From hmcleod <@t> chempath.uct.ac.za Mon Sep 1 02:38:32 2003 From: hmcleod <@t> chempath.uct.ac.za (Mcleod) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] (no subject) Message-ID: <3F52F778.C4E6D39@chempath.uct.ac.za> Dear All Has anybody used Novocastra's (new) NCL-p53-505 on rodent (mouse) tissue? Also p53 clone D-07 is not supposed to cross react with mouse tissue ........ is this actually so? I have tried it on irradiated mouse livers and got some staining. Whether it is staining all that it is supposed to stain is what is of concern because of the cross reactivity question. Thanks for all replies Heather From hmcleod <@t> chempath.uct.ac.za Mon Sep 1 02:41:27 2003 From: hmcleod <@t> chempath.uct.ac.za (Mcleod) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] p53 Message-ID: <3F52F827.E3291E81@chempath.uct.ac.za> -------------- next part -------------- An embedded message was scrubbed... From: Mcleod Subject: (no subject) Date: Mon, 01 Sep 2003 09:38:32 +0200 Size: 908 Url: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030901/98fea3e1/attachment.eml From d.a.faichney <@t> stir.ac.uk Mon Sep 1 03:32:07 2003 From: d.a.faichney <@t> stir.ac.uk (Deborah Faichney) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Block storage Message-ID: <570E2BEE7BC5A34684EE5914FCFC368C190D26@fillan.stir.ac.uk> Hello all, Does a lockable block storage cabinet exist? Our Good Laboratory Practice (GLP) Quality Assurance person has requested one to comply with GLP block archive regulations. I cannot find any in the catalogues that I have here. I have suggested that he could buy a lockable cabinet to house the existing block storage unit!! With thanks Debbie Faichney Histopathology Institute of Aquaculture University of Stirling Stirling, FK9 4LA Scotland UK daf3@stir.ac.uk -- The University of Stirling is a university established in Scotland by charter at Stirling, FK9 4LA. Privileged/Confidential Information may be contained in this message. If you are not the addressee indicated in this message (or responsible for delivery of the message to such person), you may not disclose, copy or deliver this message to anyone and any action taken or omitted to be taken in reliance on it, is prohibited and may be unlawful. In such case, you should destroy this message and kindly notify the sender by reply email. Please advise immediately if you or your employer do not consent to Internet email for messages of this kind. Opinions, conclusions and other information in this message that do not relate to the official business of the University of Stirling shall be understood as neither given nor endorsed by it. From RSRICHMOND <@t> aol.com Mon Sep 1 12:58:24 2003 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Orcein Variation for EVG to detect Elastosis Perforans Serpiginosum Message-ID: <1de.eb02e8e.2c84e2c0@aol.com> A. Kevin Williams asks: >>My pathologist has asked for the Orcein variation on the EVG (Elastin Van Gieson) or VVG as some of you know it. I have to get the Orcein so that is my first challenge, second is my protocol. There are a few out there and I would like to hear from anyone who has a favorite for this method [elastosis perforans serpiginosa].<< Here are my reading notes - never tried it - on the traditional Pinkus orcein stain used by dermatopathologists of yore. See also the short description of the stain in John Kiernan's Histological & Histochemical Methods (199), page 162. Bob Richmond Samurai Pathologist Knoxville TN ****************** Mehregan, Amir H. (Wayne State). Pinkus' Guide to Dermatohistopathology. 4th ed. Appleton-Century-Crofts, 1986. Hermann Pinkus, 1905-1985 Acid orcein and Giemsa stain: from Pinkus H, Hunter R. Simplified acid orcein and Giemsa technique for routine staining of skin sections. Arch Dermatol 82:699, 1960 Krobock E, Rahbari H, Mehregan AH. Acid orcein and Giemsa stain. Modification of a valuable stain for dermatologic specimens. J Cutan Pathol 5:37, 1978. Fix in formalin or alcohol, without chromium or mercury. 1. Stain in ORCEIN for 30 minutes. Synthetic orcein stains elastic fibers specifically, with very little background staining. The background may be decolorized by short immersion in absolute alcohol or 0.1% acid alcohol. 2. Wash in running water 10 minutes. 3. Stain overnight in GIEMSA. Krobock et al. speeded this up by staining 1 hour in 1% Giemsa solution at 60? C. 4. Wipe slides. Remove excess blue by rinsing in 95% alcohol to which a small amount of eosin has been added if necessary. Continue until the collagen of the skin looks pink. Then dehydrate. ORCEIN: dissolve 200 mg of Harleco's synthetic Orcein in 100 mL of 70% alcohol. Add 0.6 mL of concentrated HCl. The solution improves on standing and has a long shelf life. GIEMSA: one drop of stock in 20 mL of distilled water or pH 7.0 phosphate buffer. RESULTS: Collagen is rose-pink, while elastic is dark brown to black. Melanin is dark green to black. Bacteria and fungi are dark blue. Looking at the book, I would suppose that Mehregan has just about abandoned the use of this stain. From t-sherman <@t> comcast.net Mon Sep 1 15:48:23 2003 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Nos Abs In-Reply-To: <20030901170001.15109.19708.Mailman@swlx167.swmed.edu> References: <20030901170001.15109.19708.Mailman@swlx167.swmed.edu> Message-ID: <3F53B097.5010701@comcast.net> Hello Lin, Try the Histosearch Search Engine. Your request has been asked before and there are some good archived responses. I tried a quick query but the search isn't working at the moment...probably related to the new listserver changes. Nonetheless, it is a good resource for your question. http://www.histosearch.com/histonet.html Todd histonet-request@lists.utsouthwestern.edu wrote: > Today's Topics: > 2. Nos Abs (Linresearch@aol.com) > > Message: 2 > From: Linresearch@aol.com > Date: Sun, 31 Aug 2003 17:01:48 EDT > To: histonet@pathology.swmed.edu > Subject: [Histonet] Nos Abs > > > --part1_4b.3326e80e.2c83bc3c_boundary > Content-Type: text/plain; charset="US-ASCII" > Content-Transfer-Encoding: 7bit > > Hello, > Does anyone with experience using eNos. iNos and nNOS on FFPE rodent tissue > care to share their protocols and AB source/s? > Lin > > --part1_4b.3326e80e.2c83bc3c_boundary From rbrown <@t> alphahunt.com Mon Sep 1 19:33:07 2003 From: rbrown <@t> alphahunt.com (Leroy H. Brown) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] code-on stainer needed Message-ID: <200309020033.RAA12112@admin.intlgraphicsdesign.org> I am looking for a replacement code-on stainer. If you have one that is no longer in use, or know someone who has one please contact me. The code-on is the one that uses the computer linkup to control the various stain programs. Once sold be Fisher, I do not think this stainer is made any more. thanks for your help. LeRoy Brown HT(ASCP) HTL HCS 207 N. Harkness St. Everson, WA 98247 360-966-7300 www.histocs.com From g.lang <@t> bigfoot.de Tue Sep 2 06:20:42 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] special stains Message-ID: <001c01c37144$4481d4e0$0d12a8c0@SERVER> Hi! I am interested in special stains used in routine histolabs. What stains are used most frequently in your labs? We work most with: PAS, Giemsa, CAB (Chromotrop-Anilin-Blau, tri-chrome-stain), BB (=Prussian blue), Alcianblue, EVG (Resorcin), Congored, Silverimpregnation Gomori (reticular fibers), Gomori's methenamin silver for basement membranes, SFOG (=S?urefuchsin-Orange-G; =acid fuchsin, orange-g, anilinblue; trichrom-stain) Sudan Ziehl-Neelsen greetings from Austria Gundi general hospital, Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030902/d2848237/attachment.htm From histo <@t> skm.org.pk Tue Sep 2 07:57:08 2003 From: histo <@t> skm.org.pk (Histology) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] special stains Message-ID: Masson Trichrome Giemsa ZN GMS Congo Red PAS Iron Reticulm Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk -----Original Message----- From: Gudrun Lang [SMTP:g.lang@bigfoot.de] Sent: Tuesday, September 02, 2003 4:21 PM To: Histonetliste Subject: [Histonet] special stains Hi! I am interested in special stains used in routine histolabs. What stains are used most frequently in your labs? We work most with: PAS, Giemsa, CAB (Chromotrop-Anilin-Blau, tri-chrome-stain), BB (=Prussian blue), Alcianblue, EVG (Resorcin), Congored, Silverimpregnation Gomori (reticular fibers), Gomori's methenamin silver for basement membranes, SFOG (=S?urefuchsin-Orange-G; =acid fuchsin, orange-g, anilinblue; trichrom-stain) Sudan Ziehl-Neelsen greetings from Austria Gundi general hospital, Linz, Austria From cfavara <@t> niaid.nih.gov Tue Sep 2 08:44:33 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] counting positive cells in IHC slides Message-ID: I do not think this is a silly question but points out the need for a person trained in the normal microscopic evaluation of tissue to carefully review the slides, and perhaps suggest alternative stains or tests. In my work this used to be a pathologist and now I see many people wearing multiple hats and pathologists have mostly turned into managers along with the rest of the highly trained and skilled medical personnel. Probably not a good place to go the beginning of the week! Just my own thoughts! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Subratab [mailto:subratab@bdonline.com] Sent: Friday, August 29, 2003 2:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] counting positive cells in IHC slides Dear All, I am sorry to bother you with a very silly question. I am new in the field of IHC. I have stained rat renal tissue slides for macrophage (ED1) with DAKO EnVision detection system. Now I have to count number of macrophages (ED1 positive cells) per 100 glomeruli. My problem is that the all positive stained cells are not similar to look; some cells are typical cell-like with bright red color, but other cells are a bit different in shape or size or color. So I am in problem in identifying true positive cells. I like to ask you if there is any systematic way to identify true positive cells from false staining area or artefact. ED1 antibody binds with cytoplasmic lysosomal membrane-associated antigen. I treated my slides with 0.3% triton x100 to break the cell membranes for better penetration of the antibody. So I think that the shape of the positive stained cells may be changed and all of them may not look typical cell-like structure. Am I correct? Can you please explain in some detail about the change of shape/size/color of the positive stained cells in IHC slides after staining. Particularly when the antigen is cytoplasmic. Please let me know if there is any website discussing this issue. Thanks in advance Dr Subrata Biswas, MD PhD student, Nephrology Div of Internal Med, FCM, University of Campinas, SP, Brazil. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tissuearray <@t> hotmail.com Tue Sep 2 08:59:25 2003 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Tissue Arrays Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030902/2c3b2269/attachment.htm From juan.gutierrez <@t> christushealth.org Tue Sep 2 10:47:55 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] FW: Job Posting!!! Message-ID: -----Original Message----- From: GUTIERREZ, JUAN Sent: Tue 9/2/2003 10:42 AM To: histonet@utsouthwestern.edu Cc: Subject: Job Posting!!! Histo Tech/ Immuno Tech needed in beautiful San Antonio, Texas. Full-time position with great benefits. Daytime hours M-F + rotate saturday mornings. Immuno experience a must.(Ventana Benchmark exp. preferred) Small lab with about 5,000 immunos/year. Over 100 markers on list and growing. For info contact: Juan C. Gutierrez, HT(ASCP) Histology Supervisor Christus Santa Rosa Healthcare 333 N. Santa Rosa Ave. San Antonio, TX 78207 (210)704-2533 office (210)704-3180 fax juan.gutierrez@christushealth.org From hho <@t> neurology.bsd.uchicago.edu Tue Sep 2 15:21:36 2003 From: hho <@t> neurology.bsd.uchicago.edu (hho@neurology.bsd.uchicago.edu) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Help! With LEICA TP1020 Automatic Tissue Processor Message-ID: Dear Histonetters, Does anybody use LEICA TP1020 Automatic Tissue Processor? If do, do you have a good tissue process protocol I could try out for Mice brain, spinal cord, muscle...? I've been smashing my head on the wall to try to get a good block for months, so far my head hurts. If you have any pointer or suggestion please reply or e-mail me PLEASE!!! Thanks in advance =). here's the protocol I've been using for LEICA TP1020 ATP. 70% ETOH 25min NO VAC 80% ETOH 25min NO VAC 95% ETOH 25min NO VAC 100%ETOH 25min NO VAC 100%ETOH 25min NO VAC 100%ETOH 25min NO VAC Xylene 25min NO VAC Xylene 25min NO VAC Paraffin 45min (58 Degree) VAC Paraffin 45min (58 Degree) VAC Hanson a very frustrated Lab tech. From RFail <@t> Charleston.net Tue Sep 2 17:43:44 2003 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] special stains In-Reply-To: <001c01c37144$4481d4e0$0d12a8c0@SERVER> Message-ID: <000001c371a3$aae88940$a210a6a5@rena> over 3200 Masson's Trichromes a year over 3200 PAS, D-PAS, for kidneys, glycogen or fungi a year GMS Steiner Auramine Rhodamine Kinyoun's Fite's Fe Colloidal iron Movat's pentachrome Bielshowsky Congo Red Jone's VVG B&B and will be doing Gram's Twort We offer 51 special stains on the request form, and if the chemicals and dyes are available will honor requests for stains not listed on our form Rena Fail Lead Tech IHC/SS Medical University of SC Charleston, South Carolina, USA -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Tuesday, September 02, 2003 7:21 AM To: Histonetliste Subject: [Histonet] special stains Hi! I am interested in special stains used in routine histolabs. What stains are used most frequently in your labs? We work most with: PAS, Giemsa, CAB (Chromotrop-Anilin-Blau, tri-chrome-stain), BB (=Prussian blue), Alcianblue, EVG (Resorcin), Congored, Silverimpregnation Gomori (reticular fibers), Gomori's methenamin silver for basement membranes, SFOG (=S?urefuchsin-Orange-G; =acid fuchsin, orange-g, anilinblue; trichrom-stain) Sudan Ziehl-Neelsen greetings from Austria Gundi general hospital, Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030902/c4382525/attachment.htm From AnthonyH <@t> chw.edu.au Tue Sep 2 18:27:59 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Alpha-amylase for PAS w/digestion Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E01D@simba.kids> Robert, A longlife solution (10min room temp incubation) is given in: Mangan, V-M, Farago, V., Kelly, M., Henwood, A.F., (2002) "An Amylase Reagent with a Long Shelf Life for the removal of Glycogen from Tissue Sections" J. Histotechnol 25:153. The amylase reagent used is as follows: Alpha Amylase from Bacillus Subtilis (Fluka Cat No 10070,) 1g (59 U/mg Lot 12580/1 44301) Oxoid PBS Tablets (Cat No BR14a) 1 tablet Distilled water 100ml Sodium Azide 0.1g Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Lott, Robert [mailto:Robert.Lott@bhsala.com] Sent: Saturday, 30 August 2003 5:33 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alpha-amylase for PAS w/digestion For those of you that use alpha-amylase... "exactly" what kind of reagent do you order for this procedure? Thanks! Robert L. Lott, HTL(ASCP) Manager, Anatomic Pathology Baptist Health System 800 Montclair Road Birmingham, AL 35213 205-592-5388|205-592-5646 (fax) robert.lott@bhsala.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 scott.turner <@t> dnax.org Tue Sep 2 18:29:25 2003 From: scott.turner <@t> dnax.org (Turner, Scott) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] counting positive cells in IHC slides Message-ID: <29B25753F6B1D51196110002A589D4444EC048@PALMSG30.us.schp.com> I wholeheartedly agree with John Kiernan's advice on statistical considerations. You have to remember that in any counting scheme like the one you mention (counting the number of macrophages per 100 glomeruli) you're estimating cell number and you must have a method for achieving an unbiased estimation. The best way to achieve unbiased estimates is by employing the stereological methods outlined by Gundersen et al. Before you begin, you should consult the following papers: Gundersen HJ, et al. The new stereological tools: disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis. APMIS. 1988 Oct;96(10):857-81. Review. Gundersen HJ, et al. Some new, simple and efficient stereological methods and their use in pathological research and diagnosis. APMIS. 1988 May;96(5):379-94. Review. These papers are the seminal references for the most commonly practiced stereological methods. They should help in the design of your counting procedures and aid in the statistical analysis of the data. They are a must read for anyone attempting to estimate pretty much anything from microscopic sections. Scott Turner DNAX Research Institute Palo Alto, CA > Dear All, > I am sorry to bother you with a very silly question. I am new in the field > of IHC. > I have stained rat renal tissue slides for macrophage (ED1) with DAKO > EnVision detection system. Now I have to count number of macrophages (ED1 > positive cells) per 100 glomeruli. My problem is that the all positive > stained cells are not similar to look; some cells are typical cell-like with > bright red color, but other cells are a bit different in shape or size or > color. So I am in problem in identifying true positive cells. I like to ask > you if there is any systematic way to identify true positive cells from > false staining area or artefact. > ED1 antibody binds with cytoplasmic lysosomal membrane-associated antigen. I > treated my slides with 0.3% triton x100 to break the cell membranes for > better penetration of the antibody. So I think that the shape of the > positive stained cells may be changed and all of them may not look typical > cell-like structure. Am I correct? Can you please explain in some detail > about the change of shape/size/color of the positive stained cells in IHC > slides after staining. Particularly when the antigen is cytoplasmic. Please > let me know if there is any website discussing this issue. Thanks in advance > Dr Subrata Biswas, MD > PhD student, Nephrology Div of Internal Med, > FCM, University of Campinas, SP, Brazil. > ********************************************************************* 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 AnthonyH <@t> chw.edu.au Tue Sep 2 18:28:30 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Orcein Variation for EVG to detect Elastosis Perfo rans Serpiginosum Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E01E@simba.kids> Kevin, The following article may be of use: Henwood, A., (2002) "The Orcein Stain - A Versatile Stain for Histopathology" J. Histotechnol 25(1):29. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: kevin williams [mailto:akwilliams75@hotmail.com] Sent: Monday, 1 September 2003 4:15 To: MTitford@aol.com; histonet@pathology.swmed.edu Subject: [Histonet] Orcein Variation for EVG to detect Elastosis Perforans Serpiginosum Dear MT( and all you HTs out there): Thanks for the information lots of help: I am now on the quest for a procedure for Elastosis Perforans Serpiginosum. My pathologist has asked for the Orcein variation on the EVG(Elastin Van Gieson) or VVG as some of you know it. I have to get the Orcein so that is my first challenge, second is my protocol. There are a few out there and I would like to here from anyone who has a favorite for this method with this disease process. Yours faithfully A. Keivn Williams >From: MTitford@aol.com >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Alpha Amylase >Date: Fri, 29 Aug 2003 15:50:51 -0400 > >Robert Lotts asks about amylase: >We use Sigma Alpha amylase Cat # A-3176.(0.125% at 37 degrees centigrade for 30 minutes). I zap it in the microwave to get it up to temperature > >Mike Titford >USA Pathology >Mobile AL 36617 USA > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ Get MSN 8 and help protect your children with advanced parental controls. _______________________________________________ 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. ********************************************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030903/3b740a60/attachment.htm From dmccaig <@t> ckha.on.ca Wed Sep 3 05:59:35 2003 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Tamtron Users Message-ID: We are considering the implementation of Tamtron and I would like to know the pros and cons that current users have with this program. How do you use order entry? Any advise would be truly appreciated. Diana McCaig, R.T. Charge Tech, Histology Chatham Kent Health Alliance 519-352-6401 (3347) From tissuearray <@t> hotmail.com Wed Sep 3 07:37:59 2003 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Tissue Array instrument information Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030903/98f28ee4/attachment.htm From SBarnes <@t> elch.org Wed Sep 3 08:57:51 2003 From: SBarnes <@t> elch.org (Sue Barnes) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Grossing Work Station Message-ID: <3F74CB2B769A5843A0C746D3F13256FD1279EA@ELCH2> Hi I need some information on Grossing Work Stations. We are budgeting for one this year and the only information I have is from Thermo Shannon. Are there other companies out there that have the same type of station? Is anyone using a grossing station, if so how do you like it. If there are vendors please get some information to me. Thanks Sue Barnes East Liverpool City Hospital East Liverpool, Ohio From peoshel <@t> wisc.edu Wed Sep 3 08:20:59 2003 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] counting positive cells in IHC slides In-Reply-To: <29B25753F6B1D51196110002A589D4444EC048@PALMSG30.us.schp.com> References: <29B25753F6B1D51196110002A589D4444EC048@PALMSG30.us.schp.com> Message-ID: The Gundersen refs are excellent and widely used. I would also like to add a basic text in stereology: John Russ' "Practical Stereology". Note that this is for the 1st edition. The 2nd edition, recently out, has a co-author whose name I forget, sorry. Phil >I wholeheartedly agree with John Kiernan's advice on statistical >considerations. You have to remember that in any counting scheme like the >one you mention (counting the number of macrophages per 100 glomeruli) >you're estimating cell number and you must have a method for achieving an >unbiased estimation. The best way to achieve unbiased estimates is by >employing the stereological methods outlined by Gundersen et al. Before you >begin, you should consult the following papers: > >Gundersen HJ, et al. The new stereological tools: disector, fractionator, >nucleator and point sampled intercepts and their use in pathological >research and diagnosis. APMIS. 1988 Oct;96(10):857-81. Review. > >Gundersen HJ, et al. Some new, simple and efficient stereological methods >and their use in pathological research and diagnosis. APMIS. 1988 >May;96(5):379-94. Review. > >These papers are the seminal references for the most commonly practiced >stereological methods. They should help in the design of your counting >procedures and aid in the statistical analysis of the data. They are a must >read for anyone attempting to estimate pretty much anything from microscopic >sections. > >Scott Turner >DNAX Research Institute >Palo Alto, CA > > >> Dear All, >> I am sorry to bother you with a very silly question. I am new in the field >> of IHC. >> I have stained rat renal tissue slides for macrophage (ED1) with DAKO >> EnVision detection system. Now I have to count number of macrophages (ED1 >> positive cells) per 100 glomeruli. My problem is that the all positive >> stained cells are not similar to look; some cells are typical cell-like >with >> bright red color, but other cells are a bit different in shape or size or >> color. So I am in problem in identifying true positive cells. I like to >ask >> you if there is any systematic way to identify true positive cells from >> false staining area or artefact. >> ED1 antibody binds with cytoplasmic lysosomal membrane-associated antigen. >I >> treated my slides with 0.3% triton x100 to break the cell membranes for >> better penetration of the antibody. So I think that the shape of the >> positive stained cells may be changed and all of them may not look typical >> cell-like structure. Am I correct? Can you please explain in some detail >> about the change of shape/size/color of the positive stained cells in IHC >> slides after staining. Particularly when the antigen is cytoplasmic. >Please >> let me know if there is any website discussing this issue. Thanks in >advance >> Dr Subrata Biswas, MD >> PhD student, Nephrology Div of Internal Med, >> FCM, University of Campinas, SP, Brazil. >> > > >********************************************************************* >This message and any attachments are solely for the intended >recipient. If you are not the intended recipient, disclosure, >copying, use or distribution of the information included in this >message is prohibited -- Please immediately and permanently delete. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From juan.gutierrez <@t> christushealth.org Wed Sep 3 09:25:40 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Grossing Work Station Message-ID: We just had one installed this summer, and the doctors seem to like it. It is from MOPEC. You can contact them at 1-800-362-8491 or on the web at www.mopec.com. We got the MB600 with the adjustable height. When you have several docs with differing styles of grossing it comes in handy. What's nice about it is that it is self contained, including a sink with garbage disposal. It also adjust so that the doc or tech can either sit down or stand to gross. If you have any tall people grossing they're going to like it. Sorry to have rambled on. I hope this helps. Juan C. Gutierrez, HT(ASCP) -----Original Message----- From: Sue Barnes [mailto:SBarnes@elch.org] Sent: Wed 9/3/2003 8:57 AM To: 'histonet@lists.utsouthwestern.edu' Cc: Subject: [Histonet] Grossing Work Station Hi I need some information on Grossing Work Stations. We are budgeting for one this year and the only information I have is from Thermo Shannon. Are there other companies out there that have the same type of station? Is anyone using a grossing station, if so how do you like it. If there are vendors please get some information to me. Thanks Sue Barnes East Liverpool City Hospital East Liverpool, Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Lynne.Bell <@t> hitchcock.org Wed Sep 3 09:50:05 2003 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Grossing Work Station Message-ID: We currently have the ThermoShandon Grossing station and have had it for approximately 7 years. Our pathologists like it and we like it also. The stainless steel is easy to keep clean and the garbage disposal is handy. The pathologists also like the fact that it is adjustable. We have ours vented to the outside - our ventilation is so strong that you could "suck up a small child". The only drawback that we have had is that the pathologists aren't thrilled with the sensors that turn on the hot and cold water and we are in the process of getting the sensors deactivated. Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From TJJ <@t> Stowers-Institute.org Wed Sep 3 10:47:22 2003 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Ventana Discovery Message-ID: I would be interested in hearing from any users out there of the Ventana Discovery instrument. I'm thinking of taking a look at this and want to know the pros and cons you've discovered in the field. I'm much more interested in the ISH capabilities than the IHC. Thank you for your help! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From hymclab <@t> hyhc.com Wed Sep 3 11:22:33 2003 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Grossing Work Station Message-ID: <529F3A73499ED611AA9D00A0C9558E4E43EEF5@hyhcexchange.hyhc.local> We have had the Thermo Shandon (now just Thermo) grossing station for approximately 3 years and love it. We have had no problems whatsoever!!!! Dawn Schneider, HT(ASCP) Howard Young Medical Center Woodruff, WI -----Original Message----- From: Sue Barnes [mailto:SBarnes@elch.org] Sent: Wednesday, September 03, 2003 8:58 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Grossing Work Station Hi I need some information on Grossing Work Stations. We are budgeting for one this year and the only information I have is from Thermo Shannon. Are there other companies out there that have the same type of station? Is anyone using a grossing station, if so how do you like it. If there are vendors please get some information to me. Thanks Sue Barnes East Liverpool City Hospital East Liverpool, Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carolb <@t> mail.phys.mcw.edu Wed Sep 3 12:31:28 2003 From: carolb <@t> mail.phys.mcw.edu (Carol Bobrowitz) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] anti-Farnesyltransferase Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A9576@thor.phys.mcw.edu> Has anyone tried anti-Farnesyltransferase, Rat, recombinant (rabbit) on formalin fixed paraffin sections? What about perfused tissue for frozen sections? I'd like to hear from you if you have tried this antibody. 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 From g.lang <@t> bigfoot.de Wed Sep 3 12:44:23 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] specialstains Message-ID: <000201c37245$92936930$0d12a8c0@SERVER> Hi! Thanks for your answers. Although I have some difficulties about the different names and abbreviations of the specialstains, I see that the most important are used all over the world. After contacting other pathologies in Austria, I found out, that no one works with the same procedures, although the names are similar. And the pathologists are satisfied. Are there better standards in your country? Or is it the same, that each lab "cooks its own soup"? I made a list of the answer-stains. I made questionmarks to those, I never heard bevore. Perhaps someone is so friendly and explain the english terms to me. greetings from Austria Gundi general hospital, Linz, Austria Synonym Substrat German unknown for me ABPAS ? Alcian blue acid mucopolysaccharides Alzianblau Auramin Rhodamine ? Bielshowsky ? Congo Red amyloid Kongorot Diazo ? D-PAS Diastase-PAS Diastase-PAS EVG Elastica-Van-Gieson elastic fibers EVG Fite's ? Giemsa blood cells, helicobacter pylori Giemsa GMS Grocott, Methenamin Silver fungus, pneumocystis Grocott Gordon and Sweets Reticulin reticular fibers Gomori (Gitterfasern) Gram bacteria Gram Gram's Twort ? Grimelius HVG hematoxylin van gieson Iron Perls prussian blue Iron, Fe Jones's Methenamin Silver basement membrans Methenamin Kinyoun's ? Masson Fontana Melanin Masson Fontana Masson Trichrome connective tissue Masson Trichrom Movat's pentachrom connective tissue MSB ? Mucin Orcein PAS Perjod-acid-schiff glycogen, carbohydrates PAS PTAH ? Reticulum ? Steiner ? Toluidin Blue Toluidinblau WG ? ZN Ziehl-Neelsen /AFB bacteria, acid fast bacteria Ziehl-Neelsen -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030903/b2657fb0/attachment.htm From moore.chrystal <@t> cryolife.com Wed Sep 3 13:07:49 2003 From: moore.chrystal <@t> cryolife.com (moore.chrystal@cryolife.com) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] CryoLife Needs a Histotechnologist Message-ID: We currently have an opening for a Histotechnologist in NW Atlanta. Reporting to the Associate Medical Director, the main objective is to operate a laboratory providing in-house histology services to support pathology and R&D. Responsibilities: Operation and maintenance of tissue processing equipment. Preparation of histologic slides, including routine staining, special histochemical staining, and immunohistochemistry (manual and automated.) Manage inventory of histology supplies. Assist pathologist in performing gross examination of pathology sections. Qualifications: Certified HT (ASCP) or equivalent. 2 yr previous experience, supervisory level preferred. Skilled in routine histology, special histochemistry, and immunohistochemistry. Operation of histology equipment: tissue processor, embedding station, microtome, cryostat and automated stainer. Hours: 9 am to 6 pm Monday through Friday. If you know if any candidates that may be interested, please have them contact me directly. Sincerely, Chrystal Moore, PHR Human Resources Coordinator CryoLife, Inc. Direct: (678) 290-4363 Fax: (770) 590-3741 Email: moore.chrystal@cryolife.com Confidentiality Note: This e-mail message may contain information that is privileged and/or confidential. If you are not the addressee or an authorized recipient of this message, any distribution, copying, publication, or use of this information for any purpose is prohibited. Please notify the sender immediately by e-mail and then delete this message. From info <@t> instrumedics.com Wed Sep 3 16:30:06 2003 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Tissue Arrays Message-ID: <000801c37262$8bef8ec0$6401a8c0@instrumedics1> Thom Jensen invited us to visit his web site that discusses his approach to the preparation of tissue arrays. I would also suggest that those of you preparing tissue arrays might find the discussion on the Instrumedics' web site valuable. The tape-transfer system may not be the method for everyone, but for very many it has become a very important tool. It was critical for the people at the NIH who developed the tissue array technology and the first publication from the NIH that appeared in Nature Medicine in July 1998 cited the invaluable role of the paraffin tape- transfer. Bernice schiller@instrumedics.com From jessgrocki <@t> yahoo.com Wed Sep 3 16:25:04 2003 From: jessgrocki <@t> yahoo.com (Jessica Piche) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Histotechnician Responsibilities Message-ID: <20030903212504.98528.qmail@web41608.mail.yahoo.com> I just found out today that my job description has changed. The job description that I have had since I started working states, "Responsible for the preparation of routine diagnostic slides by fixation and processing of tissue blocks.........." The new job description states, "Responsible for the preparation of routine (non-biopsy) diagnostic slides by fixation etc". Are there other HT's out there that are not allowed to cut small biopsy tissue samples? Are HT's only there to cut the "big tissue"? I would appreciate other peoples input on the subject. Thanks so much. Jessica Piche-Grocki, HT --------------------------------- Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030903/52681e4e/attachment.htm From Nancy71149 <@t> aol.com Wed Sep 3 17:15:46 2003 From: Nancy71149 <@t> aol.com (Nancy71149@aol.com) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Re: Grossing stations Message-ID: <1e6.f434876.2c87c212@aol.com> We are in the process of re-locating our Histology lab from one campus to another. The area has been renovated for us. We purchased 2 gross stations from Mopec in Michigan. They build the station to your specifications. There are several options available. We got one that has the elevating unit to adjust to the height of the user, and one more simple model. I would recommend checking them out. Nancy Temple Histology Supervisor St. Francis Hospital Ctr. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030903/6e9f4aa9/attachment.htm From KHays <@t> mbhs.org Wed Sep 3 17:09:21 2003 From: KHays <@t> mbhs.org (KHays@mbhs.org) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Histotechnician Responsibilities Message-ID: Kathy Tedford-Hays HT (ASCP) Technical Specialist, Histology Dept (601)-968-3070 ext 7398 Baptist Medical Center 1225 North State Street Jackson, MS 39202 it can depend on if they are certified by ascp as a ht at my hospital. are you certified? i have all certified techs at present. one of which is half way through her exam. i would ask for the htl's job description to review. see the differences in job descriptions. maybe the supervisor has seen some cut aways coming from only the ht's and wants to limit the liability in the future. did the supervisor experience some tissues that were not cut properly recently? i have taken techs off of performing certain duties if i see a trend coming from specific ht's and then i go over the problem and they practice and hopefully when my faith is renewed i assign them more challenging cases (biopsies). Jessica Piche To: histonet@pathology.swmed.edu Sent by: cc: lelaangel@aol.com histonet-admin@lists.utsouth Fax to: western.edu Subject: [Histonet] Histotechnician Responsibilities 09/03/03 04:25 PM I just found out today that my job description has changed. The job description that I have had since I started working states, "Responsible for the preparation of routine diagnostic slides by fixation and processing of tissue blocks.........." The new job description states, "Responsible for the preparation of routine (non-biopsy) diagnostic slides by fixation etc". Are there other HT's out there that are not allowed to cut small biopsy tissue samples? Are HT's only there to cut the "big tissue"? I would appreciate other peoples input on the subject. Thanks so much. Jessica Piche-Grocki, HT Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software The information contained in this email is confidential and is intended for the recipient only. Do not forward this email without permission from its originator. From kemlo <@t> tiscali.co.uk Thu Sep 4 02:07:36 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Controls Message-ID: <001901c372b3$38f8d590$55cce150@KEMLOS> Apparently there is an issue concerning control material and organ retention in the UK. What are the recommendations please? I realise they are commercially available but can they be retained after consent? Yours truly, A recovering Cytologist Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030904/99ab3c41/attachment.htm From arme <@t> optonline.net Thu Sep 4 02:25:24 2003 From: arme <@t> optonline.net (Mark Sofferman) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] For sale - Grossing Station and VIP 3000 Message-ID: We have both items available for sale. Please contact for more information. ARME 201-833-1550 arme@optonline.net From arme <@t> optonline.net Thu Sep 4 02:26:37 2003 From: arme <@t> optonline.net (Mark Sofferman) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] For sale - Grossing Station and VIP 3000 Message-ID: We have both items available for sale. Please contact for more information. ARME 201-833-1550 arme@optonline.net From t.hacker <@t> har.mrc.ac.uk Thu Sep 4 02:39:02 2003 From: t.hacker <@t> har.mrc.ac.uk (Terry Hacker) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] unsubscribe Message-ID: <000f01c372b7$9c863a90$e0f1f682@A383R28DELL360> Terry Hacker Head of Histology and Electron Microscopy Services Medical Research Council Harwell Didcot Oxfordshire OX11 ORD Tel: 01235 841128 Fax: 01235 841200 e-mail t.hacker@har.mrc.ac.uk -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030904/463b1580/attachment.htm From stevemachinuk <@t> yahoo.co.uk Thu Sep 4 02:26:13 2003 From: stevemachinuk <@t> yahoo.co.uk (=?iso-8859-1?q?Steve=20Machin=20UK?=) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Controls In-Reply-To: <001901c372b3$38f8d590$55cce150@KEMLOS> Message-ID: <20030904072613.51911.qmail@web12406.mail.yahoo.com> In short, you can not use a sample for anything for which you do not have informed consent. Post mortem material is covered by the by the new national PM consent form and leaflets which explain the use of leftover samples for quality control. However there is no similar national consent form for biopsy/surgical material. You need to be sure that your Trust explains the possible use for samples for controls. My trust does this with a leaflet which is given to patients/parents when consent for the biopsy procedure is obtained and we operate an opt-out system. I.E. they are told if they don't opt-out of the "Non- teaching and non-research (QC) use" then the sample may be used for it. We have a pathology wide flag on the patients record alerting labs of patients who have opted out. From a patients point of view, if they tell a doctor that they want to opt-out they may well expect that all subsequent samples will also prevented from being used for QC. --- Kemlo Rogerson wrote: > Apparently there is an issue concerning control material and organ > retention in the UK. What are the recommendations please? > > I realise they are commercially available but can they be retained > after > consent? > > Yours truly, > > A recovering Cytologist > > Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS > Tel: 01270 877625 > Mob: 07830 196072 > Mobile E-Mail kemlorogerson@3mail.com > FAX & Answer Phone 0871 242 8094 > E-mail Accounts: > kemlo@tiscali.co.uk or > kemlo1@btinternet.com > Disclaimer: The information contained in this message and/or any > attachments(s) may be of a private and confidential nature, and is > intended solely for the attention of the addressee. If you have > received > this message in error or feel you should not have been the intended > recipient, please return it and any attachments to the sender > immediately. All messages relating to this communication should > then be > deleted from your system. Unauthorised usage, copying, disclosure > or > alteration of this message and/or attachment(s) is strictly > prohibited. > Barking, Havering and Redbridge Hospitals NHS Trust will not be > held > responsible for any direct or indirect damages which may arise from > alteration of this message or any attachment(s), by a third party > or > resulting from the transmission of a virus. > > > > > > ________________________________________________________________________ Want to chat instantly with your online friends? Get the FREE Yahoo! Messenger http://mail.messenger.yahoo.co.uk From davenhv <@t> hotmail.com Thu Sep 4 07:19:48 2003 From: davenhv <@t> hotmail.com (Dave McClister) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] subscribe digest Message-ID: Thanks Dave McClister davenhv@hotmail.com _________________________________________________________________ Compare Cable, DSL or Satellite plans: As low as $29.95. https://broadband.msn.com From cristiano.rumio <@t> unimi.it Thu Sep 4 07:33:28 2003 From: cristiano.rumio <@t> unimi.it (Cristiano Rumio) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] (no subject) Message-ID: <5.2.0.9.0.20030904143306.028e7868@mailserver.unimi.it> please unsubscribe From nancy.cardwell <@t> Vanderbilt.Edu Thu Sep 4 07:39:27 2003 From: nancy.cardwell <@t> Vanderbilt.Edu (Cardwell, Nancy) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Unsubscribe Message-ID: <9A5DCCC462B8934A85B1779B11AFD7959933EB@mailbe01> Nancy Cardwell Plastic Surgery Research Vanderbilt University Medical Center -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030904/88b71e62/attachment.htm From jessgrocki <@t> yahoo.com Thu Sep 4 08:19:00 2003 From: jessgrocki <@t> yahoo.com (Jessica Piche) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] histotechnician responsibilities Message-ID: <20030904131900.89165.qmail@web41612.mail.yahoo.com> I forgot to let everyone know that yes I am a certified HT, and I do not have any known problems with my cutting abilities. Thanks. --------------------------------- Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030904/f8f45061/attachment.htm From Charles.Embrey <@t> carle.com Thu Sep 4 09:31:25 2003 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Looking for lead technician Message-ID: I am currently looking for an experienced histotech to serve as my Lead technician in our hospital based histology lab. We are currently processing aprox 25,000 cases per year (about 200-250 blocks per day). We use 2 VIP processors, Leica and Microm microtomes, a Leica Autostainer XL, and a Tissue Tek automated coverslipper. We do our Immunos on a Dako autostainer. You would help supervise 1 HTL, 2 HTs, and 4 Lab assistants (3 of which are taking their HT exams this year). We also have 1 part time evening HT and an HT from the University of Illinois that fills in when needed. We are in Champaign-Urbana Illinois, home of the University of Illinois Fightin Illini. Chicago is 2 hours to the north. Indianapolis is about 2 hours to the east and ST Louis is about 3.5 hours to the southwest. Being a collage town there is lots to do and see. For more information shoot me an e-mail at charles.embrey@carle.com or give me a call at (217) 383-6621. We are on the web at www.carle.com . Sorry this listing isn't all pretty and fancy but am just a manager and not a human resource artist. Thanks, Charles R. Embrey Jr. PA(AAPA), HT(ASCP) Histology Manager Carle Clinic, Urbana Illinois From hborgeri <@t> wfubmc.edu Thu Sep 4 09:53:25 2003 From: hborgeri <@t> wfubmc.edu (Hermina Borgerink) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Measles antibody Message-ID: <9AEEF1FB6254224AA355ED285F84916501799F56@EXCHVS2.medctr.ad.wfubmc.edu> Histonetters, I have searched through various catalogs and gone on line, but so far have been unable to locate an anti-measles antibody that may be used on FFPE tissue. If anyone, including vendors, knows of the availability of such an antibody, I would be most appreciative of any information you would be willing to share with me. Thanks in advance. Hermina Hermina M. Borgerink, BA, HTL, HT(ASCP)IHQ Department of Pathology Wake Forest University Health Sciences Medical Center Blvd Winston-Salem, NC 27157 Phone (336)716-1538 Fax (336)716-1515 hborgeri@wfubmc.edu From Rcartun <@t> harthosp.org Thu Sep 4 10:49:02 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Measles antibody Message-ID: Try Chemicon International. Richard Cartun >>> "Hermina Borgerink" 09/04/03 10:53AM >>> Histonetters, I have searched through various catalogs and gone on line, but so far have been unable to locate an anti-measles antibody that may be used on FFPE tissue. If anyone, including vendors, knows of the availability of such an antibody, I would be most appreciative of any information you would be willing to share with me. Thanks in advance. Hermina Hermina M. Borgerink, BA, HTL, HT(ASCP)IHQ Department of Pathology Wake Forest University Health Sciences Medical Center Blvd Winston-Salem, NC 27157 Phone (336)716-1538 Fax (336)716-1515 hborgeri@wfubmc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mari.ann.mailhiot <@t> leica-microsystems.com Thu Sep 4 11:03:08 2003 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Test Message-ID: 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 From juan.gutierrez <@t> christushealth.org Thu Sep 4 11:40:34 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Looking for lead technician Message-ID: Try USBiological, they have both rubella and rubeola for paraffin. I have not use these Ab's, but I have used some of their other stuff with good results. Their # is 1-800-520-3011 or on the web at www.usbio.net. A good resource to find antibodies is www.sciquest.com. You can do Ab searches by name or clone, and they list the different companies that supply them. It saves a lot of time looking through all the catalogs. Good luck. Juan C. Gutierrez, HT(ASCP) -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: Thu 9/4/2003 9:31 AM To: 'histonet@lists.utsouthwestern.edu' Cc: Subject: [Histonet] Looking for lead technician I am currently looking for an experienced histotech to serve as my Lead technician in our hospital based histology lab. We are currently processing aprox 25,000 cases per year (about 200-250 blocks per day). We use 2 VIP processors, Leica and Microm microtomes, a Leica Autostainer XL, and a Tissue Tek automated coverslipper. We do our Immunos on a Dako autostainer. You would help supervise 1 HTL, 2 HTs, and 4 Lab assistants (3 of which are taking their HT exams this year). We also have 1 part time evening HT and an HT from the University of Illinois that fills in when needed. We are in Champaign-Urbana Illinois, home of the University of Illinois Fightin Illini. Chicago is 2 hours to the north. Indianapolis is about 2 hours to the east and ST Louis is about 3.5 hours to the southwest. Being a collage town there is lots to do and see. For more information shoot me an e-mail at charles.embrey@carle.com or give me a call at (217) 383-6621. We are on the web at www.carle.com . Sorry this listing isn't all pretty and fancy but am just a manager and not a human resource artist. Thanks, Charles R. Embrey Jr. PA(AAPA), HT(ASCP) Histology Manager Carle Clinic, Urbana Illinois _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kgibbon <@t> qltinc.com Thu Sep 4 12:23:32 2003 From: kgibbon <@t> qltinc.com (kgibbon@qltinc.com) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] checking for an image Message-ID: Hi Everyone, I have a small macro that sequentially goes down a stack of images converting them from one format to another. Does anyone have a line of code or a function in image pro that will report a "true" value if an image is present and a "false" value if there is none present?? I don't mind what values are used but 1 and 0 seem the easiest to use thanks Kevin Gibbon QLT Inc. This electronic transmission (including any and all attachments) is intended solely for the use of the individual or entity to whom it is addressed and may contain information that is privileged and/or confidential. If you are not the intended recipient of this electronic transmission, you are hereby notified that any disclosure, copying or distribution, or the taking of any action in reliance upon the contents of this electronic transmission, is strictly prohibited, and you are further requested to purge this electronic transmission and all copies thereof from your computer system. From philippegannon <@t> hotmail.com Thu Sep 4 12:32:25 2003 From: philippegannon <@t> hotmail.com (Philippe Gannon) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] IHC with CD25 on FFPE lymph node Message-ID: hi, I am having some difficulties with a CD25 staining on formalin-fixed paraffin embedded lymph node slides. In fact, I am not seeing any staining at all with the antibody CBL 151 from Cymbus Biotechnology LTD/Chemicon at concentration up to 1:10 in PBS. I have tried three different antigen retrieval techniques: a) Citrate 10mM pH6 (15min in pre-heated citrate at 10% power in microwave) b) EDTA 1mM pH8 (10min in pre-heated citrate at 10% power in microwave) c) Pronase: Sigma Protease P-5147. 75mg in 300ml H2O for 10min at Room Temperature. Here's the rest of my protocol: 1- Heat slides at 60oC: 30min 2- Toluene: 2 x 5min 3- Hydration with EtOH 96%, 90% 80%: 3min each 4- Water: 5min 5- 3% H2O2: 5min 6- Water: 5min 7- Pre-heated Citrate: 15min at 10% power level in microwave 8- Cooling under running water: 10min 9- PBS: 5min 10- Protein block (DAKO protein block #0909): 5min 11- CD25 @ 1:10: 60min at Room Temperature 12- PBS: 5min 13- Secondary/tertiary Antibody (DAKO LSAB2 system peroxidase): 20min each with a 5min PBS in between. Room temperature. 14- PBS: 5min 15- DAB with H2O2: 5min 16- Hematoxylin: 5secs 17- Running water: 5min 18- EtOH: 80%, 90%, 96: 3min each 19- Toluene: 2 x 5min Thank you for your suggestions, Philippe Gannon Graduate Student Laboratoire Mes-Masson (Y-4609) Institut du cancer de Montréal, Université de Montréal 2099 Alexandre de Sève Montréal, Québec, Canada H2L 2W5 _________________________________________________________________ Add photos to your messages with MSN 8. Get 2 months FREE*. http://join.msn.com/?page=features/featuredemail From davenhv <@t> hotmail.com Thu Sep 4 12:35:02 2003 From: davenhv <@t> hotmail.com (Dave McClister) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Animal Histology Texts Message-ID: I am in a new research postion working with mice and pig hearts. The histolab is not up and running yet except for an embedding center and microtome. After reviewing histonet archives, I have yet to find a good textbook dealing with animal tissue from grossing all the way to staining. Any help would be greatly appreciated. Additionally after cutting a few blocks of mouse heart with a new Microm HM 325, I can't get sections that are free of artifact. All of my experience with histology has been clinical so unless I can get some help I might have to look for a new job!! Thanks to all you can help. Thanks Dave McClister 843-789-6826 work davenhv@hotmail.com _________________________________________________________________ Express yourself with MSN Messenger 6.0 -- download now! http://www.msnmessenger-download.com/tracking/reach_general From jkiernan <@t> uwo.ca Thu Sep 4 13:00:05 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Animal Histology Texts References: Message-ID: <3F577DA5.E35C5E3C@uwo.ca> Try "Humason's Animal Tissue Techniques" 5th edn by JK Presnell & MR Schreibman. Baltimore: Johns Hopkins Univ Press, 1997. The 4th edition (by GL Humason) might be a bargain second-hand. Processing mouse and pig tissues isn't any different from doing human tissues, and there are many technical manuals aimed primarily at histopathology labs. The most comprehensive is probably "Theory and Practice of Histological Techniques" 5th ed by Bancroft & Gamble (editors), (2002). -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ _______________________________________ Dave McClister wrote: > > I am in a new research postion working with mice and pig hearts. The > histolab is not up and running yet except for an embedding center and > microtome. After reviewing histonet archives, I have yet to find a good > textbook dealing with animal tissue from grossing all the way to staining. > Any help would be greatly appreciated. > Additionally after cutting a few blocks of mouse heart with a new Microm HM > 325, I can't get sections that are free of artifact. All of my experience > with histology has been clinical so unless I can get some help I might have > to look for a new job!! > Thanks to all you can help. > > Thanks > Dave McClister > 843-789-6826 work > davenhv@hotmail.com > > _________________________________________________________________ > Express yourself with MSN Messenger 6.0 -- download now! > http://www.msnmessenger-download.com/tracking/reach_general > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Thu Sep 4 14:00:57 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Measles antibody Message-ID: Try USBiologicals. www.usbio.net. 1-800-520-3011. They have both rubella and rubeola for paraffin. I tried replying to you before, but ended up sending the info to someone else, sorry. Good luck. Juan C. Gutierrez, HT(ASCP) -----Original Message----- From: Hermina Borgerink [mailto:hborgeri@wfubmc.edu] Sent: Thu 9/4/2003 9:53 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Measles antibody Histonetters, I have searched through various catalogs and gone on line, but so far have been unable to locate an anti-measles antibody that may be used on FFPE tissue. If anyone, including vendors, knows of the availability of such an antibody, I would be most appreciative of any information you would be willing to share with me. Thanks in advance. Hermina Hermina M. Borgerink, BA, HTL, HT(ASCP)IHQ Department of Pathology Wake Forest University Health Sciences Medical Center Blvd Winston-Salem, NC 27157 Phone (336)716-1538 Fax (336)716-1515 hborgeri@wfubmc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcdonal1 <@t> ccf.org Thu Sep 4 15:35:43 2003 From: mcdonal1 <@t> ccf.org (Linda McDonald) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Zinc Formalin Message-ID: Hey Zinc formalin users, what's the pH of your buffered 10% zinc formalin? Thanks, LGMcDonald ------------------------------------------------------------------------------ Confidentiality Note: This message is intended for use only by the individual or entity to which it is addressed and may contain information that is privileged, confidential, and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please contact the sender immediately and destroy the material in its entirety, whether electronic or hard copy. Thank you. Visit us online at our award-winning www.clevelandclinic.org for a complete listing of Cleveland Clinic services, staff and locations from one of the country's leading hospitals. ============================================================================== From djcarter <@t> dallastx.net Thu Sep 4 20:22:16 2003 From: djcarter <@t> dallastx.net (Dallas Nippert) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Animal Histology Texts References: Message-ID: <002e01c3734c$284316e0$2ee8a641@djcarter> I did animal tissue for about 6 years, and the only big difference, is that the tissue is dryer. Try cutting down on the alcohol times on the processor and float your block in your water bath for about 30 seconds before putting on your ice. Dallas Nippert Ameripath, TX From slimwillie <@t> cox.net Thu Sep 4 21:45:52 2003 From: slimwillie <@t> cox.net (Jerry Wilson) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] test Message-ID: <01e501c37357$d2b2fb80$c4370e44@no.cox.net> Sorry for this, but I have to know if this thing is working ok -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030904/640c4a4f/attachment.htm From RBARNHART <@t> summithealth.org Fri Sep 5 07:09:17 2003 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Manual Message-ID: Has anyone out there in histoland put their manuals on line and done away with the paper version? This is what we are working on. We will have an icon on all computer so the manual is easily accessible. The problem I am having, I can't figure out is how to have the medical director sign the procedure electronically. I have played around some and haven't been able to figure it out yet. Any help or suggestions is appreciated. Thank you in advanced. Becky Barnhart Waynesboro Hospital rbarnhart@summithealth.org From davenhv <@t> hotmail.com Fri Sep 5 07:26:16 2003 From: davenhv <@t> hotmail.com (Dave McClister) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] (no subject) Message-ID: Thanks for all the info on the animal processing. The incubation time in the oven has been 2 hours for the animal hearts. I have always just put them in the oven (62 C) for 25 min for surgicals, 40 min for bone marrow, and 60 min for autopsy. Can this two hour incubation time be cut down as long as no water is visible? Also if paraffin is on the slides won't the xylene/histoclear remove that? Thanks Dave McClister davenhv@hotmail.com _________________________________________________________________ Fast, faster, fastest: Upgrade to Cable or DSL today! https://broadband.msn.com From fmonson <@t> wcupa.edu Fri Sep 5 07:55:03 2003 From: fmonson <@t> wcupa.edu (Monson, Frederick ) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Manual Message-ID: Morning Rebecca, Quickly, a simple method. Medical Director approves a protocol. Assistant converts editable protocol to Adobe Acrobat PDF file that is sealed with a password known only to Medical Director and his/her supervisor. All published protocols are viewable but NOT alterable unless agreed to by Medical Director. Readers can easily print a protocol in an approved format or simply read on lone. Adobe Acrobat can be used right in Word as a "Print Option", saved, then opened and pssworded, though I suspect there is an easier way. You can also password protect a Word Document, but I bet that is not as secure as a PDF file. Cheers, Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437 -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: Friday, September 05, 2003 8:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Manual Has anyone out there in histoland put their manuals on line and done away with the paper version? This is what we are working on. We will have an icon on all computer so the manual is easily accessible. The problem I am having, I can't figure out is how to have the medical director sign the procedure electronically. I have played around some and haven't been able to figure it out yet. Any help or suggestions is appreciated. Thank you in advanced. Becky Barnhart Waynesboro Hospital rbarnhart@summithealth.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From louise_renton <@t> hotmail.com Fri Sep 5 08:25:21 2003 From: louise_renton <@t> hotmail.com (louise renton) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] tape transfer Message-ID: Hi, I've been trying out cutting sections of GMA using scotch tape to keep the bits together. What do I do now - stain them on the tape and sandwich them with mounting medium under a coverslip, or is there some funky way to remove the tape and keep the section stuck on the slide? What am I missing? Or is this jsut Friday afternoon burnout? Thanks and have a great weekend Louise Renton Bone Research Unit MRC Johannesburg South Africa Tel & fax +27 11 717 2298 "Time flies like an arrow, fruit flies like a banana" _________________________________________________________________ Unsatisfied with being single? Try MSN Personals http://www.msn.co.za/personals/ From histolog <@t> fcv.unl.edu.ar Fri Sep 5 08:41:25 2003 From: histolog <@t> fcv.unl.edu.ar (Lab Histologia) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Cross Reactivity of BioGenex Antibodies In-Reply-To: Message-ID: <004a01c373b3$67bd3820$dc0cd2aa@unl.edu.ar> I would need to know if some of you has used antibodies of Genex in other species besides human. They could indicate me which. We are to buy a panel of antibodies and we would need to have that information. Thanks ------------------------------------------------------------------- Dr. Hugo H. Ortega (DMV, PhD) Departament of Cellular Biology Faculty of Veterinary Sciences Universidad Nacional del Litoral R.P. Kreder 2805 - Esperanza (3080) Santa Fe - ARGENTINA Tel. (54)3496-420639 Fax. (54)3496-426304 http://fcv.unl.edu.ar/histolog/ http://fcv.unl.edu.ar/bioterio/ From LMARGRAF <@t> childmed.dallas.tx.us Fri Sep 5 08:48:03 2003 From: LMARGRAF <@t> childmed.dallas.tx.us (LINDA MARGRAF MD) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] IMPORTANT HISTONET SERVER INFO Message-ID: Dear Histonetters: Well, we seem to have survived the transition over to the new server despite the rocky start with the viruses and worms disabling much of the internet for a while. Thanks for hanging in there despite the inconveniences (there are still> 1500 subscribers still on the list). I wanted to explain some of the changes in the new system so you know what to expect. First, as many of you have figured out, you now use different addresses to send commands to the server and to send messages to the list. These addresses are at the bottom of all Histonet messages in case you ever forget them. To subscribe, change to digest mode, put in a vacation stoppage notification etc. you need to go to http://lists.utsouthwestern.edu/mailman/listinfo/histonet Provided your current email address is on the list, any messages you want to circulate to the list go to: histonet@lists.utsouthwestern.edu If your address have changed since you subscribed (ie. by an email system upgrade) you may still receive Histonet mail (by internal routing in your email system) but not be able to post messages. If so, subscribe with your new address and let me know to remove you old address. IMPORTANT INFO: The new server automatically removes bad addresses, disabled email addresses etc after a few attempts at mailing them. (I used to do this manually). This means I will have no record or control over when you get taken off the list by the server SO, if the Histonet mail stops arriving (and there are still messages coming into the archive website) you probably got taken off the list and need to subscribe again. Also the server won't post messages if they are being sent to more than 3 recipients. This is to block spam but may impact you if you are trying to post a message to several lists at one time. You will probably receive a "too many recipients" notice when this happens. This server should be faster and hopefully more stable than the old retired Macintosh we used to run Histonet on. Let me or Herb know if you have problems. Thanks Linda M Histonet administrator From davenhv <@t> hotmail.com Fri Sep 5 09:25:25 2003 From: davenhv <@t> hotmail.com (Dave McClister) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] animal incubation time?? Message-ID: Thanks for all the info on the animal processing. The incubation time in the oven has been 2 hours for the animal hearts. I have always just put them in the oven (62 C) for 25 min for surgicals, 40 min for bone marrow, and 60 min for autopsy. Can this two hour incubation time be cut down as long as no water is visible? Also if paraffin is on the slides won't the xylene/histoclear remove that? Dave McClister davenhv@hotmail.com _________________________________________________________________ Need more e-mail storage? Get 10MB with Hotmail Extra Storage. http://join.msn.com/?PAGE=features/es From jkiernan <@t> uwo.ca Fri Sep 5 09:38:38 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Manual References: Message-ID: <3F589FEE.FF410F37@uwo.ca> For working at the bench it's surely much easier to have the instructions on a sheet of paper, or in a book, than to have to go to a computer and read it off a screen. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ________ Rebecca Barnhart wrote: > > Has anyone out there in histoland put their manuals on line and done > away with the paper version? This is what we are working on. We will > have an icon on all computer so the manual is easily accessible. The > problem I am having, I can't figure out is how to have the medical > director sign the procedure electronically. I have played around some > and haven't been able to figure it out yet. Any help or suggestions is > appreciated. Thank you in advanced. > > Becky Barnhart > Waynesboro Hospital > rbarnhart@summithealth.org From alex.knisely <@t> kcl.ac.uk Fri Sep 5 09:55:29 2003 From: alex.knisely <@t> kcl.ac.uk (Alex Knisely) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] GGT / gamma-glutamyl transpeptidase / liver / immunohistochemistry Message-ID: <3.0.6.32.20030905155529.03da9400@137.73.66.5> Hi Netters Is anyone out there staining formalin-fixed, paraffin-embedded human liver for gamma-glutamyltranspeptidase (GGT)? We have had great results using a non-commercially available antibody, but our supply is limited and we are trying to find an alternative. We have been trying an anti-GGT1/2 antibody from Santa Cruz, cat# sc-20639, and have had reasonable success, but with less staining reaction and considerably more background staining than when using our non-commercial supply. We are not using heat mediated antigen retrieval techniques or trypsin or protease pretreatments as sections treated in this way showed no staining at all. The best staining results so far have been achieved by incubating the primary antibody overnight at 4?C, diluted 1:100 with Dako ChemMate background reducing antibody diluent, followed by detection using the ChemMate HRP kit (K5001) If Netters can point us to a different technique (with the same Santa Cruz clone) that has been successful, or to any successful technique using a commercially available clone, we shall be very grateful. Many thanks in advance Alex Knisely Alex Knisely, MD Consultant Histopathologist alex.knisely@kcl.ac.uk Institute of Liver Studies King's College Hospital Denmark Hill London SE5 9RS UK +44 (0)20 - 7346 - 3125 telefax +44 (0)20 - 7346 - 4627 office From lhadley <@t> iupui.edu Fri Sep 5 10:48:05 2003 From: lhadley <@t> iupui.edu (Baldridge, Lee Ann) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] slides Message-ID: Skipped content of type multipart/alternative-------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: image/gif Size: 1115 bytes Desc: image001.gif Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030905/a3380d6f/attachment.gif From fmonson <@t> wcupa.edu Fri Sep 5 11:33:51 2003 From: fmonson <@t> wcupa.edu (Monson, Frederick ) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Re: Manual Message-ID: Morning Rebecca, Quickly, a simple method. Medical Director approves a protocol written on an MS Word template. Assistant converts editable protocol to Adobe Acrobat PDF file that is sealed as read only (with print all only permission) with a password known only to Medical Director and his/her supervisor. All published protocols are viewable but NOT alterable unless agreed to by Medical Director. Readers can easily print a protocol in an approved format or simply read on lone. Adobe Acrobat can be used right in Word as a "Print Option", saved, then opened and pssworded, though I suspect there is an easier way. You can also password protect a Word Document, but I bet that is not as secure as a PDF file. Cheers, Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437 -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: Friday, September 05, 2003 8:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Manual Has anyone out there in histoland put their manuals on line and done away with the paper version? This is what we are working on. We will have an icon on all computer so the manual is easily accessible. The problem I am having, I can't figure out is how to have the medical director sign the procedure electronically. I have played around some and haven't been able to figure it out yet. Any help or suggestions is appreciated. Thank you in advanced. Becky Barnhart Waynesboro Hospital rbarnhart@summithealth.