Mon Feb 28 13:57:22 2005 From: kelly.mcqueeney <@t> bms.com (Kelly D Mcqueeney) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] "Flying" brains !! In-Reply-To: References: Message-ID: <422377A2.6050007@bms.com> Hi Diane, There is a static guard you can spray in the cryostat. If the cryostat has a vacuum, it may be affecting your cutting because you have not set your cutting window correctly.. Dianne Holmes wrote: >HELP! a colleague is cutting rat brains on a cryostat and is having a >terrible time with static causing the sections to "jump and fly" around >in the cryo-cabinet. Has anyone got a remedy for reducing the static ? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From JWEEMS <@t> sjha.org Mon Feb 28 14:41:05 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] "Flying" brains !! Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0B8F5E@sjhaexc02.sjha.org> Gently breath into the chamber. The moisture will reduce the static. j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Dianne Holmes Sent: Mon 2/28/2005 2:34 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] "Flying" brains !! HELP! a colleague is cutting rat brains on a cryostat and is having a terrible time with static causing the sections to "jump and fly" around in the cryo-cabinet. Has anyone got a remedy for reducing the static ? _______________________________________________ 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 RCHIOVETTI <@t> aol.com Mon Feb 28 14:59:01 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] "Flying" brains !! Message-ID: <9e.212a4a47.2f54e015@aol.com> In a message dated 2/28/2005 12:44:37 PM US Mountain Standard Time, dholmes@anatomy.umsmed.edu writes: > HELP! a colleague is cutting rat brains on a cryostat and is having a > terrible time with static causing the sections to "jump and fly" around > in the cryo-cabinet. Has anyone got a remedy for reducing the static ? > > Hi, Yes, there have been several suggestions posted to Histonet re: this problem. If I recall correctly, some of the solutions have been related to the clothing and the shoes that are worn when working at the cryostat. Someone suggested gently breathing on the block face (probably a humidity effect), and someone else suggested placing a fabric softener (the sheet type) in the chamber. Placing a small beaker of 80% ethanol in the cabinet also seems to work well. You can retrieve the postings by going to: . In the search box, type the following: static electricity cryostat Good luck, hope this helps! Bob Chiovetti Robert (Bob) Chiovetti, Ph.D. Independent Consultant for The Science, Biomedical and Industrial Sectors 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From TJasper <@t> smdc.org Mon Feb 28 15:19:12 2005 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Histotechs grossing Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E450A5@harrier> Dear Heather and All, Regarding histotechs grossing, I do not have the CLIA 88 rules in front of me and certainly don't have them memorized. I am not an attorney with expertise in medico-legal matters. Having said that, as an AP Coordinator and a histotech, my understanding of histotechs grossing basically goes something like this: A standard of service/care is evolving (at least in the USA) with what our institution has defined as "minor gross dictations". "Minor gross dictations" are for the most part dermatology and endoscopy cases (occasional endometrial/cervical specimens as well). These are straightforward cases which require the accurate dictation of patient demographics, clinical history (if provided), specimen source and/or the identification of the specimen as indicated on a requisition. This is then followed by a physical description of appearance, color, size and number. We generally close with a statement such as..."all submitted, or totally submitted in a single block, two blocks"...whatever the case may be. Every institution is different and the way they run their services will vary. I do not expect my histotechs to do "minor gross dictations" very often. We have a full staff of pathologists and a PA (we are looking to obtain a 2nd). The expectation with our service is that the PA and the pathologists will handle the tissue grossing. If the situation arises when a histotech (and it is usually me) can knock down some "minor grosses" to keep "the machine running" so to speak, we've met our objective. The histotechs do not gross large or complex cases and reserve the right to hold a case to defer to a pathologist or PA. As a matter of fact this is encouraged as we do not want anyone to proceed if they are not comfortable or feel that something about a particular case needs to be observed grossly by the pathologist or PA. As I've implied this does not occur that often and when it does it is under the direct or indirect supervision of the PA or pathologist. Therefore, as far as doctors not taking the blame and having histotechs go down in flames, this type of scenario should not occur. The pathologists are ultimately responsible, even for the PAs. They carry the malpractice and receive the big bucks for making the calls. As far as doing 2 jobs for the price of one I would not necessarily agree with that mindset. The majority of grossing that histotechs are "qualified" to do (to my understanding) should not be of a complex nature. Also, the majority of the cases handled per day should not be of the type histotechs are "qualified" to do. As I've stated, all services are different, perhaps someone out there is trying to get histotechs to gross cases which should be handled only by pathologists or PAs. Or, maybe there is some type of pathology service that handles only non-complex pathology, I am unaware of either type of work scenario. What I do believe as positive for histotechs is the recognition that they have enough intelligence and responsibility to handle grossing to the limit of their qualification. Just as other areas of Histology have expanded - IHC, ISH, FISH etc. - I am viewing this grossing responsibility as a natural expansion (evolution, if you like). The educational standards are being raised and when you couple that with the vacancy rates it only makes histotechs more valuable. I am not saying that I expect histotechs to do more as far as putting in a workday. For example we've got people doing IHC etc., if there are busy working on something - whatever it is - I don't expect them to drop it and go do minor grosses. If it fits into the workday and needs to be done, fine, I do not believe in piling on unmanageable workloads just because people have additional skills. This only my opinion and maybe somebody will think I'm all wet, but this is how I see it as an administrator and a histotech. Thanks. Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org 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 JMacDonald <@t> mtsac.edu Mon Feb 28 15:53:15 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] "Flying" brains !! In-Reply-To: <422377A2.6050007@bms.com> Message-ID: I would not recommend spraying any type of aerosol into a chamber containing unfixed tissue. Jennifer MacDonald Kelly D Mcqueeney Sent by: histonet-bounces@lists.utsouthwestern.edu 02/28/2005 11:57 AM To Dianne Holmes cc Histonet@lists.utsouthwestern.edu Subject Re: [Histonet] "Flying" brains !! Hi Diane, There is a static guard you can spray in the cryostat. If the cryostat has a vacuum, it may be affecting your cutting because you have not set your cutting window correctly.. Dianne Holmes wrote: >HELP! a colleague is cutting rat brains on a cryostat and is having a >terrible time with static causing the sections to "jump and fly" around >in the cryo-cabinet. Has anyone got a remedy for reducing the static ? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SAllen <@t> exchange.hsc.mb.ca Mon Feb 28 16:09:04 2005 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] "Flying" brains !! Message-ID: <304B5A264EC9974E8121B0EA14A9C3936619EC@hsc01mx1.hsc.mb.ca> This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. From carolb <@t> mail.phys.mcw.edu Mon Feb 28 16:11:56 2005 From: carolb <@t> mail.phys.mcw.edu (Carol Bobrowitz) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] "Flying" brains !! Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A9644@thor.phys.mcw.edu> I've sprayed static guard on gauze and wiped the knife blade holder. Seem to help. Carol Medical College of Wisconsin. -----Original Message----- From: Dianne Holmes [mailto:dholmes@anatomy.umsmed.edu] Sent: Monday, February 28, 2005 1:34 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] "Flying" brains !! HELP! a colleague is cutting rat brains on a cryostat and is having a terrible time with static causing the sections to "jump and fly" around in the cryo-cabinet. Has anyone got a remedy for reducing the static ? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Gervaip <@t> aol.com Mon Feb 28 19:42:05 2005 From: Gervaip <@t> aol.com (Gervaip@aol.com) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] ASCP - lots of questions!!! Message-ID: >From what I read, those newly registered will be required to have CEUs in order to renew their registry with the ASCP. I have not ready anything that is going to require those that have been registered to acquire CEUs. I have thought that eventually we will all be required to have the CEUs in order to renew membership. Why did the ASCP not do that? I think it is a good idea. But what will happen if one does not renew their registry? What kind of regulations are going to require that the histologists renew their membership? How are they going to enforce this? And if it turns out that this will be required of all of us ... what is to keep labs from hiring unregistered (unrenewed membership). Is this really going to amount to anymore then a pile of beans? It would be nice if it did. As it is, there are some pathologists that feel a monkey can crank that microtome. No respect for our professions. This is really a true statement. Here in the state of Louisiana when Licensure came up for medical professionals, we were not included. Intentionally not included. Not just an oversight. Manicurist and hair dressers here are licensed, but not the histotechnologist! And some of this disrespect has been earned! It has taken so long for us to accept the fact that we need college. How many of us out there can say that we have kept up with new technology in our field? If you are a member of this histonet server and an NSH member you most likely have taken those giant steps forward. Think of all the folks you know in histology that think they know it all and yet you would not want them in your lab! Maybe if we do this and bit that bullet, get our education (four years of college) and CEUs CLIA might recognize us! CLIA might just realize that we are a professional group that can do "complicated" tests, such as flow, fish and probes. If they knew how complicated immunohistochemistry is, they just might take that away from us too! If they knew that we are the professionals that make that final diagnosis they might not let us work in the lab! They can do all the blood work and do all kinds of tests.... but they still need us! My apologies,if I have offended anyone. My cage just got rattled and I just can't be quiet anymore. Pearl, from Louisiana From laurie.reilly <@t> jcu.edu.au Mon Feb 28 23:10:53 2005 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Sep 16 15:24:41 2005 Subject: [Histonet] Fisher "Histomatic" 166 Processor Manual In-Reply-To: <1DB04B57B04C5747B87C3B39F3E605AA02E450A5@harrier> Message-ID: <5.1.0.14.0.20050301150729.00a42bc0@mail.jcu.edu.au> Dear All, Could anyone help me with a copy of a manual for a Fisher Histomatic Tissue Processor Model 166? Thanks in advance. Regards, Laurie.