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mhorne <@t> upei.ca Fri Sep 5 11:37:07 2003 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] liposomes Message-ID: <3F589182.12270.F65E62@localhost> Hello Histoneters, I am about to try my hand at making liposomes. I am looking for Diplasmenylcholine but Avanti and Aldrich don't carry it. Does anyone know of another company that might carry a wide variety of lipids OR know of another plasma-stable lipid or lipid combo ? 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 huffpw <@t> uleth.ca Fri Sep 5 12:26:30 2003 From: huffpw <@t> uleth.ca (Phillip Huff) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] liposomes In-Reply-To: <3F589182.12270.F65E62@localhost> References: <3F589182.12270.F65E62@localhost> Message-ID: <1499.142.66.42.15.1062782790.squirrel@webmail.uleth.ca> You might want to contact the following company (http://www.janusbio.com/index.htm ) as they develop liposomes and may know of a source for the product you are interested in. In addition, you could search the NCBI PubMed database (http://www.ncbi.nih.gov/entrez/query.fcgi) also for the product of interest and contact researchers who have obtained such a product for their research to find out where they purchased it. Hope this helps Phil Hello Histoneters, > I am about to try my hand at making liposomes. I am looking > for Diplasmenylcholine but Avanti and Aldrich don't carry it. Does > anyone know of another company that might carry a wide variety of > lipids OR know of another plasma-stable lipid or lipid combo ? > > 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 RBARNHART <@t> summithealth.org Fri Sep 5 12:45:02 2003 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Manual question update Message-ID: I want to thank everyone for their suggestions. After posting the question I continued to play. Here is what I came up with. I scanned the pathologist signature and he saved it with a password only he know. As he reviews procedures he will place the image on a table of contents beside the procedure name. That table of contents will be saved with his password. We will have a table of contents for the entire staff to use (no signatures) but will have a link to the secure table of contents (with signature). The link just makes it easier if an inspector wants to see it (we would also have a printed copy). We didn't want to put his scanned signature on any document that the staff can get to, incase some one would get the idea to do some coping and pasting. Thanks again for any help. Becky Barnhart Waynesboro Hospital From huffpw <@t> uleth.ca Fri Sep 5 13:37:56 2003 From: huffpw <@t> uleth.ca (Phillip Huff) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] liposomes post 2 In-Reply-To: <3F589182.12270.F65E62@localhost> References: <3F589182.12270.F65E62@localhost> Message-ID: <1104.142.66.42.15.1062787076.squirrel@webmail.uleth.ca> Y. Rui & D. H. Thompson, "Efficient Stereoselective Synthesis of Plasmenylcholines" Chemistry-A European Journal 1996 2, 1505-1508 You could also try Matreya. Phil Hello Histoneters, > I am about to try my hand at making liposomes. I am looking > for Diplasmenylcholine but Avanti and Aldrich don't carry it. Does > anyone know of another company that might carry a wide variety of > lipids OR know of another plasma-stable lipid or lipid combo ? > > 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 gareth.davis <@t> Vanderbilt.Edu Fri Sep 5 14:12:05 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Re: Slides Message-ID: <082C721AF78DB34983E8BA2CD085462105C64A@mailbe07> Fisher Scientific has Gold Seal Microscope slides. Gareth Davis Vanderbilt University Medical Center Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030905/551477b4/attachment.htm From gareth.davis <@t> Vanderbilt.Edu Fri Sep 5 14:17:20 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] C-fos stain Message-ID: <082C721AF78DB34983E8BA2CD085462105C64B@mailbe07> I plan to do a immunohistochemical stain on mouse brain tissue with c-fos. Some protocols I've read state that you should osmicate the sections, after mounting, to enhance the stain. I've never stained with c-fos, but I have been told it is very dark on it's own. Does anyone know if it will be necessary to osmicate. Of course, I guess I could just try and see, but any information on the staining process itself would be helpful. Thanks, Ms. Gareth Davis Vanderbilt University Medical Center Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030905/a7dfaeb2/attachment.htm From gareth.davis <@t> Vanderbilt.Edu Fri Sep 5 14:42:12 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Re: C-fos stain Message-ID: <082C721AF78DB34983E8BA2CD085462105C64C@mailbe07> Colleen, I'm so glad to find someone doing c-fos staining. I have a couple questions, if you don't mind. First, are you staining free floating sections, and if you are, what process do you use to mount and coverslip. What would be really great is if someone could send me a protocol. Thanks! Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030905/6337a8e3/attachment.htm From gsennello <@t> osip.com Fri Sep 5 14:45:53 2003 From: gsennello <@t> osip.com (Sennello, Gina) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] C-kit on mouse Message-ID: Has anyone found a C-kit antibody that works on mouse bone marrow smears? Also, do you think it would be possible to a Wright Giemsa stain before or after the c-Kit? Thanks in advance, Gina Sennello Associate Scientist/ Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 303-546-7739 From Histolady710 <@t> aol.com Fri Sep 5 14:54:48 2003 From: Histolady710 <@t> aol.com (Histolady710@aol.com) Date: Fri Sep 16 15:21:49 2005 Subject: [Histonet] Paraffin Message-ID: <6d.18752281.2c8a4408@aol.com> Our histology laboratory has used Ribbon Pro for both infiltrating and embedding for several years and our ribbons just don't seem to be as good as in the past. It's really hard to get a good section at less than 4-5 um. What are you fellow histonetters using? Your input will be greatly appreciated. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030905/82a3526c/attachment.htm From gareth.davis <@t> Vanderbilt.Edu Fri Sep 5 18:21:41 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Re: C-fos stain Message-ID: <082C721AF78DB34983E8BA2CD085462105C64F@mailbe07> Gareth, I am not doing free floating sections, mine are cut at 5u and mounted on glass. I do a steam retrievel using a 6.0pH buffer for 30 minutes followed by a 10 minute cool down. Then I rinse well, buffer rinse for 5-10 minutes and stain. I use the c-fos from Oncogene,Cat#PC38 at 1:20,000 overnight at 4 degrees followed by Signet biotinylated secondary and streptavidin third and DAB as a chromagen. I counter stain 1 min with Haematoxylin and run down and coverslip. Hope this helps. Colleen Forster U of MN Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030905/020d89a3/attachment.htm From seri_liushaohui <@t> snec.com.sg Fri Sep 5 21:50:11 2003 From: seri_liushaohui <@t> snec.com.sg (Liu Shao Hui) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Muscarinic receptor detecting References: <6d.18752281.2c8a4408@aol.com> Message-ID: <001501c37421$9b28f000$380cac0a@snec.com.sg> Dear All, I am currently trying to detect the existence of different muscarinic receptor subtypes in human tissues using immunofluorescence. My samples are OCT imbedded. I have tried fixation with 4% paraformaldehyde, acetone, acetone:methanol, 95% ethanol, neutral buffered formalin. I have used primary antibodies from R & D and Biogenesis. So far all the antibodies light up the whole cell with no intensified staining on the plasma membrane. I wonder if any one has done stainings on muscarinic receptors or other 7 transmembrane protein receptors and like to share your success with me. Thank you very much in advance. Andea Liu Ph D student -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030906/e187b1d8/attachment.htm From jkiernan <@t> uwo.ca Fri Sep 5 23:35:28 2003 From: jkiernan <@t> uwo.ca (J. A. Kiernan) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Re: Muscarinic receptor detecting Message-ID: <200309060435.h864ZRoN011978@spork.its.uwo.ca> At 10:50 AM 9/6/03 +0800, Liu Shao Hui wrote: > ...So far all the antibodies light up the whole cell > with no intensified staining on the plasma membrane. The universal false positives indicate that you are doing something wrong in every variation of the method. Do you understand all the steps of the immunohistochemical methods you are using? Can you explain the function of every ingredient of every solution that you apply to the sections? How did you determine the dilutions of the primary and secondary antisera (or antibodies)? Your "light up" phrase suggests that you use fluorescence microscopy. This introduces many other complications, notably autofluorescence of the tissue. Your controls should determine the step that introduces the false positive immunostaining. If you are doing research (= finding out something new) you should be using at least 3 control sections for every one that might possibly contain a true new result. Have you learned the principles that determine how immunohistochemistry is done, or are you following instructions that may not be appropriate to your particular needs? Nobody should do immunohistochemical staining by following a "protocol" (horrid misused word) without knowing the reason for each step of the method. John Kiernan London, Canada. _____ From adford <@t> compuserve.com Sat Sep 6 11:02:55 2003 From: adford <@t> compuserve.com (John Difford) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Van Gieson Message-ID: <200309061203_MC3-1-4C91-6232@compuserve.com> Dear All I am trying to find a Photograph of the pathologist Dr Ira Van Gieson for an article I am writting, without success. Please can someone supply me with same? Many thanks John Difford Histopathology Department Royal Free Hospital London England, UK From Salvacion.DeLovino <@t> cshs.org Sat Sep 6 13:15:46 2003 From: Salvacion.DeLovino <@t> cshs.org (DeLovino, Salvacion S.) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] IMPORTANT HISTONET SERVER INFO Message-ID: Thanks for a job well done! You guys deserve a standing ovation for the work you do to serve the Histotechs all over the world. We really appreciate everything you're doing. Thank you again!! Salvacion S. Delovino -----Original Message----- From: LINDA MARGRAF MD [mailto:LMARGRAF@childmed.dallas.tx.us] Sent: Friday, September 05, 2003 6:48 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] IMPORTANT HISTONET SERVER INFO Dear Histonetters: Well, we seem to have survived the transition over to the new server despite the rocky start with the viruses and worms disabling much of the internet for a while. Thanks for hanging in there despite the inconveniences (there are still> 1500 subscribers still on the list). I wanted to explain some of the changes in the new system so you know what to expect. First, as many of you have figured out, you now use different addresses to send commands to the server and to send messages to the list. These addresses are at the bottom of all Histonet messages in case you ever forget them. To subscribe, change to digest mode, put in a vacation stoppage notification etc. you need to go to http://lists.utsouthwestern.edu/mailman/listinfo/histonet Provided your current email address is on the list, any messages you want to circulate to the list go to: histonet@lists.utsouthwestern.edu If your address have changed since you subscribed (ie. by an email system upgrade) you may still receive Histonet mail (by internal routing in your email system) but not be able to post messages. If so, subscribe with your new address and let me know to remove you old address. IMPORTANT INFO: The new server automatically removes bad addresses, disabled email addresses etc after a few attempts at mailing them. (I used to do this manually). This means I will have no record or control over when you get taken off the list by the server SO, if the Histonet mail stops arriving (and there are still messages coming into the archive website) you probably got taken off the list and need to subscribe again. Also the server won't post messages if they are being sent to more than 3 recipients. This is to block spam but may impact you if you are trying to post a message to several lists at one time. You will probably receive a "too many recipients" notice when this happens. This server should be faster and hopefully more stable than the old retired Macintosh we used to run Histonet on. Let me or Herb know if you have problems. Thanks Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Marysia33 <@t> aol.com Sat Sep 6 14:48:10 2003 From: Marysia33 <@t> aol.com (Marysia33@aol.com) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Tampa Florida Message-ID: <39DA5727.63FF8F85.0260E07C@aol.com> I was wondering if anyone knows of any job openings in the Tampa Florida area. If so i would appreciate it if you would pass it along. Thank you, Marysia From lloyd1223 <@t> yahoo.com Sat Sep 6 15:42:16 2003 From: lloyd1223 <@t> yahoo.com (mark lloyd) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Washignton, dc Message-ID: <20030906204216.90690.qmail@web11006.mail.yahoo.com> I am a histotechnologist with 3 years of experience. I have extensive knowledge of histology including special stains, project management, Tissue Microarray, and Laser Capture Microscopy. If any of you know of a job opening in the Washington, D.C. area, I appreciate it if you would pass them on to me. Thank you all for your time, Mark Lloyd (703) 575- 7876 __________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com From djcarter <@t> dallastx.net Sat Sep 6 19:38:25 2003 From: djcarter <@t> dallastx.net (Dallas Nippert) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Tampa Florida References: <39DA5727.63FF8F85.0260E07C@aol.com> Message-ID: <001001c374d8$5cdc3840$20e8a641@djcarter> I thought I saw some on histologyjobs.com but I am not positive. Dallas Nippert From lboyd <@t> parabolagroup.com Sun Sep 7 10:16:43 2003 From: lboyd <@t> parabolagroup.com (Larry D. Boyd) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] (no subject) Message-ID: <1511.67.75.201.93.1062947803.squirrel@webmail.globalwizards.com> Hi all, Back after short period off group. I am seeking the services of a histologist to help set up a small histo lab at our Arlington WA location. Happy to pay a consulting fee and long term colaberation is possible. This could be great for a simi retired histo tech or someone generally home raising a family. We are located about 15 miles north of Everett, WA or 40 miles north of Seattle. Thanks! Larry D. Boyd Parabola Laboratory Services Research Institute for Exotic Species Microbiology 307 North Olympic Ave, Ste. 209 Arlington, WA 98223 From Linresearch <@t> aol.com Sun Sep 7 14:18:03 2003 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] C-fos Message-ID: <12f.3119bd6a.2c8cde6b@aol.com> Hello, Is there anyone doing C-fos staining on 40microns free floating sections? Could you share your protocol and let me know what counterstain if any you may be using? Lin -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030907/63d2ded6/attachment.htm From SJones <@t> cvm.tamu.edu Sun Sep 7 17:50:06 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] animal incubation time?? Message-ID: Hi Dave, I think we need more info before we can answer this question. Are you processing without an automated tissue processor? I assume you are talking about paraffin infiltration when you refer to incubating time in an oven. Is that correct? We would need to know how big the tissue is you are infiltrating to know if the times are correct. Can you tell us what the species and dimensions are? And I have no idea what you mean by "Also if paraffin is on the slides won't the xylene/histoclear remove that?" What is that refering to exactly? One thing I can tell you is infiltration is not one of the steps I like to shorten. Are you using several stations of paraffin during this 2 hour period? Thanks, Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Dave McClister" 09/05/03 09:25AM >>> Thanks for all the info on the animal processing. The incubation time in the oven has been 2 hours for the animal hearts. I have always just put them in the oven (62 C) for 25 min for surgicals, 40 min for bone marrow, and 60 min for autopsy. Can this two hour incubation time be cut down as long as no water is visible? Also if paraffin is on the slides won't the xylene/histoclear remove that? Dave McClister davenhv@hotmail.com _________________________________________________________________ Need more e-mail storage? Get 10MB with Hotmail Extra Storage. http://join.msn.com/?PAGE=features/es _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stanley.Stylli <@t> mh.org.au Sun Sep 7 19:49:24 2003 From: Stanley.Stylli <@t> mh.org.au (Stylli, Stanley) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] PECAM antibody Message-ID: <00519F9F41D2844D9C75BEE1F6AC09AF68B941@rmhmail1.ssg.org.au> Hi everyone Has anyone tried antigen retrieval from rat FFPE tissue of the Pharmigen PECAM antibody 550310 Rat CD31 Pure MAB (TLD-3A12) ? If so, which antigen retrieval solution / protocol worked best for you ? Thankyou in advance Stan Stylli Stan Stylli Department of Surgery, 5th Floor Clinical Sciences Building Royal Melbourne Hospital University of Melbourne Parkville, Australia, 3052. Tel: 61-3-93427616 Fax:61-3-9347-7695 -------------- next part -------------- A non-text attachment was scrubbed... Name: Stylli, Stanley.vcf Type: text/x-vcard Size: 230 bytes Desc: Stylli, Stanley.vcf Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030908/27aa855b/StylliStanley.vcf From nyilmaz <@t> mersin.edu.tr Mon Sep 8 03:55:07 2003 From: nyilmaz <@t> mersin.edu.tr (=?iso-8859-9?Q?Nejat_Y=FDlmaz?=) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] (no subject) Message-ID: <000801c375e6$e83a2b40$4f01a8c0@mersin.edu.tr> Hi histonetters, Is there anybody who worked with rhodizonate method for the demonstration of Pb (lead) ? and can you give the protocol and please tell the experiences about it ? Thanks in advance, -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030908/38e76ed0/attachment.htm From frouwke <@t> sci.kun.nl Mon Sep 8 03:33:05 2003 From: frouwke <@t> sci.kun.nl (Frouwke Kuijpers) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Cryoprotection Message-ID: <00c201c375e7$12a05b20$4a8aae83@sci.kun.nl> Has anybody has experience with this Cryoprotect-solution : 0,05 Sodium phosphate buffer 30% ethylen glycol 20% glycerol for storing 40 micron sections at - 20 C Has somebody experiences with this? We don't want to put them on glasses because we do FFM. F.J.Kuijpers-Kwant Dept. Cellular Animal Physiology University of Nijmegen Toernooiveld 1 6525 ED Nijmegen frouwke@sci.kun.nl From c.m.vanderloos <@t> amc.uva.nl Mon Sep 8 04:07:29 2003 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] RE: IHC with CD25 on FFPE lymph node Message-ID: Dear Philippe, The anti-CD25 Cymbus product CBL151 you indicated is in fact clone Tu69. This clone is unfortunately not working on FFPE sections. In other words, this particular anti-CD25 clone is reacting with an epitope on the CD25 molecule that is not surviving the FFPE procedure. Unfortunately, I am not aware of any anti-CD25 clones working with FFPE sections. Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands >From "Philippe Gannon" >Date Thu, 04 Sep 2003 17:32:25 +0000 >To HistoNet@pathology.swmed.edu >Subject [Histonet] IHC with CD25 on FFPE lymph node > >hi, > >I am having some difficulties with a CD25 staining on formalin-fixed >paraffin embedded lymph node slides. In fact, I am not seeing any >staining at all with the antibody CBL 151 from Cymbus Biotechnology >LTD/Chemicon at concentration up to 1:10 in PBS. > >Laboratoire Mes-Masson (Y-4609) >Institut du cancer de Montr?al, Universit? de Montr?al >2099 Alexandre de S?ve >Montr?al, Qu?bec, Canada >H2L 2W5 From nick.kirk3 <@t> btopenworld.com Mon Sep 8 04:20:22 2003 From: nick.kirk3 <@t> btopenworld.com (nick.kirk3@btopenworld.com) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] RE: IHC with CD25 on FFPE lymph node Message-ID: <7929897.1063012822968.JavaMail.root@127.0.0.1> Philippe Novocastra produce a CD25 that survives paraffin processing. It's clone number 4C9. It requires Citrate buffer heat mediated antigen retrieval and its code no is NCL-CD25-305 Online catalogue sheet is at http://www.novocastra.co.uk/data/gfr/cd25-305.pdf The Canadian distributor is Vector labs Nick Kirk Histopathology Hinchingbrooke Hiospital Huntingdon England > from: "C.M. van der Loos" > date: Mon, 08 Sep 2003 10:07:29 > to: histonet@lists.utsouthwestern.edu > cc: philippegannon@hotmail.com > subject: Re: [Histonet] RE: IHC with CD25 on FFPE lymph node > > Dear Philippe, > The anti-CD25 Cymbus product CBL151 you indicated is in fact clone > Tu69. This clone is unfortunately not working on FFPE sections. In > other words, this particular anti-CD25 clone is reacting with an > epitope on the CD25 molecule that is not surviving the FFPE procedure. > Unfortunately, I am not aware of any anti-CD25 clones working with FFPE > sections. > > Chris van der Loos > Dept. of Cardiovascular Pathology > Academical Medical Center > Amsterdam - The Netherlands > > >From "Philippe Gannon" > >Date Thu, 04 Sep 2003 17:32:25 0000 > >To HistoNet@pathology.swmed.edu > >Subject [Histonet] IHC with CD25 on FFPE lymph node > > > >hi, > > > >I am having some difficulties with a CD25 staining on formalin-fixed > >paraffin embedded lymph node slides. In fact, I am not seeing any > >staining at all with the antibody CBL 151 from Cymbus Biotechnology > >LTD/Chemicon at concentration up to 1:10 in PBS. > > > >Laboratoire Mes-Masson (Y-4609) > >Institut du cancer de Montr?al, Universit? de Montr?al > >2099 Alexandre de S?ve > >Montr?al, Qu?bec, Canada > >H2L 2W5 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From philip.bergin <@t> microbio.gu.se Mon Sep 8 00:49:03 2003 From: philip.bergin <@t> microbio.gu.se (Phil Bergin) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Gerbils Message-ID: <000001c375cc$ea2d6e90$a660f182@philb> Hi! We have recently started using Mongolian gerbils as a model- does anyone know a vendor making antibodies for use in this animal. I believe the researcher is mainly interested in defining the immune response (i.e. Th1 vs Th2), and she would rather use anti gerbil antibodies then start looking for reactivity in other antibodies, Thanks for the help, Phil ----------------------------------------------------------------- Philip Bergin G?teborg University Department of Medical Microbiology and Immunology Box 435, SE-405 30 G?teborg, Sweden Phone: +46-31-3424471 Fax: +46-31-826976 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030908/58c30b86/attachment.htm From jmitchell <@t> neurology.wisc.edu Mon Sep 8 09:03:47 2003 From: jmitchell <@t> neurology.wisc.edu (Mitchell (Jean A.)) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Cryoprotection Message-ID: <72E391EA29BCD341B0B38A2C5473524B051B44@nrl-lemur.neurology.wisc.edu> I have been using a cryoprotectant solution very similar to yours for storing 50 micron frozen sections of skin at -20C. I use 96-well clear assay plates and the cryoprotectant solution is pipetted into each well. The 50 micron frozen sections are placed directly into the appropriate well and are stored at -20C until I can begin staining procedures. If you have further questions or would like reference articles on this procedure/solution - feel free to contact me. Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI -----Original Message----- From: Frouwke Kuijpers [mailto:frouwke@sci.kun.nl] Sent: Monday, September 08, 2003 3:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cryoprotection Has anybody has experience with this Cryoprotect-solution : 0,05 Sodium phosphate buffer 30% ethylen glycol 20% glycerol for storing 40 micron sections at - 20 C Has somebody experiences with this? We don't want to put them on glasses because we do FFM. F.J.Kuijpers-Kwant Dept. Cellular Animal Physiology University of Nijmegen Toernooiveld 1 6525 ED Nijmegen frouwke@sci.kun.nl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Mon Sep 8 09:45:29 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Cryoprotection References: <00c201c375e7$12a05b20$4a8aae83@sci.kun.nl> Message-ID: <3F5C9609.157E4D51@uwo.ca> The solution you describe is not a cryoprotectant. It is an anti-freeze. A cryoprotectant is used before freezing and cutting sections, to reduce the sizes of the ice crystals in the tissue. 30% sucrose in water is a commonly used cryoprotectant for blocks of fixed tissue. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ___________________________________ Frouwke Kuijpers wrote: > > Has anybody has experience with this Cryoprotect-solution : > 0,05 Sodium phosphate buffer > 30% ethylen glycol > 20% glycerol > for storing 40 micron sections at - 20 C > > Has somebody experiences with this? > We don't want to put them on glasses because we do FFM. > > F.J.Kuijpers-Kwant > Dept. Cellular Animal Physiology > University of Nijmegen > Toernooiveld 1 > 6525 ED Nijmegen > frouwke@sci.kun.nl > > _______________________________________________ From tvedilago <@t> system1.net Mon Sep 8 10:39:22 2003 From: tvedilago <@t> system1.net (Tommy Vedilago) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Histotechs and Managers Wanted Message-ID: Hello histonetters, I noticed some postings over the weekend of people looking for jobs. I am always looking for qualified Histotechs and Managers for several locations around the country. Currently, I have positions in Atlanta (nights), Pittsburgh (mgr), Philadelphia, Michigan (nights), Milwaukee (mgr), Nashville, Tampa (eve supv), Orlando (IHC), Washington D.C. (mgr), San Antonio, Baltimore (mgr and techs), Indiana (lead tech), and 3 day shift tech positions in Dallas. Thank you so much for all of the responses to this point, we are growing in Histology very rapidly and I appreciate all of the help. I am happy to be of assistance to as many of you as possible so, please feel free to give me a call at 866-SYSTEM1 or 866-797-8361. Tommy Vedilago System 1 Search (864) 627-0012 (864) 627-0013 Fax From stevemachinuk <@t> yahoo.co.uk Mon Sep 8 11:04:13 2003 From: stevemachinuk <@t> yahoo.co.uk (=?iso-8859-1?q?Steve=20Machin=20UK?=) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] CD1a Message-ID: <20030908160413.96764.qmail@web12407.mail.yahoo.com> We are having trouble with our CD1a from DAKO. We use microwave for 20 mins and get background and very weak staining. What method do others use? Best Wishes Steve Machin UK ________________________________________________________________________ Want to chat instantly with your online friends? Get the FREE Yahoo! Messenger http://mail.messenger.yahoo.co.uk From tpmorken <@t> labvision.com Mon Sep 8 11:20:39 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Dana Dittus? Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CB3B@usca0082k03.rallansci.apogent.com> Dana, please reply to this email. thanks Tim Morken Lab Vision / NeoMarkers www.labvision.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030908/34e58dfd/attachment.htm From jmitchell <@t> neurology.wisc.edu Mon Sep 8 11:20:46 2003 From: jmitchell <@t> neurology.wisc.edu (Mitchell (Jean A.)) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Cryoprotection Message-ID: <72E391EA29BCD341B0B38A2C5473524B052209@nrl-lemur.neurology.wisc.edu> I should have also made that distinction in my reply that the solution in question is an "antifreeze solution" not a "cyroprotectant solution". Jean Mitchell University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI -----Original Message----- From: John Kiernan [mailto:jkiernan@uwo.ca] Sent: Monday, September 08, 2003 9:45 AM To: Frouwke Kuijpers Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Cryoprotection The solution you describe is not a cryoprotectant. It is an anti-freeze. A cryoprotectant is used before freezing and cutting sections, to reduce the sizes of the ice crystals in the tissue. 30% sucrose in water is a commonly used cryoprotectant for blocks of fixed tissue. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ___________________________________ Frouwke Kuijpers wrote: > > Has anybody has experience with this Cryoprotect-solution : 0,05 > Sodium phosphate buffer 30% ethylen glycol > 20% glycerol > for storing 40 micron sections at - 20 C > > Has somebody experiences with this? > We don't want to put them on glasses because we do FFM. > > F.J.Kuijpers-Kwant > Dept. Cellular Animal Physiology > University of Nijmegen > Toernooiveld 1 > 6525 ED Nijmegen > frouwke@sci.kun.nl > > _______________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rfail <@t> toolkitmail.com Mon Sep 8 11:22:35 2003 From: rfail <@t> toolkitmail.com (rfail@toolkitmail.com) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] CD1a Message-ID: <01L0F72LLTZG98BMFI@SMTP00.InfoAve.Net> Steve, We use DAKO's CD1a, HIER ph6.0 in a steamer at 99-100 degrees for 20 minutes, cool down for 10 minutes, rinse in TRIS with tween five minutes, DAKO autostainer with envision +, 30 minute incubation with Ab, 1:400 dilution. We use protein block on everything. Rena Fail We are having trouble with our CD1a from DAKO. > We use microwave for 20 mins and get background and very weak > staining. > What method do others use? > > Best Wishes > > Steve Machin UK > > ________________________________________________________________________ > Want to chat instantly with your online friends? Get the FREE Yahoo! > Messenger http://mail.messenger.yahoo.co.uk > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GDawson <@t> Milw.Dynacare.com Mon Sep 8 11:48:17 2003 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] IHC - CD25 Message-ID: Phillipe, You may want to consider trying either a different vendor or a different antibody lot # at this point. Once you get up to a 1:10 dilution, you should be getting at least some staining somewhere... Email me directly and I'll give you some suggestions that I could catch heat for posting to the general list. gdawson@milw.dynacare.com Good Luck, Glen Dawson BS, HT & QIHC (ASCP) Lead IHC Technologist Milwaukee, WI From g.lang <@t> bigfoot.de Mon Sep 8 12:12:21 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bouin Message-ID: <003f01c3762c$5db46d50$0d12a8c0@SERVER> Hi! Can anyone tell me, how long bouin's fixative can be stored? For staining we mix it always fresh, but it would be easier to have a bigger amount. greetings from Austria Gundi -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030908/163cf942/attachment.htm From kb2drkprk <@t> yahoo.com Mon Sep 8 13:40:13 2003 From: kb2drkprk <@t> yahoo.com (Kelly Booher) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bouin In-Reply-To: <003f01c3762c$5db46d50$0d12a8c0@SERVER> Message-ID: <20030908184013.49765.qmail@web20913.mail.yahoo.com> Frieda Carson (author of Histotechnology: A Self-Instructional Text) suggests a 2 month shelf-life for Bouin's solution. In our lab, we make a stock bottle of Bouin's solution up to 6 months in advance. The coplin jar of Bouin's that we use for Masson's trichrome is replaced monthly from the stock bottle. Kelly Booher, BS, HTL (ASCP) Anatomic Pathology Swedish Medical Center - Providence Campus Seattle, WA 98122 --- Gudrun Lang wrote: > Hi! > Can anyone tell me, how long bouin's fixative can be > stored? > For staining we mix it always fresh, but it would be > easier to have a bigger amount. > > greetings from Austria > Gundi > __________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com From vbaker60 <@t> yahoo.com Mon Sep 8 13:40:24 2003 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Tunel ISH on the Ventana Discovery System Message-ID: <20030908184024.92690.qmail@web12106.mail.yahoo.com> Hi! I was wondering if any Ventana Discovery users are currently doing Tunel by ISH on their units. Since they do not put out their own detection kit I've been trying to adapt our Enzo Kit with not a lot of success. Any assistance would be greatly appreciated. Sincerely Vikki Baker Institute for Cancer Prevention Valhalla, New York __________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com From g.lang <@t> bigfoot.de Mon Sep 8 14:47:16 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] histonetlist in german Message-ID: <00a901c37642$034f17a0$0d12a8c0@SERVER> Hi to all german speaking people in Histoland! Do you know something about a similar mailing list in german? Wer wei? etwas ?ber eine ?hnliche mailing liste zum Austausch ?ber Histofragen im deutschsprachigen Internet? greetings from Austria Gundi -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030908/06090cac/attachment.htm From GalbraithJ <@t> uihc.uiowa.edu Mon Sep 8 15:04:04 2003 From: GalbraithJ <@t> uihc.uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] CD1a Message-ID: <5D03ED7B9391D4119D9B0008C76B7B24030085CD@uihc-mail1.uihc.uiowa.edu> Steve: We use CD1A from Immunotech on a Dako Autostainer (15-15 program), dilution neat (ouch) with full power (600W) microwave hier for 8 minutes with a 15 min cool down then into Dako wash solution for 5 minutes prior to loading on the instrument. We get good results without background. Good luck, Joe Galbraith -----Original Message----- From: Steve Machin UK [mailto:stevemachinuk@yahoo.co.uk] Sent: Monday, September 08, 2003 11:04 AM To: histonet@pathology.swmed.edu Subject: [Histonet] CD1a We are having trouble with our CD1a from DAKO. We use microwave for 20 mins and get background and very weak staining. What method do others use? Best Wishes Steve Machin UK ________________________________________________________________________ Want to chat instantly with your online friends? Get the FREE Yahoo! Messenger http://mail.messenger.yahoo.co.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Allison_Scott <@t> hchd.tmc.edu Mon Sep 8 15:42:04 2003 From: Allison_Scott <@t> hchd.tmc.edu (Scott, Allison D) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] wooden slide boards Message-ID: Hello to everyone in histoland. Does anyone know of a company that makes the 5 position wooden slide board. I used to get them from a company called Night Owl Industries located near th dallas area. It went out of business or something. Any help would be appreciated. Allison Scott HT(ASCP) Histology Supervisor 5656 Kelley Houston, Texas 77026 From jkiernan <@t> uwo.ca Mon Sep 8 15:43:35 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bouin References: <003f01c3762c$5db46d50$0d12a8c0@SERVER> Message-ID: <3F5CE9F7.38DCB1C@uwo.ca> Certainly for 10 years, on the shelf. Probably for much longer. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ _________________________________________ > Gudrun Lang wrote: > Can anyone tell me, how long bouin's fixative can be > stored? From David.Muskett <@t> btopenworld.com Mon Sep 8 16:28:32 2003 From: David.Muskett <@t> btopenworld.com (David Muskett) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Hard Tissues Message-ID: Skipped content of type multipart/alternative-------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: image/gif Size: 530 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030908/b0c01c85/attachment.gif -------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: image/gif Size: 530 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030908/b0c01c85/attachment-0001.gif From doscwk <@t> nus.edu.sg Mon Sep 8 20:12:09 2003 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Hard Tissues Message-ID: Hi David, You did not specify what the problem was or the type of bone you're doing but since you mention clearing, our lab uses toluene for clearing most bones overnight except for fibrous bones when we'll use chloroform. Hope that helps. Julee Chan Experimental Surgery Dept. of Orthopaedic Surgery National University of Singapore -----Original Message----- From: David Muskett [mailto:David.Muskett@btopenworld.com] Sent: Tuesday, September 09, 2003 5:29 AM To: histonet Subject: [Histonet] Hard Tissues Dear Histonetters I am looking for some advice and comments as to the best way of processing hard tissues. We are currently experiencing some problems with our bone tissues. We are considering clearing in oil of cedar wood. Does anybody have any experience of this or procedures to share? I have read in the histonet archive of people softening tissues in fabric conditioners. Does anyone have any procedures to share for this method? Regards David Muskett Liverpool, UK From SJones <@t> cvm.tamu.edu Mon Sep 8 20:19:43 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Hard Tissues Message-ID: Hi David, Is the problem that the bone is too hard or not clearing well? I have used cedarwood oil for brain tissues. It penetrates very well. You can't use it on an automated tissue processor because it separates and becomes like petroleum jelly. So you have to do that step by hand. I can't remember using it on bone. If the bone is too hard, maybe it isn't decalcified enough. What bone is it? How big is the specimen? What knives are you using? How are you decaling it? How is it being trimmed in and fixed? I prefer the RDO decal, it's fast and can handle anything. Hope this helps. Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "David Muskett" 09/08/03 04:28PM >>> Dear Histonetters I am looking for some advice and comments as to the best way of processing hard tissues. We are currently experiencing some problems with our bone tissues. We are considering clearing in oil of cedar wood. Does anybody have any experience of this or procedures to share? I have read in the histonet archive of people softening tissues in fabric conditioners. Does anyone have any procedures to share for this method? Regards David Muskett Liverpool, UK From SJones <@t> cvm.tamu.edu Mon Sep 8 22:23:33 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] parched earth effect Message-ID: Luna's book states that under processing can cause the parched earth effect. Will under fixation also cause this artifact? Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 From rowani.mohdrawi <@t> student.adelaide.edu.au Mon Sep 8 23:54:07 2003 From: rowani.mohdrawi <@t> student.adelaide.edu.au (Rowani) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] looking for atlas of the prenatal mouse brain by Uta Schumbra References: Message-ID: <000c01c3768e$66ec1f50$87997f81@LaptopRowani> Hi I really need to have a copy of atlas of the prenatal mouse brain by Uta B. Schumbra. The book is out of print and very difficult to find. Let me know if anyone have a used book to sell or dispose. Regard Rowani Mohd Rawi PhD student Lysosomal Diseases Research Unit Department of Chemical Pathology Women's and Children's Hospital 72 King William Road, 5006 North Adelaide, South Australia From arme <@t> optonline.net Mon Sep 8 23:55:36 2003 From: arme <@t> optonline.net (Mark Sofferman) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] For Sale - Grossing Station and VIP 3000 In-Reply-To: <20030908170000.422.58069.Mailman@swlx167.swmed.edu> Message-ID: Email or call 201-833-1550. ARME From jkiernan <@t> uwo.ca Tue Sep 9 00:56:27 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Hard Tissues References: Message-ID: <3F5D6B8B.E055FC72@uwo.ca> Dear David Muskett Your message to Histonet contained unusual graphic decorations. My email program (Netscape, updated July 2003) could not quote your lines in a reply to the group. I typed several lines of a reply before finding that your question was unanswerable for technical reasons. Please send your emails as plain text. You should not assume that all histonetters are equipped with the latest from Silicon Valley. If your email was not readable by me (in Canada), how was it received in Bangladesh, Botswana, Papua New Guinea etc? I'll be happy to send a reply to your question, but only in a completely open forum. For an internet response it is necessary to be able to include quotations from earlier messages. There is no place for clever decorative stuff on a listserver that deals with simple questions and answers. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 From jqb7 <@t> cdc.gov Mon Sep 8 12:23:23 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bouin Message-ID: We buy ours already made and it has an expiration date on the bottle of about a year. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: Gudrun Lang [mailto:g.lang@bigfoot.de] Sent: Monday, September 08, 2003 1:12 PM To: Histonetliste Subject: [Histonet] bouin Hi! Can anyone tell me, how long bouin's fixative can be stored? For staining we mix it always fresh, but it would be easier to have a bigger amount. greetings from Austria Gundi -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030908/e877a8e6/attachment.htm From jluis.palazon <@t> icman.csic.es Tue Sep 9 03:26:31 2003 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] sudan black B Message-ID: <20030909082631.A794A47004@perceval.uca.es> Dear List members. I posted this question some weeks before but had no answer. As part of my thesis I have to study lipids. I stained some slides with sudan black B. I used 4 slides per specimen and I did (delipidized, sudan black B, Bromide-sudan black, and bromide-acetone-sudan black). I have noticed that the slides were clearly black after staining, the bromide treated the most black, but after 1-2 weeks, they tured green in color and difusing in the borders of the tissue. I used an aqueous mounting media. Have any of you working with lipids had a similar situation? Is the color of sudan black B stained slides not permanent? must the slides be read inmediately? do this change in color alter the results if I read some recently stained slides and other stained some weeks ago?. Is it possible (adequate) to put the coverlips out and re-stain the slides? I would appreciate some advice from you. Thanks in advance José Luis From g.lang <@t> bigfoot.de Tue Sep 9 07:10:25 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bouin Message-ID: <003501c376cb$5b032fc0$0d12a8c0@SERVER> Thank you, for all your answers. Histotechs use the solution between one month and "ever" and all have good experience. greetings from Austria Gundi -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/f7fc6ddd/attachment.htm From g.lang <@t> bigfoot.de Tue Sep 9 07:14:48 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] CAB-stain Message-ID: <00bb01c376cb$f6b40f20$0d12a8c0@SERVER> Hi! Does anyone know a reference for the CAB-stain (a variant of Gomori's Trichrome onestep method). CAB = Chromotrop Anilinblue, for staining collagen fibers Gundi -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/6d2ffc28/attachment.htm From Lynne.Bell <@t> hitchcock.org Tue Sep 9 08:42:07 2003 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Containers with wells for multiple specimens Message-ID: Good Morning All, I am looking for a type of specimen container that has multiple wells in it that can be filled with a fixative. Our urologists will be starting to do 11 biopsies on each patient and they would like a container such as the one I just described. My pathologist thinks that such a thing exists. I am aware of the cassettes that have six partitions for biopsies, but not of a specimen container. I appreciate any help you can give me! Thanks, Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From Jenny-Oblander <@t> omrf.ouhsc.edu Tue Sep 9 08:54:22 2003 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Hard Tissues Message-ID: Skipped content of type multipart/alternative-------------- next part -------------- A non-text attachment was scrubbed... Name: image001.gif Type: image/gif Size: 530 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/acadc39c/image001.gif From funderwood <@t> mcohio.org Tue Sep 9 09:42:34 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Paraffin Message-ID: I've used surgi-path for years and think it's great. Fred >>> 09/05/03 03:54PM >>> Our histology laboratory has used Ribbon Pro for both infiltrating and embedding for several years and our ribbons just don't seem to be as good as in the past. It's really hard to get a good section at less than 4-5 um. What are you fellow histonetters using? Your input will be greatly appreciated. From Jmrwalker <@t> aol.com Tue Sep 9 09:49:14 2003 From: Jmrwalker <@t> aol.com (Jmrwalker@aol.com) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] cleaning microtomes Message-ID: <1f0.f76bf14.2c8f426a@aol.com> I have a question that may seem silly. After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. Thanks! From jkiernan <@t> uwo.ca Tue Sep 9 09:56:35 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] [Fwd: Sorry (cedarwood oil)] Message-ID: <3F5DEA23.CBC64A82@uwo.ca> John Kiernan wrote: > > Manfred Gabe ("Histological Techniques." 1976; an > excellent big fat book) extolled the virtues of > cedarwood oil. In particular, he claimed that > it was essential when thin paraffin sections were > to be stained by classical techniques for > mitochondria, Golgi apparatus etc following > classical cytological fixatives (mixtures > containing K2Cr2O7 and OsO4). > > I've used it for decalcified rodent heads, which > can be hard in parts, and found it better than > less viscous solvents such as chloroform, xylene > etc. I think the softening action is probably > due to retention of some of the oil in the > tissue despite thorough rinsing in a thinner > solvent and several changes of wax. You can smell > the cedarwood oil while cutting the block. > > I'm not convinced that cedarwood oil is any > better than terpineol, which is less viscous and > also cheaper. Another very effective clearing > agent for large hard objects is Gabe's phenolic > benzene. This is used in double-embedding routines > after the celloidin and before the paraffin. For > biggish decalcified objects containing tissues > of varying hardness, double embedding is the > best method I know. The phenol in the clearing > agent largely prevents later extraction of the > celloidin. It may also have a softening action; > it's certainly present (smell) in the final block. > > I've no experience of fabric softener as an aid > to histological technique. If you search the > archives at http://www.histosearch.com you will > probably find some anecdotal accounts of its > use. > -- > ------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ > ________________________________________ > > Muskett David wrote: > > > > Dear John > > > > Sorry that my e-mail had some colour on the > > background. I had attempted to send it as plain text! > > > > My Questions were > > > > I am having some difficulty with processing hard > > tissues. Would the use of cedar wood oil as the > > clearing agent be of use? Or would the use of fabric > > conditioner be helpful? > > > > I am looking for protocols and methods > > > > Hope you can help. > > > > Thanks > > > > David > > > > David Muskett > > Histology > > Royal Liverpool Children's NHS Trust - Alder Hey > > Eaton Road > > Liverpool > > L12 2AP > _______________________ -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ From GauchV <@t> mail.amc.edu Tue Sep 9 10:05:55 2003 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] cleaning microtomes Message-ID: We use Paragard but it leaves such an oily residue that we also use a little alcohol afterwards... Vicki Gauch AMCH Pathology Albany, NY >>> 9/9/2003 10:49:14 AM >>> I have a question that may seem silly. After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Tue Sep 9 10:23:16 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] cleaning microtomes Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CB42@usca0082k03.rallansci.apogent.com> Personally I never liked using solvents on the microtome. I simply rub off the paraffin with a gauze pad - they seem abrasive enough to do that, but not to damage the finish. I do clean up a lot as I go along, so there is never much buildup in the first place. Tim Morken -----Original Message----- From: Jmrwalker@aol.com [mailto:Jmrwalker@aol.com] Sent: Tuesday, September 09, 2003 7:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cleaning microtomes I have a question that may seem silly. After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfavara <@t> niaid.nih.gov Tue Sep 9 10:31:37 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] cleaning microtomes Message-ID: I use paragard which is just oil of wintergreen, followed by ETOH. Do not use xylene because of health concerns. I would be interested in what Gayle Callis has to say, I believe she took a course at NSH previously and the instructor recommended not using any clearing agent. Would be interesting to have a manufacturers recommendation. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Jmrwalker@aol.com [mailto:Jmrwalker@aol.com] Sent: Tuesday, September 09, 2003 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cleaning microtomes I have a question that may seem silly. After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From suryawcn <@t> email.uc.edu Tue Sep 9 10:55:50 2003 From: suryawcn <@t> email.uc.edu (suryawcn@email.uc.edu) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] (no subject) Message-ID: <114780-22003929155550240@M2W087.mail2web.com> Does any one know the source for SQUARE cover glass slips of 40mm. For my experiment, the glass material should NOT be borosilicate. As far as I know most companies make cover slips of borosilicate glass. Soda lime should be fine. For my experiment, the material should be glass, not contain boron and the cover slip be thin. Do any companies make microscope slides of 40mm square size? AS long as the slide is not thicker than 2mm. Information would be greatly appreciated. Thanks Chetan -------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ . From lesley <@t> vancouverbc.net Tue Sep 9 11:08:03 2003 From: lesley <@t> vancouverbc.net (Lesley Weston) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Hard Tissues In-Reply-To: Message-ID: I used to use cedarwood oil for clearing hard tissues such as bone. It's very effective, but also very slow. You have to float the tissue on the oil, after dehydration of course, and wait for it to sink to the bottom of the container, which can take days. But when all else has failed, it does make a real difference. Hope this helps. Lesley Weston. on 08/09/2003 2:28 PM, David Muskett at David.Muskett@btopenworld.com wrote: > > Dear Histonetters > > I am looking for some advice and comments as to the best way of processing > hard tissues. We are currently experiencing some problems with our bone > tissues. > > We are considering clearing in oil of cedar wood. Does anybody have any > experience of this or procedures to share? > > I have read in the histonet archive of people softening tissues in fabric > conditioners. Does anyone have any procedures to share for this method? > > Regards > > David Muskett > > Liverpool, UK > From rstapf <@t> adventisthealthcare.com Tue Sep 9 11:30:12 2003 From: rstapf <@t> adventisthealthcare.com (Ross Stapf) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] cleaning microtomes Message-ID: We use Paragard to clean our Microtomes. Ross Stapf Histology Supervisor Washington Adventist Hospital Takoma Park MD -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/10c4cd5e/attachment.htm From LElby <@t> hsh.org Tue Sep 9 09:17:12 2003 From: LElby <@t> hsh.org (Elby, Lynette) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] controls Message-ID: <86EF8BDFC3E8D411BE6500508B2E176C02CFFF4C@hshmail.hsh.org> Does anyone out there have any extra mesothelioma controls and H. Pylori control blocks the we can have. Or maybe we can do a swap for some controls that you might need. Thanks in advance...... LYN Lynette Elby HT(ASCP) Histology Lead Tech. Holy Spirit Hospital 503 N. 21st Street Camphill, PA 17011 (717)763-2930 office (717)763-2947 fax Confidentiality Disclaimer: The information contained in this communication may be confidential, is intended for the use of the recipient named above, and may be legally privileged. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication, or any of its contents, is strictly prohibited. If you received this communication in error, please resend this communication to the sender and delete the original message and any copy of it from your computer system. Thank You -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/5b1d9c41/attachment.htm From RCHIOVETTI <@t> aol.com Tue Sep 9 13:15:18 2003 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] cleaning microtomes Message-ID: <7a.4744d677.2c8f72b6@aol.com> In a message dated 09/09/2003 7:51:39 AM US Mountain Standard Time, Jmrwalker@aol.com writes: << After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. Thanks! >> I see mostly Paragard being used in our customers' labs, and I agree with the other posters, if the "slick" surface is a problem after the Paragard treatment you can then wipe down with a little ethanol or isopropanol. With time xylene can affect the surfaces of the microtome's external parts, especially if used in excess. Lots of the microtome parts have anodized (plated) surfaces that are either black or clear, and you'd think they would hold up to just about any kind of abuse. But they will definitely break down and corrode with prolonged or repeated exposure to xylene. This can lead to pitting and discoloration of the surfaces. If you *do* use xylene, use it sparingly and be sure to wear gloves. I personally don't like the stuff. Try to use a substitute instead. First, use a brush or gauze pad to remove the majority of the paraffin. Then use only enough xylene or xylene substitute to barely moisten a gauze 4x4 or a tissue. Wipe off the excess immediately with another dry gauze or tissue. You can also follow this with an alcohol wipe-down like you would use with Paragard if the surface feels slick or oily after you clean it. Hope this helps! Bob Chiovetti GTI Microsystems Leica Regional Dealer Desert Southwest Region, USA From JosefaNava <@t> texashealth.org Tue Sep 9 13:30:04 2003 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] effects of decalcifying reagents on probes Message-ID: <3B1A15523787D71197170000E867C0A20120AF81@PHDEX04> Hello to all, Does anybody know if decalcifying reagents affect the tissue(bone) when in situ (kappa, lambda, RNA+) is to be done on the tissue? I did a kappa ,lambda ,RNA+ on a decaled bone and I have no staining on all of them, but the controls are good. Any suggestions ? I thank you very much. By the way I am using the Ventana in situ reagents, and have talked to their tech support but did not get good explanation . 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 have received this message in error, 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 Jackie.O'Connor <@t> abbott.com Tue Sep 9 13:35:42 2003 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Looking for Richard Ornberg Message-ID: If anyone can tell me how to get in contact with Dick Ornberg (formerly of Monsanto/Pharmacia/Pfizer) in St. Louis, please get in touch with me. Thanks. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 Fax 847.938.3266 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/9e17c74e/attachment.htm From STapper <@t> slhduluth.com Tue Sep 9 12:29:01 2003 From: STapper <@t> slhduluth.com (Tapper, Sheila) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Computer system question Message-ID: <3BFBBD68413CB443A7125A63EC0ACD1D671695@slhw2smail01.slhdomain.com> To anyone out there who uses an LIS system - especially Meditech - do you use outside order entry into your system? We are investigating the usage of this function. The downfall - accession numbers will be assigned at the time of entry by the clinic, or outside client - not at the time of receipt into the lab. Does anyone use a system, or function like this in their facility? Sheila Tapper -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/aacc643f/attachment.htm From Sw918890 <@t> aol.com Tue Sep 9 15:18:32 2003 From: Sw918890 <@t> aol.com (Sw918890@aol.com) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Re: Cleaning Microtomes Message-ID: <018F624A.325ABF6C.0082A251@aol.com> I personally wipe off all of the paraffin that I can and then follow it up with 409. I don't like using paragard or other oily residues. I take out the department credit card and stock up on the 409. It is cheap and works wonders on my microtome. Steven R. Westra University of Iowa Health Care From SJones <@t> cvm.tamu.edu Tue Sep 9 16:19:28 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] cleaning microtomes Message-ID: I just use a 3 x 3 gauze square and a little elbow grease. Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> 09/09/03 09:49AM >>> I have a question that may seem silly. After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Histolady710 <@t> aol.com Tue Sep 9 16:53:52 2003 From: Histolady710 <@t> aol.com (Histolady710@aol.com) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] cleaning microtomes Message-ID: We vacuum the loose particles, spray with paragard and then wipe dry with gauze. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/8c4d5f5c/attachment.htm From amosbrooks <@t> earthlink.net Tue Sep 9 16:51:29 2003 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] cleaning microtomes References: <20030909170000.21215.91776.Mailman@swlx167.swmed.edu> Message-ID: <004901c3771d$1a2cb3c0$30684b43@oemcomputer> Hi, I'm with Tim on this one. I hate that oily residue that paragaurd and the like leave. It often interferes with sectioning when there is any left on the blade holder too. A gauze sponge works fine. Cleaning off oil with alcohol seems redundant to me. Amos Brooks Message: 22 From: Jmrwalker@aol.com Date: Tue, 9 Sep 2003 10:49:14 EDT To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cleaning microtomes I have a question that may seem silly. After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. Thanks! From amosbrooks <@t> earthlink.net Tue Sep 9 16:54:19 2003 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Containers with wells for multiple specimens References: <20030909170000.21215.91776.Mailman@swlx167.swmed.edu> Message-ID: <004a01c3771d$1be8bba0$30684b43@oemcomputer> Lynne, Why not use 11 cassettes and one large formalin container? Amos Brooks Message: 19 Date: Tue, 9 Sep 2003 09:42:07 -0400 From: "Bell, Lynne" To: Subject: [Histonet] Containers with wells for multiple specimens Good Morning All, I am looking for a type of specimen container that has multiple wells in it that can be filled with a fixative. Our urologists will be starting to do 11 biopsies on each patient and they would like a container such as the one I just described. My pathologist thinks that such a thing exists. I am aware of the cassettes that have six partitions for biopsies, but not of a specimen container. =20 I appreciate any help you can give me! Thanks, Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From SJones <@t> cvm.tamu.edu Tue Sep 9 16:41:36 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bitchin about ASCP Message-ID: This may seem really trivial......... But I was anxiously awaiting my 20 year gold sticker to put on my certificate (I had received a gold sticker at 10 years) and all I got was the usual sticker, although a different shape now. So I called ASCP and they told me they don't give them out for 20 years, just 25 years. Well, I am not amused! I don't even know if I will be doing histology in 5 more years. Any comments? The rate we are losing histotechs, they should give them out for every 5 years..... sorry for bitchin..................... Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 From SJones <@t> cvm.tamu.edu Tue Sep 9 16:50:13 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Kimwipes Message-ID: Hi everyone, I've been having trouble with the Kimwipes EX-L with lintguard (cough-cough) leaving lint in the waterbath. I would be interested to know what other products are out there that may work better. Thanks, Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 From garygill <@t> dcla.com Tue Sep 9 17:11:44 2003 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bitchin about ASCP Message-ID: 25 years is a tombstone shaped sticker. -----Original Message----- From: Sarah Jones [mailto:SJones@cvm.tamu.edu] Sent: Tuesday, September 09, 2003 4:42 PM To: histonet@pathology.swmed.edu Subject: [Histonet] bitchin about ASCP This may seem really trivial......... But I was anxiously awaiting my 20 year gold sticker to put on my certificate (I had received a gold sticker at 10 years) and all I got was the usual sticker, although a different shape now. So I called ASCP and they told me they don't give them out for 20 years, just 25 years. Well, I am not amused! I don't even know if I will be doing histology in 5 more years. Any comments? The rate we are losing histotechs, they should give them out for every 5 years..... sorry for bitchin..................... Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Linke_Noelle <@t> Allergan.com Tue Sep 9 17:29:57 2003 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bitchin about ASCP Message-ID: <805C4A052A253B4A8E4948B13F54D12704D3D3@IRMAIL102.irvine.allergan.com> HAHAHAHA....that's too damn funny..... -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, September 09, 2003 3:12 PM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] bitchin about ASCP 25 years is a tombstone shaped sticker. -----Original Message----- From: Sarah Jones [mailto:SJones@cvm.tamu.edu] Sent: Tuesday, September 09, 2003 4:42 PM To: histonet@pathology.swmed.edu Subject: [Histonet] bitchin about ASCP This may seem really trivial......... But I was anxiously awaiting my 20 year gold sticker to put on my certificate (I had received a gold sticker at 10 years) and all I got was the usual sticker, although a different shape now. So I called ASCP and they told me they don't give them out for 20 years, just 25 years. Well, I am not amused! I don't even know if I will be doing histology in 5 more years. Any comments? The rate we are losing histotechs, they should give them out for every 5 years..... sorry for bitchin..................... Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 _______________________________________________ Histonet mailing list Histonet@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 RFail <@t> Charleston.net Tue Sep 9 17:27:54 2003 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] CAB-stain In-Reply-To: <00bb01c376cb$f6b40f20$0d12a8c0@SERVER> Message-ID: <000201c37721$9d98f8a0$d810a6a5@rena> Sometimes called Chromotrope 2 R, or Roque's. reference is from Laboratory nvestigation, vol.2 no 5 1953 pg 15-21 Title is Chromotrope Aniline Blue Method of Staining Mallory Bodies of aennec's Cirrhosis, Author Agustin L. Roque Rena Fail -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Tuesday, September 09, 2003 8:15 AM To: Histonetliste Subject: [Histonet] CAB-stain Hi! Does anyone know a reference for the CAB-stain (a variant of Gomori's Trichrome onestep method). CAB = Chromotrop Anilinblue, for staining collagen fibers Gundi -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/12e9c9bc/attachment.htm From JWEEMS <@t> sjha.org Tue Sep 9 17:38:54 2003 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bitchin about ASCP Message-ID: <9E75DAB5F369D84ABF84FAB7A0243B44B6D3B0@exch4.sjha.org> Oh is that what that was?! -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, September 09, 2003 6:12 PM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] bitchin about ASCP 25 years is a tombstone shaped sticker. -----Original Message----- From: Sarah Jones [mailto:SJones@cvm.tamu.edu] Sent: Tuesday, September 09, 2003 4:42 PM To: histonet@pathology.swmed.edu Subject: [Histonet] bitchin about ASCP This may seem really trivial......... But I was anxiously awaiting my 20 year gold sticker to put on my certificate (I had received a gold sticker at 10 years) and all I got was the usual sticker, although a different shape now. So I called ASCP and they told me they don't give them out for 20 years, just 25 years. Well, I am not amused! I don't even know if I will be doing histology in 5 more years. Any comments? The rate we are losing histotechs, they should give them out for every 5 years..... sorry for bitchin..................... Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 _______________________________________________ Histonet mailing list Histonet@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 WWmn916 <@t> aol.com Tue Sep 9 17:41:48 2003 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Bitchin about ASCP Message-ID: <136.241be3d1.2c8fb12c@aol.com> All due respect for the importance of milestones measured in years.....it's good to appreciate the humor too!!!!! So is it really a tombstone gold sticker from ASCP after 25 years?!!!! That is funny!!! Deb King, HT Sacramento, CA -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/fae994b6/attachment.htm From SJones <@t> cvm.tamu.edu Tue Sep 9 17:45:19 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bitchin about ASCP Message-ID: Gary, you made my day. Thanks for the laughs! I can envision a new advertising campaign for ASCP.............. "What do you want on your Tombstone? 25 years in ASCP!" I wish I knew how to do PhotoShop ............... Sarah if you like PhotoShop, check out http://www.worth1000.com/ great laughs >>> Gary Gill 09/09/03 05:11PM >>> 25 years is a tombstone shaped sticker. -----Original Message----- From: Sarah Jones [mailto:SJones@cvm.tamu.edu] Sent: Tuesday, September 09, 2003 4:42 PM To: histonet@pathology.swmed.edu Subject: [Histonet] bitchin about ASCP This may seem really trivial......... But I was anxiously awaiting my 20 year gold sticker to put on my certificate (I had received a gold sticker at 10 years) and all I got was the usual sticker, although a different shape now. So I called ASCP and they told me they don't give them out for 20 years, just 25 years. Well, I am not amused! I don't even know if I will be doing histology in 5 more years. Any comments? The rate we are losing histotechs, they should give them out for every 5 years..... sorry for bitchin..................... Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo <@t> tiscali.co.uk Wed Sep 10 01:39:31 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bitchin about ASCP In-Reply-To: Message-ID: <001401c37766$4e3c2580$01062850@KEMLOS> Um...... Where do you stick this sticker? I've done 33 years and I know where I'd stick mine! Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts:? ???????????? kemlo@tiscali.co.uk?or kemlo1@btinternet.com? Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. ? ? ? -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Gary Gill Sent: 09 September 2003 23:12 To: histonet@pathology.swmed.edu Subject: RE: [Histonet] bitchin about ASCP 25 years is a tombstone shaped sticker. -----Original Message----- From: Sarah Jones [mailto:SJones@cvm.tamu.edu] Sent: Tuesday, September 09, 2003 4:42 PM To: histonet@pathology.swmed.edu Subject: [Histonet] bitchin about ASCP This may seem really trivial......... But I was anxiously awaiting my 20 year gold sticker to put on my certificate (I had received a gold sticker at 10 years) and all I got was the usual sticker, although a different shape now. So I called ASCP and they told me they don't give them out for 20 years, just 25 years. Well, I am not amused! I don't even know if I will be doing histology in 5 more years. Any comments? The rate we are losing histotechs, they should give them out for every 5 years..... sorry for bitchin..................... Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 _______________________________________________ Histonet mailing list Histonet@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 m.vermehren <@t> anat.vetmed.uni-muenchen.de Wed Sep 10 02:03:00 2003 From: m.vermehren <@t> anat.vetmed.uni-muenchen.de (Margarete Vermehren) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] 409 Message-ID: <000601c37769$92ff9280$64ae548d@HistoPC3> I may have missed something, but what is 409? Margarete Vermehren LMU M?nchen / Tieranatomie II Tel: +49 89 21805857 Fax: +49 89 21802569 m.vermehren@anat.vetmed.uni-muenchen.de -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030910/5cdff943/attachment.htm From mikael.niku <@t> helsinki.fi Wed Sep 10 02:05:23 2003 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Scanner for microscope slides Message-ID: <000301c37769$e73d46d0$8c0fd680@ekk1116> Hello! I'm thinking of getting a film scanner capaple of scanning microscope slides. At least for Nikon Coolscan 4000 one can get a microscope slide adapter. Any experience of such a setup, or other recommendations? +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ + Mikael Niku + University of Helsinki, Dept. Basic Veterinary Sciences + URL: www.helsinki.fi/~mniku/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! (Gandhi) From ree3 <@t> leicester.ac.uk Wed Sep 10 04:55:11 2003 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] bitchin about ASCP Message-ID: In the UK it is number of blocks cut that are recognised as the significant milestones in ones career, after cutting 100,000 blocks the Health and Safety Executive come along and count one's fingers, if they are all present and correct with no significant scar tissue then one is presented with a metal badge in the shape of an outstretched hand, this is much prized by all who receive it, although they are often greeted on their return to the laboratory by their less fortunate colleagues with the traditional Agincourt salute. R.F.(Fingers)Edwards LEICESTER...U.K... -----Original Message----- From: Sarah Jones [mailto:SJones@cvm.tamu.edu] Sent: 09 September 2003 22:42 To: histonet@pathology.swmed.edu Subject: [Histonet] bitchin about ASCP This may seem really trivial......... But I was anxiously awaiting my 20 year gold sticker to put on my certificate (I had received a gold sticker at 10 years) and all I got was the usual sticker, although a different shape now. So I called ASCP and they told me they don't give them out for 20 years, just 25 years. Well, I am not amused! I don't even know if I will be doing histology in 5 more years. Any comments? The rate we are losing histotechs, they should give them out for every 5 years..... sorry for bitchin..................... Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ASelf <@t> gmhsc.com Wed Sep 10 05:01:41 2003 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] de-staining immunos Message-ID: Does anyone out in histoland know if Immuno stained slides and/or air-dried Diff-Quik stained slides can be de-stained? If so could you please share with me this procedure. I know that H&E's can be de-stained but I wasn't sure about immunos and diff-quiks. Thanks for the help.... Amy Self Georgetown Memorial Hospital Georgetown, SC (phone) 843-527-7179 (fax) 843-520-7882 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 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 mlm11 <@t> cornell.edu Wed Sep 10 08:16:49 2003 From: mlm11 <@t> cornell.edu (Mary Lou) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Kimwipes In-Reply-To: Message-ID: <5.2.1.1.2.20030910090424.03ad6e80@postoffice9.mail.cornell.edu> I use old phone book pages to clean my water bath. I spend a little time cutting the pages lengthwise and they fit just fine in my rectangular bath. They work better than KimWipes because they're free! Mary Lou Norman Clin Sci Histo Lab Vet School Cornell University At 04:50 PM 9/9/2003 -0500, you wrote: >Hi everyone, > I've been having trouble with the Kimwipes EX-L with lintguard > (cough-cough) leaving lint in the waterbath. I would be interested to > know what other products are out there that may work better. Thanks, Sarah > >Sarah Jones HT(ASCP) >Dept. of Vet. Anatomy & Public Health >Histology Lab >Texas A&M University >College Station, TX 77843-4458 >phone: 979-845-3177 >fax: 979-458-3499 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed Sep 10 08:37:35 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] 409 In-Reply-To: <000601c37769$92ff9280$64ae548d@HistoPC3> References: <000601c37769$92ff9280$64ae548d@HistoPC3> Message-ID: <3F5F291F.3040409@umdnj.edu> 409 is a household cleaner popular in the USA. For the auto buffs on the list it is also the displacement, in cubic inches, of a Chevrolet V-8 of the early to mid 1960's and the inspiration for a Beach Boys song. Geoff Margarete Vermehren wrote: > I may have missed something, but what is 409? > > > > Margarete Vermehren > > LMU M?nchen / Tieranatomie II > > Tel: +49 89 21805857 > > Fax: +49 89 21802569 > > m.vermehren@anat.vetmed.uni-muenchen.de > > > -- -- ********************************************** 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 ********************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030910/0ebd505e/attachment.htm From mbryhan <@t> NORTHERNHEALTH.ORG Wed Sep 10 09:37:46 2003 From: mbryhan <@t> NORTHERNHEALTH.ORG (Mary Bryhan) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] IHC systems Message-ID: Need feedback from both BioGenex I6000 and Ventana benchmark users. We currently use the BioGenex Optimax Plus, but would like to bring HPV testing in-house. Mary Bryhan From hhawkins <@t> utmb.edu Wed Sep 10 09:49:24 2003 From: hhawkins <@t> utmb.edu (Hawkins, Hal K.) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] Scanner for microscope slides Message-ID: <1874CD14866CD6118DF000D0B79E84F1016E76D8@exchange4> Meyer Instruments has modified a Microtek scanner for the purpose, see http://www.meyerinst.com/html/oem/pseiii/default.htm Hal Hawkins, UTMB Galveston (not a stockholder) -----Original Message----- From: Mikael Niku [mailto:mikael.niku@helsinki.fi] Sent: Wednesday, September 10, 2003 2:05 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Scanner for microscope slides Hello! I'm thinking of getting a film scanner capaple of scanning microscope slides. At least for Nikon Coolscan 4000 one can get a microscope slide adapter. Any experience of such a setup, or other recommendations? +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ + Mikael Niku + University of Helsinki, Dept. Basic Veterinary Sciences + URL: www.helsinki.fi/~mniku/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! (Gandhi) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mward <@t> wfubmc.edu Wed Sep 10 10:50:51 2003 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:21:50 2005 Subject: [Histonet] kappa and lambda staining Message-ID: <61135F0455D33347B5AAE209B903A30403262297@EXCHVS2.medctr.ad.wfubmc.edu> I was wondering if anyone has any tips on kappa and lambda staining on bone marrow and bone plug sections? My pathologists say we are getting inconsistant staining on these specimens. Our histology lab puts the bone marrows in B5 solution for 2 hours and then into formalin for processing. The bone plugs go into B5 for 2 hours and then into DeltaCal (from Delta Products Group). We are currently using a NexEs for our staining and a BioCare monoclonal Kappa and Lambda antibody, using antigen retrieval. I also have recently installed a Dako Autostainer. I would appreciate any feedback for either systems. Thanks in advance. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center From g.lang <@t> bigfoot.de Wed Sep 10 10:55:53 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] ASCP Message-ID: <003601c377b4$088f3f10$0d12a8c0@SERVER> Is ASCP your professional organisation in USA? Gundi -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030910/df62ecd4/attachment.htm From tpmorken <@t> labvision.com Wed Sep 10 11:06:10 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Scanner for microscope slides Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CB50@usca0082k03.rallansci.apogent.com> You might look at this scanner too: http://arrayit.com/Products/MicroarrayI/Scanner/scanner.html Tim Morken -----Original Message----- From: Mikael Niku [mailto:mikael.niku@helsinki.fi] Sent: Wednesday, September 10, 2003 12:05 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Scanner for microscope slides Hello! I'm thinking of getting a film scanner capaple of scanning microscope slides. At least for Nikon Coolscan 4000 one can get a microscope slide adapter. Any experience of such a setup, or other recommendations? +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ + Mikael Niku + University of Helsinki, Dept. Basic Veterinary Sciences + URL: www.helsinki.fi/~mniku/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! (Gandhi) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Wed Sep 10 11:12:51 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] ASCP Message-ID: Yes, it's the American Society for Clinical Pathology. www.ascp.org. -----Original Message----- From: Gudrun Lang [mailto:g.lang@bigfoot.de] Sent: Wed 9/10/2003 10:55 AM To: Histonetliste Cc: Subject: [Histonet] ASCP Is ASCP your professional organisation in USA? Gundi From Rcartun <@t> harthosp.org Wed Sep 10 11:24:58 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] kappa and lambda staining Message-ID: We published our routine method in, "Immunohistochemical detection of immunoglobulin light chain expression in B-cell non-Hodgkin lymphomas using formalin fixed, paraffin-embedded tissues and a heat-induced epitope retrieval technique" that appeared in Applied Immunohistochemistry & Molecular Morphology 10(3):258-262, 2002. However, I can tell you that we use different retrieval methods and different primary antibody dilutions when the initial results are suboptimal. Richard Cartun >>> "Martha Ward" 09/10/03 11:50AM >>> I was wondering if anyone has any tips on kappa and lambda staining on bone marrow and bone plug sections? My pathologists say we are getting inconsistant staining on these specimens. Our histology lab puts the bone marrows in B5 solution for 2 hours and then into formalin for processing. The bone plugs go into B5 for 2 hours and then into DeltaCal (from Delta Products Group). We are currently using a NexEs for our staining and a BioCare monoclonal Kappa and Lambda antibody, using antigen retrieval. I also have recently installed a Dako Autostainer. I would appreciate any feedback for either systems. Thanks in advance. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bryand <@t> netbistro.com Wed Sep 10 11:34:22 2003 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] bitchin about ASCP References: Message-ID: <005501c377b9$654b61c0$ad70c2cf@bryand> I don't know about the lack of scar tissue. I have lots on my finger tips from my base sledge days. In fact, I was told by the big boss that I wasn't really a fully qualified histology tech until I needed to have at least two stiches for a cut. Fortunately for me I stayed non-fully qualified, although I did come close a few times! Bryan Llewellyn ----- Original Message ----- From: "Edwards, R.E." To: "Sarah Jones" ; Sent: Wednesday, September 10, 2003 2:55 AM Subject: RE: [Histonet] bitchin about ASCP In the UK it is number of blocks cut that are recognised as the significant milestones in ones career, after cutting 100,000 blocks the Health and Safety Executive come along and count one's fingers, if they are all present and correct with no significant scar tissue then one is presented with a metal badge in the shape of an outstretched hand, this is much prized by all who receive it, although they are often greeted on their return to the laboratory by their less fortunate colleagues with the traditional Agincourt salute. R.F.(Fingers)Edwards LEICESTER...U.K... -----Original Message----- From: Sarah Jones [mailto:SJones@cvm.tamu.edu] Sent: 09 September 2003 22:42 To: histonet@pathology.swmed.edu Subject: [Histonet] bitchin about ASCP This may seem really trivial......... But I was anxiously awaiting my 20 year gold sticker to put on my certificate (I had received a gold sticker at 10 years) and all I got was the usual sticker, although a different shape now. So I called ASCP and they told me they don't give them out for 20 years, just 25 years. Well, I am not amused! I don't even know if I will be doing histology in 5 more years. Any comments? The rate we are losing histotechs, they should give them out for every 5 years..... sorry for bitchin..................... Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Wed Sep 10 12:11:47 2003 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] bitchin about ASCP Message-ID: At first I thought they gave the award in the shape of an outstretched hand to replace the one you lost piece by piece through years of microtomy. Maybe I'm naive, but is the Agincourt salute the same as one given in the US by angry motorists? Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 Fax 847.938.3266 "Bryan Llewellyn" Sent by: histonet-admin@lists.utsouthwestern.edu 09/10/2003 11:34 AM To: "Histonet" cc: Subject: Re: [Histonet] bitchin about ASCP I don't know about the lack of scar tissue. I have lots on my finger tips from my base sledge days. In fact, I was told by the big boss that I wasn't really a fully qualified histology tech until I needed to have at least two stiches for a cut. Fortunately for me I stayed non-fully qualified, although I did come close a few times! Bryan Llewellyn ----- Original Message ----- From: "Edwards, R.E." To: "Sarah Jones" ; Sent: Wednesday, September 10, 2003 2:55 AM Subject: RE: [Histonet] bitchin about ASCP In the UK it is number of blocks cut that are recognised as the significant milestones in ones career, after cutting 100,000 blocks the Health and Safety Executive come along and count one's fingers, if they are all present and correct with no significant scar tissue then one is presented with a metal badge in the shape of an outstretched hand, this is much prized by all who receive it, although they are often greeted on their return to the laboratory by their less fortunate colleagues with the traditional Agincourt salute. R.F.(Fingers)Edwards LEICESTER...U.K... -----Original Message----- From: Sarah Jones [mailto:SJones@cvm.tamu.edu] Sent: 09 September 2003 22:42 To: histonet@pathology.swmed.edu Subject: [Histonet] bitchin about ASCP This may seem really trivial......... But I was anxiously awaiting my 20 year gold sticker to put on my certificate (I had received a gold sticker at 10 years) and all I got was the usual sticker, although a different shape now. So I called ASCP and they told me they don't give them out for 20 years, just 25 years. Well, I am not amused! I don't even know if I will be doing histology in 5 more years. Any comments? The rate we are losing histotechs, they should give them out for every 5 years..... sorry for bitchin..................... Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 _______________________________________________ Histonet mailing list Histonet@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 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030910/6b7a1417/attachment.htm From timothy_m_coskran <@t> groton.pfizer.com Wed Sep 10 12:19:07 2003 From: timothy_m_coskran <@t> groton.pfizer.com (Coskran, Timothy M) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] MHC slow or fast muscle fibers Message-ID: <75FBD3E83B69D711A9E600080261980C83F1CA@groexmb04bak.pfizer.com> Hello, Does anyone know of a source for a polyclonal to MHC slow or MHC fast muscle fibers? We have had success with the monoclonals, but would like either of the two as a polyclonal Thanks, Tim Coskran Pfizer LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From BoozerKA <@t> pa1.ah.org Wed Sep 10 12:30:25 2003 From: BoozerKA <@t> pa1.ah.org (Kathleen Boozer) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Protocol for frozen IHC Message-ID: Could anyone fax me their protocol for frozen slides? Thanks! From bhewlett <@t> cogeco.ca Wed Sep 10 12:42:19 2003 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] bitchin about ASCP References: <005501c377b9$654b61c0$ad70c2cf@bryand> Message-ID: <004b01c377c2$e254b9b0$b6833918@bryaniwx13voft> Me too!! Although, after 45 years and literally millions of sections, I was just beginning to get the hang of it when I retired! Bryan Hewlett ----- Original Message ----- From: "Bryan Llewellyn" To: "Histonet" Sent: Wednesday, September 10, 2003 12:34 PM Subject: Re: [Histonet] bitchin about ASCP > I don't know about the lack of scar tissue. I have lots on my finger tips > from my base sledge days. In fact, I was told by the big boss that I wasn't > really a fully qualified histology tech until I needed to have at least two > stiches for a cut. Fortunately for me I stayed non-fully qualified, > although I did come close a few times! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Edwards, R.E." > To: "Sarah Jones" ; > Sent: Wednesday, September 10, 2003 2:55 AM > Subject: RE: [Histonet] bitchin about ASCP > > > In the UK it is number of blocks cut that are recognised as the > significant milestones in ones career, after cutting 100,000 blocks the > Health and Safety Executive come along and count one's fingers, if > they are all present and correct with no significant scar tissue > then one is presented with a metal badge in the shape of an > outstretched hand, this is much prized by all who receive it, although > they are often greeted on their return to the laboratory by their less > fortunate colleagues with the traditional Agincourt salute. > R.F.(Fingers)Edwards > LEICESTER...U.K... > > > > > -----Original Message----- > From: Sarah Jones [mailto:SJones@cvm.tamu.edu] > Sent: 09 September 2003 22:42 > To: histonet@pathology.swmed.edu > Subject: [Histonet] bitchin about ASCP > > > This may seem really trivial......... But I was anxiously awaiting my 20 > year gold sticker to put on my certificate (I had received a gold sticker at > 10 years) and all I got was the usual sticker, although a different shape > now. So I called ASCP and they told me they don't give them out for 20 > years, just 25 years. Well, I am not amused! I don't even know if I will > be doing histology in 5 more years. Any comments? The rate we are losing > histotechs, they should give them out for every 5 years..... > sorry for bitchin..................... Sarah > > Sarah Jones HT(ASCP) > Dept. of Vet. Anatomy & Public Health > Histology Lab > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > > > _______________________________________________ > Histonet mailing list > Histonet@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 amarusk1 <@t> FAIRVIEW.ORG Wed Sep 10 13:10:12 2003 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] IHC for neutophilic granules Message-ID: Skipped content of type multipart/alternative-------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:ANN MARUSKA EMAIL;WORK;PREF;NGW:AMARUSK1.EAST.NORTH ORG:;LAB N:MARUSKA;ANN TEL;WORK:612-273-9119 END:VCARD From tvedilago <@t> system1.net Wed Sep 10 14:06:57 2003 From: tvedilago <@t> system1.net (Tommy Vedilago) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Job in Milwaukee Message-ID: Hello again Histonetters, I am looking, still, for an AP Manager for a multiple hospital system in Milwaukee. They need a strong manager with excellent people skills and dynamic personality. The lab is functioning well but, they are looking to the future for expansions and growth. Please give me a call with any interest or send me a referral or 2 if you know of a good leader in need of a step up. Thanks, and I look forward to hearing from anyone in need of my assistance at 866-SYSTEM1. Tommy Vedilago System 1 Search (864) 627-0012 (864) 627-0013 Fax From dpconsult <@t> comcast.net Wed Sep 10 14:07:07 2003 From: dpconsult <@t> comcast.net (Dick Paulson) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] bitchin about ASCP Message-ID: Jackie - here is everything you wanted to know and then some about the Agincourt Salute. I couldn't resist. King Henry V of England's victory over the French in the Battle of Agincourt on October 25, 1415 has been attributed to the overwhelming superiority of the English longbow. Henry matched his force of approximately 5700 men against the French, some 25,000 strong, outside the village of Agincourt, south of Calais in the north of France. Prior to the battle, confident of victory due to their superior numbers, the French cockily proposed to cut off the middle finger of all captured English soldiers. English archers had the reputation of firing up to twenty rounds a minute. Evidence of the longbow's ability to prove decisive in battle had been demonstrated in the earlier battles of Crecy and Poitiers where immense numbers of knights were slaughtered before they ever got close to the English. Since it is impossible to draw the longbow without the third finger it would therefore, render the men incapable of using a longbow in the future. Word spread quickly throughout the French army and, as intended, made itself know to the English. Inspired by King Henry's passionate call to fight beside him on St. Crispin's Day, the English forces formed up in four ranks with their archers in front. They advanced, then halted and drove stakes into the ground to deter oncoming French cavalry. The ground was thick with mud, which, combined with the heavy rain of fierce arrows and the row of stakes, thwarted the French cavalry. The English foot soldiers attacked, engaging in hand-to-hand combat and defeated the first line of the French force. The English continued to advance upon the second and third lines, driving the French from the field. The French lost over 6000 men while the English marked only 1600 casualties. The famous longbow was made from the wood of the Yew tree, whereby the act of drawing the longbow was known as to 'pluck yew'. In the aftermath of the battle, the triumphant English, mocking the defeated French, began waving their middle fingers at them and shouting, "See, we can still pluck yew! PLUCK YEW!" Since 'pluck yew' is rather difficult to say the consonant blend at the beginning has gradually changed to a labiodental fricative 'F'. It is also because of the feathers on the arrows that the gesture, holding up the lone middle finger, is known as "giving the bird". After all, if you're drawing a bow and loosing an arrow at someone, you are, obviously, sending 'the bird' with a rather specific intention, not all together pleasant for the recipient. How these words, often used in conjunction with the one-finger-salute, are mistakenly thought to have something to do with an encounter of a closer proximity is another story. Statlab Medical Products Dick Paulson dpconsult@earthlink.net Phone (800) 393-6345 From mcauliff <@t> umdnj.edu Tue Sep 9 14:52:22 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] C-fos stain In-Reply-To: <082C721AF78DB34983E8BA2CD085462105C64B@mailbe07> References: <082C721AF78DB34983E8BA2CD085462105C64B@mailbe07> Message-ID: <3F5E2F76.3070301@umdnj.edu> Hi Gareth: I have not seen an answer to your question on Histonet so ......... Osmium (1% aqueous or less) will enhance the DAB reaction product. If your (brain) sections have not been defatted/delipidized osmium will also stain the lipids (myelin) in the brain. This may actually reduce the contrast of the DAB-osmium reaction product depending on where it is. Osmium will also make the sections less flexible so they should be mounted on slides before exposure. Osmium is expensive and must be used under a hood ("fume cupboard") as its vapors will fix your corneal and nasal epithelium. It is also requires some care in its disposal. There are several less dangerous/expensive ways to enhance the DAB reaction product, including nickel or nickel+cobalt ions in the DAB-peroxide incubation medium is popular. While nckel and cobalt are certainly not benign they don't have the fume problems that osmium does and they are less expensive. I can send you a nickel+cobalt recipe if you like. In my experience DAB should be 'enhanced' with something if you want the intensity of the reaction product to be permant. Also, use DPX as a mounting media. Geoff Davis, Gareth wrote: > I plan to do a immunohistochemical stain on mouse brain tissue with > c-fos. Some protocols I've read state that you should osmicate the > sections, after mounting, to enhance the stain. I've never stained > with c-fos, but I have been told it is very dark on it's own. Does > anyone know if it will be necessary to osmicate. Of course, I guess I > could just try and see, but any information on the staining process > itself would be helpful. > Thanks, > Ms. Gareth Davis > Vanderbilt University Medical Center > > > Ms. Gareth B. Davis > Research Assistant II > Neuro-magnetics Division of > the Department of Neurology > Vanderbilt University Medical Center > 615-936-3318 > > -- -- ********************************************** 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 ********************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/c4d5438e/attachment.htm From GalbraithJ <@t> uihc.uiowa.edu Tue Sep 9 15:03:05 2003 From: GalbraithJ <@t> uihc.uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] cleaning microtomes Message-ID: <5D03ED7B9391D4119D9B0008C76B7B24030085CE@uihc-mail1.uihc.uiowa.edu> Histonetters: Most people in my lab use Paraguard to clean microtomes but some prefer 409 (Misty Breeze has a nice smell). The 409 cleaner seems to do as good of a job as Paraguard and costs less and smells better. Joe Galbraith -----Original Message----- From: RCHIOVETTI@aol.com [mailto:RCHIOVETTI@aol.com] Sent: Tuesday, September 09, 2003 1:15 PM To: Jmrwalker@aol.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] cleaning microtomes In a message dated 09/09/2003 7:51:39 AM US Mountain Standard Time, Jmrwalker@aol.com writes: << After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. Thanks! >> I see mostly Paragard being used in our customers' labs, and I agree with the other posters, if the "slick" surface is a problem after the Paragard treatment you can then wipe down with a little ethanol or isopropanol. With time xylene can affect the surfaces of the microtome's external parts, especially if used in excess. Lots of the microtome parts have anodized (plated) surfaces that are either black or clear, and you'd think they would hold up to just about any kind of abuse. But they will definitely break down and corrode with prolonged or repeated exposure to xylene. This can lead to pitting and discoloration of the surfaces. If you *do* use xylene, use it sparingly and be sure to wear gloves. I personally don't like the stuff. Try to use a substitute instead. First, use a brush or gauze pad to remove the majority of the paraffin. Then use only enough xylene or xylene substitute to barely moisten a gauze 4x4 or a tissue. Wipe off the excess immediately with another dry gauze or tissue. You can also follow this with an alcohol wipe-down like you would use with Paragard if the surface feels slick or oily after you clean it. Hope this helps! Bob Chiovetti GTI Microsystems Leica Regional Dealer Desert Southwest Region, USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Tue Sep 9 14:48:17 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] cleaning microtomes Message-ID: I find that a dry gauze does the job. I have tried paragard, however, and it seems that the paraffin that accumulates on the paragard cleaned surface seems to be sticky and more difficult to clean. Has anyone else had this experience, or am I on an island here? Fred >>> "Favara, Cynthia (NIH/NIAID)" 09/09/03 11:31AM >>> I use paragard which is just oil of wintergreen, followed by ETOH. Do not use xylene because of health concerns. I would be interested in what Gayle Callis has to say, I believe she took a course at NSH previously and the instructor recommended not using any clearing agent. Would be interesting to have a manufacturers recommendation. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Jmrwalker@aol.com [mailto:Jmrwalker@aol.com] Sent: Tuesday, September 09, 2003 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cleaning microtomes I have a question that may seem silly. After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. 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 RFail <@t> Charleston.net Tue Sep 9 18:30:39 2003 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] CAB-stain/typo In-Reply-To: <000201c37721$9d98f8a0$d810a6a5@rena> Message-ID: <000001c3772a$68524620$d010a6a5@rena> Sorry for typo Laennec's cirrhosis -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Rena Fail Sent: Tuesday, September 09, 2003 6:28 PM To: 'Gudrun Lang'; 'Histonetliste' Subject: RE: [Histonet] CAB-stain Sometimes called Chromotrope 2 R, or Roque's. reference is from Laboratory nvestigation, vol.2 no 5 1953 pg 15-21 Title is Chromotrope Aniline Blue Method of Staining Mallory Bodies of aennec's Cirrhosis, Author Agustin L. Roque Rena Fail -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Tuesday, September 09, 2003 8:15 AM To: Histonetliste Subject: [Histonet] CAB-stain Hi! Does anyone know a reference for the CAB-stain (a variant of Gomori's Trichrome onestep method). CAB = Chromotrop Anilinblue, for staining collagen fibers Gundi -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/0c6c25f4/attachment.htm From Vicki.Goodman <@t> se.amedd.army.mil Wed Sep 10 14:25:01 2003 From: Vicki.Goodman <@t> se.amedd.army.mil (Goodman, Vicki W Ms BACH) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Unsubscribe Message-ID: <24832C55638FB6478091DEF8241E677F762BB9@dasmthdbl001.amedd.army.mil> -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030910/0e771132/attachment.htm From gentras <@t> vetmed.auburn.edu Wed Sep 10 14:54:52 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Fwd: Returned mail: see transcript for details Message-ID: <5.2.0.9.0.20030910145420.00a0b6f0@mailhost.vetmed.auburn.edu> >From: "Atoska S. Gentry" >Subject: tissue processors >Mime-Version: 1.0 >Content-Type: text/plain; charset="us-ascii"; format=flowed > > >Hello, please is anyone familiar with a tissue processor in today's market >that is comparable to the Autotechnicon Ultra II ? Any assistance with >sources, prices, and etc. will be greatly appreciated. Thanks, Atoska > > >Atoska S. Gentry B.S., HT(ASCP) >Research Assistant III >Scott-Ritchey Research Center >College of Veterinary Medicine >Auburn University, AL 36849 >Phone# (334)844-5579 Fax# (334)844-5850 Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From gareth.davis <@t> Vanderbilt.Edu Wed Sep 10 15:04:46 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Re: C-fos Message-ID: <082C721AF78DB34983E8BA2CD085462105C658@mailbe07> Geoff, Thanks for the information on osmium. I have used the stuff for Scanning EM and know how awful it is to use, so would love to avoid it. I was going to try staining free floating sections with c-fos, but have decided to mount the sections onto glass slides first. We have done nickel enhancement when staining cell cultures, and decided it didn't make much of a difference. But, maybe your nickel+cobalt recipe would be helpful for our sections. Thanks, Gareth Geoff replied to C-fos and osmication question: Hi Gareth: I have not seen an answer to your question on Histonet so ......... Osmium (1% aqueous or less) will enhance the DAB reaction product. If your (brain) sections have not been defatted/delipidized osmium will also stain the lipids (myelin) in the brain. This may actually reduce the contrast of the DAB-osmium reaction product depending on where it is. Osmium will also make the sections less flexible so they should be mounted on slides before exposure. Osmium is expensive and must be used under a hood ("fume cupboard") as its vapors will fix your corneal and nasal epithelium. It is also requires some care in its disposal. There are several less dangerous/expensive ways to enhance the DAB reaction product, including nickel or nickel+cobalt ions in the DAB-peroxide incubation medium is popular. While nckel and cobalt are certainly not benign they don't have the fume problems that osmium does and they are less expensive. I can send you a nickel+cobalt recipe if you like. In my experience DAB should be 'enhanced' with something if you want the intensity of the reaction product to be permant. Also, use DPX as a mounting media. Geoff Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030910/edd8583e/attachment.htm From lcardenas <@t> utmem.edu Wed Sep 10 15:22:24 2003 From: lcardenas <@t> utmem.edu (Luz Cardenas) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Re: C-fos Message-ID: <5d2d925d0fb3.5d0fb35d2d92@utnet2.utmem.edu> Hi Geoff, I am also trying C-fos in brain tissue, free floating and I would like to try nickel-cobalt recipe is you don't mind, can you send it to me also?. Thanks! Luz C?rdenas, MD Post-doctoral Research Trainee Department of Anatomy and Neurobiology University of Tennessee Health Science Center 855 Monroe Avenue, Room 309/302 Memphis, TN 38163 Lab: (901) 448-6002 / 7318 Fax: (901) 448-7193 (Dept) ----- Original Message ----- From: "Davis, Gareth" Date: Wednesday, September 10, 2003 3:04 pm Subject: [Histonet] Re: C-fos > Geoff, > Thanks for the information on osmium. I have used the stuff for > Scanning EM and know how awful it is to use, so would love to > avoid it. I was going to try staining free floating sections with > c-fos, but have decided to mount the sections onto glass slides > first. We have done nickel enhancement when staining cell > cultures, and decided it didn't make much of a difference. But, > maybe your nickel+cobalt recipe would be helpful for our sections. > > Thanks, > Gareth > > > > > Geoff replied to C-fos and osmication question: > Hi Gareth: > > I have not seen an answer to your question on Histonet so > ......... Osmium (1% aqueous or less) will enhance the DAB > reaction product. If your (brain) sections have not been > defatted/delipidized osmium will also stain the lipids (myelin) in > the brain. This may actually reduce the contrast of the DAB-osmium > reaction product depending on where it is. Osmium will also make > the sections less flexible so they should be mounted on slides > before exposure. Osmium is expensive and must be used under a hood > ("fume cupboard") as its vapors will fix your corneal and nasal > epithelium. It is also requires some care in its disposal. There > are several less dangerous/expensive ways to enhance the DAB > reaction product, including nickel or nickel+cobalt ions in the > DAB-peroxide incubation medium is popular. While nckel and cobalt > are certainly not benign they don't have the fume problems that > osmium does and they are less expensive. I can send you a > nickel+cobalt recipe if you like. In my experience DAB should be > 'enhanced' with something if you want the intensity of the > reaction product to be permant. Also, use DPX as a mounting media. > > Geoff > > > Ms. Gareth B. Davis > Research Assistant II > Neuro-magnetics Division of > the Department of Neurology > Vanderbilt University Medical Center > 615-936-3318 > > > From gareth.davis <@t> Vanderbilt.Edu Wed Sep 10 15:30:53 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Re: Protocol for frozen IHC Message-ID: <082C721AF78DB34983E8BA2CD085462105C65B@mailbe07> Kathleen, Here is a protocol from Oncogene Research. I'm not sure where you need the protocol to start, so this starts from the very beginning. Clean glass slides with 95% ethanol, treat with subbing solution and air dry (not necessary to sub, if using SuperFrost Plus slides). Cut 4-8 micron thick sections of tissue block. Allow frozen sections to come to room temp. Fix slides with cold acetone for 10 minutes (I've used 4% Formaldehyde and have gotten great results). Rinse in three changes of Buffer. Incubate for 5 to 10 minutes in 0.1% hydrogen peroxide in PBS to quench endogenous peroxidase activity. Rinse with PBS Incubate slides with blocking serum in PBS for 20 minutes, to suppress non-specific binding of immunoglobulin. Rinse with three changes of PBS, 5 minutes each rinse. Incubate with Primary antibody for one hour at room temp or overnight at 4 degrees C. Rinse three times with PBS as above Incubate with secondary antibody (usually biotinylated) for 30 minutes Rinse three times (In this protocol they use ABC reagent) Incubate with ABC Reagent for 30 minutes Wash three times with PBS Rinse with 1% Triton X-100/PBS for 30 seconds Incubate with DAB solution for 1-3 minutes Rinse with distilled water Counterstain (if desired) with Hematoxylin for 1-3 minutes Rinse with distilled water Dehydrate, Clear with Xylene and mount with ageous mounting medium. I hope this helps, if it's totally off the mark for you, sorry. Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030910/4e84a302/attachment.htm From Histolady710 <@t> aol.com Wed Sep 10 15:34:00 2003 From: Histolady710 <@t> aol.com (Histolady710@aol.com) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Forceps Message-ID: <18.3501dc77.2c90e4b8@aol.com> What are some of the methods that you Histonetters use to heat forceps for embedding? We have used propane for years but have been having problems with the tanks leaking and I also feel they are unsafe. Thanks for your help. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030910/d0e4d395/attachment.htm From mari.ann.mailhiot <@t> leica-microsystems.com Wed Sep 10 15:52:52 2003 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Cleaning microtomes Message-ID: Joe Paraguard or mineral oil have been my choice. I have tried to get away from xylene as much as possible. Any cleaner such as 409 is fine. You just don't want the 409 solution to get on the inside of the microtome. It is better to spray the solution on a piece of gauze. In fact, I always spray the paragaurd on the gauze and then wipe my microtome. If I have to much paraguard on the microtome, I just take and put some alcohol on a piece of gauze and wipe the paraguard off of the microtome. 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 From ljb <@t> medicine.wisc.edu Wed Sep 10 16:14:03 2003 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] methyl green counterstain recipe please Message-ID: Dear Histonetters, I have someone visiting our lab for two months from Poland. I have a favorite commercial Methyl Green which I use as a counterstain in IHC. She would very much appreciate a good "homemade" protocol, because it would better suit her budget at home. If anyone is willing to share we'd both be very grateful. Thanks Cindy B LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 From gentras <@t> vetmed.auburn.edu Wed Sep 10 16:23:45 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] tissue processors Message-ID: <5.2.0.9.0.20030910162317.009f9060@mailhost.vetmed.auburn.edu> >Hello, please is anyone familiar with a tissue processor in today's market >that is comparable to the Autotechnicon Ultra II ? Any assistance with >sources, prices, and etc. will be greatly appreciated. Thanks, Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From cwscouten <@t> myneurolab.com Wed Sep 10 16:53:06 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Forceps Message-ID: A hot glass bead sterilizer would excellent for that application. See the link below. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=4 15001&catdesc=Surgical+Equipment&CatThreeID=51&CatOneID=2&subcatdesc=Ste rilizers&idsubcategory=10 Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 FAX 314 522 0377 cwscouten@myneurolab.com www.myneurolab.com -----Original Message----- From: Histolady710@aol.com [mailto:Histolady710@aol.com] Sent: Wednesday, September 10, 2003 3:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Forceps What are some of the methods that you Histonetters use to heat forceps for embedding? We have used propane for years but have been having problems with the tanks leaking and I also feel they are unsafe. Thanks for your help. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030910/192a0f4c/attachment.htm From RCHIOVETTI <@t> aol.com Wed Sep 10 17:32:12 2003 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] tissue processors Message-ID: <16.34f844e8.2c91006c@aol.com> In a message dated 09/10/2003 2:26:12 PM US Mountain Standard Time, gentras@vetmed.auburn.edu writes: << >Hello, please is anyone familiar with a tissue processor in today's market >that is comparable to the Autotechnicon Ultra II ? Any assistance with >sources, prices, and etc. will be greatly appreciated. Thanks, Atoska >> Atoska, The Leica TP 1020 is a modern version of a benchtop processor. Price will vary, depending on whether you get vacuum processing, fume filtration, etc. (they're options). Call Leica Customer Service at 1-800-248-0123 for details. Bob Chiovetti GTI Microsystems Leica Exclusive Regional Dealer Desert Southwest Region USA From AnthonyH <@t> chw.edu.au Wed Sep 10 18:37:36 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] de-staining immunos Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E02B@simba.kids> Amy, Diff-quik slides can be destained. Place in 1%acetic acid in ethanol. This will remove most of the dyes. As for IP stained slides, unless an ethanol soluble substrate is used (eg AEC), it may be very difficult to remove the DAB substrate. You could try 5 min treatment with KMnO4 followed by Oxalic acid, though I can't be certain this will work. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Amy Self [mailto:ASelf@gmhsc.com] Sent: Wednesday, 10 September 2003 20:02 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] de-staining immunos Does anyone out in histoland know if Immuno stained slides and/or air-dried Diff-Quik stained slides can be de-stained? If so could you please share with me this procedure. I know that H&E's can be de-stained but I wasn't sure about immunos and diff-quiks. Thanks for the help.... Amy Self Georgetown Memorial Hospital Georgetown, SC (phone) 843-527-7179 (fax) 843-520-7882 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 agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Wed Sep 10 18:50:09 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Scanner for microscope slides Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E02C@simba.kids> We use an Epson Perfection 2450 Photo Scanner. We place the glass slide in the 35mm holder, place it on the scanner (ensure that back light cover is off) and scan as you would a 35mm slide. We also use the scanner for EM negatives, 35mm photos and picture scanning for Grand Rounds etc. I will upload some examples to the Histonet Home page for you to look at. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Hawkins, Hal K. [mailto:hhawkins@utmb.edu] Sent: Thursday, 11 September 2003 0:49 To: 'Mikael Niku'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Scanner for microscope slides Meyer Instruments has modified a Microtek scanner for the purpose, see http://www.meyerinst.com/html/oem/pseiii/default.htm Hal Hawkins, UTMB Galveston (not a stockholder) -----Original Message----- From: Mikael Niku [mailto:mikael.niku@helsinki.fi] Sent: Wednesday, September 10, 2003 2:05 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Scanner for microscope slides Hello! I'm thinking of getting a film scanner capaple of scanning microscope slides. At least for Nikon Coolscan 4000 one can get a microscope slide adapter. Any experience of such a setup, or other recommendations? +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ + Mikael Niku + University of Helsinki, Dept. Basic Veterinary Sciences + URL: www.helsinki.fi/~mniku/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! (Gandhi) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Wed Sep 10 19:24:00 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Scanner for microscope slides Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E02F@simba.kids> Stan, The pics are regularly used at meetings. I find that rather than DPI, I aim for a file size of 300kB for emailing and powerpoint presentations. As for zooming, you would be surprised the level of detail you can zoom in on. Tony -----Original Message----- From: Stylli, Stanley [mailto:Stanley.Stylli@mh.org.au] Sent: Thursday, 11 September 2003 10:20 To: Tony Henwood Subject: RE: [Histonet] Scanner for microscope slides Importance: High Hi Tony... do you get reasonable images with this scanner to be able to show at presentations ? to zoom in ??? what DPI do you set the scanner at ? thanks Stan -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Thu 11/09/2003 09:50 To: histonet@pathology.swmed.edu Cc: Subject: RE: [Histonet] Scanner for microscope slides We use an Epson Perfection 2450 Photo Scanner. We place the glass slide in the 35mm holder, place it on the scanner (ensure that back light cover is off) and scan as you would a 35mm slide. We also use the scanner for EM negatives, 35mm photos and picture scanning for Grand Rounds etc. I will upload some examples to the Histonet Home page for you to look at. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Hawkins, Hal K. [mailto:hhawkins@utmb.edu] Sent: Thursday, 11 September 2003 0:49 To: 'Mikael Niku'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Scanner for microscope slides Meyer Instruments has modified a Microtek scanner for the purpose, see http://www.meyerinst.com/html/oem/pseiii/default.htm Hal Hawkins, UTMB Galveston (not a stockholder) -----Original Message----- From: Mikael Niku [mailto:mikael.niku@helsinki.fi] Sent: Wednesday, September 10, 2003 2:05 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Scanner for microscope slides Hello! I'm thinking of getting a film scanner capaple of scanning microscope slides. At least for Nikon Coolscan 4000 one can get a microscope slide adapter. Any experience of such a setup, or other recommendations? +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ + Mikael Niku + University of Helsinki, Dept. Basic Veterinary Sciences + URL: www.helsinki.fi/~mniku/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! (Gandhi) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ This email has been scanned for all viruses by the MessageLabs Email Security System. For more information on a proactive email security service working around the clock, around the globe, visit http://www.messagelabs.com ________________________________________________________________________ From ranahay <@t> fastmail.fm Wed Sep 10 20:17:49 2003 From: ranahay <@t> fastmail.fm (Rana Hay) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] H&E Message-ID: <20030911011749.ED4113B5F7@www.fastmail.fm> Hello, I am trying to trace information on the historical aspects of H&E ,especially the use of Haematoxylin.Are you able to help me?When was it introduced & by whom.etc Thank you, Rana hay -- Rana Hay ranahay@fastmail.fm -- http://www.fastmail.fm - IMAP accessible web-mail From djcarter <@t> dallastx.net Wed Sep 10 20:34:56 2003 From: djcarter <@t> dallastx.net (Dallas Nippert) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Forceps References: <18.3501dc77.2c90e4b8@aol.com> Message-ID: <005201c37804$ec072780$0ae8a641@djcarter> Purchase an embedding center. They have forceps wells and keep everything nice and warm. ----- Original Message ----- From: Histolady710@aol.com To: histonet@lists.utsouthwestern.edu Sent: Wednesday, September 10, 2003 3:34 PM Subject: [Histonet] Forceps What are some of the methods that you Histonetters use to heat forceps for embedding? We have used propane for years but have been having problems with the tanks leaking and I also feel they are unsafe. Thanks for your help. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030910/de9fed94/attachment.htm From roy.ellis <@t> imvs.sa.gov.au Wed Sep 10 20:40:36 2003 From: roy.ellis <@t> imvs.sa.gov.au (Roy Ellis) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] H&E In-Reply-To: <20030911011749.ED4113B5F7@www.fastmail.fm> Message-ID: <002501c37805$b279c440$a88a140a@imvs.sa.gov.au> Hi Rana You will find a paragraph on the history of Haematoxylin as a dye in histology if you visit http://adam.com.au/royellis then go to the histology page. You will find the information in the first paragraph of the Chapter on Haematoxylin and counterstains. Regards Roy mailto:roy.ellis@imvs.sa.gov.au > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Rana Hay > Sent: Thursday, 11 September 2003 10:48 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] H&E > > > Hello, > I am trying to trace information on the historical aspects of H&E > ,especially the use of Haematoxylin.Are you able to help me?When was it > introduced & by whom.etc > > Thank you, > Rana hay > -- > Rana Hay > ranahay@fastmail.fm > > -- > http://www.fastmail.fm - IMAP accessible web-mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jkiernan <@t> uwo.ca Wed Sep 10 23:50:24 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Re: H&E - history of References: <20030911011749.ED4113B5F7@www.fastmail.fm> Message-ID: <3F5FFF10.6A872ACA@uwo.ca> Haematoxylin was first used by CG Reichel (1758), as unmordanted logwood, to stain vessels in plant tissues. It was next mentioned in a microtechnique book by Quekett (1848). Bohmer (1865) first used haematoxylin with alum (haemalum, or hemalum if you're an American). Numerous mixtures followed, including that of the great Paul Ehrlich (1886). The first author to recognize the nature of "ripening" (oxidation of colourless haematoxylin to yellow haematein) was Mayer (1891). The first use of a haemalum in conjunction with an eosin was Wissowzky (1876). To put things in a historical context all these discoveries and nearly all the classical descriptions of cells and tissues predate the introduction of formaldehyde as a fixative (probably Hauser 1893, for bacterial cultures on gelatin). No, I don't carry all this stuff in my head! I've got a copy of the 10th edition of "Conn's Biological Stains" (2002). Chapter 2 by B. Bracegirdle (pp.15-21) is "The History of Staining." The references cited above about haematoxylin are in the book's common bibliography. The only one I've read myself is Ehrlich (1886), a half-page paper. I'm taking the others on trust, so you'll need to check them out yourself. The Hauser reference is also one not seen by me - cited from JF Walker's "Formaldehyde," 3rd ed p.577 (1964). There is a book, "History of Staining" by G Clark & FH Kasten, 3rd ed 1983 (Williams & Wilkins). It was sponsored by the Biological Stain Commission and is out of print. I don't own a copy, and didn't drive in to the library at 11pm before typing this answer. There's more information about the early days of H and E (separately and combined) in that book. Sorry I can't provide verified answers to your questions. Bracegirdle's chapter (easily available) and the Clark & Kasten book (if you can find it) will direct you to the primary sources. Do not quote anything in this casual email as any kind of "source." For all you know, I could have made it all up, just for the fun of deceiving people. I didn't, but you have only my word for that. Even if you believe that JA Kiernan is honest, this message could be from JOL Roger, who can easily send messages apparently from other people's email addresses. In good faith, I wish you well with your historical stainology. Check out the Biological Stain Commission web site: http://www.biostains.org This may have links to other people interested in historical aspects of staining. I'm not one of them. Fred Kasten is still academically active in the field of the history of staining. ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ______________________________________ Rana Hay wrote: > > Hello, > I am trying to trace information on the historical aspects of H&E > ,especially the use of Haematoxylin.Are you able to help me?When was it > introduced & by whom.etc > > Thank you, > Rana hay > -- > Rana Hay > ranahay@fastmail.fm From arme <@t> optonline.net Thu Sep 11 00:12:17 2003 From: arme <@t> optonline.net (Mark Sofferman) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] For Sale - Grossing Station and VIP 3000 In-Reply-To: <20030908170000.422.58069.Mailman@swlx167.swmed.edu> Message-ID: Email or call 201-833-1550. ARME From aep14 <@t> leicester.ac.uk Thu Sep 11 06:46:15 2003 From: aep14 <@t> leicester.ac.uk (Prior, A.E.) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] RE: OT. Agincourt Salute Message-ID: <6728E9270FDF8D44A41AF135DBB553E870339F@sumac.cfs.le.ac.uk> As an Englishman I feel I must correct Dick Paulson. The traditional English "Agincourt Salute" is performed by forming a V with the index and middle fingers ( a reversed peace sign if you like). These two fingers were used to draw the long bow. (Incidently it is possible to draw a bow using ones index and ring finger, though at a lower poundage so the removal of the middle finger would not be sufficient to stop an English archer) The use of only the middle finger and the phrase " giving the bird" seem to be of American origin, as my first encounters of these 'rituals' were via Hollywood movies. Now that this has been cleared up, we can revert to histological discussions. Oh, and can people please send entries as plain text as MIME formats are not read properly by my univerity supplied software. Andrew Prior University of Leicester UK From ctsmolde <@t> capeheart.uct.ac.za Thu Sep 11 06:45:00 2003 From: ctsmolde <@t> capeheart.uct.ac.za (Jenny Molde) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Biltong Message-ID: Hi everyone out there Is it necessary to fix biltong in formalin before processing it. WE would like to use it as a gram control. Many thanks. Jenny Molde Cardiovascular Research Unit University of Cape Town From ctsmolde <@t> capeheart.uct.ac.za Thu Sep 11 07:14:56 2003 From: ctsmolde <@t> capeheart.uct.ac.za (Jenny Molde) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Biltong Message-ID: Sorry everyone I forgot to mention that it is called beef jerky in the States. Here in South Africa we call it biltong. Many thanks. Jenny Molde From tissuearray <@t> hotmail.com Thu Sep 11 07:37:08 2003 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Tissue Arrayer Testing Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/8a8abefb/attachment.htm From Nancy.Walker <@t> sanofi-synthelabo.com Thu Sep 11 07:37:27 2003 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] lost ISH signal, OK now Message-ID: Remember this: Help histonetter! Can you imagine it's like "now you see it now you don't"! A vanishing act that's flipping me out!! The pb appears at the photographic emulsion stage, that is the films are great. It's our silver grains that once devllopped dissappear with time (ie 1-3 weeks depending on the strength of the signal). We haven't been able to pinpoint any change to our protocol. And sometimes we don't lose the signal! Is it VOODOO! That is we've changed from Kodak NBT emulsion to Amersham LM1, it's not changed the pb. We've had no change in products that we know of (ie devellopper and fixateur are KODAK LX24 , Xylene clearing with ENTELLAN mounting medium). I would appreciate a discussion on the actual process that could explain this loss and might help pinpoint the cause. Or has anybody experienced this. My psychological health and professional reputation depends on this! Please Help. Thanks for all your suggestions; Here is what I tried : 1.more thorough washing after fixation step of emulsion devellopment - this certainly helped but 10 days after developping I was still losing silvergrains with appearance of brown deposits 2. removing Xylene clearing during coverslipping, that is after ETOH steps, I airdried before mounting with ENTELLAN - this completely eliminated silver grain loss!!! 3. I tried replacing the ENTELLAN with EUKITT, but in the absence of xylene this wasn't necessary, I didn't try an aqueous mounting medium, this suggestion came after I started my trials. I put the pictures together from an trial that I did with a moderately abundant RNA target (1 week exposition), the loss is not very dramatic probably because of longer washes but imagine with a low-expressing RNA target. I asked Herb Hagler to post the images at : ( http://pathcuri1.swmed.edu). but I myself was unable to access this adress today. If anyone is interested I can send a small JPG file. thanks again, Nancy Walker Molecular Biology Scientist Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 From carmen_loiselle <@t> hotmail.com Thu Sep 11 07:50:27 2003 From: carmen_loiselle <@t> hotmail.com (carmen loiselle) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] (no subject) Message-ID: Hello everyone, Recently, we seem to have a recurrent problem with our antibody CD138, IHC done on parafffin section. The staining pattern is nuclear ! Anybody out there experiment the same thing. If so, do you have any idea what could be the problem, that you would be willing to share. Can it be a fixation problem, how ? Any advise or opinion would be greatly appreciated , Thank you all _________________________________________________________________ MSN Messenger : discutez en direct avec vos amis ! http://messenger.fr.msn.ca/ From mcauliff <@t> umdnj.edu Thu Sep 11 07:58:15 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] H&E In-Reply-To: <20030911011749.ED4113B5F7@www.fastmail.fm> References: <20030911011749.ED4113B5F7@www.fastmail.fm> Message-ID: <3F607167.1010304@umdnj.edu> You might get a copy of "History of Staining" by George Clark and Fred Kasten. The third edition was published in 1983 by Williams and Wilkins. I don't know if there is a more recent edition but it should not matter. Geoff Rana Hay wrote: >Hello, >I am trying to trace information on the historical aspects of H&E >,especially the use of Haematoxylin.Are you able to help me?When was it >introduced & by whom.etc > >Thank you, >Rana hay > > -- -- ********************************************** 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 g.lang <@t> bigfoot.de Thu Sep 11 08:51:04 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] forceps Message-ID: <009201c3786b$c2088d40$0d12a8c0@SERVER> Hi! We use a so called "Heidelberg-Pinzette" (=forcep). It works with current and a transformer. On the place, you hold it, its isolated. You can plug in two sizes of forceps an it steadily holds temperature. Gundi Akh Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/668a6705/attachment.htm From HornHV <@t> archildrens.org Thu Sep 11 09:28:45 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Computer system question Message-ID: Sheila, We have Meditech. You don't have to have spec. numbers assigned at entry. It can just assign a req. number and you can still assign a specimen number upon receipt in the lab. I would not let it assign my spec. numbers. Hazel Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Tapper, Sheila [mailto:STapper@slhduluth.com] Sent: Tuesday, September 09, 2003 12:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Computer system question To anyone out there who uses an LIS system - especially Meditech - do you use outside order entry into your system? We are investigating the usage of this function. The downfall - accession numbers will be assigned at the time of entry by the clinic, or outside client - not at the time of receipt into the lab. Does anyone use a system, or function like this in their facility? Sheila Tapper 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. Arkansas Children's Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/1b6a26cd/attachment.htm From LMARGRAF <@t> childmed.dallas.tx.us Thu Sep 11 09:24:59 2003 From: LMARGRAF <@t> childmed.dallas.tx.us (LINDA MARGRAF MD) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Tony Henwood's scanned pictures Message-ID: Dear Histonetters: Two examples of Tony's scanned images will be available on the www.Histonet.org website soon. Remember if you have images that you want the Histonet community to be able to see, go to that website and email them the images. The Histonet email server won't allow attachments like pictures to prevent viruses from going through the list. Thanks Linda M Histonet administrator From g.lang <@t> bigfoot.de Thu Sep 11 09:30:22 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Biltong References: Message-ID: <010301c37871$3c4fdbd0$0d12a8c0@SERVER> We use pig-liver as a control for collagen fiber staining and fix it with formalin and process it in the same way as all probes. Gundi Akh Linz, Austria ----- Original Message ----- From: "Jenny Molde" To: Sent: Thursday, September 11, 2003 1:45 PM Subject: [Histonet] Biltong > Hi everyone out there > Is it necessary to fix biltong in formalin before processing it. WE > would like to use it as a gram control. Many thanks. > > Jenny Molde > Cardiovascular Research Unit > University of Cape Town > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From mari.ann.mailhiot <@t> leica-microsystems.com Wed Sep 10 15:28:44 2003 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] cleaning microtomes Message-ID: Joe Paraguard or mineral oil have been my choice. I have tried to get away from xylene as much as possible. Any cleaner such as 409 is fine. You just don't want the 409 solution to get on the inside of the microtome. It is better to spray the solution on a piece of gauze. In fact, I always spray the paragaurd on the gauze and then wipe my microtome. If I have to much paraguard on the microtome, I just take and put some alcohol on a piece of gauze and wipe the paraguard off of the microtome. 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 "Galbraith, Joe" To: "'RCHIOVETTI@aol.com'" , Jmrwalker@aol.com, Sent by: histonet@lists.utsouthwestern.edu histonet-admin@lists.utsouth cc: western.edu Subject: RE: [Histonet] cleaning microtomes 09/09/2003 03:03 PM Histonetters: Most people in my lab use Paraguard to clean microtomes but some prefer 409 (Misty Breeze has a nice smell). The 409 cleaner seems to do as good of a job as Paraguard and costs less and smells better. Joe Galbraith -----Original Message----- From: RCHIOVETTI@aol.com [mailto:RCHIOVETTI@aol.com] Sent: Tuesday, September 09, 2003 1:15 PM To: Jmrwalker@aol.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] cleaning microtomes In a message dated 09/09/2003 7:51:39 AM US Mountain Standard Time, Jmrwalker@aol.com writes: << After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. Thanks! >> I see mostly Paragard being used in our customers' labs, and I agree with the other posters, if the "slick" surface is a problem after the Paragard treatment you can then wipe down with a little ethanol or isopropanol. With time xylene can affect the surfaces of the microtome's external parts, especially if used in excess. Lots of the microtome parts have anodized (plated) surfaces that are either black or clear, and you'd think they would hold up to just about any kind of abuse. But they will definitely break down and corrode with prolonged or repeated exposure to xylene. This can lead to pitting and discoloration of the surfaces. If you *do* use xylene, use it sparingly and be sure to wear gloves. I personally don't like the stuff. Try to use a substitute instead. First, use a brush or gauze pad to remove the majority of the paraffin. Then use only enough xylene or xylene substitute to barely moisten a gauze 4x4 or a tissue. Wipe off the excess immediately with another dry gauze or tissue. You can also follow this with an alcohol wipe-down like you would use with Paragard if the surface feels slick or oily after you clean it. Hope this helps! Bob Chiovetti GTI Microsystems Leica Regional Dealer Desert Southwest Region, USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ This email has been scanned for all viruses by the MessageLabs Email Security System. For more information on a proactive email security service working around the clock, around the globe, visit http://www.messagelabs.com ________________________________________________________________________ From Noel.Clark <@t> ORTHO.UAB.EDU Thu Sep 11 10:18:40 2003 From: Noel.Clark <@t> ORTHO.UAB.EDU (Noel D. Clark) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Polyurethane processing Message-ID: <85F6C7A1330E794DB8540AFD001CC77E0D66EA@ROSCO> Any logistics comments on processing polyurethane material in either paraffin or plastics? Thanks. noel Noel Clark, M.A., HTL (ASCP) 1919 7th Ave South Orthopaedic Research Laboratory Center for Metabolic Bone Disease University of Alabama at Birmingham Birmingham, Alabama 35294 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/525469b5/attachment.htm From Ian.Bernard <@t> LACKLAND.AF.MIL Thu Sep 11 10:14:47 2003 From: Ian.Bernard <@t> LACKLAND.AF.MIL (Bernard Ian R SSgt 59 CRES/MSROP) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. Message-ID: <588C513CC306D611A2910003479604F9065917F2@fsmpls17.whmc.af.mil> Paint brushes that we use at the microtome. Digital desk top scale to weigh whole animal parts, organs, rats etc., to be used in necropsy room. Thanks SSgt Bernard . -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/3c05d83c/attachment.htm From jqb7 <@t> cdc.gov Thu Sep 11 10:27:46 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. Message-ID: Fisher sells the brushes. Just go to www.fishersci.com and do a search for camel hair brushes. You can then look at the different sizes and select what works for you. They also have Ohaus brand digital desk top scales. -----Original Message----- From: Bernard Ian R SSgt 59 CRES/MSROP [mailto:Ian.Bernard@LACKLAND.AF.MIL] Sent: Thursday, September 11, 2003 11:15 AM To: Histonet (E-mail) Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. Paint brushes that we use at the microtome. Digital desk top scale to weigh whole animal parts, organs, rats etc., to be used in necropsy room. Thanks SSgt Bernard . -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/cdbb55a9/attachment.htm From tpmorken <@t> labvision.com Thu Sep 11 10:34:16 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CB5C@usca0082k03.rallansci.apogent.com> SSgt Bernard needs: "Paint brushes that we use at the microtome." The best source I can think of for good a good brush is to go to the Leica booth at the NSH meeting and pick one up for free - just one benefit of going to the meeting. I get a new one every year! "Digital desk top scale to weigh whole animal parts, organs, rats etc., to be used in necropsy room." Fisher Scientific, VWR (or whatever it is now), CMS, etc all have such scales. Tim Morken . -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/d668b8ac/attachment.htm From algranth <@t> u.arizona.edu Thu Sep 11 10:38:20 2003 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. In-Reply-To: <588C513CC306D611A2910003479604F9065917F2@fsmpls17.whmc.af. mil> Message-ID: <4.3.2.7.2.20030911082928.00c8da78@algranth.inbox.email.arizona.edu> Ian, Call Mopec for the scale - they should have something that you can use. Necropsy stuff is their line of business. I just got a catalog yesterday from them and they have several scales in it on special! Brushes...what do you want to do with them? For cleaning I use a brush I get at the office supply store. It is called "Crayola So Big" No. 208. It makes a great brush for sweeping away those paraffin shavings. You can't put it in xylene since the handle is plastic but for a dry brush it is wonderful - and cheap. If you are using the brush to brush down ribbons or sections then Crayola makes smaller brushes that can be used for this too. If you have an office supply catalog browse through it and see if they carry them. If not...go to Michaels or a JoAnn Fabric and Crafts. They have great brushes for painting on fabric that I love for frozens but they have larger brushes for cleaning too. Good luck. Andi Grantham At 10:14 AM 9/11/2003 -0500, you wrote: >Paint brushes that we use at the microtome. >Digital desk top scale to weigh whole animal parts, organs, rats etc., to >be used in necropsy room. > > >Thanks >SSgt Bernard > > >. ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From jqb7 <@t> cdc.gov Thu Sep 11 10:40:44 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. Message-ID: Do you mean the small type of brush for picking up sections on the waterbath or the type for cleaning around the microtome? -----Original Message----- From: Bernard Ian R SSgt 59 CRES/MSROP [mailto:Ian.Bernard@LACKLAND.AF.MIL] Sent: Thursday, September 11, 2003 11:15 AM To: Histonet (E-mail) Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. Paint brushes that we use at the microtome. Digital desk top scale to weigh whole animal parts, organs, rats etc., to be used in necropsy room. Thanks SSgt Bernard . -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/96017416/attachment.htm From SHargrove <@t> urhcs.org Thu Sep 11 10:47:54 2003 From: SHargrove <@t> urhcs.org (SHargrove@urhcs.org) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] mail Message-ID: How do I stop getting all this mail. I tried to go to the site listed but it said that it did not have my address in it. Susie Hargrove Histology 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 intented 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 abright <@t> brightinstruments.com Thu Sep 11 10:58:20 2003 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. Message-ID: We sell microtome brushes also anti-static ones too. Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Bernard Ian R SSgt 59 CRES/MSROP [mailto:Ian.Bernard@LACKLAND.AF.MIL] Sent: 11 September 2003 16:15 To: Histonet (E-mail) Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. Paint brushes that we use at the microtome. Digital desk top?scale to weigh whole?animal parts, organs, rats etc., to be used in necropsy room. ? ? Thanks SSgt Bernard ? ? . From juan.gutierrez <@t> christushealth.org Thu Sep 11 11:41:07 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. Message-ID: Home Depot has the brushes with the natural fiber that you can put in xylene, the ones with the wooden handles. They cost about $00.59./ea For the scale I too would try either Fisher or MOPEC. Good luck. Juan C. Gutierrez, HT(ASCP) -----Original Message----- From: Bernard Ian R SSgt 59 CRES/MSROP [mailto:Ian.Bernard@LACKLAND.AF.MIL] Sent: Thu 9/11/2003 10:14 AM To: Histonet (E-mail) Cc: Subject: [Histonet] Looking to buy following Histo item: Need source, tel# etc. Paint brushes that we use at the microtome. Digital desk top scale to weigh whole animal parts, organs, rats etc., to be used in necropsy room. Thanks SSgt Bernard . From g.lang <@t> bigfoot.de Thu Sep 11 12:44:45 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] haematoxylin Message-ID: <000c01c3788c$637a2650$0d12a8c0@SERVER> In our lab we have the discussion, if staining with Mayer's or Harris' or Weigert's hematoxylin leads to an endpoint-staining. I have read, that only Weigert stains to an endpoint. The others can be overstained. Who can explain and help? Gundi Akh, Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/6b069209/attachment.htm From gareth.davis <@t> Vanderbilt.Edu Thu Sep 11 12:54:54 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] haematoxylin Message-ID: <082C721AF78DB34983E8BA2CD085462105C663@mailbe07> I'm trying to find a good Hematoxylin for good H&E results, is that what your discussion was about? Gill's is best for counterstaining IHC sections, in my opinion. Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -----Original Message----- From: Gudrun Lang [mailto:g.lang@bigfoot.de] Sent: Thu 9/11/2003 12:44 PM To: Histonetliste Cc: Subject: [Histonet] haematoxylin In our lab we have the discussion, if staining with Mayer's or Harris' or Weigert's hematoxylin leads to an endpoint-staining. I have read, that only Weigert stains to an endpoint. The others can be overstained. Who can explain and help? Gundi Akh, Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/92eecf63/attachment.htm From bryand <@t> netbistro.com Thu Sep 11 13:19:57 2003 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] H&E References: <20030911011749.ED4113B5F7@www.fastmail.fm> Message-ID: <007f01c37891$58b2a6c0$9c70c2cf@bryand> There is a very interesting article on hematoxylin and the other logwoods at http://waynesword.palomar.edu/ecoph4.htm. The article describes how logwood dyes were introduced to Europe, and how they resulted in the creation of two countries. Photos of the trees are included. Bryan Llewellyn ----- Original Message ----- From: "Rana Hay" To: Sent: Wednesday, September 10, 2003 6:17 PM Subject: [Histonet] H&E > Hello, > I am trying to trace information on the historical aspects of H&E > ,especially the use of Haematoxylin.Are you able to help me?When was it > introduced & by whom.etc > > Thank you, > Rana hay > -- > Rana Hay > ranahay@fastmail.fm > > -- > http://www.fastmail.fm - IMAP accessible web-mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From cfavara <@t> niaid.nih.gov Thu Sep 11 13:32:11 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] ASCP Message-ID: Gundi, American Society of Clinical Pathologists. They like to put their stamp of approval on lots of people sometimes beneficial sometimes not! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Gudrun Lang [mailto:g.lang@bigfoot.de] Sent: Wednesday, September 10, 2003 9:56 AM To: Histonetliste Subject: [Histonet] ASCP Is ASCP your professional organisation in USA? Gundi -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/e354954c/attachment.htm From dmccaig <@t> ckha.on.ca Thu Sep 11 14:39:18 2003 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Auto stainer program schedules Message-ID: We use a LEICA Autostainer XL and Surgipath Stains (Gil III haematoxylin and Eosin). The stains used to be filtered daily but it rendered the slides too dark so the filtering was reduced to weekly. I want to revert to daily filtering but am getting some resistance. We use 90 seconds in the haematoxylin followed by 1 minute in Scott's Tap Water Substitute. Can anyone share their time schedule for routine H&E's. Do you dilute your stains? How frequently are they changed or are they just topped up? Thanks in advance Diana McCaig, R.T. Charge Tech, Histology Chatham Kent Health Alliance 519-352-6401 (3347) From Don.Birgerson <@t> leica-microsystems.com Thu Sep 11 15:12:36 2003 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Polyurethane processing Message-ID: Hi Noel, Most of the time materials such as plastics are cut without embedding. The heat and chemical interaction will interfere with pol or IR examination of the material. I am not sure why your cutting this material but if you can give me a call we can discuss the options. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Noel D. Clark" To: "'histonet@lists.utsouthwestern.edu'" Sent by: histonet-admin@lists.utsouth cc: western.edu Subject: [Histonet] Polyurethane processing 09/11/2003 10:18 AM Any logistics comments on processing polyurethane material in either paraffin or plastics? Thanks. noel Noel Clark, M.A., HTL (ASCP) 1919 7th Ave South Orthopaedic Research Laboratory Center for Metabolic Bone Disease Universityof Alabamaat Birmingham Birmingham, Alabama35294 From rusalced <@t> utmb.edu Thu Sep 11 15:04:52 2003 From: rusalced <@t> utmb.edu (Salcedo, Rudy) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Seal Whiskers Message-ID: <2456B984836CD611B61500D0B79E87E00B76D750@exchange2> Good afternoon, I'm looking for information on how to process seal whiskers, for paraffin embedding any information would be appreciated. Thanks in advance Rudy Salcedo rusalced@utmb.edu -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/ff329221/attachment.htm From jdecker <@t> vectorlabs.com Thu Sep 11 15:30:12 2003 From: jdecker <@t> vectorlabs.com (Judy Decker) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Antibody Availability Message-ID: <007601c378a3$80c878d0$0601a8c0@pacbell.net> Dear Histonetters: We would like to clarify some misinformation circulating recently regarding the availability of Novocastra products. A Histonet posting by Elliot G. Osterhaus a few weeks ago incorrectly indicated Novocastra products were no longer available through Vector Laboratories. We attempted to contact this individual privately to correct this misinformation. To date, he has not responded to us. Vector Laboratories continues to be the authorized distributor for all Novocastra antibodies and products in the United States, Canada, and the UK, as we have been for over the last 10 years. Sincerely, Judy Decker -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/a9fa8813/attachment.htm From cfranci <@t> rigel.com Thu Sep 11 15:31:45 2003 From: cfranci <@t> rigel.com (Christian Franci) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Axl IHC for frozen sections Message-ID: I got several questions and would much appreciate any help you can give me. 1) Heard that pickling tissues over night (in 30% sucrose at 4 degrees C) prior to freezing them works well to limit the size of ice crystals forming in the specimens. MY concern is over degradation of my tissue during this time. Since, I'm working with precious tumor xenographs, should I be concerned that this might happen? Any way to prevent this? Also, after the sucrose bath, is there a difference as to how one freezes the samples? 2)Lastly, I need to find antibodies specific for staining MOUSE fibroblasts as well as for HUMAN Axl (aka: UFO,ARK) Anyone have any experience with either and, can recommend a good supplier? The less species cross-reactivity the better but, I'd settle for less non specific staining. Would prefer monoclonals but, if there are good polyclonals I'll gladly try them out so long as I can get an idea as to how much to use to stain my sections. Thank you all for all your help with the CD31 staining problems I was having. Got some great replies and sage advice and I'm happy to report that it all work out well. Thanks again, Cheers Chris From gentras <@t> vetmed.auburn.edu Thu Sep 11 15:48:39 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] counterstain for ABC-HRP Message-ID: <5.2.0.9.0.20030911154133.009fab00@mailhost.vetmed.auburn.edu> Hello, Is anyone using a counterstain other than methyl green that stains neurons induced from bone marrow stem cells with the Vectastain ABC-HRP system? Thanks, Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From jsdiallo <@t> yahoo.com Thu Sep 11 15:52:09 2003 From: jsdiallo <@t> yahoo.com (Jean-Simon Diallo) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Antibody against activated JunD Message-ID: <20030911205209.90286.qmail@web11506.mail.yahoo.com> Hello, my name is Jean-Sim. I was just wondering if anyone has come across a good anti-human activated (phosphorylated) JunD for use in immunohistochemistry on formalin-fixed paraffin embedded tissues. Thanks in advance. __________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com From SJones <@t> cvm.tamu.edu Thu Sep 11 17:56:34 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Seal Whiskers Message-ID: Hi Rudy, Do these whiskers have any soft tissue attached or are they just the whiskers? How are things at UTMB, my old stomping grounds? Are you working in Linda's lab? Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Salcedo, Rudy" 09/11/03 03:04PM >>> Good afternoon, I'm looking for information on how to process seal whiskers, for paraffin embedding any information would be appreciated. Thanks in advance Rudy Salcedo rusalced@utmb.edu From Mauricio.Morais <@t> tufts.edu Thu Sep 11 18:24:38 2003 From: Mauricio.Morais <@t> tufts.edu (Mauricio S. Morais) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Manipulation time of human muscle biopsies and formation of freezing artifacts. Message-ID: <3F610436.B10ACDE3@tufts.edu> Hello Histonet community. Because I'm new into the histology "word" I've been facing some unexpected (to me) challenges in my actual Laboratory work. My background is originally from Organic and Analytical Chemistry, Teaching, and a bit of Biochemistry (my MSc degree). I'm kind of new in the country too (been here less than 2 years) and English is not my primary language, so please accept my apologies for any errors within this text. I'm assign to reorganize and analyze human muscle biopsies, stored in boxes distributed among 4 racks inside a Cryoplus cryofreezer. Seeking for information on how to work safely and at the same time not compromise the integrity of the stored samples, I found the Histonet Histosearch archives. They were very helpful providing answer about most of the common issues related with formation of freezing artifacts and possible solutions to the problem at different stages the samples can generate it, from the biopsy to the cryostat slice machine. Because of the enormous amount of samples that'll need to be handle at one time during the reorganization of our cryofreezer, an issue about "time" became critical (in my opinion) raising some questions: 1-How long can a box (full of biopsies), or the hole rack be kept out of the cryofreezer, while I'm updating data and re-arranging boxes between racks, without compromise the biopsies and generate (and/or enhancing the chances of creating) freezing artifacts? 2-Does any damage may occur if I have several biopsies floating inside a dewar container while I'm making notes or grouping them into other dewar containers to a new order? A website information (from www.asymptote.co.uk) says that tissue inside straws immersed in liquid Nitrogen when exposed to air at room temperature have their temperature raised to -50C in 40 seconds. Thawing may start around -10C or early. I was told they (racks, boxes) could be out for about 5 minutes before some damage can start to affect the biopsies inside it. None of them been hold by hands during all this time, but on the bench top (boxes) or floor (racks) while the work is done. I was also told that 5 minutes is away too long... For me it seems impossible to safely remove a rack (containing a dozen boxes) from the cryofreezer, make notes or, for example, remove one box from the rack to add a new sample and return it to the cryofreezer in 40 seconds. Reminding that I'm working with cryogloves over my lab. gloves... I was never able to open a box without remove the cryogloves first. Maybe I need more training... I would greatly appreciate any comments from this expert community to help me solve my immediate problems (above) and improve my knowledge and safety at work place. Thank you all. Mauricio S. Morais Senior Research Technician Nutrition Exercise Physiology and Sarcopenia Laboratory (NEPS) TUFTS University Jean Mayer USDA Human Nutrition Research Center on Aging 711 Washington Street, Rm 436 Boston, MA 02111 (617) 556-3226 -------------- next part -------------- A non-text attachment was scrubbed... Name: Mauricio.Morais.vcf Type: text/x-vcard Size: 1220 bytes Desc: Card for Mauricio S. Morais Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030911/9d4058ec/Mauricio.Morais.vcf From j.browne <@t> auckland.ac.nz Thu Sep 11 20:53:05 2003 From: j.browne <@t> auckland.ac.nz (Jeremy Browne) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] validating oil red O Message-ID: Anyone out there with an idea of oil red O staining could be validated? We need to set up a range of fat concentrations on slides, stain and then check to see that there is a linear response relationship between fat concentration and staining intensity but because fat melts and slides off slides this is not easy. However, I imagine that someone out there is histoworld has solved this problem - perhaps with an intermediate substance to which the fat adheres? Thanks Jeremy Browne Liggins Institute University of Auckland NZ (09) 373 7599 extn 86444 j.browne@auckland.ac.nz -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/c0a2643e/attachment.htm From jkiernan <@t> uwo.ca Thu Sep 11 23:43:23 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] Re: counterstain for ABC-HRP (Neurons) References: <5.2.0.9.0.20030911154133.009fab00@mailhost.vetmed.auburn.edu> Message-ID: <3F614EEB.19F9F3FF@uwo.ca> If the HRP is detected with regular DAB, giving a brown product, you can use a blue cationic dye to counterstain neurons. A thiazine dye is easier to use than "methyl green." (That dye that has been deliberately sold under a wrong name for 30+ years; it's really ethyl green, which is much better; old timers can tell you why - but I'm getting off-topic). Toluidine blue is my favourite thiazine; use it at 0.5%, pH 3 to 4. Azure A is just as good. So, probably, are the other azures and thionine, but I've not used them myself for this purpose. >From your summary of the research, I assume that you are working with monolayers of cultured cells. If the cells are on plastic, you probably won't be able to use a resinous mounting medium. (Its solvent is likely to dissolve the plastic.) Aqueous mounting media generally extract basic dyes ("bleeding"), so you will need to use a trick to render the blue dye insoluble in water. It's an old one that dates back to Paul Ehrlich in the 1880s. When the counterstain looks right under a X10 objective, in a preparation that has been rinsed in distilled water after staining, pour on a 5% aqueous solution of ammonium molybdate and leave it there for 5 minutes. Wash in water and mount. The molybdate complexes (salts?) of the thiazine dyes (methylene blue, toluidine blue, azures etc) are insoluble in water, acids, alcohols etc. I have used this strategy to do Van Gieson (pH 1) after Nissl staining with toluidine blue at pH 4. If the neurons are not stained by your immunohistochemical method, they will have strongly blue cytoplasm. Cells other than neurons will, for the most part, have only their nuclei blue. If you have cells with high rRNA content that are not neurons, they will have blue cytoplasm, but their shapes will be very different. If your neurons are immunopositive, they will be blue+brown - not a lovely combination. You asked for a counterstain, so I assume that the immunostain is for cells other than neurons. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ___________________________ "Atoska S. Gentry" wrote: > > Hello, Is anyone using a counterstain other than methyl green that stains > neurons induced from bone marrow stem cells with the Vectastain ABC-HRP > system? Thanks, Atoska > > Atoska S. Gentry B.S., HT(ASCP) > Research Assistant III > Scott-Ritchey Research Center > College of Veterinary Medicine > Auburn University, AL 36849 > Phone# (334)844-5579 Fax# (334)844-5850 ____________________________ From jluis.palazon <@t> icman.csic.es Fri Sep 12 02:50:23 2003 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] sudan black staining Message-ID: <20030912075023.375C247E4E@perceval.uca.es> Dear List-members Thanks for the answers and advise about sudan black staining. I will cut and stain the samples again (fortunately I kept the blocks in the freezer¡) and read the slides immediately to avoid fading. I will also restain the faded slides and see what happens. José Luis From jluis.palazon <@t> icman.csic.es Fri Sep 12 03:08:31 2003 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] fading times Message-ID: <20030912080831.46E7A46DE9@perceval.uca.es> Dear Histonetters After my bad experience with sudan black staining, a question that I think will be of value for everyone that as me are new to the histochemistry world. I have found the list very instructive and of much value for me, and everyday I learn new things with the questions and answers you post to the list. My question is, In your experience in the different histochemistry methods, i.e. for carbohidrate, proteins, lipids, enzimes) what is the maximun period of time a slide must be read before any change in the staining (fading) can alter the results. Maybe each member can share his(her) experience with different staining methods (PAS, Alcian blue, lisine, and other aminoacids, different lipid reactions, phosphatases, etc). greetings José Luis From lpwenk <@t> mail.netquest.com Fri Sep 12 04:22:18 2003 From: lpwenk <@t> mail.netquest.com (Lee & Peggy Wenk) Date: Fri Sep 16 15:21:51 2005 Subject: [Histonet] ASCP References: Message-ID: <006301c3790f$5d4b6b80$8732fea9@hppav> ASCP has changed it's name. It is now the American Society for Clinical Pathology. The goal of changing it from "of . . . Pathologists" to "of . . . Pathology" was to reflect that this organization is made up of many different people from pathology - pathologists, residents, and all laboratory technical staff. This organization does the certification exams for many of the laboratory professionals (histotechs, med techs, cytotech, phlebotomists, etc.). In addition, each group has their own organization that is more specific to their field, such as the National Society for Histotechnology (NSH) is specifically for histotechs working in the field of histotechnology. ASCP and NSH often work together on projects, and histotechs often belong to both groups. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI ----- Original Message ----- From: Favara, Cynthia (NIH/NIAID) To: 'Gudrun Lang' ; Histonetliste Sent: Thursday, September 11, 2003 2:32 PM Subject: RE: [Histonet] ASCP Gundi, American Society of Clinical Pathologists. They like to put their stamp of approval on lots of people sometimes beneficial sometimes not! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Gudrun Lang [mailto:g.lang@bigfoot.de] Sent: Wednesday, September 10, 2003 9:56 AM To: Histonetliste Subject: [Histonet] ASCP Is ASCP your professional organisation in USA? Gundi -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/d6e66c39/attachment.htm From g.lang <@t> bigfoot.de Fri Sep 12 05:13:39 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] validating oil red O References: Message-ID: <003b01c37916$8b2205d0$0d12a8c0@SERVER> Don't you use frozen cuts with the fat-staining? ----- Original Message ----- From: Jeremy Browne To: 'histonet@lists.utsouthwestern.edu' Sent: Friday, September 12, 2003 3:53 AM Subject: [Histonet] validating oil red O Anyone out there with an idea of oil red O staining could be validated? We need to set up a range of fat concentrations on slides, stain and then check to see that there is a linear response relationship between fat concentration and staining intensity but because fat melts and slides off slides this is not easy. However, I imagine that someone out there is histoworld has solved this problem - perhaps with an intermediate substance to which the fat adheres? Thanks Jeremy Browne Liggins Institute University of Auckland NZ (09) 373 7599 extn 86444 j.browne@auckland.ac.nz -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/9d9c5003/attachment.htm From histo <@t> skm.org.pk Fri Sep 12 08:05:07 2003 From: histo <@t> skm.org.pk (Histology) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] test Message-ID: Test Message Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk From ttroyer <@t> pcllab.com Fri Sep 12 08:07:26 2003 From: ttroyer <@t> pcllab.com (Travis Troyer) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] TESTICULAR BIOSY Message-ID: <004501c3792e$d0957c60$6601010a@Peterson.local> Our lab is receiving a testicular biopsy that will be received in Bouin's Solution. We routinely place specimans in 10% NBF after grossing. So, my question is how long should the biopsy stay in the Bouin's solution for proper fixation before placing into 10% NBF? Thanks! -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/799037fc/attachment.htm From carolb <@t> mail.phys.mcw.edu Fri Sep 12 08:18:00 2003 From: carolb <@t> mail.phys.mcw.edu (Carol Bobrowitz) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Y-1 antibody Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A957C@thor.phys.mcw.edu> Has anyone out there worked with Y-1? If so, please contact me. 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 From mari.ann.mailhiot <@t> leica-microsystems.com Fri Sep 12 08:40:54 2003 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] (no subject) Message-ID: Diana How often you change solutions or stains on any stainer depends on the amount of slides you are staining. It is never a good idea to top off or add to a stain more than once. It is much better to completely change the stain out whenever possible. If you keep on adding fresh stain to stain already used, and never completely change the stain, your stain is always diluted. It becomes a situation of "it is not really new and not it is not really old", it is just diluted. Sometimes small institutions only change stains once a week. In that situation you will probably add some new stain to the old once a week. Just make sure at the end of a week you throw the stain away and start with fresh stain on Monday. It is always a good idea to run a couple of control slides before you start a staining run everyday. Check your slides and see how your stain really looks. As far as filtering stains, it depends on the manufacturer. It also depends on how long you use that stain on the stainer and how many slides you stain. If have any questions just send me an email. 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 From rusalced <@t> utmb.edu Fri Sep 12 08:37:20 2003 From: rusalced <@t> utmb.edu (Salcedo, Rudy) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Seal Whiskers Message-ID: <2456B984836CD611B61500D0B79E87E00B76D753@exchange2> Hi Sara, Yes I work with Linda, at UTMB- research histopathology Core/Sealy Center for environmental health & medicine. Yes the whiskers have a white soft tissue around it we are just not sure how to process and Linda mention your name. She is sending you an e-mail direct. Rudy Salcedo UTMB-Galveston 409-747-0735 -----Original Message----- From: Sarah Jones [mailto:SJones@cvm.tamu.edu] Sent: Thursday, September 11, 2003 5:57 PM To: histonet@pathology.swmed.edu; rusalced@utmb.edu Subject: Re: [Histonet] Seal Whiskers Hi Rudy, Do these whiskers have any soft tissue attached or are they just the whiskers? How are things at UTMB, my old stomping grounds? Are you working in Linda's lab? Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Salcedo, Rudy" 09/11/03 03:04PM >>> Good afternoon, I'm looking for information on how to process seal whiskers, for paraffin embedding any information would be appreciated. Thanks in advance Rudy Salcedo rusalced@utmb.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dwalker <@t> selway.umt.edu Fri Sep 12 09:06:51 2003 From: dwalker <@t> selway.umt.edu (David Walker) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Used microtome Message-ID: Hi all- I'm looking for a good used microtome to add to our department- does any one have an extra just sitting around? Please email me direct with any information. Thanks- Dave David Walker, Staff Scientist CEHS/CSFNS School of Pharmacy and Biomedical Science University of Montana, Missoula Missoula MT From Lynne.Bell <@t> hitchcock.org Fri Sep 12 09:17:52 2003 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] TESTICULAR BIOSY Message-ID: Travis, In our lab we fix testicular biopsies in Bouin's solution for 2-3 hours, wash in several changes of 70% alcohol to remove the picric acid and then place in 10% NBF for processing. The removal of picric acid from the tissue is necessary to insure proper staining. Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/7a7ff1bd/attachment.htm From jkiernan <@t> uwo.ca Fri Sep 12 09:58:28 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] TESTICULAR BIOSY References: <004501c3792e$d0957c60$6601010a@Peterson.local> Message-ID: <3F61DF14.BFC860DD@uwo.ca> It doesn't make sense to transfer a specimen from Bouin to a neutral formaldehyde solution. The usual way to use Bouin is to fix overnight and then transfer the specimens to 70% or 95% alcohol - several changes, to extract as much yellow colour (picric acid) as is reasonably possible. Then complete the dehydration, clear, infiltrate with wax and embed in the usual way. The tissue will be properly fixed after only a few hours in Bouin - a major difference from neutral formaldehyde. The nuclear morphology will be clearer than you'll ever see after formaldehyde alone, and connective tissue stains (trichromes etc) will give really bright colours. ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ____________________________ > Travis Troyer wrote: > > Our lab is receiving a testicular biopsy that will be > received in Bouin's Solution. We routinely place > specimans in 10% NBF after grossing. So, my question > is how long should the biopsy stay in the Bouin's > solution for proper fixation before placing into 10% > NBF? > Thanks! From gentras <@t> vetmed.auburn.edu Fri Sep 12 10:15:22 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] nuclear counterstain Message-ID: <5.2.0.9.0.20030912100039.00a08df0@mailhost.vetmed.auburn.edu> Hello again, guess I need to rephrase my initial inquiry on the counterstain to use with the Vectastain ABC-HRP kit. But, first special thanks to Dr. John Kiernan. He always replies with such informative and thorough remarks. Please pardon my partial entry of facts. This inquiry is on behalf of a couple of colleagues of mine. And yes their work was with monolayers of cultured bone marrow cells. However, the HRP was detected with the Vector VIP substrate ( for which Vector Labs does not reveal the contents) and yielded neurons that are purple in color. Therefore, we need something like acridine orange which stains DNA. Has anyone had any experience/success using acridine orange or something similar with this system that yields the same results? Thanks, Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From g.lang <@t> bigfoot.de Fri Sep 12 10:19:11 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] compliment Message-ID: <002801c37941$47026400$0d12a8c0@SERVER> Dear histonetters! I have to pay you a compliment. I am so happy, that I found some people with the same interest in acquireing the knowledge about histotechnic. (and histotechs with such a profound knowledge). The Histo-instructions at my medical school were a little poor. So if I want to become a good histotech, I have to do it on my own. The colleges in my lab consider me a little bit of mad, when I deal with the job-issues at home. So I have to moan about it a little bit. (Do you understand me?) Please don't mind my bad English. Again, thank you Gudrun Lang General Hospital, Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/e4a1c9b3/attachment.htm From amarusk1 <@t> FAIRVIEW.ORG Fri Sep 12 10:18:59 2003 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] anti-human 6Ckine Message-ID: Skipped content of type multipart/alternative-------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:ANN MARUSKA EMAIL;WORK;PREF;NGW:AMARUSK1.EAST.NORTH ORG:;LAB N:MARUSKA;ANN TEL;WORK:612-273-9119 END:VCARD From straussj <@t> upstate.edu Fri Sep 12 10:16:59 2003 From: straussj <@t> upstate.edu ( Judy Strauss) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] R2 Message-ID: If anyone has experience with Santa Cruz antibody R2(E-16): sc-10846 on fixed tissue please contact me. Thanks, Judy Judith Strauss SUNY Upstate Medical University Department of Orthopedic Surgery IHP room 3118 505 Irving Avenue phone: (315) 464-9960 fax: (315) 464-6638 Syracuse, NY 13120 From thoward <@t> unm.edu Fri Sep 12 11:02:02 2003 From: thoward <@t> unm.edu (Tamara Howard) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Methacarn/Carnoy's storage? Message-ID: Hi - For how long can methacarn or Carnoy's be stored, once made up? Or should I make a fresh batch every time I need some? We'll be using it for a few slides (5-10), maybe once a week, and it seems a huge waste to dump it every time. Thanks! Tamara |--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward@unm.edu |--------------------------------------------------| From gcallis <@t> montana.edu Fri Sep 12 11:25:23 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Trying to help a colleague and find these antibodies Message-ID: <3.0.6.32.20030912102523.00b781a8@gemini.msu.montana.edu> Dear All, Does anyone have a source of Glucagon-biotinylated and Somatostatin-biotinylated? A lady is trying to locate these antibodies and I do not work with them. Just trying to help a friend. 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 g.lang <@t> bigfoot.de Fri Sep 12 11:44:44 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] SFOG Message-ID: <006f01c37950$1a7e5f60$0d12a8c0@SERVER> Hi! Do you know, what pH the dye-solution of SFOG has to have? SFOG (= S?urefuchsin-Orange-G-Anilinblau) is similar to Cason's onestep trichrome staining solution. It must be something about 1,09 - 1,9. thanks, Gudrun -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/75b59a8b/attachment.htm From kgibbon <@t> qltinc.com Fri Sep 12 13:06:47 2003 From: kgibbon <@t> qltinc.com (kgibbon@qltinc.com) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] nuclear counterstain Message-ID: I have used Vectors VIP kit and found it excellent. I have used both Nuclear fast red and Mayer's haematoxylin for a counterstain and find that the blue of the Mayer's is best for my work. That includes image analysis. Kevin Gibbon QLT Inc. "Atoska S. Gentry" To: Histonet Sent by: cc: histonet-admin@lists.utsouth Subject: [Histonet] nuclear counterstain western.edu 09/12/03 08:15 AM Hello again, guess I need to rephrase my initial inquiry on the counterstain to use with the Vectastain ABC-HRP kit. But, first special thanks to Dr. John Kiernan. He always replies with such informative and thorough remarks. Please pardon my partial entry of facts. This inquiry is on behalf of a couple of colleagues of mine. And yes their work was with monolayers of cultured bone marrow cells. However, the HRP was detected with the Vector VIP substrate ( for which Vector Labs does not reveal the contents) and yielded neurons that are purple in color. Therefore, we need something like acridine orange which stains DNA. Has anyone had any experience/success using acridine orange or something similar with this system that yields the same results? Thanks, Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfavara <@t> niaid.nih.gov Fri Sep 12 13:08:25 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] nuclear counterstain Message-ID: Atoska, I have used the Methylgreen nuclear counter stain from Chris van der Loos with VIP. It is a s follows: Methylgreen, Crystal Violet-free [Sigma M6776] 0.1% methylgreen in sodium acetate buffer [ 100mM, pH 5.2] Solution stable @4C for 1 month Cover section with sodium acetate buffer [10mM, pH5.2] 15 min Blot off do not wash Cover with methyl green solution 5 min Wash briefly with distilled water Dehydrate and mount organically This is out of his book Immunoenzyme Multiple Staining Methods Fabulous resource c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Atoska S. Gentry [mailto:gentras@vetmed.auburn.edu] Sent: Friday, September 12, 2003 9:15 AM To: Histonet Subject: [Histonet] nuclear counterstain Hello again, guess I need to rephrase my initial inquiry on the counterstain to use with the Vectastain ABC-HRP kit. But, first special thanks to Dr. John Kiernan. He always replies with such informative and thorough remarks. Please pardon my partial entry of facts. This inquiry is on behalf of a couple of colleagues of mine. And yes their work was with monolayers of cultured bone marrow cells. However, the HRP was detected with the Vector VIP substrate ( for which Vector Labs does not reveal the contents) and yielded neurons that are purple in color. Therefore, we need something like acridine orange which stains DNA. Has anyone had any experience/success using acridine orange or something similar with this system that yields the same results? Thanks, Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Fri Sep 12 13:10:02 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] hematoxylin article Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CB63@usca0082k03.rallansci.apogent.com> Bryan, that is an interesting article. Here is a site about Campeche, Mexico, where the tree was discovered (hence "Haematoxylum campechianum"). http://www.travelyucatan.com/campeche.htm But now I am wondering how this name originally pronounced? It seems the genus name would be pronounced "Haemato-xylum, considering the dye source which is the xylum of the tree. And if that is the original plant name pronunciation, how was the name of the dye originally pronounced? We now (at least in the US) say "haema-tox-ylin" but maybe it was originally "haemato-xylin." Tim Morken -----Original Message----- From: Bryan Llewellyn [mailto:bryand@netbistro.com] Sent: Thursday, September 11, 2003 11:20 AM To: Histonet Subject: Re: [Histonet] H&E There is a very interesting article on hematoxylin and the other logwoods at http://waynesword.palomar.edu/ecoph4.htm. The article describes how logwood dyes were introduced to Europe, and how they resulted in the creation of two countries. Photos of the trees are included. Bryan Llewellyn ----- Original Message ----- From: "Rana Hay" To: Sent: Wednesday, September 10, 2003 6:17 PM Subject: [Histonet] H&E > Hello, > I am trying to trace information on the historical aspects of H&E > ,especially the use of Haematoxylin.Are you able to help me?When was it > introduced & by whom.etc > > Thank you, > Rana hay > -- > Rana Hay > ranahay@fastmail.fm > > -- > http://www.fastmail.fm - IMAP accessible web-mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Fri Sep 12 14:48:24 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] SFOG References: <006f01c37950$1a7e5f60$0d12a8c0@SERVER> Message-ID: <3F622307.8E9EF7FA@uwo.ca> pH between 1 and 2 should be about right for a trichrome. The acidity is due to the phosphotungstic or phosphomolybdic acid, which is an essential ingredient. Checking the pH is not part of the routine of making up these solutions. It is important to rinse in acidified water after staining, and I advise taking the slides directly from the acidified water to the first of 3 lots of 100% alcohol. The dyes are easily extracted from the sections by non-acidified water or by water-alcohol mixtures. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ___________________________ > Gudrun Lang wrote: > > Hi! > Do you know, what pH the dye-solution of SFOG has to have? > SFOG (= S?urefuchsin-Orange-G-Anilinblau) is similar to Cason's onestep trichrome staining solution. > It must be something about 1,09 - 1,9. > > thanks, Gudrun From jkiernan <@t> uwo.ca Fri Sep 12 14:58:51 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] hematoxylin article References: <8CD0CDC791DCC343ABB1F15298E4F3C417CB63@usca0082k03.rallansci.apogent.com> Message-ID: <3F62257B.60764EC5@uwo.ca> The waynesword web page on logwood referred to by Bryan is indeed a good read! The Oxford Dictionary says the name is from haemato (blood) and Greek xylon (wood) rather than from xylem, the vascular tissue. They give the pronunciation with emphasis on the tox. Wayne spells the generic name of the tree Haematoxylum, with Haematoxylon as a common synonym. I've never seen it written with an -um ending elsewhere, in the dictionary or in botany books. Wayne is a botanist, so he must have his reasons. There are international rules that govern the scientific names of plants - not the same rules that are used for animals. "Morken, Tim - Labvision" wrote: > > Bryan, that is an interesting article. > > Here is a site about Campeche, Mexico, where the tree was discovered (hence > "Haematoxylum campechianum"). > http://www.travelyucatan.com/campeche.htm > > But now I am wondering how this name originally pronounced? It seems the > genus name would be pronounced "Haemato-xylum, considering the dye source > which is the xylum of the tree. > > And if that is the original plant name pronunciation, how was the name of > the dye originally pronounced? We now (at least in the US) say > "haema-tox-ylin" but maybe it was originally "haemato-xylin." > > Tim Morken > > -----Original Message----- > From: Bryan Llewellyn [mailto:bryand@netbistro.com] > Sent: Thursday, September 11, 2003 11:20 AM > To: Histonet > Subject: Re: [Histonet] H&E > > There is a very interesting article on hematoxylin and the other logwoods at > http://waynesword.palomar.edu/ecoph4.htm. The article describes how logwood > dyes were introduced to Europe, and how they resulted in the creation of two > countries. Photos of the trees are included. > > Bryan Llewellyn > > ----- Original Message ----- > From: "Rana Hay" > To: > Sent: Wednesday, September 10, 2003 6:17 PM > Subject: [Histonet] H&E > > > Hello, > > I am trying to trace information on the historical aspects of H&E > > ,especially the use of Haematoxylin.Are you able to help me?When was it > > introduced & by whom.etc > > > > Thank you, > > Rana hay > > -- > > Rana Hay > > ranahay@fastmail.fm > > > > -- > > http://www.fastmail.fm - IMAP accessible web-mail > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ From rlemieux <@t> planibureauplus.com Fri Sep 12 10:00:38 2003 From: rlemieux <@t> planibureauplus.com (Roland Lemieux) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] hydrolic microscope table Message-ID: <41D816D7CF0BD5118B5300105A85836982ED54@FOCUSMAIL> i saw your request on the web for a microscope table ( oct.15, 1999 ) Just for your information and 4 years later a Canadian Company based in Montr?al launch few months ago a table or a station specialy for microscope Roland Lemieux Plani Bureau Plus Director of operation ( 514-644-5553 ) Fax ( 514-332-4400 ) <> <> www.planibureauplus.com -------------- next part -------------- A non-text attachment was scrubbed... Name: poste microscopie2.jpg Type: image/jpeg Size: 79062 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/5fc6d13f/postemicroscopie2.jpg -------------- next part -------------- A non-text attachment was scrubbed... Name: poste microscopie1.jpg Type: image/jpeg Size: 128921 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/5fc6d13f/postemicroscopie1.jpg From briany <@t> ufl.edu Fri Sep 12 11:14:10 2003 From: briany <@t> ufl.edu (Brian Hatcher) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Masson Trichrome stain Message-ID: <3F61F0D2.3040607@ufl.edu> An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/7b9db288/attachment.htm From LMARGRAF <@t> childmed.dallas.tx.us Fri Sep 12 15:25:32 2003 From: LMARGRAF <@t> childmed.dallas.tx.us (LINDA MARGRAF MD) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] meeting announcement Message-ID: September 9, 2003 -- 3rd Announcement == DEADLINES EXTENDED: for Registration, Late Fees, and low Hotel Rates == Subject: Molecular Morphology Conference and Workshops, Santa Fe, New Mexico, USA, October 12-17, 2003 Dear Fellow Histotechnologists and Colleagues: The purpose of this letter is to bring to your attention the "Conference and Workshops on Molecular Morphology: Santa Fe 2003," to be held at the La Fonda Hotel, Santa Fe, New Mexico, October 12 - 17, 2003. This meeting is sponsored by the International Society for Analytical and Molecular Morphology (ISAMM). There will be two (2) Symposia and six (6) Workshops. The six (6) Workshops are: * RNAi Workshops: Part-1& 2. * cDNA Array Demonstrations. * Practical Applications of Real-Time qPCR. * Proteomics Demonstrations: 1& 2. * Automated In Situ Procedures Part-1 & 2. * Nanodots / Nanocrysts and Confocal Microscopy. Deadline for Pre-Registration for the Workshops is EXTENDED UNTIL ALL SPACES ARE FILLED. Call Dr. DeBault at (405) 271-7300 or E-mail him at "ldebault@ouhsc.edu" to reserve a space... as all Workshop space is limited. Deadline for "Late Fees" is EXTENDED to Wednesday, October 1, 2003. For further details on the Conference and Workshops, please visit the WebPage: http://www.isamm.org/Prog2003/isamm_preliminary_program.htm. To register on-line for the Conference and Workshops, please visit the WebPage: http://www.isamm.org/Registration_2003.htm Deadline for room reservations at the reduced conference rate has been EXTENDED to Friday, September 19, 2003. Individual reservations should be made by calling the La Fonda Hotel's Reservation Department at 1-800-523-5002 and then choosing extension #2. To receive the conference rate ($158.00 US + tax) be sure to mention that you are attending the Molecular Morphology Conference and Workshops sponsored by ISAMM.. DEADLINE for Poster Abstracts has been extended to October 6, 2003 and will be printed in the Conference Program Booklet, only. These late Poster abstracts should be E-mailed as attached Microsoft-Word files or as plan text, to: L.E. De Bault at "ldebault@ouhsc.edu". For additional information on the La Fonda Hotel visit: http://www.travelbase.com/destinations/santa-fe/la-fonda/. For additional information on Santa Fe and the surrounding area visit: http://www.santafe.org/. If you wish a hardcopy of the Program and Registration Form, they can be FAXed to you. Please contact L.E. De Bault, Treasurer, ISAMM, via e-mail, at Ldebault@ouhsc.edu, and provide your name and FAX number. Looking forward to seeing you in Santa Fe. Sincerely yours, Raymond Tubbs, President International Society for Analytical and Molecular Morphology Chairman, Department of Clinical Pathology The Cleveland Clinic Foundation 9500 Euclid Avenue Cleveland, OH 44195 Tele: (405) 271-7300 FAX: (405) 271-1107 E-mail: (ldebault@ouhsc.edu) From garygill <@t> dcla.com Fri Sep 12 15:04:01 2003 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] hematoxylin article Message-ID: Then too, of course, there's always: Gill GW, Frost JK, Miller KA. A new formula for a half-oxidized hematoxylin solution that neither overstains nor requires differentiation. Acta Cytol. 1974 Jul-Aug;18(4):300-11. ;-) Gary Gill -----Original Message----- From: John Kiernan [mailto:jkiernan@uwo.ca] Sent: Friday, September 12, 2003 2:59 PM To: Morken, Tim - Labvision Cc: Histonet (histonet@pathology.swmed.edu) Subject: Re: [Histonet] hematoxylin article The waynesword web page on logwood referred to by Bryan is indeed a good read! The Oxford Dictionary says the name is from haemato (blood) and Greek xylon (wood) rather than from xylem, the vascular tissue. They give the pronunciation with emphasis on the tox. Wayne spells the generic name of the tree Haematoxylum, with Haematoxylon as a common synonym. I've never seen it written with an -um ending elsewhere, in the dictionary or in botany books. Wayne is a botanist, so he must have his reasons. There are international rules that govern the scientific names of plants - not the same rules that are used for animals. "Morken, Tim - Labvision" wrote: > > Bryan, that is an interesting article. > > Here is a site about Campeche, Mexico, where the tree was discovered (hence > "Haematoxylum campechianum"). > http://www.travelyucatan.com/campeche.htm > > But now I am wondering how this name originally pronounced? It seems the > genus name would be pronounced "Haemato-xylum, considering the dye source > which is the xylum of the tree. > > And if that is the original plant name pronunciation, how was the name of > the dye originally pronounced? We now (at least in the US) say > "haema-tox-ylin" but maybe it was originally "haemato-xylin." > > Tim Morken > > -----Original Message----- > From: Bryan Llewellyn [mailto:bryand@netbistro.com] > Sent: Thursday, September 11, 2003 11:20 AM > To: Histonet > Subject: Re: [Histonet] H&E > > There is a very interesting article on hematoxylin and the other logwoods at > http://waynesword.palomar.edu/ecoph4.htm. The article describes how logwood > dyes were introduced to Europe, and how they resulted in the creation of two > countries. Photos of the trees are included. > > Bryan Llewellyn > > ----- Original Message ----- > From: "Rana Hay" > To: > Sent: Wednesday, September 10, 2003 6:17 PM > Subject: [Histonet] H&E > > > Hello, > > I am trying to trace information on the historical aspects of H&E > > ,especially the use of Haematoxylin.Are you able to help me?When was it > > introduced & by whom.etc > > > > Thank you, > > Rana hay > > -- > > Rana Hay > > ranahay@fastmail.fm > > > > -- > > http://www.fastmail.fm - IMAP accessible web-mail > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GalbraithJ <@t> uihc.uiowa.edu Fri Sep 12 16:17:02 2003 From: GalbraithJ <@t> uihc.uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Manipulation time of human muscle biopsies and for mation of freezing artifacts. Message-ID: <5D03ED7B9391D4119D9B0008C76B7B24030085D2@uihc-mail1.uihc.uiowa.edu> Mauricio: If you have access to a walk-in freezer as we do, you may be able to transfer the racks/boxes to the walk-in using styrofoam shipping containers or a large plastic cooler (like those used for camping) containing dry ice. Once in the walk-in freezer you would be able to work with the specimens for some time without having them thaw out (handle with gloves to insulate your hands from the cold and the specimens from your body heat. However, most walk-in freezers (-20 C) are not as cold as the cryofreezers commonly used here to store muscles (-70 C). But the closet-sized sized freezer might offer enough working time to allow you to sort and label without much loss or damage. Good luck, Joe Galbraith -----Original Message----- From: Mauricio S. Morais [mailto:Mauricio.Morais@tufts.edu] Sent: Thursday, September 11, 2003 6:25 PM To: Histonet Subject: [Histonet] Manipulation time of human muscle biopsies and formation of freezing artifacts. Hello Histonet community. Because I'm new into the histology "word" I've been facing some unexpected (to me) challenges in my actual Laboratory work. My background is originally from Organic and Analytical Chemistry, Teaching, and a bit of Biochemistry (my MSc degree). I'm kind of new in the country too (been here less than 2 years) and English is not my primary language, so please accept my apologies for any errors within this text. I'm assign to reorganize and analyze human muscle biopsies, stored in boxes distributed among 4 racks inside a Cryoplus cryofreezer. Seeking for information on how to work safely and at the same time not compromise the integrity of the stored samples, I found the Histonet Histosearch archives. They were very helpful providing answer about most of the common issues related with formation of freezing artifacts and possible solutions to the problem at different stages the samples can generate it, from the biopsy to the cryostat slice machine. Because of the enormous amount of samples that'll need to be handle at one time during the reorganization of our cryofreezer, an issue about "time" became critical (in my opinion) raising some questions: 1-How long can a box (full of biopsies), or the hole rack be kept out of the cryofreezer, while I'm updating data and re-arranging boxes between racks, without compromise the biopsies and generate (and/or enhancing the chances of creating) freezing artifacts? 2-Does any damage may occur if I have several biopsies floating inside a dewar container while I'm making notes or grouping them into other dewar containers to a new order? A website information (from www.asymptote.co.uk) says that tissue inside straws immersed in liquid Nitrogen when exposed to air at room temperature have their temperature raised to -50C in 40 seconds. Thawing may start around -10C or early. I was told they (racks, boxes) could be out for about 5 minutes before some damage can start to affect the biopsies inside it. None of them been hold by hands during all this time, but on the bench top (boxes) or floor (racks) while the work is done. I was also told that 5 minutes is away too long... For me it seems impossible to safely remove a rack (containing a dozen boxes) from the cryofreezer, make notes or, for example, remove one box from the rack to add a new sample and return it to the cryofreezer in 40 seconds. Reminding that I'm working with cryogloves over my lab. gloves... I was never able to open a box without remove the cryogloves first. Maybe I need more training... I would greatly appreciate any comments from this expert community to help me solve my immediate problems (above) and improve my knowledge and safety at work place. Thank you all. Mauricio S. Morais Senior Research Technician Nutrition Exercise Physiology and Sarcopenia Laboratory (NEPS) TUFTS University Jean Mayer USDA Human Nutrition Research Center on Aging 711 Washington Street, Rm 436 Boston, MA 02111 (617) 556-3226 From GalbraithJ <@t> uihc.uiowa.edu Fri Sep 12 16:27:29 2003 From: GalbraithJ <@t> uihc.uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] cleaning microtomes Message-ID: <5D03ED7B9391D4119D9B0008C76B7B24030085D3@uihc-mail1.uihc.uiowa.edu> Mariann: I totally agree and should have mentioned in my post that one should never spray a cleaner directly onto the microtome. My oversight. Thanks. Joe Galbraith -----Original Message----- From: mari.ann.mailhiot@leica-microsystems.com [mailto:mari.ann.mailhiot@leica-microsystems.com] Sent: Wednesday, September 10, 2003 3:29 PM To: Galbraith, Joe Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cleaning microtomes Joe Paraguard or mineral oil have been my choice. I have tried to get away from xylene as much as possible. Any cleaner such as 409 is fine. You just don't want the 409 solution to get on the inside of the microtome. It is better to spray the solution on a piece of gauze. In fact, I always spray the paragaurd on the gauze and then wipe my microtome. If I have to much paraguard on the microtome, I just take and put some alcohol on a piece of gauze and wipe the paraguard off of the microtome. 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 "Galbraith, Joe" To: "'RCHIOVETTI@aol.com'" , Jmrwalker@aol.com, Sent by: histonet@lists.utsouthwestern.edu histonet-admin@lists.utsouth cc: western.edu Subject: RE: [Histonet] cleaning microtomes 09/09/2003 03:03 PM Histonetters: Most people in my lab use Paraguard to clean microtomes but some prefer 409 (Misty Breeze has a nice smell). The 409 cleaner seems to do as good of a job as Paraguard and costs less and smells better. Joe Galbraith -----Original Message----- From: RCHIOVETTI@aol.com [mailto:RCHIOVETTI@aol.com] Sent: Tuesday, September 09, 2003 1:15 PM To: Jmrwalker@aol.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] cleaning microtomes In a message dated 09/09/2003 7:51:39 AM US Mountain Standard Time, Jmrwalker@aol.com writes: << After you are finished cutting on your microtome, what do you use to clean it? Our lab in the past always used xylene or alcohol to clean it. A few new people have been hired in the lab and now Paragard and baby oil have been used. I'm just curious what the majority of people in histoland are using. Thanks! >> I see mostly Paragard being used in our customers' labs, and I agree with the other posters, if the "slick" surface is a problem after the Paragard treatment you can then wipe down with a little ethanol or isopropanol. With time xylene can affect the surfaces of the microtome's external parts, especially if used in excess. Lots of the microtome parts have anodized (plated) surfaces that are either black or clear, and you'd think they would hold up to just about any kind of abuse. But they will definitely break down and corrode with prolonged or repeated exposure to xylene. This can lead to pitting and discoloration of the surfaces. If you *do* use xylene, use it sparingly and be sure to wear gloves. I personally don't like the stuff. Try to use a substitute instead. First, use a brush or gauze pad to remove the majority of the paraffin. Then use only enough xylene or xylene substitute to barely moisten a gauze 4x4 or a tissue. Wipe off the excess immediately with another dry gauze or tissue. You can also follow this with an alcohol wipe-down like you would use with Paragard if the surface feels slick or oily after you clean it. Hope this helps! Bob Chiovetti GTI Microsystems Leica Regional Dealer Desert Southwest Region, USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ This email has been scanned for all viruses by the MessageLabs Email Security System. For more information on a proactive email security service working around the clock, around the globe, visit http://www.messagelabs.com ________________________________________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ander093 <@t> gold.tc.umn.edu Fri Sep 12 16:29:50 2003 From: ander093 <@t> gold.tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] golgi Message-ID: <5.2.0.9.0.20030912162531.009f9850@ander093.email.umn.edu> Hi, Has anyone out there in histo-land had experience with the Rapid GolgiStain Kit from FD NeuroTechnologies? We are looking into it......but any info would be appreciated. I have checked the archives, but nothing turned up. Thanks in advance. LuAnn Anderson HT(ASCP) Division of Neuropathology University of Minnesota From gcallis <@t> montana.edu Fri Sep 12 18:05:02 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Reply #3 Manipulation time of human muscle biopsies and formation of freezing artifacts. In-Reply-To: <5D03ED7B9391D4119D9B0008C76B7B24030085D2@uihc-mail1.uihc.u iowa.edu> Message-ID: <3.0.6.32.20030912170502.00b99338@gemini.msu.montana.edu> To sustain a lengthy sort of biopsies, buy a big block(s) of dry ice and pulverize it so you can lay things on top of ice and/or surround boxes with mutiple smaller dry ice chunks and still be able to handle the vials while sorting. Use good insulation of hands and forceps (I have some LONG Russian forceps with large rounded serrated tips - shoved into dry ice to keep cold or long, grippy hemostats to pick up vials. Vials can be laid on or shoved into pulverized dry ice and certainly stay out of a freezer for more than 5 minutes as long as they are surrounded by dry ice (hmmm approx -70 to -90C). BE sure to use a styrofoam container to hold the dry ice, open boxes can be worked with at RT for as long as you need. You could still work in a cold room with dry ice, however close quarters could be a problem - see very last comment. If possible, try to recycle some dry ice pellets (Sigma et al use to ship biologicals, antibodies, etc in styrofoam containers and if possible stockpile pellets in -80C freezer in a styrofoam box, some means for this purpose (hope you have some space!) We do this for snap freezing purposes and rarely buy dry ice blocks. We sort large 50 ml tubes w/frozen tissue blocks using dry ice method frequently and the biggest problem is cold hands, take care to insulate your fingers from snap freezing! I have seen people use thinner silk glove/liners from Winter Silks inside latex gloves and forceps to handle (juggle?) tubes. We find the dry ice easier to work with than colder, fog creating liquid nitrogen where I end up going on a frustrating fishing for tubes somewhere in bottom of a dewar. Be sure you have good ventilation, you don't want to be found passed out on the floor due to oxygen depletion by CO2 or N2 fumes. Good luck Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 At 04:17 PM 9/12/2003 -0500, you wrote: >Mauricio: > >If you have access to a walk-in freezer as we do, you may be able to >transfer the racks/boxes to the walk-in using styrofoam shipping containers >or a large plastic cooler (like those used for camping) containing dry ice. >Once in the walk-in freezer you would be able to work with the specimens for >some time without having them thaw out (handle with gloves to insulate your >hands from the cold and the specimens from your body heat. However, most >walk-in freezers (-20 C) are not as cold as the cryofreezers commonly used >here to store muscles (-70 C). But the closet-sized sized freezer might >offer enough working time to allow you to sort and label without much loss >or damage. > >Good luck, > >Joe Galbraith > >-----Original Message----- >From: Mauricio S. Morais [mailto:Mauricio.Morais@tufts.edu] >Sent: Thursday, September 11, 2003 6:25 PM >To: Histonet >Subject: [Histonet] Manipulation time of human muscle biopsies and >formation of freezing artifacts. > > >Hello Histonet community. > >Because I'm new into the histology "word" I've been facing some >unexpected (to me) challenges in my actual Laboratory work. >My background is originally from Organic and Analytical Chemistry, >Teaching, and a bit of Biochemistry (my MSc degree). >I'm kind of new in the country too (been here less than 2 years) and >English is not my primary language, so please accept my apologies for >any errors within this text. > >I'm assign to reorganize and analyze human muscle biopsies, stored in >boxes distributed among 4 racks inside a Cryoplus cryofreezer. >Seeking for information on how to work safely and at the same time not >compromise the integrity of the stored samples, I found the Histonet >Histosearch archives. >They were very helpful providing answer about most of the common issues >related with formation of freezing artifacts and possible solutions to >the problem at different stages the samples can generate it, from the >biopsy to the cryostat slice machine. > >Because of the enormous amount of samples that'll need to be handle at >one time during the reorganization of our cryofreezer, an issue about >"time" became critical (in my opinion) raising some questions: > >1-How long can a box (full of biopsies), or the hole rack be kept out of >the cryofreezer, while I'm updating data and re-arranging boxes between >racks, without compromise the biopsies and generate (and/or enhancing >the chances of creating) freezing artifacts? > >2-Does any damage may occur if I have several biopsies floating inside a >dewar container while I'm making notes or grouping them into other dewar >containers to a new order? > >A website information (from www.asymptote.co.uk) says that tissue inside >straws immersed in liquid Nitrogen when exposed to air at room >temperature have their temperature raised to -50C in 40 seconds. Thawing >may start around -10C or early. > >I was told they (racks, boxes) could be out for about 5 minutes before >some damage can start to affect the biopsies inside it. None of them >been hold by hands during all this time, but on the bench top (boxes) or >floor (racks) while the work is done. >I was also told that 5 minutes is away too long... > >For me it seems impossible to safely remove a rack (containing a dozen >boxes) from the cryofreezer, make notes or, for example, remove one box >from the rack to add a new sample and return it to the cryofreezer in 40 >seconds. Reminding that I'm working with cryogloves over my lab. >gloves... I was never able to open a box without remove the cryogloves >first. >Maybe I need more training... > >I would greatly appreciate any comments from this expert community to >help me solve my immediate problems (above) and improve my knowledge and >safety at work place. > > >Thank you all. > > >Mauricio S. Morais >Senior Research Technician >Nutrition Exercise Physiology and Sarcopenia Laboratory (NEPS) >TUFTS University >Jean Mayer USDA Human Nutrition Research Center on Aging >711 Washington Street, Rm 436 >Boston, MA 02111 >(617) 556-3226 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From gcallis <@t> montana.edu Fri Sep 12 18:14:11 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] another reply on cleaning microtomes In-Reply-To: <5D03ED7B9391D4119D9B0008C76B7B24030085D3@uihc-mail1.uihc.u iowa.edu> Message-ID: <3.0.6.32.20030912171411.00b99338@gemini.msu.montana.edu> I was taught by a microtomy expert, Jan Minshew, to never use solvents on a microtome - they remove necessry lubricants on screws and other critical microtome areas. We just use an extremely rough terry cloth towel or coarse gauze to rub most of the paraffin off with a bit of extreme "elbow grease" but remove knife to do this. One can always get the last of the paraffin "film" off with Windex or 409 cleaner on a cloth - I tend to wipe this off with a damp cloth. 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 tigersnake <@t> ecybermind.net Fri Sep 12 21:56:35 2003 From: tigersnake <@t> ecybermind.net (Paul Howard Lockwood) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] validating oil red O Message-ID: <200309130053.h8D0rUA09499@ginsberg.ecybermind.net> Dear Jeremy, I would suggest using frozen sections, and mounting them on slides coated with poly-l lysine. Also, do not expose the slides to heat above room tempeture. I hope this is of some help to you. Sincerely, Paul Lockwood 9/11/03 6:53:05 PM, Jeremy Browne wrote: > > > > We need to set up a range of fat concentrations on slides, stain and then > check to see that there is a linear response relationship between fat > concentration and staining intensity but because fat melts and slides off > histoworld has solved this problem - perhaps with an intermediatere is > substance to which the fat adheres? > Thanks > Jeremy Browne > Liggins Institute > NZ (09) 373 7599 extn 86444 > j.browne@auckland.ac.nz From tjey <@t> hotmail.com Fri Sep 12 20:32:25 2003 From: tjey <@t> hotmail.com (Tanya Ewing) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Temporary Agencies Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030912/8aa4e2c3/attachment.htm From jkiernan <@t> uwo.ca Sat Sep 13 00:26:12 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] golgi References: <5.2.0.9.0.20030912162531.009f9850@ander093.email.umn.edu> Message-ID: <3F62AA74.5AAF4A7A@uwo.ca> In the literature of neuroanatomy, the rapid Golgi method is a distinct technique, different from the original Golgi method and also different from other variants, which have informal names like Golgi-Cox and Golgi-Sholl. All these techniques blacken a small proportion the cells in a block or slice of CNS, but they have different emphases. The choice of a Golgi method is determined by the requirements of the investigation. For example, the Cox, Sholl and related methods are preferred for quantitative studies of dendrites. These techniques do not use silver nitrate and they have little in common with the techniques invented by Camillo Golgi. The traditional rapid Golgi method, which uses osmium tetroxide as well as the silver nitrate and potassium dichromate of the original method, has been used to impregnate occasional whole interneurons (including the axon and its terminal branches), especially in the fetal CNS, where most of the axons are not yet myelinated. I think it's still true to declare that no Golgi variant will reliably deposit black material in myelinated axons. Your email asks about a commercial rapid Golgi staining kit. Does the company's "rapid" relate to traditional rapid Golgi methods, or is it selling something quite different? None of the Golgi techniques are rapid when compared with ordinary staining techniques. They all take weeks, and rapid means 2 weeeks rather than 6. It's important for you to know that a 2-week classical rapid Golgi method will not provide the same kind of stained sections as as traditional Golgi or a Golgi-Cox. A company selling a kit for Golgi staining must provide a full explanation of which chemicals are used and why. The company should also tell you how its product relates to the various traditional Golgi techniques. All the Golgi variants are technically easy and they use chemicals available in any lab that does neuro-anything. Production of good Golgi preparations (any variant) involves plenty of disappointment. The randomness of the chromate precipitation causes all sorts of strongly positive artifacts. Golgi preparations require educated interpretation. Do not trust anyone. Do the tests yourself before maqking recommendations. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ _______________________________________- LuAnn Anderson wrote: > > Hi, > Has anyone out there in histo-land had experience with the Rapid GolgiStain > Kit from FD NeuroTechnologies? We are looking into it......but any info > would be appreciated. I have checked the archives, but nothing turned up. > Thanks in advance. > > LuAnn Anderson HT(ASCP) > Division of Neuropathology > University of Minnesota > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peptolab <@t> hamptons.com Sat Sep 13 08:32:16 2003 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Bouin's fixation Message-ID: <000b01c379fb$7567e080$e176bd18@JEFF> A 3-5 mm testicular biopsy should be fixed in one to two hours. Be sure to rinse out as much yellow picric acid as possible (you can use running water or 70% alcohol ). The basophilia of chromatin in the stored block will deteriorate after some time if there is picric acid residue. Jeff Silverman HT HTL QIHC (ASCP) Southside Hospital Bay Shore NY From brett_connolly <@t> merck.com Sat Sep 13 09:27:16 2003 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] rhodanine for copper Message-ID: <58E3A5F2EFDB224F95D4C62DDDCCA2BE05036110@uswpmx08.merck.com> 2 questions about this histochemical stain.... First, does rhodanine bind directly to the copper or does it bind to another moiety which, in turn, is bound to the copper. Second, is anyone aware of any fluorescent properties of rhodanine (not rhodamine). Third, (OK, I lied) does rhodanine detect any other metals in addition to copper, specifically zinc or iron. Any references would be much appreciated! Thanks, 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 ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (Whitehouse Station, New Jersey, USA), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD) that may be confidential, proprietary copyrighted and/or legally privileged, and 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 immediately return this by e-mail and then delete it. ------------------------------------------------------------------------------ From kacw <@t> citlink.net Sat Sep 13 15:32:18 2003 From: kacw <@t> citlink.net (Kristen&Chuck) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] disaster planning Message-ID: Hi! I was wondering if anyone has any info on emergency procedures. I'm helping our administration revise our emergency procedures, any suggestions? kacw@citlink.net From Histolady710 <@t> aol.com Sat Sep 13 15:56:57 2003 From: Histolady710 <@t> aol.com (Histolady710@aol.com) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] HT, ASCP Exam Message-ID: <15.18bafedf.2c94de99@aol.com> After many months of studying, the last two being intense, I sat for the HT exam and felt as if I was taking the exam in a foreign language !!! I felt very confident going into this exam having studied from Carson's Histotechnology, BOR Study Guide and NSH Self Assessment guides with computer disks and with 14 years histology under my belt. In no way did I expect this to be easy but I did expect to recognize more than 20% of the questions asked. So, my question is - has anyone else had this experience and if so what did you do to correct it. I'm at a loss and have no idea WHAT to study should I decide to take the exam again. Thank you. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030913/595a3dcb/attachment.htm From Histolady710 <@t> aol.com Sun Sep 14 07:35:15 2003 From: Histolady710 <@t> aol.com (Histolady710@aol.com) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Peggy Wenk Message-ID: <1e8.f83dce9.2c95ba83@aol.com> Peggy, Please contact me again. My reply to you was returned MAILER-DAEMON. Thanks, DDietz -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030914/c531824b/attachment.htm From HornHV <@t> archildrens.org Sun Sep 14 16:27:34 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] CAP Question anp.23075 Message-ID: I'm having my CAP inspection soon and wondered what is meant by this question. Is there EVIDENCE of ongoing evaluation of results of instrument maintenance a nd function for all devices? The 2 questions before this one are about a schedule for checking instruments (yes) and are the service and repair records promptly available. (yes) So......What does the above question pertain too?? HELP! Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Arkansas Children's Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030914/24152a8c/attachment.htm From j.browne <@t> auckland.ac.nz Sun Sep 14 16:34:43 2003 From: j.browne <@t> auckland.ac.nz (Jeremy Browne) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] validating oil red o staining Message-ID: Clarification, Thanks for all your responses. Its clear that I haven't explained myself very well at all: We would like to use oil red O to quantify lipid localisation in frozen sections of hepatic and muscle tissue. However, we must first demonstrate that the amount of staining we get (density) is in fact equal to the amount of lipid present in the sections and to do this we need to set up some titrations using positive controls with known and different concentrations of lipid to show a linear response between staining intensity and lipid concentration. However, a fat smear is difficult to use as a positive control in this way because it simply washes off the slides. Therefore, we would like to know what others have done to demonstrate linearity with the oil red O staining procedure. Is there a way we can treat the fat to overcome its liquid nature (that is to say not the lipid in the muscle and liver but the actual 'fat' being used as a positive control)? Thanks Jeremy Jeremy Browne Liggins Institute University of Auckland NZ (09) 373 7599 extn 86444 j.browne@auckland.ac.nz -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/9b350829/attachment.htm From mayhew <@t> vaxxine.com Sun Sep 14 16:59:55 2003 From: mayhew <@t> vaxxine.com (D. Mayhew) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Computer system question Message-ID: <012501c37b0b$891f6a60$2ad705d1@dave> Hi Sheila, We are currently in the middle of implementing Meditech 4.9 and decided not to use Order Entry into the Pathology Module. Instead we have the nurses enter the Pathology specimens into the Lab module in order to get a barcoded label with the date and time of collection on it. In the OE Module there are mandatory queries regarding diagnosis and operative procedure, as well as type of tissue that are attached to the category of Path. The type of tissue is pulled on to the label as well. We had Meditech create a priority of P in order to keep these specimens from cluttering up the outstanding lists for regular Lab specimens. When the specimens arrive at the Histology Lab, they are electronically received, and that "Lab" test is automatically resulted as NP (no Print). The specimen is then entered in to the Pathology module by the Histo techs, and of course this date and time is recorded. This allows us to put the specimens in the order we prefer and avoids the problem of the Breast Screening clinic entering 3 breast biopsies in a row, for example. Lee Mayhew MLT Niagara Health System St. Catharines Ont. From SJones <@t> cvm.tamu.edu Sun Sep 14 18:07:17 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Bouin's fixation/removal of picric acid Message-ID: I've always used saturated lithium carbonate in 70% ethanol to remove the picric acid. I believe it would be faster and more complete than 70% alone. Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "peptolab" 09/13/03 08:32AM >>> A 3-5 mm testicular biopsy should be fixed in one to two hours. Be sure to rinse out as much yellow picric acid as possible (you can use running water or 70% alcohol ). The basophilia of chromatin in the stored block will deteriorate after some time if there is picric acid residue. Jeff Silverman HT HTL QIHC (ASCP) Southside Hospital Bay Shore NY _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Sun Sep 14 19:28:02 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] TESTICULAR BIOSY Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E03B@simba.kids> Beware, Protein picrates are believed to be quite soluble in water. I would recommend storing in 80% or more ethanol prior to processing, not NBF. Fixation time 4-8 hours for a small biopsy (up to 4mm in diameter) Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Travis Troyer [mailto:ttroyer@pcllab.com] Sent: Friday, 12 September 2003 23:07 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] TESTICULAR BIOSY Our lab is receiving a testicular biopsy that will be received in Bouin's Solution. We routinely place specimans in 10% NBF after grossing. So, my question is how long should the biopsy stay in the Bouin's solution for proper fixation before placing into 10% NBF? Thanks! ********************************************************************** 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. ********************************************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/7b9b6e86/attachment.htm From Stanley.Stylli <@t> mh.org.au Sun Sep 14 21:46:19 2003 From: Stanley.Stylli <@t> mh.org.au (Stylli, Stanley) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] tumour vascularization Message-ID: <00519F9F41D2844D9C75BEE1F6AC09AF68B9B1@rmhmail1.ssg.org.au> Hi Histonetters Is there anyone out there who has looked at vascularity of rat tumours with antibodies such as VEGF or alternatively looking at metalloproteinase expression with something like MMP2 ? I will be doing this work on FFPE tissue sections, so I would mind if someone had a recommendation on something that works on FFPE sections (even with antigen retrieval). Any recommendations and/or protocols would be appreciated. Thankyou for your assistance. regards Stan Stylli Stan Stylli Department of Surgery, 5th Floor Clinical Sciences Building Royal Melbourne Hospital University of Melbourne Parkville, Australia, 3052. Tel: 61-3-93427616 Fax:61-3-9347-7695 -------------- next part -------------- A non-text attachment was scrubbed... Name: Stylli, Stanley.vcf Type: text/x-vcard Size: 230 bytes Desc: Stylli, Stanley.vcf Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/45696cdd/StylliStanley.vcf From jkiernan <@t> uwo.ca Sun Sep 14 23:54:25 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Bouin's fixation/removal of picric acid References: Message-ID: <3F654601.3863CD1C@uwo.ca> Sarah, do you use the saturated lithium carbonate in 70% ethanol on blocks of tissue or on hydrated paraffin sections that are yellow? If it's for blocks, how do you remove lithium carbonate from the decolorized specimens? John Kiernan -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ___________ Sarah Jones wrote: > > I've always used saturated lithium carbonate in 70% ethanol to remove > the picric acid. I believe it would be faster and more complete than > 70% alone. > > Sarah Jones HT(ASCP) > Dept. of Vet. Anatomy & Public Health > Histology Lab > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > >>> "peptolab" 09/13/03 08:32AM >>> > A 3-5 mm testicular biopsy should be fixed in one to two hours. Be sure > to > rinse out as much yellow picric acid as possible (you can use running > water > or 70% alcohol ). The basophilia of chromatin in the stored block will > deteriorate after some time if there is picric acid residue. > > Jeff Silverman HT HTL QIHC (ASCP) > Southside Hospital > Bay Shore NY > From rowani.mohdrawi <@t> student.adelaide.edu.au Mon Sep 15 00:08:17 2003 From: rowani.mohdrawi <@t> student.adelaide.edu.au (Rowani) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Mice embryonic brain - EM Poor tissue quality - need help Message-ID: <00a201c37b47$60163b30$87997f81@LaptopRowani> Hi My electron microscopy sections of brain of mice foetuses (18.5 dpc) are difficult to interpret because of disrupted and lysed cells. The connective tissue surrounding the cells appears disintegrated with empty spaces/vacuoles like artifact ?. The brain was processed according to the following protocols: brain was dissected and immersed in ice cold fixative (glutaryldehyde 2.5%, paraformaldehyde 4%, sucrose 2% in PBS pH 7.2) for 1-2 hrs to firm it up before cutting them to smaller fragments (2 cumm). After an overnight fixation, re-fixed in osmium tetroxide, washed in PBS, dehydrated through a series of ethanol (70%, 90%, 95% and 100%) and embedded in araldyte resin. The brain of adult mice processed using similiar protocol was excellent. COuld someone suggest ways to improve the tissue quality ? TQ Rowani From m.vermehren <@t> anat.vetmed.uni-muenchen.de Mon Sep 15 05:36:46 2003 From: m.vermehren <@t> anat.vetmed.uni-muenchen.de (Margarete Vermehren) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] ISH on murine embryos Message-ID: <000301c37b75$44baf8c0$64ae548d@HistoPC3> Attention Mikael Niku, University of Helsinki Dept. Basic Veterinary Sciences I am starting to work on mouse ISH (paraffin). The animals are young (~ 15 days) but no embryos. Could you give me a hint on the protocol to use? Which proteinase, heat or not Very grateful for help, M. Vermehren Margarete Vermehren LMU M?nchen / Tieranatomie II Tel: +49 89 21805857 Fax: +49 89 21802569 m.vermehren@anat.vetmed.uni-muenchen.de -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/43e7692d/attachment.htm From kemlo <@t> tiscali.co.uk Mon Sep 15 05:46:52 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] compliment In-Reply-To: <002801c37941$47026400$0d12a8c0@SERVER> Message-ID: <000001c37b76$b2677d70$06c4e150@KEMLOS> This is an American board (mostly), so 'bad English' is common. Yours seems fine! Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: 12 September 2003 16:19 To: Histonetliste Subject: [Histonet] compliment Dear histonetters! I have to pay you a compliment. I am so happy, that I found some people with the same interest in acquireing the knowledge about histotechnic. (and histotechs with such a profound knowledge). The Histo-instructions at my medical school were a little poor. So if I want to become a good histotech, I have to do it on my own. The colleges in my lab consider me a little bit of mad, when I deal with the job-issues at home. So I have to moan about it a little bit. (Do you understand me?) Please don't mind my bad English. Again, thank you Gudrun Lang General Hospital, Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/e9d95ad1/attachment.htm From Tbarnhart <@t> primecare.org Mon Sep 15 06:25:59 2003 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] CAP Question anp.23075 Message-ID: <1779904B5E82D511914C00D0B793339220540B@exchangent> Hazel, We struggled with this one too. In fact my last inspector didn't know what to make of this question, either. I gave examples of: when we brake down the cryostat for cleaning (or maintenance) we always use a chuck of OTC to realign the roll plate and make sure the instrument is cutting OK, or when our stainer has maintenance work done we run a test batch of slides to make sure everything is working properly. He seemed to like those for examples of this standard. Make sure you have written procedures and documented evidence that you did these checks. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND tbarnhart@primecare.org -----Original Message----- From: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: Sunday, September 14, 2003 4:28 PM To: HISTONET (E-mail) Subject: [Histonet] CAP Question anp.23075 I'm having my CAP inspection soon and wondered what is meant by this question. Is there EVIDENCE of ongoing evaluation of results of instrument maintenance a nd function for all devices? The 2 questions before this one are about a schedule for checking instruments (yes) and are the service and repair records promptly available. (yes) So......What does the above question pertain too?? HELP! Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 _____ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Arkansas Children's Hospital Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/2b89bd0d/attachment.htm From Tony.Fearn <@t> cd.burnleyhc-tr.nwest.nhs.uk Mon Sep 15 08:17:08 2003 From: Tony.Fearn <@t> cd.burnleyhc-tr.nwest.nhs.uk (Fearn Tony) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Picric acid crystals Message-ID: <8C3DE9069A06D611B8950002A550C15943BC95@BHC_MAIL02> Recently I have been trying to find a UK supplier of picric acid crystals and have failed. I wonder if anyone could suggest a source. I'm looking for about 500g. Thanks Tony Fearn Anthony J Fearn Chief Biomedical Scientist Histopathology Burnley U.K. From gareth.davis <@t> Vanderbilt.Edu Mon Sep 15 08:59:33 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Mice embryonic brain - EM Poor tissue quality - need help Message-ID: <082C721AF78DB34983E8BA2CD085462105C66C@mailbe07> I'm not sure about EM procedures, but I've been told, and do find, perfusion fixation to be the best for fixing brain tissue. If you need a protocol for that I have one. Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -----Original Message----- From: Rowani [mailto:rowani.mohdrawi@student.adelaide.edu.au] Sent: Mon 9/15/2003 12:08 AM To: histonet@pathology.swmed.edu Cc: Subject: [Histonet] Mice embryonic brain - EM Poor tissue quality - need help Hi My electron microscopy sections of brain of mice foetuses (18.5 dpc) are difficult to interpret because of disrupted and lysed cells. The connective tissue surrounding the cells appears disintegrated with empty spaces/vacuoles like artifact ?. The brain was processed according to the following protocols: brain was dissected and immersed in ice cold fixative (glutaryldehyde 2.5%, paraformaldehyde 4%, sucrose 2% in PBS pH 7.2) for 1-2 hrs to firm it up before cutting them to smaller fragments (2 cumm). After an overnight fixation, re-fixed in osmium tetroxide, washed in PBS, dehydrated through a series of ethanol (70%, 90%, 95% and 100%) and embedded in araldyte resin. The brain of adult mice processed using similiar protocol was excellent. COuld someone suggest ways to improve the tissue quality ? TQ Rowani _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/13b3c450/attachment.htm From gareth.davis <@t> Vanderbilt.Edu Mon Sep 15 09:02:07 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] haematoxylin Message-ID: <082C721AF78DB34983E8BA2CD085462105C66D@mailbe07> Thanks for all the responses to the Hematoxylin problem. I'm going to Ehrlich's and see how that goes. Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -----Original Message----- From: damien desmond [mailto:vanlochemstraat@yahoo.com] Sent: Fri 9/12/2003 7:04 AM To: Davis, Gareth Cc: Subject: RE: [Histonet] haematoxylin Harris and Weigert are regressive stains, so the stains overstain and then you decolourize to the desired effect, Mayers is progressive so no use of a differentiator and this will stain to an end point in a certain time period to be determined by testing and microscopic interpretation. David --- "Davis, Gareth" wrote: > I'm trying to find a good Hematoxylin for good H&E > results, is that what your discussion was about? > Gill's is best for counterstaining IHC sections, in > my opinion. > Gareth > > Ms. Gareth B. Davis > Research Assistant II > Neuro-magnetics Division of > the Department of Neurology > Vanderbilt University Medical Center > 615-936-3318 > > > > -----Original Message----- > From: Gudrun Lang [mailto:g.lang@bigfoot.de] > Sent: Thu 9/11/2003 12:44 PM > To: Histonetliste > Cc: > Subject: [Histonet] haematoxylin > > > In our lab we have the discussion, if staining with > Mayer's or Harris' or Weigert's hematoxylin leads to > an endpoint-staining. > I have read, that only Weigert stains to an > endpoint. The others can be overstained. > Who can explain and help? > > Gundi > Akh, Linz, Austria > > __________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/eb821ef7/attachment.htm From mcauliff <@t> umdnj.edu Mon Sep 15 09:55:25 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Mice embryonic brain - EM Poor tissue quality - need help In-Reply-To: <00a201c37b47$60163b30$87997f81@LaptopRowani> References: <00a201c37b47$60163b30$87997f81@LaptopRowani> Message-ID: <3F65D2DD.2040508@umdnj.edu> Hi Rowani: Immersing the whole brain in fixative will not allow good ultrastructure, no matter what the fixative is. Even immersing small (1mm cubed) pieces will not allow optimal ultrastructure. There is no advantage to using cold fixative, in fact it may make your results worse due to slowing of the reaction of fixative with the tissue. Finally, embryonic tissue is more difficult to fix (well) than adult tissue. I would recommend vascular perfusion with a glutaraldehyde or glut-paraformaldehyde fixative. Consult the literature for one or more exact recommendations for the part of the brain you are interested in and the age of the embryo. The concentration of the buffer you use and the presence of additives (sucrose) will be more important than the exact concentrations of glut. or para. Geoff Rowani wrote: >Hi >My electron microscopy sections of brain of mice foetuses (18.5 dpc) are >difficult to interpret because of disrupted and lysed cells. The connective >tissue surrounding the cells appears disintegrated with empty >spaces/vacuoles like artifact ?. The brain was processed according to the >following protocols: brain was dissected and immersed in ice cold fixative >(glutaryldehyde 2.5%, paraformaldehyde 4%, sucrose 2% in PBS pH 7.2) for 1-2 >hrs to firm it up before cutting them to smaller fragments (2 cumm). After >an overnight fixation, re-fixed in osmium tetroxide, washed in PBS, >dehydrated through a series of ethanol (70%, 90%, 95% and 100%) and embedded >in araldyte resin. The brain of adult mice processed using similiar >protocol was excellent. COuld someone suggest ways to improve the tissue >quality ? > >TQ > >Rowani > > > > > > -- -- ********************************************** 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 mcauliff <@t> umdnj.edu Mon Sep 15 10:07:04 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] validating oil red o staining In-Reply-To: References: Message-ID: <3F65D598.3090606@umdnj.edu> Hi Jeremy: Interesting question. First, my experience with lipid staining is minimal so intrepret my suggestions accordingly. Jeremy Browne wrote: > Clarification, > > Thanks for all your responses. Its clear that I haven't explained > myself very well at all: > > We would like to use oil red O to quantify lipid localisation in > frozen sections of hepatic and muscle tissue. However, we must first > demonstrate that the amount of staining we get (density) is in fact > equal to the amount of lipid present in the sections and to do this we > need to set up some titrations using positive controls with known and > different concentrations of lipid to show a linear response between > staining intensity and lipid concentration. > Is it known that Oil Red O has a linear reaction with the lipids in question (do you know the identity of the lipids in question)? What does the literature say on this topic? Have you tried a model system in which varing amounts of the lipids in question are spotted on paper or gels and the linearity of Oil Red O staining is investigated? > However, a fat smear is difficult to use as a positive control in > this way because it simply washes off the slides. Therefore, we would > like to know what others have done to demonstrate linearity with the > oil red O staining procedure. Is there a way we can treat the fat to > overcome its liquid nature (that is to say not the lipid in the muscle > and liver but the actual 'fat' being used as a positive control)? > How about cutting serial sections and staining one with Oil Red O and using some biochemical method to quantify the lipids you extract/wash off from the adjacent section? Spectrophotmetery? Of course, the method you use to extract the lipids would have to be specific to the Oil Red O stained lipids or those lipids would have to be a very arge majority of the lipids present. This project may keep you busy for awhile. Geoff -- -- ********************************************** 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 mcauliff <@t> umdnj.edu Mon Sep 15 10:14:21 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Masson Trichrome stain In-Reply-To: <3F61F0D2.3040607@ufl.edu> References: <3F61F0D2.3040607@ufl.edu> Message-ID: <3F65D74D.9040300@umdnj.edu> Hi Brian: Using colors to identify tissues is not the way to go. Muscle (skeletal, cardiac or smooth) and collagen have very different morphologies, that alone should be the criteria for identification. A good hematoxylin and eosin or even toluidine blue should tell you what you need to know. Geoff Brian Hatcher wrote: > I am attempting to use Masson's trichrome stain on some rat bone > marrow stem cells cultured on fibrous contstructs. Following staining > with the acid fuchsin, a large amount of tissue growing in between the > fibers is staining red. Following the treatment with aniline blue, > however, this tissue is no longer visible. In some areas it appears > as though it has torn away from the fibers where it was previously > spread between (some arease of floating tissue are visible). My > initial thoughts were that perhaps this tissue was muscle, although > this was a bit supprising as these cells should be differentiating > into osteoblasts in the presence of these fibers. I had also read > somewhere that the collagen would stain red with acid fuchsin, > however, subsequent staining with aniline blue should result in > collagen appearing blue. The only problem is I am seeing neither blue > nor red cells in these specific areas following aniline blue > staining. Any suggestions??? > Thanks > Brian > -- > Brian Hatcher > Graduate Research Assistant > Department of Biomedical Engineering > University of Florida > PO Box 116400 > Gainesville, FL 32611-6400 > Ph: 352-392-6656 > Fax: 352-392-3771 > email: briany@ufl.edu > -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From tpmorken <@t> labvision.com Mon Sep 15 10:31:44 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] CAP Question anp.23075 Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CB67@usca0082k03.rallansci.apogent.com> My interpretation is that they want to see whether someone has actively reviewed the instrument/equipment maintenance sheets and signed them. For instance, does the histology or lab supervisor review freezer temperature log sheets weekly and sign the sheet to prove they have? That would show "evidence of ongoing evaluation." Even better, has the supervisor made any notes on the sheet as to modifications in temp settings? That would be icing on the cake - showing that someone was actually paying attention. Have fun! Tim Morken Lab Vision / NeoMarkers www.labvision.com -----Original Message----- From: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: Sunday, September 14, 2003 2:28 PM To: HISTONET (E-mail) Subject: [Histonet] CAP Question anp.23075 I'm having my CAP inspection soon and wondered what is meant by this question. Is there EVIDENCE of ongoing evaluation of results of instrument maintenance a nd function for all devices? The 2 questions before this one are about a schedule for checking instruments (yes) and are the service and repair records promptly available. (yes) So......What does the above question pertain too?? HELP! Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 _____ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Arkansas Children's Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/b8226730/attachment.htm From gareth.davis <@t> Vanderbilt.Edu Mon Sep 15 11:03:39 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Perfusion Protocol Message-ID: <082C721AF78DB34983E8BA2CD085462105C670@mailbe07> Perfusion Protocol Preparation for Perfusion: Fixative =4% PFA Prepare fresh on day of use or the night before and keep at 4oC. 4% Paraformaldehyde 3.8% Sodium Borate Add 40g of paraformaldehyde to 800ml of water and heat to 60oC while stirring. Add 38g of sodium borate, paraformaldehyde will not dissolve until it is added. Cool off to 4oC. Adjust pH to 9.5 with glacial acetic acid. 4% PFA with 10% sucrose= In a 100ml cylinder add 10g of Sucrose, than add 4% PFA to 100ml Perfusion 1. Anesthetize animal, open thoracic cavity, expose heart and visualize ascending aorta. Insert 18-gauge needle (on vacutainer collection tubing) through left ventricle into ascending aorta- if possible, hard to tell sometimes. Clamp cannula into place with would clip or small clamp and then snip right atrium. (I find it's easier - on mice- to hold needle in place, because clamp gets in the way). 2. Slowly perfuse animal with about 40-50ml room temperature saline (it's recommended to use a pump here, but I use 50 ml syringe, and perfuse slowly) until fluid is clear. 3. Change to cold 4% PFA and slowly perfuse animal. When the animal?s forelimbs are fully extended reduce perfusion speed, use about 50ml of fixative (for a 30g mouse). 4. After perfusion is complete, decapitate the animal and quickly remove the brain using a caudal approach. Place brain in 4% PFA with 10% sucrose at 4oC for overnight. (The 4%PFA with sucrose will post-fix brain and cryoprotect tissue at the same time.) Some recommend to post-fix in 4% PFA-only, overnight, then cryoprotect with 30% sucrose for another 24 hours. Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/6c87bf01/attachment.htm From Kathy.Johnston <@t> CLS.ab.ca Mon Sep 15 11:45:30 2003 From: Kathy.Johnston <@t> CLS.ab.ca (Johnston, Kathy) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Bad Plants/Yarrow Message-ID: <30C050525B881C4AAFF41E6D16543E68015D02A1@mail3.cls.ab.ca> While we are talking "Bad Plants" I would like everyone's opinion on Yarrow. I bought 2 plants this spring and they have grown into to very large although quite stunning plants. I can't remember it's exact name right now, but each stem is a different pastel color. My only concern with is is that is started out in a 2" pot, and right now I would need a 5 gallon pail to put it in if I dug it out! I am understanding that is seems quite prolific. But is it easy to control. Your honest opinions please!!!! Thanks! Kathy -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/70c058fa/attachment.htm From cogold13 <@t> yahoo.com Mon Sep 15 14:43:52 2003 From: cogold13 <@t> yahoo.com (sara goldston) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Bad Plants/Yarrow In-Reply-To: <30C050525B881C4AAFF41E6D16543E68015D02A1@mail3.cls.ab.ca> Message-ID: <20030915194352.7762.qmail@web41507.mail.yahoo.com> I use to be a naturalist in high school and yarrrow is suppose to have medicinal purposes (hopefully you and I are talking about the same plant). If you crush either the leaves or blooms, cannot recall which, under you nose you will smell something similar to Vick's vapor rub. Yarrow has the same effect as Vick's by opening up the nasal passages. But more to your question at hand. Yarrow is a wild bush so I would assume like any wild and un-domesticated plant, it will grow quite freely if given the opportunity to. If you are from anywhere east of the Mississippi, there has been an unusual amount of rain not to mention cooler than normal temperatures. If your plant was kept outside, it would have loved the weather this season and flourished! You may have to break out the pruining shears. Sorry, I don't have any other helpful advice. Try going to a nature reserve, greenhouse or horticulturalist, and perhaps someone along those lines will have some more concrete advise. Sara "Johnston, Kathy" wrote: While we are talking "Bad Plants" I would like everyone's opinion on Yarrow. I bought 2 plants this spring and they have grown into to very large although quite stunning plants. I can't remember it's exact name right now, but each stem is a different pastel color. My only concern with is is that is started out in a 2" pot, and right now I would need a 5 gallon pail to put it in if I dug it out! I am understanding that is seems quite prolific. But is it easy to control. Your honest opinions please!!!! Thanks! Kathy --------------------------------- Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030915/6bc49d9b/attachment.htm From Jacqueline.Miller <@t> UTSouthwestern.edu Mon Sep 15 15:02:24 2003 From: Jacqueline.Miller <@t> UTSouthwestern.edu (Jacqueline Miller) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] background problem Message-ID: Hi, We're having trouble with high background when using Rat IgG1 antibodies (4 ?g/ml) on paraffin-embedded sections of human tissues, with or without retrieval procedures. Does anyone have any advice? Thank you, Jacqueline Miller Research Asst. I OB/GYN Dept. University of Texas Southwestern Medical Center Dallas, TX From SJones <@t> cvm.tamu.edu Mon Sep 15 15:57:36 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Bouin's fixation/removal of picric acid Message-ID: I use it on wet tissue, it does not appear that the lithium needs to be removed. Maybe I'm not understanding your question. Sarah Ref. Theory and Practice of Histotechnology, Sheehan, Hrapchak, page 47. >>> John Kiernan 09/14/03 11:54PM >>> Sarah, do you use the saturated lithium carbonate in 70% ethanol on blocks of tissue or on hydrated paraffin sections that are yellow? If it's for blocks, how do you remove lithium carbonate from the decolorized specimens? John Kiernan -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ___________ Sarah Jones wrote: > > I've always used saturated lithium carbonate in 70% ethanol to remove > the picric acid. I believe it would be faster and more complete than > 70% alone. > > Sarah Jones HT(ASCP) > Dept. of Vet. Anatomy & Public Health > Histology Lab > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > >>> "peptolab" 09/13/03 08:32AM >>> > A 3-5 mm testicular biopsy should be fixed in one to two hours. Be sure > to > rinse out as much yellow picric acid as possible (you can use running > water > or 70% alcohol ). The basophilia of chromatin in the stored block will > deteriorate after some time if there is picric acid residue. > > Jeff Silverman HT HTL QIHC (ASCP) > Southside Hospital > Bay Shore NY > From MGondo <@t> popmail.opt.uh.edu Mon Sep 15 16:28:30 2003 From: MGondo <@t> popmail.opt.uh.edu (Margaret Gondo) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] eye morphology advice Message-ID: <200309151628.AA14942812@popmail.opt.uh.edu> Hi All - I have a grad student who is interested in preserving the morphology of a monkey eyeball once it is removed. Does anyone have any suggestions? Everything we have tried always results in the posterior part of the eyeball caving in. Thanks in advance, Margaret From lpwenk <@t> mail.netquest.com Mon Sep 15 18:10:59 2003 From: lpwenk <@t> mail.netquest.com (Lee & Peggy Wenk) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] ASCP BOR results Message-ID: <006101c37bde$a0817260$8732fea9@hppav> For those interested in the pass rates of the HT and HTL and SLS ASCP exams Jan. - June 2003, please read on. (For those not interested, please press the DELETE key now.) Scaled score of 400 is required to pass the exam. For HT and HTL, both the written (MCQ = Multiple Choice Question) and the practical portions must be passed. NAACLS = National Accrediting Agency for Clinical Laboratory Sciences, which is the accrediting agency for lab training programs. Total Number non-NAACLS candidates = Total Number taking exam - Total Number NAACLS students HISTOTECHNICIAN HT MCQ Mean = 428 Range of Scores = 100-761 Total Number Taking Exam = 206 Total Pass = 111 (54%) Total Number NAACLS students = 24 Total Pass NAACLS students = 18 (75%) Total Number non-NAACLS candidates = 182 Total Pass non-NAACLS candidates = 93 (51%) HT Practical Mean = 446 Range of Scores = 201-889 Total Number Taking Exam = 217 Total Pass = 144 (66%) Total Number NAACLS students = 24 Total Pass NAACLS students = 20 (83%) Total Number non-NAACLS candidates = 193 Total Pass non-NAACLS candidates = 20 (64%) HT COMBINED Total Number Taking Exam = 349 Total Pass = 141 (40%) Total Number NAACLS students = 23 Total Pass NAACLS students = 15 (65%) Total Number non-NAACLS candidates = 326 Total Pass non-NAACLS candidates = 126 (39%) HT exam first given in 1948. Certified to date = 18,556 HISTOTECHNOLOGIST HTL MCQ Mean = 451 Range of Scores = 100-646 Total Number Taking Exam = 43 Total Pass = 32 (74%) Total Number NAACLS students = 0 HTL PRACTICAL Mean = 487 Range of Scores = 244-700 Total Number Taking Exam = 54 Total Pass = 43 (80%) Total Number NAACLS students = 0 HTL COMBINED Total Number Taking Exam = 78 Total Pass = 39 (50%) Total Number NAACLS students = 0 HTL exam first given in 1980. Certified to date = 2,070 SPECIALIST IN LABORATORY SAFETY Mean = 422 Range of Scores = 363-511 Total Number Taking Exam = 7 Total Pass = 3 (43%) SLS exam first given in 2000. Certified to date = 138 Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From lpwenk <@t> mail.netquest.com Mon Sep 15 18:19:56 2003 From: lpwenk <@t> mail.netquest.com (Lee & Peggy Wenk) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] wage and salary survey Message-ID: <006f01c37bdf$e08f0a60$8732fea9@hppav> FYI - The September 2003 issue of "Laboratory Medicine" from ASCP has Part I of the latest Wage and Salary Survey. This survey is done every other year. This article shows national averages of all lab disciplines, and then breaks it down into regional areas. Please remember, your state, or even your part of the state, may pay higher or lower than these averages. Things of interest that I found: - HT and HTL still have the highest vacancy rates of all laboratory personnel. This is the same as the survey done 2 years ago. - HTL and MT now have the same salary ranges, on average nationwide. In other words, on national average, HTL are now being paid the same rate as MT. A few of the charts on the national averages can be found on the ASCP Board of Registry website: http://www.ascp.org/bor/center/center_research.asp Thought some people might like to find their issue, or borrow one from a colleague (or maybe even join ASCP). Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From lenaspencer <@t> insightbb.com Mon Sep 15 19:58:58 2003 From: lenaspencer <@t> insightbb.com (lena spencer) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Colloidal Iron Message-ID: <005801c37bed$b65e3a00$21b6dc0c@insightbb.com> Hi All: One of my Pathologist's has requested a Hale's Colloidal Iron for renal cell carcinoma chromophobes. I am familiar with colloidal iron for mucosubstances, but he feel's that this is not the same stain. The article stated that the diagnosis is made from the colloidal iron stain or from electron microscopy . This Pathologist is requesting that I post on the histonet for additional information or for someone out in histoland who is performing this procedure. Your is insight would be greatly appreciated. Lena From jkiernan <@t> uwo.ca Mon Sep 15 23:24:20 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Bouin's fixation/removal of picric acid References: Message-ID: <3F669073.682F479E@uwo.ca> I didn't phrase my question very clearly, and should have explained the reason for asking. Previously I've encountered lithium carbonate for removing picric from sections. It's the traditional thing to do after Bouin fixation when the tissue is still yellow in the wax and after hydration of the sections. You can easily see when all the yellow has gone. With a block you can't see the middle, and I was wondering how long it takes until no more yellow comes out into the liquid. With 70% and higher alcohols alone it seems to take for ever (at least with 5 mm and larger specimens; I haven't used tiny biopsies after Bouin fixation). It would be nice to have an idea how much more quickly the picric acid is extracted when the 70% alc is saturated with Li2CO3. I asked about removing the Li2CO3 because it's almost insoluble in alcohol (Merck Index). That would be 95-100% alc. I guess you rinse the specimens in more 70% after extracting the picric acid, and that removes the Li2CO3 and you don't get damage from crystals forming in the tissue. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ _________________________ Sarah Jones wrote: > > I use it on wet tissue, it does not appear that the lithium needs to be > removed. Maybe I'm not understanding your question. Sarah > Ref. Theory and Practice of Histotechnology, Sheehan, Hrapchak, page > 47. > > >>> John Kiernan 09/14/03 11:54PM >>> > Sarah, do you use the saturated lithium > carbonate in 70% ethanol on blocks of > tissue or on hydrated paraffin sections > that are yellow? > > If it's for blocks, how do you remove > lithium carbonate from the decolorized > specimens? > John Kiernan > -- > ------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ > ___________ > Sarah Jones wrote: > > > > I've always used saturated lithium carbonate in 70% ethanol to > remove > > the picric acid. I believe it would be faster and more complete > than > > 70% alone. > > > > Sarah Jones HT(ASCP) > > Dept. of Vet. Anatomy & Public Health > > Histology Lab > > Texas A&M University > > College Station, TX 77843-4458 > > phone: 979-845-3177 > > fax: 979-458-3499 > > > > >>> "peptolab" 09/13/03 08:32AM >>> > > A 3-5 mm testicular biopsy should be fixed in one to two hours. Be > sure > > to > > rinse out as much yellow picric acid as possible (you can use > running > > water > > or 70% alcohol ). The basophilia of chromatin in the stored block > will > > deteriorate after some time if there is picric acid residue. > > > > Jeff Silverman HT HTL QIHC (ASCP) > > Southside Hospital > > Bay Shore NY > > From jkiernan <@t> uwo.ca Tue Sep 16 00:22:32 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Colloidal Iron References: <005801c37bed$b65e3a00$21b6dc0c@insightbb.com> Message-ID: <3F669E18.C5B0A4E2@uwo.ca> There's just the one Hale's (1946) method, also called the dialysed iron technique because if you make the original reagent yourself you have to dialyse the ferric hydroxide sol. Detailed instructions are given in Pearse's Histochemistry (any edition). There have been several variants of the principal reagent. In all the techniques the bound ferric iron is made visible by a Prussian blue reaction with potassium ferrocyanide. In Chapter 10 of the latest (2002) edition of Bancroft & Gamble's "Theory and Practice ..." by Barbara Totty, the method attributed to Hale (1946) uses a more easily made colloidal iron sol that does not need to be dialysed. A more detailed reading of Pearse (3rd ed) shows that this is close to the reagent of G.Muller (3 papers in Acta Histochem vol 2, 1955-56, not seen by me; they'll be in German). This colloidal iron sol quickly became more popular than Hale's reagent not only because it was easier to make but also because it was more specific for acid mucosubstances. The not-dialysed colloidal iron solution, unlike Hale's, does not stain nuclei and there is no "background" cytoplasmic blue colour in renal tubules. Both Pearse and Totty point out that colloidal iron staining does the same job as alcian blue (pH 2.5) but provides a darker blue product and also some nonspecific staining. It's more sensitive and less clean. "Hale's colloidal iron" (subject of the original enquiry) is probably not an appropriate name for this technique, which has been significantly improved since 1946. Pearse seriously questioned its histochemical specificity despite recognizing the improvements made by Muller and several others. Alcian blue is easier to use and can be incorporated into many other rational mixed staining procedures for mucosubstances. I know nothing about renal carcinoma chromophobes. If they can be stained with colloidal iron they must be basophilic, not "chromophobic." There are very few genuinely chromophobic (unstainable) cytoplasms. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ __________________________________ lena spencer wrote: > > Hi All: > One of my Pathologist's has requested a Hale's Colloidal Iron for renal cell > carcinoma chromophobes. I am familiar with colloidal iron for > mucosubstances, but he feel's that this is not the same stain. The article > stated that the diagnosis is made from the colloidal iron stain or from > electron microscopy . This Pathologist is requesting that I post on the > histonet for additional information or for someone out in histoland who is > performing this procedure. > Your is insight would be greatly appreciated. > Lena From kemlo <@t> tiscali.co.uk Tue Sep 16 01:33:44 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] eye morphology advice In-Reply-To: <200309151628.AA14942812@popmail.opt.uh.edu> Message-ID: <000001c37c1c$7e35f620$48d4e150@KEMLOS> Nitrocellulose and celloidin technique. Page 91 of Histopathologic Technic and Practical Histochemistry. RD Lillie and Harold M. Fullmer.1976, 1965 McGrew Inc. 1234567890 KPKP 7832109876 48hr Form fixation dehydrate with successive baths of 35, 50, 65 & 80% alc. 24 hours each. Open eye Return to 80% alc. 24 hours 2x 100% alc 24 hours 6 hour bath in equal vols 100% alc and ether Infiltrate 5-7 days in 10% nitrocellulose in 100% alc. Infiltrate 5-7 days in 20% nitrocellulose in 100% alc Place under bell jar and when surface is solid, examine daily, but lower portion soft flood with chloroform 16-24 hours. Pour off chloroform and let dry. Attach to block with 20% nitrocellulose Let dry and immerse in chloroform for several hours. 24 hour baths of 3:1 chloroform: cedar wood oil, 1:1 of the same, then pure cedar wood oil. Cut sections and store in 80% alc. Dangerous, smelly, time consuming but extremely good fun; definitely a non-PC technique that works; done it myself as a pup. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts:? ???????????? kemlo@tiscali.co.uk?or kemlo1@btinternet.com? Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. ? ? ? -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Margaret Gondo Sent: 15 September 2003 22:29 To: histonet@pathology.swmed.edu Subject: [Histonet] eye morphology advice Hi All - I have a grad student who is interested in preserving the morphology of a monkey eyeball once it is removed. Does anyone have any suggestions? Everything we have tried always results in the posterior part of the eyeball caving in. Thanks in advance, Margaret _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bosborn <@t> hsc.usf.edu Tue Sep 16 07:10:58 2003 From: bosborn <@t> hsc.usf.edu (Barbara Osborn) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] Re: Methacarn/Carnoy's Storage Message-ID: <200309161210.h8GCAwO04890@hsc.usf.edu> Tamara, You can store methacarn or Carnoy's for up to a couple of years as long as you cap it very, very tightly to avoid evaporation of chloroform and alcohol. So you can store a large batch of fixative and only take out what is needed each week. After use, however, it must be discarded. From Jackie.O'Connor <@t> abbott.com Tue Sep 16 07:30:32 2003 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:21:52 2005 Subject: [Histonet] IHC on Blood Smears Message-ID: A colleague of mine would like to stain murine blood smears for Bcl2. Does anyone have any experience doing IHC on blood smears which have been fixed in Methanol? Thanks Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 Jackie.O'Connor@abbott.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030916/a51742e3/attachment.htm From funderwood <@t> mcohio.org Tue Sep 16 09:14:27 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] eye morphology advice Message-ID: I think this was discussed in the past. If my memory serves me right, injecting the eye with a gelatin solution before processing will help retain the shape. A search of the archives may be worth while. Fred >>> "Margaret Gondo" 09/15/03 05:28PM >>> Hi All - I have a grad student who is interested in preserving the morphology of a monkey eyeball once it is removed. Does anyone have any suggestions? Everything we have tried always results in the posterior part of the eyeball caving in. Thanks in advance, Margaret _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Tue Sep 16 09:14:27 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] eye morphology advice Message-ID: I think this was discussed in the past. If my memory serves me right, injecting the eye with a gelatin solution before processing will help retain the shape. A search of the archives may be worth while. Fred >>> "Margaret Gondo" 09/15/03 05:28PM >>> Hi All - I have a grad student who is interested in preserving the morphology of a monkey eyeball once it is removed. Does anyone have any suggestions? Everything we have tried always results in the posterior part of the eyeball caving in. Thanks in advance, Margaret _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Tue Sep 16 09:26:51 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] IHC on Blood Smears Message-ID: If the smears are quite thick and bloody, you may want to use a more dilute hydrogen peroxide solution (1% or so) for quenching. The vigorous reaction can strip material from the slide. Prolonged buffer wahes are a good idea as well. Fred >>> 09/16/03 08:30AM >>> A colleague of mine would like to stain murine blood smears for Bcl2. Does anyone have any experience doing IHC on blood smears which have been fixed in Methanol? Thanks Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 Jackie.O'Connor@abbott.com From v.lamanna <@t> abdn.ac.uk Tue Sep 16 09:48:43 2003 From: v.lamanna <@t> abdn.ac.uk (Vincenzo La Manna) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] immunohistochemistry and frozen sections Message-ID: <2429.139.133.23.42.1063723723.squirrel@www.abdn.ac.uk> Hi Histonetters, I've been struggling for 4 months on my 5 microns frozen sections trying to find estrogen receptors. I've tried both immunoperoxidase kit from Vector laboratories either indirect immunofluorecsence (FITC Goat anti mouse Secondary Ab from Sigma) but I've not yet obtained good results. My primary Ab is a mouse monoclonal to bovine Estrogen receptor from Cymbus biotechnology. Sections come from tissue that is supposed to be a non-target tissue (mid laminar region from cow's hooves)blocking with 2% goat normal serum (sigma) whash in tbs buffer All different methods for antigen retrieval have been tested but sections look poor regarding histologic quality and I still have very big problems with non specific binding an staining (indirect immunofluorescence). It looks like frozen sections are too tender to efford the whole processing. Every suggestion or reference is welcome, thank U La Manna Vincenzo Department of Agriculture and Forestry, University of Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. Telephone; 01224 274259 Fax; 01224 273731 e-mail v.lamanna@abdn.ac.uk From huffpw <@t> uleth.ca Tue Sep 16 10:29:40 2003 From: huffpw <@t> uleth.ca (Phillip Huff) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] immunohistochemistry and frozen sections In-Reply-To: <2429.139.133.23.42.1063723723.squirrel@www.abdn.ac.uk> References: <2429.139.133.23.42.1063723723.squirrel@www.abdn.ac.uk> Message-ID: <2073.142.66.42.15.1063726180.squirrel@webmail.uleth.ca> First, I would recommend that you run some Western Blots to ensure that your Antigen of interest is being expressed within your tissues. Once your Western blots are positive, you can definetly say that your immunoreaction is what needs to be optimized. Second, have you applied your immunoreaction technique to a known positive tissue source, that has also been sectioned at 5 um? It may be a good confirmation that your tissue may need to be processed differently or cut thicker. Finally, if your end goal is immunolocalization and the immunohistochem is not working, you may want to look into in situ hybridization. In your tissue samples, the receptor may be quite sensitive to processing and you may be losing a great deal of your antigen. I know very little about your receptor, but does it have a high turnover rate? Perhaps it has been degraded over time when frozen. Has the reaction been done in freshly harvested tissues? Perhaps localizing the mRNA in the tissues may provide you with the results you are looking for. Before doing this though, I would confirm that your tissues express the mRNA of interest by doing some RT-PCR. This will both confirm that your tissues have your target mRNA, and if you use an internal housekeeping primer in the PCR, you can determine the approximate copy number of your mRNA to help you in determining how sensitive your in situ reaction must be. Hope this helps you out, Phil > Hi Histonetters, > > I've been struggling for 4 months on my 5 microns > frozen sections trying to find estrogen receptors. > I've tried both immunoperoxidase kit from Vector > laboratories either indirect immunofluorecsence (FITC > Goat anti mouse Secondary Ab from Sigma) but I've not > yet obtained good results. > My primary Ab is a mouse monoclonal to bovine > Estrogen receptor from Cymbus biotechnology. > Sections come from tissue that is supposed to be a > non-target tissue (mid laminar region from cow's > hooves)blocking with 2% goat normal serum (sigma) > whash in tbs buffer > All different methods for antigen retrieval have been > tested but sections look poor regarding histologic > quality and I still have very big problems with non > specific binding an staining (indirect > immunofluorescence). > It looks like frozen sections are too tender to > efford the whole processing. > Every suggestion or reference is welcome, > thank U > > La Manna Vincenzo > Department of Agriculture and Forestry, University of > Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. > Telephone; 01224 274259 > Fax; 01224 273731 > e-mail v.lamanna@abdn.ac.uk > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From tvedilago <@t> system1.net Tue Sep 16 10:31:16 2003 From: tvedilago <@t> system1.net (Tommy Vedilago) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Talented Histotechs wanted Message-ID: Hello Histonetters, If you have ever wanted to work for a lab that is truly a family atmosphere where your skills and abilities are concretely appreciated, I have just the job for you. I have 3 positions for Histotechs in Dallas, TX with a progressive lab that has been in place for 36 years now. They are offering outstanding pay, benefits, and atmosphere. They are also offering relocation and hire bonus. If this holds interest for you or if you know of someone in the job market, please give me a call at 866-SYSTEM1. Tommy Vedilago System 1 Search (864) 627-0012 (864) 627-0013 Fax From nick.kirk3 <@t> btopenworld.com Tue Sep 16 10:58:35 2003 From: nick.kirk3 <@t> btopenworld.com (nick.kirk3@btopenworld.com) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] immunohistochemistry and frozen sections Message-ID: <123866.1063727915417.JavaMail.root@127.0.0.1> If it's not an inpolite question, why are you trying to demonstrate estrogen receptors on frozen tissue? Is this absolutely neccessary as estrogen receptors are very easily demonstrated on paraffin processed tissue, which maintains better morphology and gets almost guaranteed results. We use the DAKO 1D5 clone on FFPE tissue using Vector Unmasking fluid for Antigen Retrieval and the DAKO ChemMate detection system and get excellent results every time. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England > from: Vincenzo La Manna > date: Tue, 16 Sep 2003 15:48:43 > to: histonet@lists.utsouthwestern.edu > subject: Re: [Histonet] immunohistochemistry and frozen sections > > > > Hi Histonetters, > > I've been struggling for 4 months on my 5 microns > frozen sections trying to find estrogen receptors. > I've tried both immunoperoxidase kit from Vector > laboratories either indirect immunofluorecsence (FITC > Goat anti mouse Secondary Ab from Sigma) but I've not > yet obtained good results. > My primary Ab is a mouse monoclonal to bovine > Estrogen receptor from Cymbus biotechnology. > Sections come from tissue that is supposed to be a > non-target tissue (mid laminar region from cow's > hooves)blocking with 2% goat normal serum (sigma) > whash in tbs buffer > All different methods for antigen retrieval have been > tested but sections look poor regarding histologic > quality and I still have very big problems with non > specific binding an staining (indirect > immunofluorescence). > It looks like frozen sections are too tender to > efford the whole processing. > Every suggestion or reference is welcome, > thank U > > La Manna Vincenzo > Department of Agriculture and Forestry, University of > Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. > Telephone; 01224 274259 > Fax; 01224 273731 > e-mail v.lamanna@abdn.ac.uk > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dobbin <@t> upei.ca Tue Sep 16 11:58:43 2003 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] immunohistochemistry and frozen sections In-Reply-To: <2429.139.133.23.42.1063723723.squirrel@www.abdn.ac.uk> Message-ID: <3F671714.30069.1205637@localhost> Vincenzo, You did not mention whether or not the tissues were fixed, so assuming you are cutting fresh frozen sections, why are you applying atigen retreival methods? Antigen retreival methods are only for formalin fixed tissues. Greg Date sent: Tue, 16 Sep 2003 15:48:43 +0100 (BST) From: Vincenzo La Manna Subject: [Histonet] immunohistochemistry and frozen sections To: histonet@lists.utsouthwestern.edu > > > Hi Histonetters, > > I've been struggling for 4 months on my 5 microns > frozen sections trying to find estrogen receptors. > I've tried both immunoperoxidase kit from Vector > laboratories either indirect immunofluorecsence (FITC > Goat anti mouse Secondary Ab from Sigma) but I've not > yet obtained good results. > My primary Ab is a mouse monoclonal to bovine > Estrogen receptor from Cymbus biotechnology. > Sections come from tissue that is supposed to be a > non-target tissue (mid laminar region from cow's > hooves)blocking with 2% goat normal serum (sigma) > whash in tbs buffer > All different methods for antigen retrieval have been > tested but sections look poor regarding histologic > quality and I still have very big problems with non > specific binding an staining (indirect > immunofluorescence). > It looks like frozen sections are too tender to > efford the whole processing. > Every suggestion or reference is welcome, > thank U > > La Manna Vincenzo > Department of Agriculture and Forestry, University of > Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. > Telephone; 01224 274259 > Fax; 01224 273731 > e-mail v.lamanna@abdn.ac.uk > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Pathology Lab Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Making a living is getting; making a life is giving. From weipingren <@t> yahoo.com Tue Sep 16 12:13:55 2003 From: weipingren <@t> yahoo.com (weiping Ren) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] questions regarding to a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate stain(non specific esterase) Message-ID: <20030916171355.43299.qmail@web41603.mail.yahoo.com> Hi, Histonetters: I need to set up non specific esterase staining method on bone marrow smear samples. My questions are: 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate stain(non specific esterase) are the same stain? 2. If these are two different stains but have the same clinical diagnosis significance, which one is better regarding to the time and accuracy? 3. Are there any protocols for these stains? Where I can get these references? Thank you! Any suggestions and advice welcome. Weiping Wayne State University Detroit, MI --------------------------------- Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030916/8ff18437/attachment.htm From Sue.Kapoor <@t> uhsi.org Tue Sep 16 13:22:12 2003 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] buying used lab equipment Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F54D5@khmcexch.uhsi.org> Can anyone tell me their experience with purchasing used lab equipment...did the equipment last? how long? if you had any problems with it, was it resolved to your satisfaction? did you purchase from a company dealing in used equipment? from another lab? and anything else you care to share. I'm looking for a used slide dryer, the type you open the lid and set the rack of slides in. One source only had a very old model and another that was recommended on histonet was similar to ebay....I just want a simple and affordable slide drying oven. Thanks, Sue Kapoor, HT (ASCP) Histology Supervisor Kenosha Medical Center Kenosha, WI sue.kapoor@uhsi.org From g.lang <@t> bigfoot.de Tue Sep 16 14:23:10 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] eye morphology advice References: <200309151628.AA14942812@popmail.opt.uh.edu> Message-ID: <000c01c37c87$f7007f60$0d12a8c0@SERVER> look at this histologic-site from 1976. http://206.137.77.9/Sakura/pdf/6n10176.pdf greetings, Gundi Lang ----- Original Message ----- From: "Margaret Gondo" To: Sent: Monday, September 15, 2003 11:28 PM Subject: [Histonet] eye morphology advice > Hi All - > > I have a grad student who is interested in preserving the > morphology of a monkey eyeball once it is removed. Does anyone > have any suggestions? Everything we have tried always results in > the posterior part of the eyeball caving in. > > > > Thanks in advance, > Margaret > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From rosemarie_g_behan <@t> groton.pfizer.com Tue Sep 16 14:57:42 2003 From: rosemarie_g_behan <@t> groton.pfizer.com (Behan, Rosemarie G) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] RE: Histonet digest, Vol 1 #49 - 19 msgs Message-ID: <9DF8DDFE3736D3119D6D0008C79148D60A5BFD21@groexmbcr17.pfizer.com> I am looking for a recipe for acid cleaning glassware to do silver stains, can anyone help me? -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, September 16, 2003 1:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet digest, Vol 1 #49 - 19 msgs 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-admin@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: Bad Plants/Yarrow (sara goldston) 2. background problem (Jacqueline Miller) 3. Re: Bouin's fixation/removal of picric acid (Sarah Jones) 4. eye morphology advice (Margaret Gondo) 5. ASCP BOR results (Lee & Peggy Wenk) 6. wage and salary survey (Lee & Peggy Wenk) 7. Colloidal Iron (lena spencer) 8. Re: Bouin's fixation/removal of picric acid (John Kiernan) 9. Re: Colloidal Iron (John Kiernan) 10. RE: eye morphology advice (Kemlo Rogerson) 11. Re: Methacarn/Carnoy's Storage (Barbara Osborn) 12. IHC on Blood Smears (Jackie.O'Connor@abbott.com) 13. Re: eye morphology advice (Fred Underwood) 14. Re: eye morphology advice (Fred Underwood) 15. Re: IHC on Blood Smears (Fred Underwood) 16. immunohistochemistry and frozen sections (Vincenzo La Manna) 17. Re: immunohistochemistry and frozen sections (Phillip Huff) 18. Talented Histotechs wanted (Tommy Vedilago) 19. Re: immunohistochemistry and frozen sections (nick.kirk3@btopenworld.com) --__--__-- Message: 1 Date: Mon, 15 Sep 2003 12:43:52 -0700 (PDT) From: sara goldston To: "Johnston, Kathy" , "Histonet \(E-mail\)" Subject: Re: [Histonet] Bad Plants/Yarrow --0-349928651-1063655032=:6512 Content-Type: text/plain; charset=us-ascii I use to be a naturalist in high school and yarrrow is suppose to have medicinal purposes (hopefully you and I are talking about the same plant). If you crush either the leaves or blooms, cannot recall which, under you nose you will smell something similar to Vick's vapor rub. Yarrow has the same effect as Vick's by opening up the nasal passages. But more to your question at hand. Yarrow is a wild bush so I would assume like any wild and un-domesticated plant, it will grow quite freely if given the opportunity to. If you are from anywhere east of the Mississippi, there has been an unusual amount of rain not to mention cooler than normal temperatures. If your plant was kept outside, it would have loved the weather this season and flourished! You may have to break out the pruining shears. Sorry, I don't have any other helpful advice. Try going to a nature reserve, greenhouse or horticulturalist, and perhaps someone along those lines will have some more concrete advise. Sara "Johnston, Kathy" wrote: While we are talking "Bad Plants" I would like everyone's opinion on Yarrow. I bought 2 plants this spring and they have grown into to very large although quite stunning plants. I can't remember it's exact name right now, but each stem is a different pastel color. My only concern with is is that is started out in a 2" pot, and right now I would need a 5 gallon pail to put it in if I dug it out! I am understanding that is seems quite prolific. But is it easy to control. Your honest opinions please!!!! Thanks! Kathy --------------------------------- Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software --0-349928651-1063655032=:6512 Content-Type: text/html; charset=us-ascii
I use to be a naturalist in high school and yarrrow is suppose to have medicinal purposes (hopefully you and I are talking about the same plant).  If you crush either the leaves or blooms, cannot recall which, under you nose you will smell something similar to Vick's vapor rub.  Yarrow has the same effect as Vick's by opening up the nasal passages.  But more to your question at hand.  Yarrow is a wild bush so I would assume like any wild and un-domesticated plant, it will grow quite freely if given the opportunity to.  If you are from anywhere east of the Mississippi, there has been an unusual amount of rain not to mention cooler than normal temperatures.  If your plant was kept outside, it would have loved the weather this season and flourished!  You may have to break out the pruining shears.  Sorry, I don't have any other helpful advice.  Try going to a nature reserve, greenhouse or horticulturalist, and perhaps so meone along those lines will have some more concrete advise.
Sara 

"Johnston, Kathy" <Kathy.Johnston@CLS.ab.ca> wrote:
While we are talking "Bad Plants"  I would like everyone's opinion on Yarrow.  I bought 2 plants this spring and they have grown into to very large although quite stunning plants.  I can't remember it's exact name right now, but each stem is a different pastel color.  My only concern with is is that is started out in a 2" pot, and right now I would need a 5 gallon pail to put it in if I dug it out!
 
I am understanding that is seems quite prolific.  But is it easy to control.
 
Your honest opinions please!!!!
 
Thanks!
 
Kathy

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Yahoo! SiteBuilder - Free, easy-to-use web site design software --0-349928651-1063655032=:6512-- --__--__-- Message: 2 Date: Mon, 15 Sep 2003 15:02:24 -0500 From: "Jacqueline Miller" To: Subject: [Histonet] background problem Hi, We're having trouble with high background when using Rat IgG1 antibodies = (4 =B5g/ml) on paraffin-embedded sections of human tissues, with or = without retrieval procedures. Does anyone have any advice? Thank you, Jacqueline Miller Research Asst. I OB/GYN Dept. University of Texas Southwestern Medical Center Dallas, TX --__--__-- Message: 3 Date: Mon, 15 Sep 2003 15:57:36 -0500 From: "Sarah Jones" To: Cc: , Subject: Re: [Histonet] Bouin's fixation/removal of picric acid I use it on wet tissue, it does not appear that the lithium needs to be removed. Maybe I'm not understanding your question. Sarah Ref. Theory and Practice of Histotechnology, Sheehan, Hrapchak, page 47. >>> John Kiernan 09/14/03 11:54PM >>> Sarah, do you use the saturated lithium carbonate in 70% ethanol on blocks of tissue or on hydrated paraffin sections that are yellow? If it's for blocks, how do you remove lithium carbonate from the decolorized specimens? John Kiernan -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ___________ Sarah Jones wrote: > > I've always used saturated lithium carbonate in 70% ethanol to remove > the picric acid. I believe it would be faster and more complete than > 70% alone. > > Sarah Jones HT(ASCP) > Dept. of Vet. Anatomy & Public Health > Histology Lab > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > >>> "peptolab" 09/13/03 08:32AM >>> > A 3-5 mm testicular biopsy should be fixed in one to two hours. Be sure > to > rinse out as much yellow picric acid as possible (you can use running > water > or 70% alcohol ). The basophilia of chromatin in the stored block will > deteriorate after some time if there is picric acid residue. > > Jeff Silverman HT HTL QIHC (ASCP) > Southside Hospital > Bay Shore NY > --__--__-- Message: 4 Date: Mon, 15 Sep 2003 16:28:30 -0500 From: "Margaret Gondo" Reply-To: To: Subject: [Histonet] eye morphology advice Hi All - I have a grad student who is interested in preserving the morphology of a monkey eyeball once it is removed. Does anyone have any suggestions? Everything we have tried always results in the posterior part of the eyeball caving in. Thanks in advance, Margaret --__--__-- Message: 5 From: "Lee & Peggy Wenk" To: "Histonet" Cc: Date: Mon, 15 Sep 2003 19:10:59 -0400 Subject: [Histonet] ASCP BOR results For those interested in the pass rates of the HT and HTL and SLS ASCP exams Jan. - June 2003, please read on. (For those not interested, please press the DELETE key now.) Scaled score of 400 is required to pass the exam. For HT and HTL, both the written (MCQ = Multiple Choice Question) and the practical portions must be passed. NAACLS = National Accrediting Agency for Clinical Laboratory Sciences, which is the accrediting agency for lab training programs. Total Number non-NAACLS candidates = Total Number taking exam - Total Number NAACLS students HISTOTECHNICIAN HT MCQ Mean = 428 Range of Scores = 100-761 Total Number Taking Exam = 206 Total Pass = 111 (54%) Total Number NAACLS students = 24 Total Pass NAACLS students = 18 (75%) Total Number non-NAACLS candidates = 182 Total Pass non-NAACLS candidates = 93 (51%) HT Practical Mean = 446 Range of Scores = 201-889 Total Number Taking Exam = 217 Total Pass = 144 (66%) Total Number NAACLS students = 24 Total Pass NAACLS students = 20 (83%) Total Number non-NAACLS candidates = 193 Total Pass non-NAACLS candidates = 20 (64%) HT COMBINED Total Number Taking Exam = 349 Total Pass = 141 (40%) Total Number NAACLS students = 23 Total Pass NAACLS students = 15 (65%) Total Number non-NAACLS candidates = 326 Total Pass non-NAACLS candidates = 126 (39%) HT exam first given in 1948. Certified to date = 18,556 HISTOTECHNOLOGIST HTL MCQ Mean = 451 Range of Scores = 100-646 Total Number Taking Exam = 43 Total Pass = 32 (74%) Total Number NAACLS students = 0 HTL PRACTICAL Mean = 487 Range of Scores = 244-700 Total Number Taking Exam = 54 Total Pass = 43 (80%) Total Number NAACLS students = 0 HTL COMBINED Total Number Taking Exam = 78 Total Pass = 39 (50%) Total Number NAACLS students = 0 HTL exam first given in 1980. Certified to date = 2,070 SPECIALIST IN LABORATORY SAFETY Mean = 422 Range of Scores = 363-511 Total Number Taking Exam = 7 Total Pass = 3 (43%) SLS exam first given in 2000. Certified to date = 138 Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 --__--__-- Message: 6 From: "Lee & Peggy Wenk" To: "Histonet" Cc: Date: Mon, 15 Sep 2003 19:19:56 -0400 Subject: [Histonet] wage and salary survey FYI - The September 2003 issue of "Laboratory Medicine" from ASCP has Part I of the latest Wage and Salary Survey. This survey is done every other year. This article shows national averages of all lab disciplines, and then breaks it down into regional areas. Please remember, your state, or even your part of the state, may pay higher or lower than these averages. Things of interest that I found: - HT and HTL still have the highest vacancy rates of all laboratory personnel. This is the same as the survey done 2 years ago. - HTL and MT now have the same salary ranges, on average nationwide. In other words, on national average, HTL are now being paid the same rate as MT. A few of the charts on the national averages can be found on the ASCP Board of Registry website: http://www.ascp.org/bor/center/center_research.asp Thought some people might like to find their issue, or borrow one from a colleague (or maybe even join ASCP). Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 --__--__-- Message: 7 From: "lena spencer" To: Date: Mon, 15 Sep 2003 20:58:58 -0400 Subject: [Histonet] Colloidal Iron Hi All: One of my Pathologist's has requested a Hale's Colloidal Iron for renal cell carcinoma chromophobes. I am familiar with colloidal iron for mucosubstances, but he feel's that this is not the same stain. The article stated that the diagnosis is made from the colloidal iron stain or from electron microscopy . This Pathologist is requesting that I post on the histonet for additional information or for someone out in histoland who is performing this procedure. Your is insight would be greatly appreciated. Lena --__--__-- Message: 8 Date: Tue, 16 Sep 2003 00:24:20 -0400 From: John Kiernan Reply-To: jkiernan@uwo.ca To: Sarah Jones CC: histonet@pathology.swmed.edu Subject: Re: [Histonet] Bouin's fixation/removal of picric acid I didn't phrase my question very clearly, and should have explained the reason for asking. Previously I've encountered lithium carbonate for removing picric from sections. It's the traditional thing to do after Bouin fixation when the tissue is still yellow in the wax and after hydration of the sections. You can easily see when all the yellow has gone. With a block you can't see the middle, and I was wondering how long it takes until no more yellow comes out into the liquid. With 70% and higher alcohols alone it seems to take for ever (at least with 5 mm and larger specimens; I haven't used tiny biopsies after Bouin fixation). It would be nice to have an idea how much more quickly the picric acid is extracted when the 70% alc is saturated with Li2CO3. I asked about removing the Li2CO3 because it's almost insoluble in alcohol (Merck Index). That would be 95-100% alc. I guess you rinse the specimens in more 70% after extracting the picric acid, and that removes the Li2CO3 and you don't get damage from crystals forming in the tissue. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ _________________________ Sarah Jones wrote: > > I use it on wet tissue, it does not appear that the lithium needs to be > removed. Maybe I'm not understanding your question. Sarah > Ref. Theory and Practice of Histotechnology, Sheehan, Hrapchak, page > 47. > > >>> John Kiernan 09/14/03 11:54PM >>> > Sarah, do you use the saturated lithium > carbonate in 70% ethanol on blocks of > tissue or on hydrated paraffin sections > that are yellow? > > If it's for blocks, how do you remove > lithium carbonate from the decolorized > specimens? > John Kiernan > -- > ------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ > ___________ > Sarah Jones wrote: > > > > I've always used saturated lithium carbonate in 70% ethanol to > remove > > the picric acid. I believe it would be faster and more complete > than > > 70% alone. > > > > Sarah Jones HT(ASCP) > > Dept. of Vet. Anatomy & Public Health > > Histology Lab > > Texas A&M University > > College Station, TX 77843-4458 > > phone: 979-845-3177 > > fax: 979-458-3499 > > > > >>> "peptolab" 09/13/03 08:32AM >>> > > A 3-5 mm testicular biopsy should be fixed in one to two hours. Be > sure > > to > > rinse out as much yellow picric acid as possible (you can use > running > > water > > or 70% alcohol ). The basophilia of chromatin in the stored block > will > > deteriorate after some time if there is picric acid residue. > > > > Jeff Silverman HT HTL QIHC (ASCP) > > Southside Hospital > > Bay Shore NY > > --__--__-- Message: 9 Date: Tue, 16 Sep 2003 01:22:32 -0400 From: John Kiernan Reply-To: jkiernan@uwo.ca To: lena spencer CC: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Colloidal Iron There's just the one Hale's (1946) method, also called the dialysed iron technique because if you make the original reagent yourself you have to dialyse the ferric hydroxide sol. Detailed instructions are given in Pearse's Histochemistry (any edition). There have been several variants of the principal reagent. In all the techniques the bound ferric iron is made visible by a Prussian blue reaction with potassium ferrocyanide. In Chapter 10 of the latest (2002) edition of Bancroft & Gamble's "Theory and Practice ..." by Barbara Totty, the method attributed to Hale (1946) uses a more easily made colloidal iron sol that does not need to be dialysed. A more detailed reading of Pearse (3rd ed) shows that this is close to the reagent of G.Muller (3 papers in Acta Histochem vol 2, 1955-56, not seen by me; they'll be in German). This colloidal iron sol quickly became more popular than Hale's reagent not only because it was easier to make but also because it was more specific for acid mucosubstances. The not-dialysed colloidal iron solution, unlike Hale's, does not stain nuclei and there is no "background" cytoplasmic blue colour in renal tubules. Both Pearse and Totty point out that colloidal iron staining does the same job as alcian blue (pH 2.5) but provides a darker blue product and also some nonspecific staining. It's more sensitive and less clean. "Hale's colloidal iron" (subject of the original enquiry) is probably not an appropriate name for this technique, which has been significantly improved since 1946. Pearse seriously questioned its histochemical specificity despite recognizing the improvements made by Muller and several others. Alcian blue is easier to use and can be incorporated into many other rational mixed staining procedures for mucosubstances. I know nothing about renal carcinoma chromophobes. If they can be stained with colloidal iron they must be basophilic, not "chromophobic." There are very few genuinely chromophobic (unstainable) cytoplasms. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ __________________________________ lena spencer wrote: > > Hi All: > One of my Pathologist's has requested a Hale's Colloidal Iron for renal cell > carcinoma chromophobes. I am familiar with colloidal iron for > mucosubstances, but he feel's that this is not the same stain. The article > stated that the diagnosis is made from the colloidal iron stain or from > electron microscopy . This Pathologist is requesting that I post on the > histonet for additional information or for someone out in histoland who is > performing this procedure. > Your is insight would be greatly appreciated. > Lena --__--__-- Message: 10 From: "Kemlo Rogerson" To: , Date: Tue, 16 Sep 2003 07:33:44 +0100 Subject: RE: [Histonet] eye morphology advice Nitrocellulose and celloidin technique. Page 91 of Histopathologic Technic and Practical Histochemistry. RD Lillie and Harold M. Fullmer.1976, 1965 McGrew Inc. 1234567890 KPKP 7832109876 48hr Form fixation dehydrate with successive baths of 35, 50, 65 & 80% alc. 24 hours each. Open eye Return to 80% alc. 24 hours 2x 100% alc 24 hours 6 hour bath in equal vols 100% alc and ether Infiltrate 5-7 days in 10% nitrocellulose in 100% alc. Infiltrate 5-7 days in 20% nitrocellulose in 100% alc Place under bell jar and when surface is solid, examine daily, but lower portion soft flood with chloroform 16-24 hours. Pour off chloroform and let dry. Attach to block with 20% nitrocellulose Let dry and immerse in chloroform for several hours. 24 hour baths of 3:1 chloroform: cedar wood oil, 1:1 of the same, then pure cedar wood oil. Cut sections and store in 80% alc. Dangerous, smelly, time consuming but extremely good fun; definitely a non-PC technique that works; done it myself as a pup. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 Mob: 07830 196072=20 Mobile E-Mail kemlorogerson@3mail.com =20 FAX & Answer Phone 0871 242 8094 E-mail Accounts:=A0=20 =A0=A0=A0=A0=A0=A0=A0=A0=A0=A0=A0=A0 kemlo@tiscali.co.uk=A0or=20 kemlo1@btinternet.com=A0 Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. =A0 =A0 =A0 -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Margaret Gondo Sent: 15 September 2003 22:29 To: histonet@pathology.swmed.edu Subject: [Histonet] eye morphology advice Hi All - I have a grad student who is interested in preserving the=20 morphology of a monkey eyeball once it is removed. Does anyone=20 have any suggestions? Everything we have tried always results in=20 the posterior part of the eyeball caving in. =20 Thanks in advance, Margaret =20 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 11 Date: Tue, 16 Sep 2003 08:10:58 -0400 (EDT) From: "Barbara Osborn" To: "Histonet" Subject: [Histonet] Re: Methacarn/Carnoy's Storage Tamara, You can store methacarn or Carnoy's for up to a couple of years as long as you cap it very, very tightly to avoid evaporation of chloroform and alcohol. So you can store a large batch of fixative and only take out what is needed each week. After use, however, it must be discarded. --__--__-- Message: 12 To: "Histonet" From: Jackie.O'Connor@abbott.com Date: Tue, 16 Sep 2003 07:30:32 -0500 Subject: [Histonet] IHC on Blood Smears This is a multipart message in MIME format. --=_alternative 0044E0DC86256DA3_= Content-Type: text/plain; charset="us-ascii" A colleague of mine would like to stain murine blood smears for Bcl2. Does anyone have any experience doing IHC on blood smears which have been fixed in Methanol? Thanks Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 Jackie.O'Connor@abbott.com --=_alternative 0044E0DC86256DA3_= Content-Type: text/html; charset="us-ascii"
A colleague of mine would like to stain murine blood smears for Bcl2.  Does anyone have any experience doing IHC on blood smears which have been fixed in Methanol?

Thanks

Jacqueline M. O'Connor HT(ASCP)
Abbott Laboratories
Global Pharmaceutical Research and Development
Discovery Chemotheraputics
847.938.4919
Jackie.O'Connor@abbott.com --=_alternative 0044E0DC86256DA3_=-- --__--__-- Message: 13 Date: Tue, 16 Sep 2003 10:14:27 -0400 From: "Fred Underwood" To: Subject: Re: [Histonet] eye morphology advice I think this was discussed in the past. If my memory serves me right, injecting the eye with a gelatin solution before processing will help retain the shape. A search of the archives may be worth while. Fred >>> "Margaret Gondo" 09/15/03 05:28PM >>> Hi All - I have a grad student who is interested in preserving the morphology of a monkey eyeball once it is removed. Does anyone have any suggestions? Everything we have tried always results in the posterior part of the eyeball caving in. Thanks in advance, Margaret _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 14 Date: Tue, 16 Sep 2003 10:14:27 -0400 From: "Fred Underwood" To: Subject: Re: [Histonet] eye morphology advice I think this was discussed in the past. If my memory serves me right, injecting the eye with a gelatin solution before processing will help retain the shape. A search of the archives may be worth while. Fred >>> "Margaret Gondo" 09/15/03 05:28PM >>> Hi All - I have a grad student who is interested in preserving the morphology of a monkey eyeball once it is removed. Does anyone have any suggestions? Everything we have tried always results in the posterior part of the eyeball caving in. Thanks in advance, Margaret _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 15 Date: Tue, 16 Sep 2003 10:26:51 -0400 From: "Fred Underwood" To: Subject: Re: [Histonet] IHC on Blood Smears If the smears are quite thick and bloody, you may want to use a more dilute hydrogen peroxide solution (1% or so) for quenching. The vigorous reaction can strip material from the slide. Prolonged buffer wahes are a good idea as well. Fred >>> 09/16/03 08:30AM >>> A colleague of mine would like to stain murine blood smears for Bcl2. Does anyone have any experience doing IHC on blood smears which have been fixed in Methanol? Thanks Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 Jackie.O'Connor@abbott.com --__--__-- Message: 16 Date: Tue, 16 Sep 2003 15:48:43 +0100 (BST) From: "Vincenzo La Manna" To: Subject: [Histonet] immunohistochemistry and frozen sections Hi Histonetters, I've been struggling for 4 months on my 5 microns frozen sections trying to find estrogen receptors. I've tried both immunoperoxidase kit from Vector laboratories either indirect immunofluorecsence (FITC Goat anti mouse Secondary Ab from Sigma) but I've not yet obtained good results. My primary Ab is a mouse monoclonal to bovine Estrogen receptor from Cymbus biotechnology. Sections come from tissue that is supposed to be a non-target tissue (mid laminar region from cow's hooves)blocking with 2% goat normal serum (sigma) whash in tbs buffer All different methods for antigen retrieval have been tested but sections look poor regarding histologic quality and I still have very big problems with non specific binding an staining (indirect immunofluorescence). It looks like frozen sections are too tender to efford the whole processing. Every suggestion or reference is welcome, thank U La Manna Vincenzo Department of Agriculture and Forestry, University of Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. Telephone; 01224 274259 Fax; 01224 273731 e-mail v.lamanna@abdn.ac.uk --__--__-- Message: 17 Date: Tue, 16 Sep 2003 09:29:40 -0600 (MDT) From: Phillip Huff To: "Vincenzo La Manna" Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] immunohistochemistry and frozen sections First, I would recommend that you run some Western Blots to ensure that your Antigen of interest is being expressed within your tissues. Once your Western blots are positive, you can definetly say that your immunoreaction is what needs to be optimized. Second, have you applied your immunoreaction technique to a known positive tissue source, that has also been sectioned at 5 um? It may be a good confirmation that your tissue may need to be processed differently or cut thicker. Finally, if your end goal is immunolocalization and the immunohistochem is not working, you may want to look into in situ hybridization. In your tissue samples, the receptor may be quite sensitive to processing and you may be losing a great deal of your antigen. I know very little about your receptor, but does it have a high turnover rate? Perhaps it has been degraded over time when frozen. Has the reaction been done in freshly harvested tissues? Perhaps localizing the mRNA in the tissues may provide you with the results you are looking for. Before doing this though, I would confirm that your tissues express the mRNA of interest by doing some RT-PCR. This will both confirm that your tissues have your target mRNA, and if you use an internal housekeeping primer in the PCR, you can determine the approximate copy number of your mRNA to help you in determining how sensitive your in situ reaction must be. Hope this helps you out, Phil > Hi Histonetters, > > I've been struggling for 4 months on my 5 microns > frozen sections trying to find estrogen receptors. > I've tried both immunoperoxidase kit from Vector > laboratories either indirect immunofluorecsence (FITC > Goat anti mouse Secondary Ab from Sigma) but I've not > yet obtained good results. > My primary Ab is a mouse monoclonal to bovine > Estrogen receptor from Cymbus biotechnology. > Sections come from tissue that is supposed to be a > non-target tissue (mid laminar region from cow's > hooves)blocking with 2% goat normal serum (sigma) > whash in tbs buffer > All different methods for antigen retrieval have been > tested but sections look poor regarding histologic > quality and I still have very big problems with non > specific binding an staining (indirect > immunofluorescence). > It looks like frozen sections are too tender to > efford the whole processing. > Every suggestion or reference is welcome, > thank U > > La Manna Vincenzo > Department of Agriculture and Forestry, University of > Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. > Telephone; 01224 274259 > Fax; 01224 273731 > e-mail v.lamanna@abdn.ac.uk > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --__--__-- Message: 18 Date: Tue, 16 Sep 2003 08:31:16 -0700 From: "Tommy Vedilago" To: "Histonet (E-mail)" Subject: [Histonet] Talented Histotechs wanted Hello Histonetters, If you have ever wanted to work for a lab that is truly a family = atmosphere where your skills and abilities are concretely appreciated, I = have just the job for you. I have 3 positions for Histotechs in Dallas, = TX with a progressive lab that has been in place for 36 years now. They = are offering outstanding pay, benefits, and atmosphere. They are also = offering relocation and hire bonus. If this holds interest for you or if = you know of someone in the job market, please give me a call at = 866-SYSTEM1. Tommy Vedilago System 1 Search (864) 627-0012 (864) 627-0013 Fax --__--__-- Message: 19 Date: Tue, 16 Sep 2003 16:58:35 +0100 (BST) From: nick.kirk3@btopenworld.com To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] immunohistochemistry and frozen sections If it's not an inpolite question, why are you trying to demonstrate estrogen receptors on frozen tissue? Is this absolutely neccessary as estrogen receptors are very easily demonstrated on paraffin processed tissue, which maintains better morphology and gets almost guaranteed results. We use the DAKO 1D5 clone on FFPE tissue using Vector Unmasking fluid for Antigen Retrieval and the DAKO ChemMate detection system and get excellent results every time. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England > from: Vincenzo La Manna > date: Tue, 16 Sep 2003 15:48:43 > to: histonet@lists.utsouthwestern.edu > subject: Re: [Histonet] immunohistochemistry and frozen sections > > > > Hi Histonetters, > > I've been struggling for 4 months on my 5 microns > frozen sections trying to find estrogen receptors. > I've tried both immunoperoxidase kit from Vector > laboratories either indirect immunofluorecsence (FITC > Goat anti mouse Secondary Ab from Sigma) but I've not > yet obtained good results. > My primary Ab is a mouse monoclonal to bovine > Estrogen receptor from Cymbus biotechnology. > Sections come from tissue that is supposed to be a > non-target tissue (mid laminar region from cow's > hooves)blocking with 2% goat normal serum (sigma) > whash in tbs buffer > All different methods for antigen retrieval have been > tested but sections look poor regarding histologic > quality and I still have very big problems with non > specific binding an staining (indirect > immunofluorescence). > It looks like frozen sections are too tender to > efford the whole processing. > Every suggestion or reference is welcome, > thank U > > La Manna Vincenzo > Department of Agriculture and Forestry, University of > Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. > Telephone; 01224 274259 > Fax; 01224 273731 > e-mail v.lamanna@abdn.ac.uk > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest 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 tigersnake <@t> ecybermind.net Tue Sep 16 18:49:05 2003 From: tigersnake <@t> ecybermind.net (Paul Howard Lockwood) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] RE: Histonet digest, Vol 1 #49 - 19 msgs Message-ID: <200309162145.h8GLjwC27288@ginsberg.ecybermind.net> Dear Rosemarie, This is the procedure from Sheehan and Hrapchak: 200gm Potassium dichromate 1 liter Distilled water 750ml Concentrated sulfuric acid Use heat resistant glassware, as heat will be genenated when adding sulfuric acid to mixutre. The solution is dark red brown, and may be continued to be reused for soak until it turns dark green. Do your stirring with a glass rod, and use all safety precautions due to the highly corrosive nature of the mixture. I hope this will be of some help to you. Sincerely, Paul Lockwood 9/16/03 12:57:42 PM, "Behan, Rosemarie G" wrote: >I am looking for a recipe for acid cleaning glassware to do silver stains, >can anyone help me? > >-----Original Message----- >From: histonet-request@lists.utsouthwestern.edu >[mailto:histonet-request@lists.utsouthwestern.edu] >Sent: Tuesday, September 16, 2003 1:00 PM >To: histonet@lists.utsouthwestern.edu >Subject: Histonet digest, Vol 1 #49 - 19 msgs > > >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-admin@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: Bad Plants/Yarrow (sara goldston) > 2. background problem (Jacqueline Miller) > 3. Re: Bouin's fixation/removal of picric acid (Sarah Jones) > 4. eye morphology advice (Margaret Gondo) > 5. ASCP BOR results (Lee & Peggy Wenk) > 6. wage and salary survey (Lee & Peggy Wenk) > 7. Colloidal Iron (lena spencer) > 8. Re: Bouin's fixation/removal of picric acid (John Kiernan) > 9. Re: Colloidal Iron (John Kiernan) > 10. RE: eye morphology advice (Kemlo Rogerson) > 11. Re: Methacarn/Carnoy's Storage (Barbara Osborn) > 12. IHC on Blood Smears (Jackie.O'Connor@abbott.com) > 13. Re: eye morphology advice (Fred Underwood) > 14. Re: eye morphology advice (Fred Underwood) > 15. Re: IHC on Blood Smears (Fred Underwood) > 16. immunohistochemistry and frozen sections (Vincenzo La Manna) > 17. Re: immunohistochemistry and frozen sections (Phillip Huff) > 18. Talented Histotechs wanted (Tommy Vedilago) > 19. Re: immunohistochemistry and frozen sections >(nick.kirk3@btopenworld.com) > >--__--__-- > >Message: 1 >Date: Mon, 15 Sep 2003 12:43:52 -0700 (PDT) >From: sara goldston >To: "Johnston, Kathy" , > "Histonet \(E-mail\)" >Subject: Re: [Histonet] Bad Plants/Yarrow > >--0-349928651-1063655032=:6512 >Content-Type: text/plain; charset=us-ascii > >I use to be a naturalist in high school and yarrrow is suppose to have >medicinal purposes (hopefully you and I are talking about the same plant). >If you crush either the leaves or blooms, cannot recall which, under you >nose you will smell something similar to Vick's vapor rub. Yarrow has the >same effect as Vick's by opening up the nasal passages. But more to your >question at hand. Yarrow is a wild bush so I would assume like any wild and >un-domesticated plant, it will grow quite freely if given the opportunity >to. If you are from anywhere east of the Mississippi, there has been an >unusual amount of rain not to mention cooler than normal temperatures. If >your plant was kept outside, it would have loved the weather this season and >flourished! You may have to break out the pruining shears. Sorry, I don't >have any other helpful advice. Try going to a nature reserve, greenhouse or >horticulturalist, and perhaps someone along those lines will have some more >concrete advise. >Sara > >"Johnston, Kathy" wrote: >While we are talking "Bad Plants" I would like everyone's opinion on >Yarrow. I bought 2 plants this spring and they have grown into to very >large although quite stunning plants. I can't remember it's exact name >right now, but each stem is a different pastel color. My only concern with >is is that is started out in a 2" pot, and right now I would need a 5 gallon >pail to put it in if I dug it out! > >I am understanding that is seems quite prolific. But is it easy to control. > >Your honest opinions please!!!! > >Thanks! > >Kathy > > > > >--------------------------------- >Do you Yahoo!? >Yahoo! SiteBuilder - Free, easy-to-use web site design software >--0-349928651-1063655032=:6512 >Content-Type: text/html; charset=us-ascii > >
I use to be a naturalist in high school and yarrrow is suppose to have >medicinal purposes (hopefully you and I are talking about the same >plant).  If you crush either the leaves or blooms, cannot recall which, >under you nose you will smell something similar to Vick's vapor rub.  >Yarrow has the same effect as Vick's by opening up the nasal passages.  >But more to your question at hand.  Yarrow is a wild bush so I would >assume like any wild and un-domesticated plant, it will grow quite freely if >given the opportunity to.  If you are from anywhere east of the >Mississippi, there has been an unusual amount of rain not to mention cooler >than normal temperatures.  If your plant was kept outside, it would >have loved the weather this season and flourished!  You may have to >break out the pruining shears.  Sorry, I don't have any other helpful >advice.  Try going to a nature reserve, >greenhouse or horticulturalist, and perhaps so > meone > along those lines will have some more concrete advise.
>
Sara 

"Johnston, Kathy" ><Kathy.Johnston@CLS.ab.ca> wrote:
>
> >
While we are talking "Bad >Plants"  I would like everyone's opinion on Yarrow.  I bought 2 >plants this spring and they have grown into to very large although quite >stunning plants.  I can't remember it's exact name right now, but each >stem is a different pastel color.  My only concern with is is that is >started out in a 2" pot, and right now I would need a 5 gallon pail to put >it in if I dug it out!
>
 
>
I am understanding that is >seems quite prolific.  But is it easy to control.
>
 
>
Your honest opinions >please!!!!
>
 
>
Thanks!
>
 
>
Kathy
>

>Do you Yahoo!?
>href="http://us.rd.yahoo.com/evt=10469/*http://sitebuilder.yahoo.com">Yahoo! >SiteBuilder - Free, easy-to-use web site design software >--0-349928651-1063655032=:6512-- > > >--__--__-- > >Message: 2 >Date: Mon, 15 Sep 2003 15:02:24 -0500 >From: "Jacqueline Miller" >To: >Subject: [Histonet] background problem > >Hi, > >We're having trouble with high background when using Rat IgG1 antibodies = >(4 =B5g/ml) on paraffin-embedded sections of human tissues, with or = >without retrieval procedures. Does anyone have any advice? > >Thank you, > >Jacqueline Miller >Research Asst. I >OB/GYN Dept. >University of Texas Southwestern Medical Center >Dallas, TX > > > > > >--__--__-- > >Message: 3 >Date: Mon, 15 Sep 2003 15:57:36 -0500 >From: "Sarah Jones" >To: >Cc: , >Subject: Re: [Histonet] Bouin's fixation/removal of picric acid > >I use it on wet tissue, it does not appear that the lithium needs to be >removed. Maybe I'm not understanding your question. Sarah >Ref. Theory and Practice of Histotechnology, Sheehan, Hrapchak, page >47. > > >>>> John Kiernan 09/14/03 11:54PM >>> >Sarah, do you use the saturated lithium >carbonate in 70% ethanol on blocks of >tissue or on hydrated paraffin sections >that are yellow? > >If it's for blocks, how do you remove >lithium carbonate from the decolorized >specimens? > John Kiernan >-- >------------------------- >John A. Kiernan >Department of Anatomy and Cell Biology >The University of Western Ontario >London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ >___________ >Sarah Jones wrote: >> >> I've always used saturated lithium carbonate in 70% ethanol to >remove >> the picric acid. I believe it would be faster and more complete >than >> 70% alone. >> >> Sarah Jones HT(ASCP) >> Dept. of Vet. Anatomy & Public Health >> Histology Lab >> Texas A&M University >> College Station, TX 77843-4458 >> phone: 979-845-3177 >> fax: 979-458-3499 >> >> >>> "peptolab" 09/13/03 08:32AM >>> >> A 3-5 mm testicular biopsy should be fixed in one to two hours. Be >sure >> to >> rinse out as much yellow picric acid as possible (you can use >running >> water >> or 70% alcohol ). The basophilia of chromatin in the stored block >will >> deteriorate after some time if there is picric acid residue. >> >> Jeff Silverman HT HTL QIHC (ASCP) >> Southside Hospital >> Bay Shore NY >> > > >--__--__-- > >Message: 4 >Date: Mon, 15 Sep 2003 16:28:30 -0500 >From: "Margaret Gondo" >Reply-To: >To: >Subject: [Histonet] eye morphology advice > >Hi All - > >I have a grad student who is interested in preserving the >morphology of a monkey eyeball once it is removed. Does anyone >have any suggestions? Everything we have tried always results in >the posterior part of the eyeball caving in. > > > >Thanks in advance, >Margaret > > > > > > >--__--__-- > >Message: 5 >From: "Lee & Peggy Wenk" >To: "Histonet" >Cc: >Date: Mon, 15 Sep 2003 19:10:59 -0400 >Subject: [Histonet] ASCP BOR results > >For those interested in the pass rates of the HT and HTL and SLS ASCP exams >Jan. - June 2003, please read on. (For those not interested, please press >the DELETE key now.) > >Scaled score of 400 is required to pass the exam. For HT and HTL, both the >written (MCQ = Multiple Choice Question) and the practical portions must be >passed. > >NAACLS = National Accrediting Agency for Clinical Laboratory Sciences, which >is the accrediting agency for lab training programs. > >Total Number non-NAACLS candidates = Total Number taking exam - Total Number >NAACLS students > >HISTOTECHNICIAN > >HT MCQ >Mean = 428 >Range of Scores = 100-761 >Total Number Taking Exam = 206 >Total Pass = 111 (54%) >Total Number NAACLS students = 24 >Total Pass NAACLS students = 18 (75%) >Total Number non-NAACLS candidates = 182 >Total Pass non-NAACLS candidates = 93 (51%) > >HT Practical >Mean = 446 >Range of Scores = 201-889 >Total Number Taking Exam = 217 >Total Pass = 144 (66%) >Total Number NAACLS students = 24 >Total Pass NAACLS students = 20 (83%) >Total Number non-NAACLS candidates = 193 >Total Pass non-NAACLS candidates = 20 (64%) > >HT COMBINED >Total Number Taking Exam = 349 >Total Pass = 141 (40%) >Total Number NAACLS students = 23 >Total Pass NAACLS students = 15 (65%) >Total Number non-NAACLS candidates = 326 >Total Pass non-NAACLS candidates = 126 (39%) > >HT exam first given in 1948. Certified to date = 18,556 > > >HISTOTECHNOLOGIST > >HTL MCQ >Mean = 451 >Range of Scores = 100-646 >Total Number Taking Exam = 43 >Total Pass = 32 (74%) >Total Number NAACLS students = 0 > >HTL PRACTICAL >Mean = 487 >Range of Scores = 244-700 >Total Number Taking Exam = 54 >Total Pass = 43 (80%) >Total Number NAACLS students = 0 > >HTL COMBINED >Total Number Taking Exam = 78 >Total Pass = 39 (50%) >Total Number NAACLS students = 0 > >HTL exam first given in 1980. Certified to date = 2,070 > > >SPECIALIST IN LABORATORY SAFETY >Mean = 422 >Range of Scores = 363-511 >Total Number Taking Exam = 7 >Total Pass = 3 (43%) > >SLS exam first given in 2000. Certified to date = 138 > >Peggy A. Wenk, HTL(ASCP)SLS >William Beaumont Hospital >Royal Oak, MI 48073 > > > >--__--__-- > >Message: 6 >From: "Lee & Peggy Wenk" >To: "Histonet" >Cc: >Date: Mon, 15 Sep 2003 19:19:56 -0400 >Subject: [Histonet] wage and salary survey > >FYI - > >The September 2003 issue of "Laboratory Medicine" from ASCP has Part I of >the latest Wage and Salary Survey. This survey is done every other year. >This article shows national averages of all lab disciplines, and then breaks >it down into regional areas. Please remember, your state, or even your part >of the state, may pay higher or lower than these averages. > >Things of interest that I found: > >- HT and HTL still have the highest vacancy rates of all laboratory >personnel. This is the same as the survey done 2 years ago. > >- HTL and MT now have the same salary ranges, on average nationwide. In >other words, on national average, HTL are now being paid the same rate as >MT. > >A few of the charts on the national averages can be found on the ASCP Board >of Registry website: >http://www.ascp.org/bor/center/center_research.asp > >Thought some people might like to find their issue, or borrow one from a >colleague (or maybe even join ASCP). > >Peggy A. Wenk, HTL(ASCP)SLS >William Beaumont Hospital >Royal Oak, MI 48073 > > > >--__--__-- > >Message: 7 >From: "lena spencer" >To: >Date: Mon, 15 Sep 2003 20:58:58 -0400 >Subject: [Histonet] Colloidal Iron > >Hi All: >One of my Pathologist's has requested a Hale's Colloidal Iron for renal cell >carcinoma chromophobes. I am familiar with colloidal iron for >mucosubstances, but he feel's that this is not the same stain. The article >stated that the diagnosis is made from the colloidal iron stain or from >electron microscopy . This Pathologist is requesting that I post on the >histonet for additional information or for someone out in histoland who is >performing this procedure. >Your is insight would be greatly appreciated. >Lena > > > > >--__--__-- > >Message: 8 >Date: Tue, 16 Sep 2003 00:24:20 -0400 >From: John Kiernan >Reply-To: jkiernan@uwo.ca >To: Sarah Jones >CC: histonet@pathology.swmed.edu >Subject: Re: [Histonet] Bouin's fixation/removal of picric acid > >I didn't phrase my question very clearly, and >should have explained the reason for asking. > >Previously I've encountered lithium carbonate >for removing picric from sections. It's the >traditional thing to do after Bouin fixation >when the tissue is still yellow in the wax >and after hydration of the sections. >You can easily see when all the yellow has >gone. > >With a block you can't see the middle, and >I was wondering how long it takes until no >more yellow comes out into the liquid. With >70% and higher alcohols alone it seems to take >for ever (at least with 5 mm and larger specimens; >I haven't used tiny biopsies after Bouin fixation). >It would be nice to have an idea how much more >quickly the picric acid is extracted when the >70% alc is saturated with Li2CO3. > >I asked about removing the Li2CO3 because it's >almost insoluble in alcohol (Merck Index). That >would be 95-100% alc. I guess you rinse the >specimens in more 70% after extracting the >picric acid, and that removes the Li2CO3 and >you don't get damage from crystals forming in >the tissue. >-- >------------------------- >John A. Kiernan >Department of Anatomy and Cell Biology >The University of Western Ontario >London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ >_________________________ >Sarah Jones wrote: >> >> I use it on wet tissue, it does not appear that the lithium needs to be >> removed. Maybe I'm not understanding your question. Sarah >> Ref. Theory and Practice of Histotechnology, Sheehan, Hrapchak, page >> 47. >> >> >>> John Kiernan 09/14/03 11:54PM >>> >> Sarah, do you use the saturated lithium >> carbonate in 70% ethanol on blocks of >> tissue or on hydrated paraffin sections >> that are yellow? >> >> If it's for blocks, how do you remove >> lithium carbonate from the decolorized >> specimens? >> John Kiernan >> -- >> ------------------------- >> John A. Kiernan >> Department of Anatomy and Cell Biology >> The University of Western Ontario >> London, Canada N6A 5C1 >> kiernan@uwo.ca >> http://publish.uwo.ca/~jkiernan/ >> ___________ >> Sarah Jones wrote: >> > >> > I've always used saturated lithium carbonate in 70% ethanol to >> remove >> > the picric acid. I believe it would be faster and more complete >> than >> > 70% alone. >> > >> > Sarah Jones HT(ASCP) >> > Dept. of Vet. Anatomy & Public Health >> > Histology Lab >> > Texas A&M University >> > College Station, TX 77843-4458 >> > phone: 979-845-3177 >> > fax: 979-458-3499 >> > >> > >>> "peptolab" 09/13/03 08:32AM >>> >> > A 3-5 mm testicular biopsy should be fixed in one to two hours. Be >> sure >> > to >> > rinse out as much yellow picric acid as possible (you can use >> running >> > water >> > or 70% alcohol ). The basophilia of chromatin in the stored block >> will >> > deteriorate after some time if there is picric acid residue. >> > >> > Jeff Silverman HT HTL QIHC (ASCP) >> > Southside Hospital >> > Bay Shore NY >> > > > >--__--__-- > >Message: 9 >Date: Tue, 16 Sep 2003 01:22:32 -0400 >From: John Kiernan >Reply-To: jkiernan@uwo.ca >To: lena spencer >CC: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Colloidal Iron > >There's just the one Hale's (1946) method, also called >the dialysed iron technique because if you make >the original reagent yourself you have to dialyse >the ferric hydroxide sol. Detailed instructions are >given in Pearse's Histochemistry (any edition). >There have been several variants of the principal >reagent. In all the techniques the bound ferric >iron is made visible by a Prussian blue reaction >with potassium ferrocyanide. > >In Chapter 10 of the latest (2002) edition of >Bancroft & Gamble's "Theory and Practice ..." by >Barbara Totty, the method attributed to Hale (1946) >uses a more easily made colloidal iron sol that >does not need to be dialysed. A more detailed >reading of Pearse (3rd ed) shows that this is close >to the reagent of G.Muller (3 papers in Acta Histochem >vol 2, 1955-56, not seen by me; they'll be in >German). This colloidal iron sol quickly became more >popular than Hale's reagent not only because it was >easier to make but also because it was more specific >for acid mucosubstances. The not-dialysed colloidal >iron solution, unlike Hale's, does not stain nuclei >and there is no "background" cytoplasmic blue colour >in renal tubules. > >Both Pearse and Totty point out that colloidal iron >staining does the same job as alcian blue (pH 2.5) >but provides a darker blue product and also some >nonspecific staining. It's more sensitive and less >clean. > >"Hale's colloidal iron" (subject of the original >enquiry) is probably not an appropriate name for this >technique, which has been significantly improved >since 1946. Pearse seriously questioned its >histochemical >specificity despite recognizing the improvements made >by Muller and several others. > >Alcian blue is easier to use and can be incorporated >into many other rational mixed staining procedures >for mucosubstances. I know nothing about renal >carcinoma >chromophobes. If they can be stained with colloidal >iron >they must be basophilic, not "chromophobic." There are >very few genuinely chromophobic (unstainable) >cytoplasms. >-- >------------------------- >John A. Kiernan >Department of Anatomy and Cell Biology >The University of Western Ontario >London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ >__________________________________ >lena spencer wrote: >> >> Hi All: >> One of my Pathologist's has requested a Hale's Colloidal Iron for renal >cell >> carcinoma chromophobes. I am familiar with colloidal iron for >> mucosubstances, but he feel's that this is not the same stain. The >article >> stated that the diagnosis is made from the colloidal iron stain or from >> electron microscopy . This Pathologist is requesting that I post on the >> histonet for additional information or for someone out in histoland who is >> performing this procedure. >> Your is insight would be greatly appreciated. >> Lena > > >--__--__-- > >Message: 10 >From: "Kemlo Rogerson" >To: , > >Date: Tue, 16 Sep 2003 07:33:44 +0100 >Subject: RE: [Histonet] eye morphology advice > >Nitrocellulose and celloidin technique. Page 91 of Histopathologic >Technic and Practical Histochemistry. RD Lillie and Harold M. >Fullmer.1976, 1965 McGrew Inc. 1234567890 KPKP 7832109876 > >48hr Form fixation >dehydrate with successive baths of 35, 50, 65 & 80% alc. 24 hours each. >Open eye >Return to 80% alc. 24 hours >2x 100% alc 24 hours >6 hour bath in equal vols 100% alc and ether >Infiltrate 5-7 days in 10% nitrocellulose in 100% alc. >Infiltrate 5-7 days in 20% nitrocellulose in 100% alc >Place under bell jar and when surface is solid, examine daily, but lower >portion soft flood with chloroform 16-24 hours. >Pour off chloroform and let dry. >Attach to block with 20% nitrocellulose >Let dry and immerse in chloroform for several hours. >24 hour baths of 3:1 chloroform: cedar wood oil, 1:1 of the same, then >pure cedar wood oil. >Cut sections and store in 80% alc. > >Dangerous, smelly, time consuming but extremely good fun; definitely a >non-PC technique that works; done it myself as a pup. > >Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS >Tel: 01270 877625 >Mob: 07830 196072=20 >Mobile E-Mail kemlorogerson@3mail.com =20 >FAX & Answer Phone 0871 242 8094 >E-mail Accounts:=A0=20 >=A0=A0=A0=A0=A0=A0=A0=A0=A0=A0=A0=A0 kemlo@tiscali.co.uk=A0or=20 > kemlo1@btinternet.com=A0 >Disclaimer: The information contained in this message and/or any >attachments(s) may be of a private and confidential nature, and is >intended solely for the attention of the addressee. If you have received >this message in error or feel you should not have been the intended >recipient, please return it and any attachments to the sender >immediately. All messages relating to this communication should then be >deleted from your system. Unauthorised usage, copying, disclosure or >alteration of this message and/or attachment(s) is strictly prohibited. >Barking, Havering and Redbridge Hospitals NHS Trust will not be held >responsible for any direct or indirect damages which may arise from >alteration of this message or any attachment(s), by a third party or >resulting from the transmission of a virus. > >=A0 >=A0 >=A0 > > >-----Original Message----- >From: histonet-admin@lists.utsouthwestern.edu >[mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Margaret >Gondo >Sent: 15 September 2003 22:29 >To: histonet@pathology.swmed.edu >Subject: [Histonet] eye morphology advice > >Hi All - > >I have a grad student who is interested in preserving the=20 >morphology of a monkey eyeball once it is removed. Does anyone=20 >have any suggestions? Everything we have tried always results in=20 >the posterior part of the eyeball caving in. =20 > > > >Thanks in advance, >Margaret > > >=20 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >--__--__-- > >Message: 11 >Date: Tue, 16 Sep 2003 08:10:58 -0400 (EDT) >From: "Barbara Osborn" >To: "Histonet" >Subject: [Histonet] Re: Methacarn/Carnoy's Storage > > >Tamara, > >You can store methacarn or Carnoy's for up to a couple of years as long >as you cap it very, very tightly to avoid evaporation of chloroform and >alcohol. So you can store a large batch of fixative and only take out >what is needed each week. After use, however, it must be discarded. > > >--__--__-- > >Message: 12 >To: "Histonet" >From: Jackie.O'Connor@abbott.com >Date: Tue, 16 Sep 2003 07:30:32 -0500 >Subject: [Histonet] IHC on Blood Smears > >This is a multipart message in MIME format. >--=_alternative 0044E0DC86256DA3_= >Content-Type: text/plain; charset="us-ascii" > >A colleague of mine would like to stain murine blood smears for Bcl2. Does >anyone have any experience doing IHC on blood smears which have been fixed >in Methanol? > >Thanks > >Jacqueline M. O'Connor HT(ASCP) >Abbott Laboratories >Global Pharmaceutical Research and Development >Discovery Chemotheraputics >847.938.4919 >Jackie.O'Connor@abbott.com >--=_alternative 0044E0DC86256DA3_= >Content-Type: text/html; charset="us-ascii" > > >
A colleague of mine would like to stain murine >blood smears for Bcl2.  Does anyone have any experience doing IHC on >blood smears which have been fixed in Methanol? >
>
Thanks >
>
Jacqueline M. O'Connor HT(ASCP)
>Abbott Laboratories
>Global Pharmaceutical Research and Development
>Discovery Chemotheraputics
>847.938.4919
>Jackie.O'Connor@abbott.com >--=_alternative 0044E0DC86256DA3_=-- > > >--__--__-- > >Message: 13 >Date: Tue, 16 Sep 2003 10:14:27 -0400 >From: "Fred Underwood" >To: >Subject: Re: [Histonet] eye morphology advice > >I think this was discussed in the past. If my memory serves me right, >injecting the eye with a gelatin solution before processing will help >retain the shape. A search of the archives may be worth while. > >Fred > >>>> "Margaret Gondo" 09/15/03 05:28PM >>> >Hi All - > >I have a grad student who is interested in preserving the >morphology of a monkey eyeball once it is removed. Does anyone >have any suggestions? Everything we have tried always results in >the posterior part of the eyeball caving in. > > > >Thanks in advance, >Margaret > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >--__--__-- > >Message: 14 >Date: Tue, 16 Sep 2003 10:14:27 -0400 >From: "Fred Underwood" >To: >Subject: Re: [Histonet] eye morphology advice > >I think this was discussed in the past. If my memory serves me right, >injecting the eye with a gelatin solution before processing will help >retain the shape. A search of the archives may be worth while. > >Fred > >>>> "Margaret Gondo" 09/15/03 05:28PM >>> >Hi All - > >I have a grad student who is interested in preserving the >morphology of a monkey eyeball once it is removed. Does anyone >have any suggestions? Everything we have tried always results in >the posterior part of the eyeball caving in. > > > >Thanks in advance, >Margaret > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >--__--__-- > >Message: 15 >Date: Tue, 16 Sep 2003 10:26:51 -0400 >From: "Fred Underwood" >To: >Subject: Re: [Histonet] IHC on Blood Smears > >If the smears are quite thick and bloody, you may want to use a more >dilute hydrogen peroxide solution (1% or so) for quenching. The >vigorous reaction can strip material from the slide. Prolonged buffer >wahes are a good idea as well. > >Fred > >>>> 09/16/03 08:30AM >>> >A colleague of mine would like to stain murine blood smears for Bcl2. >Does >anyone have any experience doing IHC on blood smears which have been >fixed >in Methanol? > >Thanks > >Jacqueline M. O'Connor HT(ASCP) >Abbott Laboratories >Global Pharmaceutical Research and Development >Discovery Chemotheraputics >847.938.4919 >Jackie.O'Connor@abbott.com > > >--__--__-- > >Message: 16 >Date: Tue, 16 Sep 2003 15:48:43 +0100 (BST) >From: "Vincenzo La Manna" >To: >Subject: [Histonet] immunohistochemistry and frozen sections > > > >Hi Histonetters, > >I've been struggling for 4 months on my 5 microns >frozen sections trying to find estrogen receptors. >I've tried both immunoperoxidase kit from Vector >laboratories either indirect immunofluorecsence (FITC >Goat anti mouse Secondary Ab from Sigma) but I've not >yet obtained good results. >My primary Ab is a mouse monoclonal to bovine >Estrogen receptor from Cymbus biotechnology. >Sections come from tissue that is supposed to be a >non-target tissue (mid laminar region from cow's >hooves)blocking with 2% goat normal serum (sigma) >whash in tbs buffer >All different methods for antigen retrieval have been >tested but sections look poor regarding histologic >quality and I still have very big problems with non >specific binding an staining (indirect >immunofluorescence). >It looks like frozen sections are too tender to >efford the whole processing. >Every suggestion or reference is welcome, >thank U > >La Manna Vincenzo >Department of Agriculture and Forestry, University of >Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. >Telephone; 01224 274259 >Fax; 01224 273731 >e-mail v.lamanna@abdn.ac.uk > > > > > >--__--__-- > >Message: 17 >Date: Tue, 16 Sep 2003 09:29:40 -0600 (MDT) >From: Phillip Huff >To: "Vincenzo La Manna" >Cc: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] immunohistochemistry and frozen sections > >First, I would recommend that you run some Western Blots to ensure that >your Antigen of interest is being expressed within your tissues. Once your >Western blots are positive, you can definetly say that your immunoreaction >is what needs to be optimized. > >Second, have you applied your immunoreaction technique to a known positive >tissue source, that has also been sectioned at 5 um? It may be a good >confirmation that your tissue may need to be processed differently or cut >thicker. > >Finally, if your end goal is immunolocalization and the immunohistochem is >not working, you may want to look into in situ hybridization. In your >tissue samples, the receptor may be quite sensitive to processing and you >may be losing a great deal of your antigen. I know very little about your >receptor, but does it have a high turnover rate? Perhaps it has been >degraded over time when frozen. Has the reaction been done in freshly >harvested tissues? Perhaps localizing the mRNA in the tissues may provide >you with the results you are looking for. Before doing this though, I >would confirm that your tissues express the mRNA of interest by doing some >RT-PCR. This will both confirm that your tissues have your target mRNA, >and if you use an internal housekeeping primer in the PCR, you can >determine the approximate copy number of your mRNA to help you in >determining how sensitive your in situ reaction must be. > >Hope this helps you out, > >Phil > > > >> Hi Histonetters, >> >> I've been struggling for 4 months on my 5 microns >> frozen sections trying to find estrogen receptors. >> I've tried both immunoperoxidase kit from Vector >> laboratories either indirect immunofluorecsence (FITC >> Goat anti mouse Secondary Ab from Sigma) but I've not >> yet obtained good results. >> My primary Ab is a mouse monoclonal to bovine >> Estrogen receptor from Cymbus biotechnology. >> Sections come from tissue that is supposed to be a >> non-target tissue (mid laminar region from cow's >> hooves)blocking with 2% goat normal serum (sigma) >> whash in tbs buffer >> All different methods for antigen retrieval have been >> tested but sections look poor regarding histologic >> quality and I still have very big problems with non >> specific binding an staining (indirect >> immunofluorescence). >> It looks like frozen sections are too tender to >> efford the whole processing. >> Every suggestion or reference is welcome, >> thank U >> >> La Manna Vincenzo >> Department of Agriculture and Forestry, University of >> Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. >> Telephone; 01224 274259 >> Fax; 01224 273731 >> e-mail v.lamanna@abdn.ac.uk >> >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > >--__--__-- > >Message: 18 >Date: Tue, 16 Sep 2003 08:31:16 -0700 >From: "Tommy Vedilago" >To: "Histonet (E-mail)" >Subject: [Histonet] Talented Histotechs wanted > >Hello Histonetters, > > If you have ever wanted to work for a lab that is truly a family = >atmosphere where your skills and abilities are concretely appreciated, I = >have just the job for you. I have 3 positions for Histotechs in Dallas, = >TX with a progressive lab that has been in place for 36 years now. They = >are offering outstanding pay, benefits, and atmosphere. They are also = >offering relocation and hire bonus. If this holds interest for you or if = >you know of someone in the job market, please give me a call at = >866-SYSTEM1. >Tommy Vedilago >System 1 Search >(864) 627-0012 >(864) 627-0013 Fax > > >--__--__-- > >Message: 19 >Date: Tue, 16 Sep 2003 16:58:35 +0100 (BST) >From: nick.kirk3@btopenworld.com >To: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] immunohistochemistry and frozen sections > >If it's not an inpolite question, why are you trying to demonstrate estrogen >receptors on frozen tissue? >Is this absolutely neccessary as estrogen receptors are very easily >demonstrated on paraffin processed tissue, which maintains better morphology >and gets almost guaranteed results. > >We use the DAKO 1D5 clone on FFPE tissue using Vector Unmasking fluid for >Antigen Retrieval and the DAKO ChemMate detection system and get excellent >results every time. > >Nick Kirk >Histopathology >Hinchingbrooke Hospital >Huntingdon >England > >> from: Vincenzo La Manna >> date: Tue, 16 Sep 2003 15:48:43 >> to: histonet@lists.utsouthwestern.edu >> subject: Re: [Histonet] immunohistochemistry and frozen sections >> >> >> >> Hi Histonetters, >> >> I've been struggling for 4 months on my 5 microns >> frozen sections trying to find estrogen receptors. >> I've tried both immunoperoxidase kit from Vector >> laboratories either indirect immunofluorecsence (FITC >> Goat anti mouse Secondary Ab from Sigma) but I've not >> yet obtained good results. >> My primary Ab is a mouse monoclonal to bovine >> Estrogen receptor from Cymbus biotechnology. >> Sections come from tissue that is supposed to be a >> non-target tissue (mid laminar region from cow's >> hooves)blocking with 2% goat normal serum (sigma) >> whash in tbs buffer >> All different methods for antigen retrieval have been >> tested but sections look poor regarding histologic >> quality and I still have very big problems with non >> specific binding an staining (indirect >> immunofluorescence). >> It looks like frozen sections are too tender to >> efford the whole processing. >> Every suggestion or reference is welcome, >> thank U >> >> La Manna Vincenzo >> Department of Agriculture and Forestry, University of >> Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. >> Telephone; 01224 274259 >> Fax; 01224 273731 >> e-mail v.lamanna@abdn.ac.uk >> >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >--__--__-- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >End of Histonet Digest > > >LEGAL NOTICE >Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From tigersnake <@t> ecybermind.net Tue Sep 16 18:59:27 2003 From: tigersnake <@t> ecybermind.net (Paul Howard Lockwood) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] CAP Question anp.23075 Message-ID: <200309162156.h8GLuJC28852@ginsberg.ecybermind.net> Dear Hazel, I am more familiar with Good Laboratory Practices than CAP, but the question seems to me to ask: Is there paperwork people make out for the instruments on a daily basis? All the years I've worked in labs, I know how much the Feds love to see tons of paperwork. (This despite the claim that they want to reduce everybody's paperwork. Go fig, eh.) Take care. Sincerely, Paul Lockwood 9/14/03 2:27:34 PM, "Horn, Hazel V" wrote: > > > > I'm having my CAP inspection soon and wondered what is meant by this > question. > > Is there EVIDENCE of ongoing evaluation of results of instrument > > maintenance a nd function for all devices? > > The 2 questions before this one are about a schedule for checking > instruments (yes) and are the service and repair records promptly > available. > > (yes) > > So......What does the above question pertain too?? HELP! > > > > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > > Phone - 501.364.4240 > > Fax - 501.364.3912 > > > The information contained in this message may be privileged and > confidential and protected from disclosure. If the reader of this message > is not the intended recipient, or an employee or agent responsible for > delivering this message to the intended recipient, you are hereby notified > that any dissemination, distribution or copying of this communication is > strictly prohibited. If you have received this communication in error, > please notify us immediately by replying to the message and deleting it > from your computer. Thank you. Arkansas Children's Hospital From Kathy.Johnston <@t> CLS.ab.ca Tue Sep 16 16:06:23 2003 From: Kathy.Johnston <@t> CLS.ab.ca (Johnston, Kathy) Date: Fri Sep 16 15:21:53 2005 Subject: FW: [Histonet] Bad Plants/Yarrow Message-ID: <30C050525B881C4AAFF41E6D16543E68015D02A3@mail3.cls.ab.ca> Oops!!! My apologies histonetters! Need to watch my mouse finger! I sent this message to the wrong person! Kathy -----Original Message----- From: Johnston, Kathy [mailto:Kathy.Johnston@CLS.ab.ca] Sent: Monday, September 15, 2003 10:46 AM To: Histonet (E-mail) Subject: [Histonet] Bad Plants/Yarrow While we are talking "Bad Plants" I would like everyone's opinion on Yarrow. I bought 2 plants this spring and they have grown into to very large although quite stunning plants. I can't remember it's exact name right now, but each stem is a different pastel color. My only concern with is is that is started out in a 2" pot, and right now I would need a 5 gallon pail to put it in if I dug it out! I am understanding that is seems quite prolific. But is it easy to control. Your honest opinions please!!!! Thanks! Kathy -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030916/66673ddf/attachment.htm From g.lang <@t> bigfoot.de Tue Sep 16 16:14:28 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] microwave Message-ID: <000e01c37c97$83d66e40$0d12a8c0@SERVER> Hi Histonetters! I have a question about microwave oven in histolabs. We did introduce such a household thing for IHC, but it was not very successfull. Now we heat our tea with it. IHC works with heat and steam. Which equipment do you use for special staining? Are there specific apparats or also the houshold ones? And has it become really common to work with it? thanks for your answers. Gundi Lang general hospital Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030916/1120196f/attachment.htm From amarusk1 <@t> FAIRVIEW.ORG Tue Sep 16 16:25:05 2003 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] S100 frozen skin Message-ID: Skipped content of type multipart/alternative-------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:ANN MARUSKA EMAIL;WORK;PREF;NGW:AMARUSK1.EAST.NORTH ORG:;LAB N:MARUSKA;ANN TEL;WORK:612-273-9119 END:VCARD From RFail <@t> Charleston.net Tue Sep 16 16:24:36 2003 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] microwave In-Reply-To: <000e01c37c97$83d66e40$0d12a8c0@SERVER> Message-ID: <007101c37c98$f0b8ec30$d211a6a5@rena> We use a 1000 watt microwave manufactured for household use. We use it every day for Special stains. Half power.. For mordanting, heating silvers, Rhodanine, Acid fast stains.colloidal irons, and more Rena Fail ---Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Tuesday, September 16, 2003 5:14 PM To: Histonetliste Subject: [Histonet] microwave Hi Histonetters! I have a question about microwave oven in histolabs. We did introduce such a household thing for IHC, but it was not very successfull. Now we heat our tea with it. IHC works with heat and steam. Which equipment do you use for special staining? Are there specific apparats or also the houshold ones? And has it become really common to work with it? thanks for your answers. Gundi Lang general hospital Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030916/b1501d5e/attachment.htm From SJones <@t> cvm.tamu.edu Tue Sep 16 17:08:09 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] microwave Message-ID: Hello Gudrun, I'm glad to see you have posted a question. I don't have a lot of experience with IHC and the microwave, but I can tell you that the microwave can shorten the times of many special stains. Many steps that usually take one hour can be shortened to just a few minutes. Silver stains work very well in the microwave. There is a great book on microwaving special stains called "Microwave Magic" by Billie Beck, University of Texas Southwestern Medical Center in Dallas Texas. It's old, from 1987, so I don't know if it is still available. Some of the stains listed are, Grocott's Methenamine Silver, Jones Methenamine Silver, Fontana-Masson, Dieterle, Grimelius, Steiner and Steiner, Seiver-Munger, Bielschowsky's, PAS, Mucicarmine, Alcian Blue, Colloidal Iron, Rhodanine, Iron Stains, Trichrome, Acid Fast, Giemsa, Luxol Fast Blue, Congo Red, PTAH, Movat's, Dunn Thompson, Bouins pre-treatment. You can use a household microwave, but you have to know the wattage of your oven. Most of them range from 700-900 watts, but if it is lower wattage, you would just leave it in longer. Usually you would want to determine how long it would take a coplin jar of water to reach 60 degrees C. Also determine how long it would take different ethanol to reach 60 degrees C. You can also make a temperature conversion chart that shows various solutions, times and power levels and what temperature is reached. Use plastic coplin jars, the glass ones will break, stir solutions immediately as they come out of the oven (I use a wooden stick), and cover the coplin jars with a gauze square when microwaving. If you go back to using chemicals in the microwave, I wouldn't continue to heat tea in it, just for safety reasons. Happy Microwaving! Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Gudrun Lang" 09/16/03 04:14PM >>> Hi Histonetters! I have a question about microwave oven in histolabs. We did introduce such a household thing for IHC, but it was not very successfull. Now we heat our tea with it. IHC works with heat and steam. Which equipment do you use for special staining? Are there specific apparats or also the houshold ones? And has it become really common to work with it? thanks for your answers. Gundi Lang general hospital Linz, Austria From WWmn916 <@t> aol.com Tue Sep 16 18:02:11 2003 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Acetylcholonesterase Message-ID: <1d1.10ef4df6.2c98f073@aol.com> Anyone have an easy recipe (or refer me to a prepared kit that can be purchased) for the acetylcholonesterase stain? Doctor is looking for Hirschsprungs disease. Any help is appreciated. Sincerely, Deb King, HT (ASCP) Sacramento, CA -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030916/e4504890/attachment.htm From amosbrooks <@t> earthlink.net Tue Sep 16 21:19:36 2003 From: amosbrooks <@t> earthlink.net (amosbrooks@earthlink.net) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Re: Histonet digest, Vol 1 #50 - 11 msgs In-Reply-To: <20030916212900.4484.71643.Mailman@swlx167.swmed.edu> Message-ID: <3F678C78.15775.3465041@localhost> Hi, We use a Black & Decker vegtable steamer. I bored a hole into the top of the steamer and we put a thermometer directly into the solution to get a Real Time measurement of the temperature. (BTW CAP loved it) We also use a waterbath as is indicated by the Herceptest (DAKO) instructions. I strongly dislike the use of household microwaves for any laboratory purposes, especially something as finiky as antigen retrieval. The temperature over time spiles up to an overheated state then drops rapidly to a point that it is not retrieving at all only to be reheated in the same way. It is impossible to have any uniformity this way. You're much better off with a steady heat source for a specific amount of time. Haste makes waste, Amos Brooks Hi Histonetters! I have a question about microwave oven in histolabs. We did introduce such a household thing for IHC, but it was not very successfull. Now we heat our tea with it. IHC works with heat and steam. Which equipment do you use for special staining? Are there specific apparats or also the houshold ones? And has it become really common to work with it? thanks for your answers. Gundi Lang general hospital Linz, Austria From jkiernan <@t> uwo.ca Wed Sep 17 00:28:14 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Glass cleaning for silver References: <9DF8DDFE3736D3119D6D0008C79148D60A5BFD21@groexmbcr17.pfizer.com> Message-ID: <3F67F0ED.77A2DCF5@uwo.ca> Rosemary Behan asked about cleaning glassware for silver staining. A reply follows. It's necessary to remove traces of metallic silver, which are not necessarily visible. The only common acid that does this is nitric. Here is what I do, with rationale. Let's assume the vessel is a Coplin jar. Rinse with one or two changes of distilled water. [Tap water contains chloride ions and silver chloride precipitation must be avoided.] Pour a few ml of concentrated nitric acid into the vessel and carefully let it wet all the inside surface. For a Coplin jar it's important to dissolve silver from all corners and from the slots for supporting slides. It takes 15-30 seconds to make black deposits or mirrors disappear. A minute of turning and tilting has to be enough to remove all the visible and invisible silver. Safely discard the nitric acid and fill up the vessel with distilled or otherwise purified water, three times. [Pure water must be used because tap water will precipitate traces of silver chloride from the residual nitric acid - which contains dissolved silver nitrate.] Wash in tap water, detergent etc as for any other dirty lab glassware, and don't spare the brush. [The nitric acid treatment does not remove all types of dirt. Bits of detached section, stuck to the glass, are made yellow by nitric acid.] Rinse in tap water, repeatedly, to get rid of the detergent (no more froth with shaking) and then in 3 generous changes of pure water to dilute out residual chloride from the tap water. Let the vessel dry by drainage and evaporation, then keep it in a closed cupboard, with its lid on (if it has a lid; and don't forget to clean the lid). If glassware is contaminated by insoluble silver compounds such as silver chloride, 5% sodium thiosulphate (10 minutes) will remove the silver. [The thiosulphate ion strongly complexes silver ions and will remove them from solid silver halides. This is the "fixation" of photographers.] In the above remarks I have not given detailed safety and disposal instructions. Conc. nitric acid is nasty stuff but becomes harmless when diluted with water. Do not use hydrochloric acid or an HCl-alcohol mixture for "acid washing" of glassware that will contain silver nitrate or protargol. [Reason is obvious from above discussion.] There are silver solvents less noxious than concentrated nitric acid. The best known one is Farmer's reducer. This is used in black & white photography for controlled removal of darkness (= silver) from negatives or prints. It is a solution containing potassium ferricyanide and sodium thiosulphate. Its actions on photo media take several minutes, but it takes much longer to weaken silver deposits on glassware and in overstained sections (my unpublished anecdotal observations). For what it's worth, I think concentrated nitric acid is the best cleaner of glassware used for silver methods. I also think that poor glass-hygiene (dishwashers etc etc etc) often causes failure or poor results with many staining techniques. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ -- (Rosemary Behan: I've deleted many irrelevant messages from the tail of your email, which contained a megabyte of unrelated stuff. Please be careful about what to quote!) _____________________________________ "Behan, Rosemarie G" wrote: > > I am looking for a recipe for acid cleaning glassware to do silver stains, > can anyone help me? __________________________________________________ From c.m.vanderloos <@t> amc.uva.nl Wed Sep 17 02:50:57 2003 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] RE: immunohistochemistry and frozen sections Message-ID: <153dd371540679.1540679153dd37@amc.uva.nl> Dear Vincenzo, You didn't describe the fixative used for your frozen sections, but I assume that it was acetone. However, as you are trying to stain a nuclear marker it is strongly advised to use either Zamboni, 4% paraformaldehyde or just buffered formalin for 5 min at room temperature. Then wash with PBS (or TBS) and start your IHC procedure. Most nuclear markers get diffuse after acetone fixation. Furthermore, you described the application of different antigen retrieval methods. To my opinion, antigen retrieval is not needed for frozen sections because antigens/epitopes are not hidden due to fixation or something like with FFPE sections. Even if buffered formalin is used as fixative for frozen sections there is nothing to retrieve, simply because a 5 min fixation time is far too short to cause cross-linking of proteins. And indeed as you described, the tissue morphology of frozen sections will heavily suffer from antigen retrieval procedures. Good luck, Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands >Original Message ----- >From "Vincenzo La Manna" >Date Tue, 16 Sep 2003 15:48:43 +0100 (BST) >To >Subject [Histonet] immunohistochemistry and frozen sections > >Hi Histonetters, > >I've been struggling for 4 months on my 5 microns >frozen sections trying to find estrogen receptors. >I've tried both immunoperoxidase kit from Vector >laboratories either indirect immunofluorecsence (FITC >Goat anti mouse Secondary Ab from Sigma) but I've not >yet obtained good results. >My primary Ab is a mouse monoclonal to bovine >Estrogen receptor from Cymbus biotechnology. >Sections come from tissue that is supposed to be a >non-target tissue (mid laminar region from cow's >hooves)blocking with 2% goat normal serum (sigma) >whash in tbs buffer >All different methods for antigen retrieval have been >tested but sections look poor regarding histologic >quality and I still have very big problems with non >specific binding an staining (indirect >immunofluorescence). >It looks like frozen sections are too tender to >efford the whole processing. >Every suggestion or reference is welcome, >thank U > >La Manna Vincenzo >Department of Agriculture and Forestry, University of >Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. >Telephone; 01224 274259 >Fax; 01224 273731 >e-mail v.lamanna@abdn.ac.uk From georgecole <@t> ev1.net Wed Sep 17 03:02:42 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] acid cleaning slides for silver preps Message-ID: <000001c37cf2$132d94b0$014dbad0@hppav> Gray's Formulary contains a reliable cleaning procedure for slides: 40 parts saturated aqueous potassium dichromate Add, with due precautions, 60 parts sulfuric acid. I always just put excess dichromate in 60 parts water so that much solid dichromate remained in the bottom of a coverable Container. Add the 60 parts of Sulfuric Acid, with care. The quantity of cleaner may be a gallon or so, mixed in a container kept for that purpose. Put the coplin jars or other glass ware in the mixture for a few minutes or over night. Rinse the glass well with deionized water. This mixture can be used for a long time. But the best procedure is when the solid dichromate is gone, a fresh batch is mixed. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030917/3a28272d/attachment.htm From Mandy <@t> serotec.co.uk Wed Sep 17 03:22:38 2003 From: Mandy <@t> serotec.co.uk (Mandy Townsend) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] S100 frozen skin Message-ID: Serotec are able to supply a polyclonal antibody against S100 that is suitable for use on both cryostat and paraffin sections. Please do not hesitate to contact me if you require further information. Mandy Mandy Townsend MSc Technical Services Supervisor Serotec Ltd 22 Bankside Station Approach Kidlington Oxfordshire OX5 1JE Tel: +44 1865 852736 Fax: +44 1865 852739 email: mandy@serotec.co.uk URL: www.serotec.com Serotec-Your first choice for antibodies! IMPORTANT NOTICE: This message and any attachments may be confidential. If this has been sent to you in error, please contact the sender as soon as possible. Serotec Ltd. Registered in England No.1604642 Registered Office: Boswell House, 1-5 Broad Street, Oxford, OX1 SAW. UK -----Original Message----- From: ANN MARUSKA [mailto:amarusk1@FAIRVIEW.ORG] Sent: Tuesday, September 16, 2003 10:25 PM To: histonet@pathology.swmed.EDU Subject: [Histonet] S100 frozen skin Hi histonetters, I have a researcher who is interested in doing an S100 on frozen skin samples for melanoma....is anyone out there doing this? Do you know of an S100 that works on frozen tissue? As always, thanks for your help. Ann Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 ________________________________________________________________________ This e-mail has been scanned for all viruses by Star Internet. The service is powered by MessageLabs. For more information on a proactive anti-virus service working around the clock, around the globe, visit: http://www.star.net.uk ________________________________________________________________________ -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030917/bff8900e/attachment.htm From peptolab <@t> hamptons.com Wed Sep 17 03:58:06 2003 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Yarrow Message-ID: <002e01c37cf9$d13ca7a0$e176bd18@JEFF> Common yarrow, or Achillea millefolium, spreads rapidly from underground rhizomes - plants placed two feet apart will fill in within one year. The mat-like, dark green finely divided ferny foliaged plants will take over any ground available and likes sunny average soil not partcularly rich. It is relatively drought tolerant. It is not a good border plant but is very useful for sunny dry waste places where a flowering groundcover would be useful. This achillea and A. ptarmica are invasive while the other species such as A. filipendulina, grandifolia, and tomentosa are well behaved clump formers, often with attractive glaucous (blue gray) and fuzzy foliage. Achillea has been used as a toothache remedy in Europe (1440) and was mixed in ale instead of hops to increase inebriation. It was said to grow in churchyards as a a reproach to the dead who "might not have died had they taken their daily yarrow", and was used to heal wounds- Achilles (Achillea) supposedly used it to stauch the wound of his soldiers. I am unaware of any histological use for the herb, except perhaps to staunch the wounds of disposable blades or broken lurking coverslip fragments. Jeff Silverman- Plantsman Southside Hospital Bay Shore NY From mezey <@t> ana.sote.hu Wed Sep 17 06:53:58 2003 From: mezey <@t> ana.sote.hu (Mezey Szilvia) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] BrdU and EM? Message-ID: <3F686775.17391.128EBA1@localhost> Hello All, I'm trying to do BrdU staining (in paraformaldehyde fixed, free- floating sections) and preserve the ultrastructure of the tissue for EM at the same time. HCl works fine for denaturing DNA for BrdU- ICC but doesn't leave much of the tissue for EM. DNase would be more EM-friendly but doesn't penetrate the tissue enough. Does anybody know a method that could provide a fair enough compromise between BrdU and EM? Maybe by increasing the penetration of DNase? Best regards to you all, Szilvi Szilvia Mezey PhD student Semmelweis University Dept. of Anatomy, Histology and Embryology Tuzolto u. 58. Budapest, 1094, Hungary T.: +36-12156920/3687 F.: +36-12155158 E-mail: mezey@ana.sote.hu From pdelvent <@t> wyoming.com Wed Sep 17 08:00:18 2003 From: pdelvent <@t> wyoming.com (Priscilla Delventhal) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] job openings in Dallas Message-ID: Hi Tom - I will be in the San Antonio area starting next week for about a month. I know Dallas is across the state but thought I'd tell you what my plans are. I am retiring from full time work this week and plan on taking a vacation for the rest of the month. After that I only want to do temp openings - a couple a year. I have 35+ years experience in histology, both supervisory and bench. I have my BA, and HT and HTL. Let me know if you could use some temporary help this Fall. My cell phone is 307-851-5595. I will be checking messages a couple of times a day for the rest of this week and then will have it on full time as I travel down to San Antonio. Hoping to hear from you. You make your lab sound wonderful. Priscilla Delventhal From joyce.christopher <@t> bayercropscience.com Wed Sep 17 08:10:56 2003 From: joyce.christopher <@t> bayercropscience.com (Joyce Christopher) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] KnifeMaker Message-ID: Our Reichert-Jung Knifemaker II has developed a problem. It no longer has the strength to break the knives. In other words its spring needs replacement. Problem - the spring part is no longer available. Does anyone have a knifemaker they are no longer using or has one that is no longer working but does have a working spring? Joyce Christopher Bayer CropScience LP 17745 S. Metcalf Stilwell, KS 66085 913-433-5244 joyce.christopher@bayercropscience.com From ASelf <@t> gmhsc.com Wed Sep 17 08:36:14 2003 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] receiving un-registered specimens Message-ID: Hi all Netters, I was wandering how everyone rec'd specimens in the histology lab? Are they registered into the hospital system before they are grossed in or are they brought straight to the histo lab, grossed and then registered. We have an on-going problem with specimens that are coming to the histology lab before they are getting registered in the hospital system. This causes a lot of confusion with everyone. Also does anyone have a specific policy for handling formalin spills that they could share with me? We have a general policy for lab spills but I don't think that it is detailed enough for the histology lab. Thanks for all the help that you all send out not only to me but to everyone.....Much appreciated. Amy Self Georgetown Hospital Systems 843-527-7179 (home) 843-520-7882 (fax) 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 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 mezey <@t> ana.sote.hu Wed Sep 17 08:44:09 2003 From: mezey <@t> ana.sote.hu (Mezey Szilvia) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] BrdU and EM? In-Reply-To: Message-ID: <3F688148.19369.18DCFE3@localhost> Animal tissue (domestic chicks), transcardially perfused. Szilvi Subject: RE: [Histonet] BrdU and EM? Date sent: Wed, 17 Sep 2003 08:03:03 -0500 From: "Charles W. Scouten, Ph.D." To: "Mezey Szilvia" > Animal tissue (perfused?) or human tissue? > > Charles W.? Scouten, Ph.D. > myNeuroLab.com > 5918 Evergreen Blvd. > St. Louis, MO 63134 > Ph: 314 522 0300? > FAX? 314 522 0377 > cwscouten@myneurolab.com > www.myneurolab.com > > > -----Original Message----- > From: Mezey Szilvia [mailto:mezey@ana.sote.hu] > Sent: Wednesday, September 17, 2003 6:54 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] BrdU and EM? > > Hello All, > > I'm trying to do BrdU staining (in paraformaldehyde fixed, free- > floating sections) and preserve the ultrastructure of the tissue for > EM at the same time. HCl works fine for denaturing DNA for BrdU- > ICC but doesn't leave much of the tissue for EM. DNase would be > more EM-friendly but doesn't penetrate the tissue enough. > > Does anybody know a method that could provide a fair enough > compromise between BrdU and EM? Maybe by increasing the > penetration of DNase? > > Best regards to you all, > > Szilvi > > > Szilvia Mezey > PhD student > Semmelweis University > Dept. of Anatomy, Histology and Embryology > Tuzolto u. 58. Budapest, 1094, Hungary > T.: +36-12156920/3687 > F.: +36-12155158 > E-mail: mezey@ana.sote.hu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Szilvia Mezey PhD student Semmelweis University Dept. of Anatomy, Histology and Embryology Tuzolto u. 58. Budapest, 1094, Hungary T.: +36-12156920/3687 F.: +36-12155158 E-mail: mezey@ana.sote.hu From GDawson <@t> Milw.Dynacare.com Wed Sep 17 08:42:50 2003 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] wage/vacancy survey Message-ID: All, Does anyone know the release date for the official 2002 ASCP wage/vacancy survey? I have seen the preliminary survey and I seem to remember someone mentioning that the complete survey would be released in September. Any info would be appreciated. Thanx, Glen Dawson From DJStashev <@t> aol.com Wed Sep 17 08:45:16 2003 From: DJStashev <@t> aol.com (DJStashev@aol.com) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Used JB-4 Microtome? Message-ID: <79.1928d8d5.2c99bf6c@aol.com> I am trying to locate a used JB-4 Sorvall microtome to buy. Does anyone out in Histo-land have this type of microtome that they are no longer using??? Thanks, Jen Stashevsky I.U. Medical Center 317-576-0338 From gcallis <@t> montana.edu Wed Sep 17 09:09:27 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] KnifeMaker In-Reply-To: Message-ID: <3.0.6.32.20030917080927.00b82038@gemini.msu.montana.edu> See if you can get a local metalworking shop (we have a fellow who does scientific instruments) make one for you or out of some type of springlike material and get several backups. Hopefully, Reichert Jung (did you try contacting Leica Microsystems about the problem?) might have one in a drawer somewhere. Contact Don.Birgerson@leica-microsystems.com, he is very knowledgable about these instruments, and he has always been graciously helpful with plights like this. 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 funderwood <@t> mcohio.org Wed Sep 17 09:28:59 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] questions regarding to a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate stain(no Message-ID: I have used the kits put together by Sigma. The results were good. However, strict adherence to expiration dates, temperatures, times and solution prepatation is necessary. Otherwise the results can be variable and less than optimal. Fred >>> weiping Ren 09/16/03 01:13PM >>> Hi, Histonetters: I need to set up non specific esterase staining method on bone marrow smear samples. My questions are: 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate stain(non specific esterase) are the same stain? 2. If these are two different stains but have the same clinical diagnosis significance, which one is better regarding to the time and accuracy? 3. Are there any protocols for these stains? Where I can get these references? Thank you! Any suggestions and advice welcome. Weiping Wayne State University Detroit, MI --------------------------------- Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software From CCLYATT <@t> mail.mcg.edu Wed Sep 17 09:32:22 2003 From: CCLYATT <@t> mail.mcg.edu (Claye Clyatt) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] microwave Message-ID: Personally, I find that microwaving in general creates more problems than they solve. We no longer use one for our laboratory procedures. Claye Claye Clyatt Chief Histotechnologist Department of Pathology Room #BF119 Medical College of Georgia Augusta, Ga 30912 office (706) 721-3630 pager (706) 721-7243-1132 e-mail: cclyatt@mail.mcg.edu >>> "Gudrun Lang" 09/16/03 05:14PM >>> Hi Histonetters! I have a question about microwave oven in histolabs. We did introduce such a household thing for IHC, but it was not very successfull. Now we heat our tea with it. IHC works with heat and steam. Which equipment do you use for special staining? Are there specific apparats or also the houshold ones? And has it become really common to work with it? thanks for your answers. Gundi Lang general hospital Linz, Austria From CCLYATT <@t> mail.mcg.edu Wed Sep 17 09:34:37 2003 From: CCLYATT <@t> mail.mcg.edu (Claye Clyatt) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Acetylcholonesterase Message-ID: Try Sigma. I've used their kits in the past and they work very nice and are easy to use. Claye >>> 09/16/03 07:02PM >>> Anyone have an easy recipe (or refer me to a prepared kit that can be purchased) for the acetylcholonesterase stain? Doctor is looking for Hirschsprungs disease. Any help is appreciated. Sincerely, Deb King, HT (ASCP) Sacramento, CA From pmarcum <@t> polysciences.com Wed Sep 17 09:38:48 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no In-Reply-To: Message-ID: <001e01c37d29$67e79ff0$4800a8c0@PMARCUM2K> and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From pmarcum <@t> polysciences.com Wed Sep 17 09:38:48 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no In-Reply-To: Message-ID: <001e01c37d29$67e79ff0$4800a8c0@PMARCUM2K> and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From pmarcum <@t> polysciences.com Wed Sep 17 09:38:48 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no In-Reply-To: Message-ID: <001e01c37d29$67e79ff0$4800a8c0@PMARCUM2K> and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From GauchV <@t> mail.amc.edu Wed Sep 17 10:26:33 2003 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] receiving un-registered specimens Message-ID: Amy, We receive specimens that are already registered into the Hospital System and also ones that need to be registered. The first set are easily handled as they are just accessioned into the system and processed as normal. The second set are sent down the hall to registration to receive a medical record and account number and then we pick the reqs up and bring them back to our lab to be accessioned and processed. As for the formalin spill policy, we use Green Z on small spills and have a procedure for that....large spills we would call our Environmental Health and Safety Unit to come and clean up the spill and we have a procedure in place for that. Hope that helps, Vicki Gauch AMCH Pathology Albany, NY >>> Amy Self 9/17/2003 9:36:14 AM >>> Hi all Netters, I was wandering how everyone rec'd specimens in the histology lab? Are they registered into the hospital system before they are grossed in or are they brought straight to the histo lab, grossed and then registered. We have an on-going problem with specimens that are coming to the histology lab before they are getting registered in the hospital system. This causes a lot of confusion with everyone. Also does anyone have a specific policy for handling formalin spills that they could share with me? We have a general policy for lab spills but I don't think that it is detailed enough for the histology lab. Thanks for all the help that you all send out not only to me but to everyone.....Much appreciated. Amy Self Georgetown Hospital Systems 843-527-7179 (home) 843-520-7882 (fax) 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 agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Wed Sep 17 10:30:16 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] receiving un-registered specimens Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CB77@usca0082k03.rallansci.apogent.com> I have never received a gross specimen in histology that was not from a registered patient. We did receive blocks and slides for referral cases that we had to put into the system. Tim Morken -----Original Message----- From: Amy Self [mailto:ASelf@gmhsc.com] Sent: Wednesday, September 17, 2003 6:36 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] receiving un-registered specimens Hi all Netters, I was wandering how everyone rec'd specimens in the histology lab? Are they registered into the hospital system before they are grossed in or are they brought straight to the histo lab, grossed and then registered. We have an on-going problem with specimens that are coming to the histology lab before they are getting registered in the hospital system. This causes a lot of confusion with everyone. Also does anyone have a specific policy for handling formalin spills that they could share with me? We have a general policy for lab spills but I don't think that it is detailed enough for the histology lab. Thanks for all the help that you all send out not only to me but to everyone.....Much appreciated. Amy Self Georgetown Hospital Systems 843-527-7179 (home) 843-520-7882 (fax) 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 agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> polysciences.com Wed Sep 17 09:38:48 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no In-Reply-To: Message-ID: <001e01c37d29$67e79ff0$4800a8c0@PMARCUM2K> and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Rcartun <@t> harthosp.org Wed Sep 17 11:24:46 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Ret IHC Message-ID: We are do "Ret" immunoperoxidase staining on thryoid tissue using a rabbit polyclonal antibody from Santa Cruz (sc-167). Does anyone have experience with this antibody on formalin-fixed, paraffin-embedded tissue? Thank you. Richard Cartun From tmhpath <@t> amigo.net Wed Sep 17 11:30:47 2003 From: tmhpath <@t> amigo.net (Michelle D. Moore) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Underprocessed tissue Message-ID: <004501c37d39$0dadaec0$4000a8c0@ever> Hello fellow histonetters, hope everyone is having a good day so far! I am in a bind and I know you can help me, I was embedding this morning and having a very difficult time with my tissues, so I finally finished embedding and when I started to cut, nothing and I mean nothing would section UUGH. Well in the process I thought maybe it was some new paraffin I was trying so I didn't get to excited until I couldn't cut anything. Needless to say what I need from you wonderful people is help on reprocessing my tissue I know I need to remove the paraffin that's in the tissue and clear the xylene out but I have no procedure for doing this and it has been years since I have had to do it. Any help you could direct my way or any procedure that you would be willing to share would be greatly appreciated!! THANK YOU in advance for saving my rear I do appreciate it a lot. That's the beauty of the histonet people helping people. Michelle D. Moore HT (ASCP) The Memorial Hospital Craig, CO -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030917/1331106a/attachment.htm From afedanov <@t> rmy.emory.edu Wed Sep 17 11:42:20 2003 From: afedanov <@t> rmy.emory.edu (Andrew Fedanov) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Clot sections Message-ID: <3F688EEC.7070203@rmy.emory.edu> Hi Histonetters, Does anybody has an experience in preparation of clot sections? Thanks -- Andrew Fedanov Ph.D. Emory Vaccine Center at Yerkes 954 Gatewood Rd. NE Atlanta, GA 30329 Phone: 404-727-3043 Fax: 404-727-8199 From DDittus787 <@t> aol.com Wed Sep 17 11:59:04 2003 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Ret IHC Message-ID: <6727FC6A.644D1F0C.0A1F969F@aol.com> Rich: we currently use c-ret , we do antigen retrieval in citrate, control slides must be fresh and incubate 32 minutes at 42 degrees Dana From DDittus787 <@t> aol.com Wed Sep 17 12:02:32 2003 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] IHC on Blood Smears Message-ID: <776598B2.2B5159B4.0A1F969F@aol.com> we use 10% nbf for bloody slides, the water content gets rid of the red cells and the 10%nbf fixes tumor cells to the slides, this has also allowed us to retrieve when needed for specific markers. we have had good results. Dana From asmith <@t> mail.barry.edu Wed Sep 17 12:19:17 2003 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Colloidal Iron Message-ID: <23CA849AB9FD7E4A9D0CD8013214437A2CCB72@bumail01.barrynet.barry.edu> The original Hale's colloidal iron is considered unreliable (Pearse, HISTOCHEMISTRY, 3rd ed. 1968; Zugibe, DIAGNOSTIC HISTOCHEMISTRY, 1970). Several more reliable modifications have appeared. Mowry's (1958, J. Clin. Invest. 7:566-576) is the easiest to use, but is erratic in my hands. Rinehart and Abul-Haj's method (1951, Arch. Pathol. 52:189-194) is terribly time-consuming but consistent. I have used both Mowry's method and Rinehart and Abul-Haj's method on prostate carcinoma, extramammary Paget's disease, Barrett's esophagus, and the sialomucins of the intercellular canaliculi of eccrine sweat glands. -----Original Message----- From: lena spencer [mailto:lenaspencer@insightbb.com] Sent: Monday, September 15, 2003 8:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Colloidal Iron Hi All: One of my Pathologist's has requested a Hale's Colloidal Iron for renal cell carcinoma chromophobes. I am familiar with colloidal iron for mucosubstances, but he feel's that this is not the same stain. The article stated that the diagnosis is made from the colloidal iron stain or from electron microscopy . This Pathologist is requesting that I post on the histonet for additional information or for someone out in histoland who is performing this procedure. Your is insight would be greatly appreciated. Lena _______________________________________________ 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 gcallis <@t> montana.edu Wed Sep 17 12:30:22 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Reply on blood clot sections In-Reply-To: <3F688EEC.7070203@rmy.emory.edu> Message-ID: <3.0.6.32.20030917113022.00b95958@gemini.msu.montana.edu> We let 10 mls blood clot (let it sit for a time, 1 hour was minimal, longer gave a firmer clot) and using razor blade, cut it into sections, froze the clot "slabs" using the heat extractor and did frozen sections fixed the sections with NBF and did oil red o staining for fat embolism in blunt truama/broken bones from accident patients. It was not the easiest thing to cut and experience with large blood clots inside distended blood vessels or areas of hemorrhage found within FFPE embedded weren't fun to section after routine tissue processing. Generally, the tissue block had to be soaked in ice water for optimal sectioning. At 12:42 PM 9/17/2003 -0400, you wrote: >Hi Histonetters, >Does anybody has an experience in preparation of clot sections? > >Thanks >Andrew Fedanov Ph.D. > 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 georgecole <@t> ev1.net Wed Sep 17 12:40:58 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] All those muscvle and nerve packets Message-ID: <000001c37d42$db9ae1a0$0b4dbad0@hppav> It has been a wonderful experience---your responses to the offer of the muscle and nerve packets----57 have gone out across the states and to 6 countries around the world. Now then---what now? Are none, some or many labs going to adopt none, some or all the methods presented in the DVD's and pages? To turn the stats around a bit---NOT adopting the procedures---sticking to the pip-on-a-cork-one-section-per-slide-procedures as shown in the texts amounts to being 5 to 50 times less likely to find the information in muscle and nerves sent to you. As I am retired, outside the labs looking in, I will be very grateful for any response from you histotechs as to just what your response to the new methods presented on the 3 DVD's and 18 pages of procedures is to be. Greetings to you all---I know I am lucky to be retired and all that, but if anyone of you would like to take on my 75 years and let me have your youth----I'm willin'!!" georgecole@ev1.net -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030917/08bcf9be/attachment.htm From juan.gutierrez <@t> christushealth.org Wed Sep 17 12:44:04 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Acetylcholonesterase Message-ID: Give me your fax number and I'll fax it to you. Once you get everything made up, it is easy to do. Juan C. Gutierrez, HT(ASCP) juan.gutierrez@christushealth.org -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Tue 9/16/2003 6:02 PM To: histonet@pathology.swmed.edu Cc: Subject: [Histonet] Acetylcholonesterase Anyone have an easy recipe (or refer me to a prepared kit that can be purchased) for the acetylcholonesterase stain? Doctor is looking for Hirschsprungs disease. Any help is appreciated. Sincerely, Deb King, HT (ASCP) Sacramento, CA From asmith <@t> mail.barry.edu Wed Sep 17 12:58:01 2003 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Masson Trichrome stain Message-ID: <23CA849AB9FD7E4A9D0CD8013214437A2CCB73@bumail01.barrynet.barry.edu> While color should not be the sole criterion of tissue identification, the color imparted by a trichrome stain is often very helpful in identifying tissues in an unfamiliar organ or animal. Smooth muscle and tendon often have similar microscopic morphology. That is why trichrome stains were developed. Personally, I prefer Gabe's trichrome (Kiernan's HISTOLOGICAL AND HISTOCHEMICAL METHODS, 3rd ed., pp. 158-159) to Masson's. -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Monday, September 15, 2003 11:14 AM To: Brian Hatcher Cc: mhanna@histosearch.com; HISTONET@pathology.swmed.edu Subject: Re: [Histonet] Masson Trichrome stain Hi Brian: Using colors to identify tissues is not the way to go. Muscle (skeletal, cardiac or smooth) and collagen have very different morphologies, that alone should be the criteria for identification. A good hematoxylin and eosin or even toluidine blue should tell you what you need to know. Geoff Brian Hatcher wrote: > I am attempting to use Masson's trichrome stain on some rat bone > marrow stem cells cultured on fibrous contstructs. Following staining > with the acid fuchsin, a large amount of tissue growing in between the > fibers is staining red. Following the treatment with aniline blue, > however, this tissue is no longer visible. In some areas it appears > as though it has torn away from the fibers where it was previously > spread between (some arease of floating tissue are visible). My > initial thoughts were that perhaps this tissue was muscle, although > this was a bit supprising as these cells should be differentiating > into osteoblasts in the presence of these fibers. I had also read > somewhere that the collagen would stain red with acid fuchsin, > however, subsequent staining with aniline blue should result in > collagen appearing blue. The only problem is I am seeing neither blue > nor red cells in these specific areas following aniline blue > staining. Any suggestions??? > Thanks > Brian > -- > Brian Hatcher > Graduate Research Assistant > Department of Biomedical Engineering > University of Florida > PO Box 116400 > Gainesville, FL 32611-6400 > Ph: 352-392-6656 > Fax: 352-392-3771 > email: briany@ufl.edu > -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** _______________________________________________ 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 jqb7 <@t> cdc.gov Wed Sep 17 13:03:46 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Masson Trichrome stain Message-ID: And I like the Gomori's One-Step trichrome. I use a kit and you can have a choice of green or blue for the collagen. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: Smith, Allen [mailto:asmith@mail.barry.edu] Sent: Wednesday, September 17, 2003 1:58 PM To: Geoff McAuliffe Cc: histonet@lists.utsouthwestern.edu; mhanna@histosearch.com Subject: RE: [Histonet] Masson Trichrome stain While color should not be the sole criterion of tissue identification, the color imparted by a trichrome stain is often very helpful in identifying tissues in an unfamiliar organ or animal. Smooth muscle and tendon often have similar microscopic morphology. That is why trichrome stains were developed. Personally, I prefer Gabe's trichrome (Kiernan's HISTOLOGICAL AND HISTOCHEMICAL METHODS, 3rd ed., pp. 158-159) to Masson's. -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Monday, September 15, 2003 11:14 AM To: Brian Hatcher Cc: mhanna@histosearch.com; HISTONET@pathology.swmed.edu Subject: Re: [Histonet] Masson Trichrome stain Hi Brian: Using colors to identify tissues is not the way to go. Muscle (skeletal, cardiac or smooth) and collagen have very different morphologies, that alone should be the criteria for identification. A good hematoxylin and eosin or even toluidine blue should tell you what you need to know. Geoff Brian Hatcher wrote: > I am attempting to use Masson's trichrome stain on some rat bone > marrow stem cells cultured on fibrous contstructs. Following staining > with the acid fuchsin, a large amount of tissue growing in between the > fibers is staining red. Following the treatment with aniline blue, > however, this tissue is no longer visible. In some areas it appears > as though it has torn away from the fibers where it was previously > spread between (some arease of floating tissue are visible). My > initial thoughts were that perhaps this tissue was muscle, although > this was a bit supprising as these cells should be differentiating > into osteoblasts in the presence of these fibers. I had also read > somewhere that the collagen would stain red with acid fuchsin, > however, subsequent staining with aniline blue should result in > collagen appearing blue. The only problem is I am seeing neither blue > nor red cells in these specific areas following aniline blue staining. > Any suggestions??? Thanks > Brian > -- > Brian Hatcher > Graduate Research Assistant > Department of Biomedical Engineering > University of Florida > PO Box 116400 > Gainesville, FL 32611-6400 > Ph: 352-392-6656 > Fax: 352-392-3771 > email: briany@ufl.edu > -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Wed Sep 17 13:29:37 2003 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Carcinogenic materials Message-ID: <0BE6ADFAE4E7E04496BF21ABD34662801D0C46@EXCHANGE1.huntingtonhospital.com> Does anyone know of some kind of reference book that lists which chemicals are carcinogenic? Even in checking specific MSDS I find conflicting information. What does it mean on the MSDS when it says "N/A" or "Not Available" next to the carcinogenicity data - is the chemical NOT carcinogenic or maybe it is and it's not listed? Laurie Colbert Huntington Hospital Pasadena, CA From mcauliff <@t> umdnj.edu Wed Sep 17 13:48:52 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Masson Trichrome stain In-Reply-To: References: Message-ID: <3F68AC94.4020300@umdnj.edu> Since Brian is working with bone marrow stem cells trying to identify them, their derivitives or their products based on colors adult tissues give with a certain stain is someplace between risky and bad science. As for "Smooth muscle and tendon often have similar microscopic morphology" they do not. Smooth muscle is composed of smooth muscle cells, tendon is composed largely of collagen fibers. Yes, a good trichrome can be useful (and beautiful) but it is not a substitute for knowing what you are looking at. Geoff -----Original Message----- >From: Smith, Allen [mailto:asmith@mail.barry.edu] >Sent: Wednesday, September 17, 2003 1:58 PM >To: Geoff McAuliffe >Cc: histonet@lists.utsouthwestern.edu; mhanna@histosearch.com >Subject: RE: [Histonet] Masson Trichrome stain > > >While color should not be the sole criterion of tissue identification, the >color imparted by a trichrome stain is often very helpful in identifying >tissues in an unfamiliar organ or animal. Smooth muscle and tendon often >have similar microscopic morphology. That is why trichrome stains were >developed. Personally, I prefer Gabe's trichrome (Kiernan's HISTOLOGICAL >AND HISTOCHEMICAL METHODS, 3rd ed., pp. 158-159) to Masson's. > >-----Original Message----- >From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] >Sent: Monday, September 15, 2003 11:14 AM >To: Brian Hatcher >Cc: mhanna@histosearch.com; HISTONET@pathology.swmed.edu >Subject: Re: [Histonet] Masson Trichrome stain > > >Hi Brian: > > Using colors to identify tissues is not the way to go. Muscle >(skeletal, cardiac or smooth) and collagen have very different >morphologies, that alone should be the criteria for identification. A >good hematoxylin and eosin or even toluidine blue should tell you what >you need to know. > >Geoff > >Brian Hatcher wrote: > > > >>I am attempting to use Masson's trichrome stain on some rat bone >>marrow stem cells cultured on fibrous contstructs. Following staining >>with the acid fuchsin, a large amount of tissue growing in between the >>fibers is staining red. Following the treatment with aniline blue, >>however, this tissue is no longer visible. In some areas it appears >>as though it has torn away from the fibers where it was previously >>spread between (some arease of floating tissue are visible). My >>initial thoughts were that perhaps this tissue was muscle, although >>this was a bit supprising as these cells should be differentiating >>into osteoblasts in the presence of these fibers. I had also read >>somewhere that the collagen would stain red with acid fuchsin, >>however, subsequent staining with aniline blue should result in >>collagen appearing blue. The only problem is I am seeing neither blue >>nor red cells in these specific areas following aniline blue staining. >>Any suggestions??? Thanks >>Brian >>-- >>Brian Hatcher >>Graduate Research Assistant >>Department of Biomedical Engineering >>University of Florida >>PO Box 116400 >>Gainesville, FL 32611-6400 >>Ph: 352-392-6656 >>Fax: 352-392-3771 >>email: briany@ufl.edu >> >> >> >-- >********************************************** >Geoff McAuliffe, Ph.D. >Neuroscience and Cell Biology >Robert Wood Johnson Medical School >675 Hoes Lane, Piscataway, NJ 08854 >voice: (732)-235-4583; fax: -4029 >mcauliff@umdnj.edu >********************************************** > > > > ********************************************** 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 ********************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030917/69579d48/attachment.htm From Gillian.Barlow <@t> cshs.org Wed Sep 17 15:21:45 2003 From: Gillian.Barlow <@t> cshs.org (Barlow, Gillian) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Need detailed protocols for mouse heart fixation and embedding Message-ID: <3CFAA0108952D111A5BF00805FA6FB0F0464D434@PEDSNTAS.csmc.edu> Dear All We are having terrible trouble with the fixation and embedding of our mouse heart tissues - the tissues are dry and brittle, with separation of the cardaic fibers. Where can I get a detailed protocol for mouse heart fixation and embedding (the latter done by hand not by machine)? Many thanks Gillian Gillian Barlow, PhD Postdoctoral Fellow Laboratory of Julie Korenberg, PhD, MD Cedars-Sinai Medical Center Davis Bldg, Lab 2007 110 George Burns Rd Los Angeles, CA 90048 Phone: (310) 423 7650 Fax: (310) 423 0302 From SJones <@t> cvm.tamu.edu Wed Sep 17 16:05:19 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Underprocessed tissue Message-ID: Hi Michelle, Have you determined if the processor malfunctioned? I was wondering why they are underprocessed. Are you planning to run them back through the same program and the same solutions? Unless you determine where the underprocessing happened, ie, not enough water taken out, not enough alcohol cleared, not enough infiltration, you might end up with the same problem. You only need to run them back to the step where the processing problem occurred. Are you planning to change solutions if they were old or increase the times? What do the tissues look like? Are they soft and mushy or just difficult to cut. What do they smell like? Can you smell xylene or alcohol? There is an article in Histologic that you can get online about reprocessing. It looked interesting, but I have not had to try it, knock on wood. Usually, I put them back into hot paraffin to melt the paraffin and put them into xylene while they are still hot. The xylene will usually be enough as it will tolerate some water, but you may have to go back into absolute if they still have a lot of water in them. I use all fresh solutions. You have to be able to look at the wet tissue and know what it still needs. White areas indicate water present. When they are in xylene, they should be "cleared" of alcohol and that has a certain look to it. It is one of the advantages of having processed tissue by hand. Maybe that is something they should add to the HT exam. Also, you can do a search on the Histonet Archives, because I know this has been discussed recently. Good luck, Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Michelle D. Moore" 09/17/03 11:30AM >>> Hello fellow histonetters, hope everyone is having a good day so far! I am in a bind and I know you can help me, I was embedding this morning and having a very difficult time with my tissues, so I finally finished embedding and when I started to cut, nothing and I mean nothing would section UUGH. Well in the process I thought maybe it was some new paraffin I was trying so I didn't get to excited until I couldn't cut anything. Needless to say what I need from you wonderful people is help on reprocessing my tissue I know I need to remove the paraffin that's in the tissue and clear the xylene out but I have no procedure for doing this and it has been years since I have had to do it. Any help you could direct my way or any procedure that you would be willing to share would be greatly appreciated!! THANK YOU in advance for saving my rear I do appreciate it a lot. That's the beauty of the histonet people helping people. Michelle D. Moore HT (ASCP) The Memorial Hospital Craig, CO From gentras <@t> vetmed.auburn.edu Wed Sep 17 16:14:11 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] mouse heart fixation and processing Message-ID: <5.2.0.9.0.20030917160046.00a12e00@mailhost.vetmed.auburn.edu> I usually fix all mouse tissues in either 10% formalin or 4%PFA for at least 24 hours. And then process on an automated tissue processor for a total of 4 hours using heat and vacumn. The process cycle used is as follows: #1 Wash in running tap H20 for 30 min. to remove fixative. #2 70% ETOH 40 min. #3 80% ETOH 10 min. #4 95% ETOH 10 min. #5 95% ETOH 10 min. #6 95% ETOH 20 min. #7 100% ETOH 20 min. #8 100% ETOH 20 min. #9 100% 20 min. #10 Clearing agent 15 min. #11 Clearing agent 25 min. #12 Paraffin 20 min. and #13 Paraffin 30 min. Best wishes. Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From Gillian.Barlow <@t> cshs.org Wed Sep 17 16:15:53 2003 From: Gillian.Barlow <@t> cshs.org (Barlow, Gillian) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Details of existing protocol Message-ID: <3CFAA0108952D111A5BF00805FA6FB0F0464D437@PEDSNTAS.csmc.edu> Dear All Several of you have replied asking for details of my protocol, so I thought I would post it and ask for comments. Please note we do not have a processor, everything is done by hand. Many thanks for your help, this seems to be a really nice group. Fixation (day 1): Fixative is 4% paraformaldehyde, made fresh each time and pH'd to 7.3-7.4 and chilled on ice. Hearts are harvested whole, dropped directly into fixative on ice (25-30ml each) and fixed overnight at 4 degrees. Overnight is usually 16 to 20 hours. I have also tried cutting the hearts in half before fixing, and substituting 10% phosphate-buffered formalin (Allegiance) for 4% paraformaldehyde. Dehydration and embedding (day 2): 10% Ethanol, 2 hours, 4 degrees 20% Ethanol, 2 hours, 4 degrees 30% Ethanol, 2 hours, 4 degrees 50% Ethanol, 1 hour, 4 degrees 70% Ethanol, overnight or until we are ready to process the tissue, 4 degrees Dehydration and embedding (day 3): 80% Ethanol, 1 hour, 4 degrees 95% Ethanol,1 hour, 4 degrees 100% Ethanol, 1 hour, 4 degrees Xylene Bath 1, 1 hour, room temp. Xylene Bath 2, 1 hour, 65 degrees (usually but not always in vacuum oven) Xylene Bath 3, 1 hour, 65 degrees (usually but not always in vacuum oven) Paraffin Bath 1, 1 hour, 65 degrees (usually but not always in vacuum oven). Paraffin is Paraplast Plus. Paraffin Bath 2, 1 hour, 65 degrees (usually but not always in vacuum oven) At this point we usually take the casettes out of the paraffin and allow them to solidify at room temperature overnight. The following day, we put them back in to a third paraffin bath and leave them there for 2 to three hours (65 degrees with or without vacuum again). Then we embed, which for us is the difficult part. The embedding machine we have access to is three floors away, so we take the jar of paraffin containing our cassettes downstairs (transporting in a larger container containing some hot water to stop the paraffin from solidifying), transfer the casettes into the holding tank of the embedding machine, give them 10 to 20 minutes to equilibrate, and then embed. Gillian Barlow, PhD Postdoctoral Fellow Laboratory of Julie Korenberg, PhD, MD Cedars-Sinai Medical Center Davis Bldg, Lab 2007 110 George Burns Rd Los Angeles, CA 90048 Phone: (310) 423 7650 Fax: (310) 423 0302 -------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: application/ms-tnef Size: 2813 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030917/5684cbe1/attachment.bin From laurie.colbert <@t> huntingtonhospital.com Wed Sep 17 17:10:07 2003 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Re: Carcinogenic Materials Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BBBF@EXCHANGE1.huntingtonhospital.com> Mary North from Oregon forwarded me some information from an NSH publication, "Health and Safety Committee Information Packet." This was exactly what I was looking for. It was very informative and even listed carcinogens in the Histology Lab. Thanks again, Mary! I also have the hazourdous materials manual from Dick and Janet Dapson that Vinnie suggested. I would recommend this manual for every Histology lab. Laurie Colbert Huntington Memorial Hospital From jschuma1 <@t> FAIRVIEW.ORG Wed Sep 17 17:24:57 2003 From: jschuma1 <@t> FAIRVIEW.ORG (JENNIFER SCHUMACHER) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Re: Carcinogenic Materials Message-ID: Are these items that could be sent to the list as a resource for everyone? Sounds like a handy thing to have on hand. Thanks. Jen >>> "Laurie Colbert" 09/17/03 05:10PM >>> Mary North from Oregon forwarded me some information from an NSH publication, "Health and Safety Committee Information Packet." This was exactly what I was looking for. It was very informative and even listed carcinogens in the Histology Lab. Thanks again, Mary! I also have the hazourdous materials manual from Dick and Janet Dapson that Vinnie suggested. I would recommend this manual for every Histology lab. Laurie Colbert Huntington Memorial Hospital _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030917/ba1562b7/attachment.htm From roy.ellis <@t> imvs.sa.gov.au Wed Sep 17 17:43:45 2003 From: roy.ellis <@t> imvs.sa.gov.au (Roy Ellis) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Carcinogenic chemicals Message-ID: <001601c37d6d$2714fce0$a88a140a@imvs.sa.gov.au> G'day all A useful resource for members of the list is 'The ABC of Safe Practices in the Biological Sciences Laboratory'. It was written by histologists with Histology in mind and contains a considerable amount of useful safety information including a section on carcinogenic chemicals. The complete text can be found at http://www.hoslink.com/aaindex.html or http://www.hoslink.com/welcome.html Regards Roy Roy Ellis Laboratory Manager IMVS Division of Pathology The Queen Elizabeth Hospital Woodville, South Australia 5011 mailto:roy.ellis@imvs.sa.gov.au http://www.adam.com.au/royellis From terribraud <@t> msn.com Wed Sep 17 18:01:34 2003 From: terribraud <@t> msn.com (TERRI BRAUD) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] eye morphology preservation Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030917/5ca1fa76/attachment.htm From tigersnake <@t> ecybermind.net Wed Sep 17 21:32:12 2003 From: tigersnake <@t> ecybermind.net (Paul Howard Lockwood) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] KnifeMaker Message-ID: <200309180029.h8I0T3L01131@ginsberg.ecybermind.net> Dear Joyce, You would be surprised where you can find the kind of spring you need for something like that. I would first suggest an auto parts store. One of the big ones would have something like. Also check out a farmer's store, like Tractor Supply. Some of the mill supply stores may have the appropriate spring. The yellow pages for your local area would help you pinpoint what store may have what. You may even want to give a lumberyard, or building supplier retailer like Lowes or Home Depot a try. From all the years I've spent working on old cars, I have found that when a particular supplier (like dealer's only parts) no longer has the required item, other places will have it, or at least a part similar enough that the original may be substituted with confindence. I hope this helps. Sincerely, Paul Lockwood 9/17/03 6:10:56 AM, Joyce Christopher wrote: > > > > >Our Reichert-Jung Knifemaker II has developed a problem. It no longer has the >strength to break the knives. In other words its spring needs replacement. >Problem - the spring part is no longer available. Does anyone have a knifemaker >they are no longer using or has one that is no longer working but does have a >working spring? > >Joyce Christopher >Bayer CropScience LP >17745 S. Metcalf >Stilwell, KS 66085 >913-433-5244 >joyce.christopher@bayercropscience.com > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From james.zimmerman <@t> pharma.novartis.com Wed Sep 17 19:09:02 2003 From: james.zimmerman <@t> pharma.novartis.com (james.zimmerman@pharma.novartis.com) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] CD4 and CD 8 Message-ID: Can anyonr recommend an antibody or have a procedure for CD4 and CD8 on formalin fixed, parrafin embedded monkey (Cynomologus) tissue ? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030917/ea02e7e4/attachment.htm From micro <@t> superlink.net Wed Sep 17 20:38:22 2003 From: micro <@t> superlink.net (Markus Meyenhofer) Date: Fri Sep 16 15:21:53 2005 Subject: [Histonet] Re: Used JB-4 Microtome? In-Reply-To: <79.1928d8d5.2c99bf6c@aol.com> References: <79.1928d8d5.2c99bf6c@aol.com> Message-ID: <20030918013822.21611.qmail@mail1.superlink.net> We sell reconditioned JB-4 and other EM, Histo and lab equipment. Equipment list available on request. Regards, Markus F. Meyenhofer Microscopy Labs Box 338 61 West Street Red Bank, NJ 07701 732 747 6228 fax 732 758 9142 DJStashev@aol.com writes: > I am trying to locate a used JB-4 Sorvall microtome to buy. Does anyone out > in Histo-land have this type of microtome that they are no longer using??? > > Thanks, > > Jen Stashevsky > I.U. Medical Center > 317-576-0338 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micro <@t> superlink.net Wed Sep 17 20:38:22 2003 From: micro <@t> superlink.net (Markus Meyenhofer) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Re: Used JB-4 Microtome? In-Reply-To: <79.1928d8d5.2c99bf6c@aol.com> References: <79.1928d8d5.2c99bf6c@aol.com> Message-ID: <20030918013822.21611.qmail@mail1.superlink.net> We sell reconditioned JB-4 and other EM, Histo and lab equipment. Equipment list available on request. Regards, Markus F. Meyenhofer Microscopy Labs Box 338 61 West Street Red Bank, NJ 07701 732 747 6228 fax 732 758 9142 DJStashev@aol.com writes: > I am trying to locate a used JB-4 Sorvall microtome to buy. Does anyone out > in Histo-land have this type of microtome that they are no longer using??? > > Thanks, > > Jen Stashevsky > I.U. Medical Center > 317-576-0338 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Thu Sep 18 00:07:59 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] ABC of Safe Practices (Gripe!) References: <001601c37d6d$2714fce0$a88a140a@imvs.sa.gov.au> Message-ID: <3F693DAF.92772E77@uwo.ca> Dear Roy, The ABC of Safe Practices web site that you recommend is impossibly slow! After 20 minutes I had not collected all the decorated titles, and I didn't collect a single word of actual information. Why do some writers of web sites use complicated graphics instead of simple words? Simple HTML provides for all sizes, fonts and colours, and allows rapid presentation of real graphics (diagrams, photographs etc), with the option to make pictures available by clicking on a link. The www.hoslink.com site is an abomination. It should be plain text, not modern art. End of growl. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ __________________ Roy Ellis wrote: > A useful resource for members of the list is 'The ABC of Safe Practices in the Biological Sciences Laboratory' ... written by histologists ... > can be found at http://www.hoslink.com/aaindex.html From L.Driessen <@t> orthop.umcn.nl Thu Sep 18 00:28:38 2003 From: L.Driessen <@t> orthop.umcn.nl (Driessen, L.) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] RE: Acetylcholonesterase Message-ID: <5B31B9F59E138A409E8F7C2183FF36F00DF486@umcnet13.umcn.nl> This recipe works well 1. Prepare the incubation-medium. 2. Incubate for 60-90 minutes at 37oC. 3. 3 min. Saturated NaSO4-solution. 4. 2 min. 1% ammoniumsulfideoplossing (fumehead!!!). 5. Rinse with tapwater. 6. 20 sec. Mayer-haematoxylin. 7. Rinse with tapwater for 5 min. 8. Mount in in glycerin-gelatin. 1. Cu-complexsolution: (Dissolve in this order) - 85 ml. Dist. water. - 34 g. NaSO4.10 H2O. - 150 mg. CuSO4.5H2O. - 187 mg. glycin. - 500 mg. MgCl2 - 875 mgr. maleinacid. - Adjust pH to 5,6-6,0 with ?15 ml. 1N NaOH. (Store in refrigerator; crystalyses when cold). 2. Incubation-medium: Dissolve 10 mg. acetylthiocholinejodid in 5 ml. Cu-complexsolution. 3. 1% ammoniumsulfide: Ad 4 ml. ammoniumsulfide (20%) to 76 ml. dist. water. Leon Driessen ORL/UMC-Nijmegen, The Netherlands l.driessen@orthop.umcn.nl -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: woensdag 17 september 2003 19:00 To: histonet@lists.utsouthwestern.edu Subject: Histonet digest, Vol 1 #51 - 30 msgs 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-admin@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: microwave (Sarah Jones) 2. Acetylcholonesterase (WWmn916@aol.com) 3. Re: Histonet digest, Vol 1 #50 - 11 msgs (amosbrooks@earthlink.net) 4. Glass cleaning for silver (John Kiernan) 5. RE: immunohistochemistry and frozen sections (C.M. van der Loos) 6. acid cleaning slides for silver preps (George Cole) 7. RE: S100 frozen skin (Mandy Townsend) 8. Yarrow (peptolab) 9. BrdU and EM? (Mezey Szilvia) 10. job openings in Dallas (Priscilla Delventhal) 11. KnifeMaker (Joyce Christopher) 12. receiving un-registered specimens (Amy Self) 13. RE: BrdU and EM? (Mezey Szilvia) 14. wage/vacancy survey (Dawson, Glen) 15. Used JB-4 Microtome? (DJStashev@aol.com) 16. Re: KnifeMaker (Gayle Callis) 17. Re: questions regarding to a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate stain(no (Fred Underwood) 18. Re: microwave (Claye Clyatt) 19. Re: Acetylcholonesterase (Claye Clyatt) 20. RE: questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no (Pamela Marcum) 21. RE: questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no (Pamela Marcum) 22. RE: questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no (Pamela Marcum) 23. Re: receiving un-registered specimens (Vicki Gauch) 24. RE: receiving un-registered specimens (Morken, Tim - Labvision) 25. RE: questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no (Pamela Marcum) 26. Ret IHC (Richard Cartun) 27. Underprocessed tissue (Michelle D. Moore) 28. Clot sections (Andrew Fedanov) 29. Re: Ret IHC (DDittus787@aol.com) --__--__-- Message: 1 Date: Tue, 16 Sep 2003 17:08:09 -0500 From: "Sarah Jones" To: , Subject: Re: [Histonet] microwave Hello Gudrun, I'm glad to see you have posted a question. I don't have a lot of experience with IHC and the microwave, but I can tell you that the microwave can shorten the times of many special stains. Many steps that usually take one hour can be shortened to just a few minutes. Silver stains work very well in the microwave. There is a great book on microwaving special stains called "Microwave Magic" by Billie Beck, University of Texas Southwestern Medical Center in Dallas Texas. It's old, from 1987, so I don't know if it is still available. Some of the stains listed are, Grocott's Methenamine Silver, Jones Methenamine Silver, Fontana-Masson, Dieterle, Grimelius, Steiner and Steiner, Seiver-Munger, Bielschowsky's, PAS, Mucicarmine, Alcian Blue, Colloidal Iron, Rhodanine, Iron Stains, Trichrome, Acid Fast, Giemsa, Luxol Fast Blue, Congo Red, PTAH, Movat's, Dunn Thompson, Bouins pre-treatment. You can use a household microwave, but you have to know the wattage of your oven. Most of them range from 700-900 watts, but if it is lower wattage, you would just leave it in longer. Usually you would want to determine how long it would take a coplin jar of water to reach 60 degrees C. Also determine how long it would take different ethanol to reach 60 degrees C. You can also make a temperature conversion chart that shows various solutions, times and power levels and what temperature is reached. Use plastic coplin jars, the glass ones will break, stir solutions immediately as they come out of the oven (I use a wooden stick), and cover the coplin jars with a gauze square when microwaving. If you go back to using chemicals in the microwave, I wouldn't continue to heat tea in it, just for safety reasons. Happy Microwaving! Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Gudrun Lang" 09/16/03 04:14PM >>> Hi Histonetters! I have a question about microwave oven in histolabs. We did introduce such a household thing for IHC, but it was not very successfull. Now we heat our tea with it. IHC works with heat and steam. Which equipment do you use for special staining? Are there specific apparats or also the houshold ones? And has it become really common to work with it? thanks for your answers. Gundi Lang general hospital Linz, Austria --__--__-- Message: 2 From: WWmn916@aol.com Date: Tue, 16 Sep 2003 19:02:11 EDT To: histonet@pathology.swmed.edu Subject: [Histonet] Acetylcholonesterase --part1_1d1.10ef4df6.2c98f073_boundary Content-Type: text/plain; charset="US-ASCII" Content-Transfer-Encoding: 7bit Anyone have an easy recipe (or refer me to a prepared kit that can be purchased) for the acetylcholonesterase stain? Doctor is looking for Hirschsprungs disease. Any help is appreciated. Sincerely, Deb King, HT (ASCP) Sacramento, CA --part1_1d1.10ef4df6.2c98f073_boundary Content-Type: text/html; charset="US-ASCII" Content-Transfer-Encoding: quoted-printable Anyone have an easy recipe (or refer me to a prepared=20= kit that can be purchased) for the acetylcholonesterase stain?  Doctor=20= is looking for Hirschsprungs disease.  Any help is appreciated.

Sincerely,
Deb King, HT (ASCP)
Sacramento, CA --part1_1d1.10ef4df6.2c98f073_boundary-- --__--__-- Message: 3 From: amosbrooks@earthlink.net To: histonet@lists.utsouthwestern.edu Date: Tue, 16 Sep 2003 22:19:36 -0400 Subject: [Histonet] Re: Histonet digest, Vol 1 #50 - 11 msgs Hi, We use a Black & Decker vegtable steamer. I bored a hole into the top of the steamer and we put a thermometer directly into the solution to get a Real Time measurement of the temperature. (BTW CAP loved it) We also use a waterbath as is indicated by the Herceptest (DAKO) instructions. I strongly dislike the use of household microwaves for any laboratory purposes, especially something as finiky as antigen retrieval. The temperature over time spiles up to an overheated state then drops rapidly to a point that it is not retrieving at all only to be reheated in the same way. It is impossible to have any uniformity this way. You're much better off with a steady heat source for a specific amount of time. Haste makes waste, Amos Brooks Hi Histonetters! I have a question about microwave oven in histolabs. We did introduce such a household thing for IHC, but it was not very successfull. Now we heat our tea with it. IHC works with heat and steam. Which equipment do you use for special staining? Are there specific apparats or also the houshold ones? And has it become really common to work with it? thanks for your answers. Gundi Lang general hospital Linz, Austria --__--__-- Message: 4 Date: Wed, 17 Sep 2003 01:28:14 -0400 From: John Kiernan Reply-To: jkiernan@uwo.ca To: "Behan, Rosemarie G" CC: "'histonet@lists.utsouthwestern.edu'" Subject: [Histonet] Glass cleaning for silver Rosemary Behan asked about cleaning glassware for silver staining. A reply follows. It's necessary to remove traces of metallic silver, which are not necessarily visible. The only common acid that does this is nitric. Here is what I do, with rationale. Let's assume the vessel is a Coplin jar. Rinse with one or two changes of distilled water. [Tap water contains chloride ions and silver chloride precipitation must be avoided.] Pour a few ml of concentrated nitric acid into the vessel and carefully let it wet all the inside surface. For a Coplin jar it's important to dissolve silver from all corners and from the slots for supporting slides. It takes 15-30 seconds to make black deposits or mirrors disappear. A minute of turning and tilting has to be enough to remove all the visible and invisible silver. Safely discard the nitric acid and fill up the vessel with distilled or otherwise purified water, three times. [Pure water must be used because tap water will precipitate traces of silver chloride from the residual nitric acid - which contains dissolved silver nitrate.] Wash in tap water, detergent etc as for any other dirty lab glassware, and don't spare the brush. [The nitric acid treatment does not remove all types of dirt. Bits of detached section, stuck to the glass, are made yellow by nitric acid.] Rinse in tap water, repeatedly, to get rid of the detergent (no more froth with shaking) and then in 3 generous changes of pure water to dilute out residual chloride from the tap water. Let the vessel dry by drainage and evaporation, then keep it in a closed cupboard, with its lid on (if it has a lid; and don't forget to clean the lid). If glassware is contaminated by insoluble silver compounds such as silver chloride, 5% sodium thiosulphate (10 minutes) will remove the silver. [The thiosulphate ion strongly complexes silver ions and will remove them from solid silver halides. This is the "fixation" of photographers.] In the above remarks I have not given detailed safety and disposal instructions. Conc. nitric acid is nasty stuff but becomes harmless when diluted with water. Do not use hydrochloric acid or an HCl-alcohol mixture for "acid washing" of glassware that will contain silver nitrate or protargol. [Reason is obvious from above discussion.] There are silver solvents less noxious than concentrated nitric acid. The best known one is Farmer's reducer. This is used in black & white photography for controlled removal of darkness (= silver) from negatives or prints. It is a solution containing potassium ferricyanide and sodium thiosulphate. Its actions on photo media take several minutes, but it takes much longer to weaken silver deposits on glassware and in overstained sections (my unpublished anecdotal observations). For what it's worth, I think concentrated nitric acid is the best cleaner of glassware used for silver methods. I also think that poor glass-hygiene (dishwashers etc etc etc) often causes failure or poor results with many staining techniques. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ -- (Rosemary Behan: I've deleted many irrelevant messages from the tail of your email, which contained a megabyte of unrelated stuff. Please be careful about what to quote!) _____________________________________ "Behan, Rosemarie G" wrote: > > I am looking for a recipe for acid cleaning glassware to do silver stains, > can anyone help me? __________________________________________________ --__--__-- Message: 5 Date: Wed, 17 Sep 2003 09:50:57 +0200 From: "C.M. van der Loos" To: histonet@lists.utsouthwestern.edu Cc: v.lamanna@abdn.ac.uk Subject: [Histonet] RE: immunohistochemistry and frozen sections Dear Vincenzo, You didn't describe the fixative used for your frozen sections, but I assume that it was acetone. However, as you are trying to stain a nuclear marker it is strongly advised to use either Zamboni, 4% paraformaldehyde or just buffered formalin for 5 min at room temperature. Then wash with PBS (or TBS) and start your IHC procedure. Most nuclear markers get diffuse after acetone fixation. Furthermore, you described the application of different antigen retrieval methods. To my opinion, antigen retrieval is not needed for frozen sections because antigens/epitopes are not hidden due to fixation or something like with FFPE sections. Even if buffered formalin is used as fixative for frozen sections there is nothing to retrieve, simply because a 5 min fixation time is far too short to cause cross-linking of proteins. And indeed as you described, the tissue morphology of frozen sections will heavily suffer from antigen retrieval procedures. Good luck, Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands >Original Message ----- >From "Vincenzo La Manna" >Date Tue, 16 Sep 2003 15:48:43 +0100 (BST) >To >Subject [Histonet] immunohistochemistry and frozen sections > >Hi Histonetters, > >I've been struggling for 4 months on my 5 microns >frozen sections trying to find estrogen receptors. >I've tried both immunoperoxidase kit from Vector >laboratories either indirect immunofluorecsence (FITC >Goat anti mouse Secondary Ab from Sigma) but I've not >yet obtained good results. >My primary Ab is a mouse monoclonal to bovine >Estrogen receptor from Cymbus biotechnology. >Sections come from tissue that is supposed to be a >non-target tissue (mid laminar region from cow's >hooves)blocking with 2% goat normal serum (sigma) >whash in tbs buffer >All different methods for antigen retrieval have been >tested but sections look poor regarding histologic >quality and I still have very big problems with non >specific binding an staining (indirect >immunofluorescence). >It looks like frozen sections are too tender to >efford the whole processing. >Every suggestion or reference is welcome, >thank U > >La Manna Vincenzo >Department of Agriculture and Forestry, University of >Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. >Telephone; 01224 274259 >Fax; 01224 273731 >e-mail v.lamanna@abdn.ac.uk --__--__-- Message: 6 From: "George Cole" To: Date: Wed, 17 Sep 2003 01:02:42 -0700 Subject: [Histonet] acid cleaning slides for silver preps This is a multi-part message in MIME format. ------=_NextPart_000_0001_01C37CB7.66CEBCB0 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: 7bit Gray's Formulary contains a reliable cleaning procedure for slides: 40 parts saturated aqueous potassium dichromate Add, with due precautions, 60 parts sulfuric acid. I always just put excess dichromate in 60 parts water so that much solid dichromate remained in the bottom of a coverable Container. Add the 60 parts of Sulfuric Acid, with care. The quantity of cleaner may be a gallon or so, mixed in a container kept for that purpose. Put the coplin jars or other glass ware in the mixture for a few minutes or over night. Rinse the glass well with deionized water. This mixture can be used for a long time. But the best procedure is when the solid dichromate is gone, a fresh batch is mixed. ------=_NextPart_000_0001_01C37CB7.66CEBCB0 Content-Type: text/html; charset="us-ascii" Content-Transfer-Encoding: quoted-printable

Gray’s Formulary contains a reliable cleaning = procedure for slides:

40 parts saturated aqueous potassium = dichromate

Add, with due precautions, 60 parts sulfuric = acid.

I always just put excess dichromate in 60 parts water = so that much solid dichromate remained in the bottom of a coverable =

 Container.

Add the 60 parts of Sulfuric Acid, with = care.

The quantity of cleaner may be a gallon or so, mixed = in a container kept for that purpose.

Put the coplin jars or = other glass ware in the mixture for a few minutes or over night.  Rinse the glass well with deionized water.

This mixture can be used for a long time.  But the best procedure is when = the solid dichromate is gone, a fresh batch is mixed.  

------=_NextPart_000_0001_01C37CB7.66CEBCB0-- --__--__-- Message: 7 Date: Wed, 17 Sep 2003 09:22:38 +0100 From: "Mandy Townsend" To: "ANN MARUSKA" , Subject: RE: [Histonet] S100 frozen skin This is a multi-part message in MIME format. ------_=_NextPart_001_01C37CF4.DADB2C8C Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Serotec are able to supply a polyclonal antibody against S100 that is = suitable for use on both cryostat and paraffin sections. Please do not = hesitate to contact me if you require further information. =20 Mandy =20 Mandy Townsend MSc=20 Technical Services Supervisor=20 Serotec Ltd=20 22 Bankside=20 Station Approach=20 Kidlington=20 Oxfordshire=20 OX5 1JE=20 Tel: +44 1865 852736=20 Fax: +44 1865 852739=20 email: mandy@serotec.co.uk=20 URL: www.serotec.com=20 Serotec-Your first choice for antibodies!=20 IMPORTANT NOTICE: This message and any attachments may be confidential. = If this has been sent to you in error, please contact the sender as soon = as possible. Serotec Ltd. Registered in England No.1604642=20 Registered Office: Boswell House, 1-5 Broad Street, Oxford, OX1 SAW. UK = -----Original Message----- From: ANN MARUSKA [mailto:amarusk1@FAIRVIEW.ORG] Sent: Tuesday, September 16, 2003 10:25 PM To: histonet@pathology.swmed.EDU Subject: [Histonet] S100 frozen skin Hi histonetters, =20 I have a researcher who is interested in doing an S100 on frozen skin = samples for melanoma....is anyone out there doing this? Do you know of = an S100 that works on frozen tissue? As always, thanks for your help. =20 Ann =20 Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 ________________________________________________________________________ This e-mail has been scanned for all viruses by Star Internet. The service is powered by MessageLabs. For more information on a proactive anti-virus service working around the clock, around the globe, visit: http://www.star.net.uk ________________________________________________________________________ ------_=_NextPart_001_01C37CF4.DADB2C8C Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
Serotec are able to supply a polyclonal antibody against = S100 that=20 is suitable for use on both cryostat and paraffin sections.  Please = do not=20 hesitate to contact me if you require further = information.
 
Mandy
 

Mandy Townsend MSc =
Technical Services Supervisor =
Serotec Ltd =
22 Bankside
Station Approach
Kidlington
Oxfordshire
OX5 1JE=20

Tel: +44 1865 852736 =
Fax: +44 1865 852739
email: mandy@serotec.co.uk =
URL: www.serotec.com
Serotec-Your = first choice for=20 antibodies!

IMPORTANT NOTICE: This message = and any=20 attachments may be confidential.  If this has been sent to you in = error,=20 please contact the sender as soon as possible.

Serotec Ltd. Registered in = England=20 No.1604642
Registered = Office:=20 Boswell House, 1-5  Broad Street, Oxford, OX1 SAW. UK=20

-----Original Message-----
From: ANN MARUSKA=20 [mailto:amarusk1@FAIRVIEW.ORG]
Sent: Tuesday, September 16, = 2003 10:25=20 PM
To: histonet@pathology.swmed.EDU
Subject: = [Histonet] S100=20 frozen skin

Hi histonetters,
 
I have a researcher who is interested in doing an S100 on frozen = skin=20 samples for melanoma....is anyone out there doing this?  Do you = know of an=20 S100 that works on frozen tissue?
As always, thanks for your help.
 
Ann
 
Ann Maruska
Fairview-University Medical Center
Mpls. MN  = 55454
amarusk1@fairview.org
612-27= 3-9119

_________________________________________________________= _______________
This=20 e-mail has been scanned for all viruses by Star Internet. The
service = is=20 powered by MessageLabs. For more information on a = proactive
anti-virus=20 service working around the clock, around the globe, visit:
http://www.star.net.uk
___________= _____________________________________________________________
<= /HTML> ------_=_NextPart_001_01C37CF4.DADB2C8C-- --__--__-- Message: 8 From: "peptolab" To: Date: Wed, 17 Sep 2003 04:58:06 -0400 Subject: [Histonet] Yarrow Common yarrow, or Achillea millefolium, spreads rapidly from underground rhizomes - plants placed two feet apart will fill in within one year. The mat-like, dark green finely divided ferny foliaged plants will take over any ground available and likes sunny average soil not partcularly rich. It is relatively drought tolerant. It is not a good border plant but is very useful for sunny dry waste places where a flowering groundcover would be useful. This achillea and A. ptarmica are invasive while the other species such as A. filipendulina, grandifolia, and tomentosa are well behaved clump formers, often with attractive glaucous (blue gray) and fuzzy foliage. Achillea has been used as a toothache remedy in Europe (1440) and was mixed in ale instead of hops to increase inebriation. It was said to grow in churchyards as a a reproach to the dead who "might not have died had they taken their daily yarrow", and was used to heal wounds- Achilles (Achillea) supposedly used it to stauch the wound of his soldiers. I am unaware of any histological use for the herb, except perhaps to staunch the wounds of disposable blades or broken lurking coverslip fragments. Jeff Silverman- Plantsman Southside Hospital Bay Shore NY --__--__-- Message: 9 From: "Mezey Szilvia" To: histonet@lists.utsouthwestern.edu Date: Wed, 17 Sep 2003 13:53:58 +0200 Subject: [Histonet] BrdU and EM? Hello All, I'm trying to do BrdU staining (in paraformaldehyde fixed, free- floating sections) and preserve the ultrastructure of the tissue for EM at the same time. HCl works fine for denaturing DNA for BrdU- ICC but doesn't leave much of the tissue for EM. DNase would be more EM-friendly but doesn't penetrate the tissue enough. Does anybody know a method that could provide a fair enough compromise between BrdU and EM? Maybe by increasing the penetration of DNase? Best regards to you all, Szilvi Szilvia Mezey PhD student Semmelweis University Dept. of Anatomy, Histology and Embryology Tuzolto u. 58. Budapest, 1094, Hungary T.: +36-12156920/3687 F.: +36-12155158 E-mail: mezey@ana.sote.hu --__--__-- Message: 10 From: "Priscilla Delventhal" To: histonet@lists.utsouthwestern.edu Date: Wed, 17 Sep 2003 07:00:18 -0600 Subject: [Histonet] job openings in Dallas Hi Tom - I will be in the San Antonio area starting next week for about a month. I know Dallas is across the state but thought I'd tell you what my plans are. I am retiring from full time work this week and plan on taking a vacation for the rest of the month. After that I only want to do temp openings - a couple a year. I have 35+ years experience in histology, both supervisory and bench. I have my BA, and HT and HTL. Let me know if you could use some temporary help this Fall. My cell phone is 307-851-5595. I will be checking messages a couple of times a day for the rest of this week and then will have it on full time as I travel down to San Antonio. Hoping to hear from you. You make your lab sound wonderful. Priscilla Delventhal --__--__-- Message: 11 To: histonet@lists.utsouthwestern.edu From: Joyce Christopher Date: Wed, 17 Sep 2003 08:10:56 -0500 Subject: [Histonet] KnifeMaker Our Reichert-Jung Knifemaker II has developed a problem. It no longer has the strength to break the knives. In other words its spring needs replacement. Problem - the spring part is no longer available. Does anyone have a knifemaker they are no longer using or has one that is no longer working but does have a working spring? Joyce Christopher Bayer CropScience LP 17745 S. Metcalf Stilwell, KS 66085 913-433-5244 joyce.christopher@bayercropscience.com --__--__-- Message: 12 From: Amy Self To: "'histonet@lists.utsouthwestern.edu'" Date: Wed, 17 Sep 2003 09:36:14 -0400 Subject: [Histonet] receiving un-registered specimens Hi all Netters, I was wandering how everyone rec'd specimens in the histology lab? Are they registered into the hospital system before they are grossed in or are they brought straight to the histo lab, grossed and then registered. We have an on-going problem with specimens that are coming to the histology lab before they are getting registered in the hospital system. This causes a lot of confusion with everyone. Also does anyone have a specific policy for handling formalin spills that they could share with me? We have a general policy for lab spills but I don't think that it is detailed enough for the histology lab. Thanks for all the help that you all send out not only to me but to everyone.....Much appreciated. Amy Self Georgetown Hospital Systems 843-527-7179 (home) 843-520-7882 (fax) 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 agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. --__--__-- Message: 13 From: "Mezey Szilvia" To: "Charles W. Scouten, Ph.D." , histonet@lists.utsouthwestern.edu Date: Wed, 17 Sep 2003 15:44:09 +0200 Subject: RE: [Histonet] BrdU and EM? Animal tissue (domestic chicks), transcardially perfused. Szilvi Subject: RE: [Histonet] BrdU and EM? Date sent: Wed, 17 Sep 2003 08:03:03 -0500 From: "Charles W. Scouten, Ph.D." To: "Mezey Szilvia" > Animal tissue (perfused?) or human tissue? > > Charles W.=A0 Scouten, Ph.D. > myNeuroLab.com > 5918 Evergreen Blvd. > St. Louis, MO 63134 > Ph: 314 522 0300=A0 > FAX=A0 314 522 0377 > cwscouten@myneurolab.com > www.myneurolab.com > > > -----Original Message----- > From: Mezey Szilvia [mailto:mezey@ana.sote.hu] > Sent: Wednesday, September 17, 2003 6:54 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] BrdU and EM? > > Hello All, > > I'm trying to do BrdU staining (in paraformaldehyde fixed, free- > floating sections) and preserve the ultrastructure of the tissue for > EM at the same time. HCl works fine for denaturing DNA for BrdU- > ICC but doesn't leave much of the tissue for EM. DNase would be > more EM-friendly but doesn't penetrate the tissue enough. > > Does anybody know a method that could provide a fair enough > compromise between BrdU and EM? Maybe by increasing the > penetration of DNase? > > Best regards to you all, > > Szilvi > > > Szilvia Mezey > PhD student > Semmelweis University > Dept. of Anatomy, Histology and Embryology > Tuzolto u. 58. Budapest, 1094, Hungary > T.: +36-12156920/3687 > F.: +36-12155158 > E-mail: mezey@ana.sote.hu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Szilvia Mezey PhD student Semmelweis University Dept. of Anatomy, Histology and Embryology Tuzolto u. 58. Budapest, 1094, Hungary T.: +36-12156920/3687 F.: +36-12155158 E-mail: mezey@ana.sote.hu --__--__-- Message: 14 From: "Dawson, Glen" To: histonet@lists.utsouthwestern.edu Date: Wed, 17 Sep 2003 08:42:50 -0500 Subject: [Histonet] wage/vacancy survey All, Does anyone know the release date for the official 2002 ASCP wage/vacancy survey? I have seen the preliminary survey and I seem to remember someone mentioning that the complete survey would be released in September. Any info would be appreciated. Thanx, Glen Dawson --__--__-- Message: 15 From: DJStashev@aol.com Date: Wed, 17 Sep 2003 09:45:16 EDT To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Used JB-4 Microtome? I am trying to locate a used JB-4 Sorvall microtome to buy. Does anyone out in Histo-land have this type of microtome that they are no longer using??? Thanks, Jen Stashevsky I.U. Medical Center 317-576-0338 --__--__-- Message: 16 Date: Wed, 17 Sep 2003 08:09:27 -0600 To: Joyce Christopher , Histonet@lists.utsouthwestern.edu From: Gayle Callis Subject: Re: [Histonet] KnifeMaker See if you can get a local metalworking shop (we have a fellow who does scientific instruments) make one for you or out of some type of springlike material and get several backups. Hopefully, Reichert Jung (did you try contacting Leica Microsystems about the problem?) might have one in a drawer somewhere. Contact Don.Birgerson@leica-microsystems.com, he is very knowledgable about these instruments, and he has always been graciously helpful with plights like this. 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: 17 Date: Wed, 17 Sep 2003 10:28:59 -0400 From: "Fred Underwood" To: Subject: Re: [Histonet] questions regarding to a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate stain(no I have used the kits put together by Sigma. The results were good. However, strict adherence to expiration dates, temperatures, times and solution prepatation is necessary. Otherwise the results can be variable and less than optimal. Fred >>> weiping Ren 09/16/03 01:13PM >>> Hi, Histonetters: I need to set up non specific esterase staining method on bone marrow smear samples. My questions are: 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate stain(non specific esterase) are the same stain? 2. If these are two different stains but have the same clinical diagnosis significance, which one is better regarding to the time and accuracy? 3. Are there any protocols for these stains? Where I can get these references? Thank you! Any suggestions and advice welcome. Weiping Wayne State University Detroit, MI --------------------------------- Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software --__--__-- Message: 18 Date: Wed, 17 Sep 2003 10:32:22 -0400 From: "Claye Clyatt" To: , Subject: Re: [Histonet] microwave Personally, I find that microwaving in general creates more problems than = they solve. We no longer use one for our laboratory procedures. Claye Claye Clyatt Chief Histotechnologist Department of Pathology=20 Room #BF119 Medical College of Georgia Augusta, Ga 30912 office (706) 721-3630 pager (706) 721-7243-1132 e-mail: cclyatt@mail.mcg.edu >>> "Gudrun Lang" 09/16/03 05:14PM >>> Hi Histonetters! I have a question about microwave oven in histolabs. We did introduce such = a household thing for IHC, but it was not very successfull. Now we heat = our tea with it. IHC works with heat and steam. Which equipment do you use for special staining? Are there specific = apparats or also the houshold ones? And has it become really common to = work with it? thanks for your answers. Gundi Lang general hospital Linz, Austria --__--__-- Message: 19 Date: Wed, 17 Sep 2003 10:34:37 -0400 From: "Claye Clyatt" To: , Subject: Re: [Histonet] Acetylcholonesterase Try Sigma. I've used their kits in the past and they work very nice and = are easy to use. Claye >>> 09/16/03 07:02PM >>> Anyone have an easy recipe (or refer me to a prepared kit that can be=20 purchased) for the acetylcholonesterase stain? Doctor is looking for = Hirschsprungs=20 disease. Any help is appreciated. Sincerely, Deb King, HT (ASCP) Sacramento, CA --__--__-- Message: 20 From: "Pamela Marcum" To: "Fred Underwood" , Date: Wed, 17 Sep 2003 10:38:48 -0400 Subject: RE: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --__--__-- Message: 21 From: "Pamela Marcum" To: "Fred Underwood" , Date: Wed, 17 Sep 2003 10:38:48 -0400 Subject: RE: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --__--__-- Message: 22 From: "Pamela Marcum" To: "Fred Underwood" , Date: Wed, 17 Sep 2003 10:38:48 -0400 Subject: RE: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --__--__-- Message: 23 Date: Wed, 17 Sep 2003 11:26:33 -0400 From: "Vicki Gauch" To: , Subject: Re: [Histonet] receiving un-registered specimens Amy, We receive specimens that are already registered into the Hospital System and also ones that need to be registered. The first set are easily handled as they are just accessioned into the system and processed as normal. The second set are sent down the hall to registration to receive a medical record and account number and then we pick the reqs up and bring them back to our lab to be accessioned and processed. As for the formalin spill policy, we use Green Z on small spills and have a procedure for that....large spills we would call our Environmental Health and Safety Unit to come and clean up the spill and we have a procedure in place for that. Hope that helps, Vicki Gauch AMCH Pathology Albany, NY >>> Amy Self 9/17/2003 9:36:14 AM >>> Hi all Netters, I was wandering how everyone rec'd specimens in the histology lab? Are they registered into the hospital system before they are grossed in or are they brought straight to the histo lab, grossed and then registered. We have an on-going problem with specimens that are coming to the histology lab before they are getting registered in the hospital system. This causes a lot of confusion with everyone. Also does anyone have a specific policy for handling formalin spills that they could share with me? We have a general policy for lab spills but I don't think that it is detailed enough for the histology lab. Thanks for all the help that you all send out not only to me but to everyone.....Much appreciated. Amy Self Georgetown Hospital Systems 843-527-7179 (home) 843-520-7882 (fax) 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 agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 24 From: "Morken, Tim - Labvision" To: 'Amy Self' , "'histonet@lists.utsouthwestern.edu'" Date: Wed, 17 Sep 2003 11:30:16 -0400 Subject: RE: [Histonet] receiving un-registered specimens I have never received a gross specimen in histology that was not from a registered patient. We did receive blocks and slides for referral cases that we had to put into the system. Tim Morken -----Original Message----- From: Amy Self [mailto:ASelf@gmhsc.com] Sent: Wednesday, September 17, 2003 6:36 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] receiving un-registered specimens Hi all Netters, I was wandering how everyone rec'd specimens in the histology lab? Are they registered into the hospital system before they are grossed in or are they brought straight to the histo lab, grossed and then registered. We have an on-going problem with specimens that are coming to the histology lab before they are getting registered in the hospital system. This causes a lot of confusion with everyone. Also does anyone have a specific policy for handling formalin spills that they could share with me? We have a general policy for lab spills but I don't think that it is detailed enough for the histology lab. Thanks for all the help that you all send out not only to me but to everyone.....Much appreciated. Amy Self Georgetown Hospital Systems 843-527-7179 (home) 843-520-7882 (fax) 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 agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 25 From: "Pamela Marcum" To: "Fred Underwood" , Date: Wed, 17 Sep 2003 10:38:48 -0400 Subject: RE: [Histonet] questions regarding to a-Naphthyl ButyrateEsterase stain and a-Naphthyl Acetate stain(no and filter paper requirements should be followed use exactly what they say or risk the reaction hydrolyzing in the filter. Pam Marcum > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Fred > Underwood > Sent: Wednesday, September 17, 2003 10:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] questions regarding to a-Naphthyl > ButyrateEsterase stain and a-Naphthyl Acetate stain(no > > > I have used the kits put together by Sigma. The results were good. > However, strict adherence to expiration dates, temperatures, times and > solution prepatation is necessary. Otherwise the results can be > variable and less than optimal. > > Fred > > >>> weiping Ren 09/16/03 01:13PM >>> > Hi, Histonetters: > I need to set up non specific esterase staining method on bone marrow > smear samples. > > My questions are: > 1. Are a-Naphthyl Butyrate Esterase stain and a-Naphthyl Acetate > stain(non specific esterase) are the same stain? > 2. If these are two different stains but have the same clinical > diagnosis significance, which one is better regarding to the time and > accuracy? > 3. Are there any protocols for these stains? Where I can get these > references? > > Thank you! > > Any suggestions and advice welcome. > > Weiping > Wayne State University > Detroit, MI > > > > --------------------------------- > Do you Yahoo!? > Yahoo! SiteBuilder - Free, easy-to-use web site design software > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --__--__-- Message: 26 Date: Wed, 17 Sep 2003 12:24:46 -0400 From: "Richard Cartun" To: Subject: [Histonet] Ret IHC We are do "Ret" immunoperoxidase staining on thryoid tissue using a rabbit polyclonal antibody from Santa Cruz (sc-167). Does anyone have experience with this antibody on formalin-fixed, paraffin-embedded tissue? Thank you. Richard Cartun --__--__-- Message: 27 From: "Michelle D. Moore" To: "Histonet" Date: Wed, 17 Sep 2003 10:30:47 -0600 Organization: The Memorial Hospital Subject: [Histonet] Underprocessed tissue This is a multi-part message in MIME format. ------=_NextPart_000_0042_01C37D06.C21779A0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Hello fellow histonetters, hope everyone is having a good day so far! I = am in a bind and I know you can help me, I was embedding this morning = and having a very difficult time with my tissues, so I finally finished = embedding and when I started to cut, nothing and I mean nothing would = section UUGH. Well in the process I thought maybe it was some new = paraffin I was trying so I didn't get to excited until I couldn't cut = anything. Needless to say what I need from you wonderful people is help = on reprocessing my tissue I know I need to remove the paraffin that's in = the tissue and clear the xylene out but I have no procedure for doing = this and it has been years since I have had to do it. Any help you could = direct my way or any procedure that you would be willing to share would = be greatly appreciated!! THANK YOU in advance for saving my rear I do = appreciate it a lot. That's the beauty of the histonet people helping = people. Michelle D. Moore HT (ASCP) The Memorial Hospital Craig, CO ------=_NextPart_000_0042_01C37D06.C21779A0 Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
Hello fellow histonetters, hope = everyone is having=20 a good day so far! I am in a bind and I know you can help me, I was = embedding=20 this morning and having a very difficult time with my tissues, so I = finally=20 finished embedding and when I started to cut, nothing and I mean nothing = would=20 section UUGH. Well in the process I thought maybe it was some new = paraffin I was=20 trying so I didn't get to excited until I couldn't cut anything. = Needless to say=20 what I need from you wonderful people is help on reprocessing my tissue = I know I=20 need to remove the paraffin that's in the tissue and clear the xylene = out but I=20 have no procedure for doing this and it has been years since I have had = to do=20 it. Any help you could direct my way or any procedure that you would be = willing=20 to share would be greatly appreciated!! THANK YOU in advance for saving = my rear=20 I do appreciate it a lot. That's the beauty of the histonet people = helping=20 people.
 
Michelle D. Moore HT = (ASCP)
The Memorial Hospital
Craig, CO
------=_NextPart_000_0042_01C37D06.C21779A0-- --__--__-- Message: 28 Date: Wed, 17 Sep 2003 12:42:20 -0400 From: Andrew Fedanov To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Clot sections Hi Histonetters, Does anybody has an experience in preparation of clot sections? Thanks -- Andrew Fedanov Ph.D. Emory Vaccine Center at Yerkes 954 Gatewood Rd. NE Atlanta, GA 30329 Phone: 404-727-3043 Fax: 404-727-8199 --__--__-- Message: 29 Date: Wed, 17 Sep 2003 12:59:04 -0400 From: DDittus787@aol.com To: Rcartun@harthosp.org, histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Ret IHC Rich: we currently use c-ret , we do antigen retrieval in citrate, control slides must be fresh and incubate 32 minutes at 42 degrees Dana --__--__-- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest From nick.kirk3 <@t> btopenworld.com Thu Sep 18 00:41:33 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:21:54 2005 Subject: FW: [Histonet] ABC of Safe Practices (Gripe!) Message-ID: Try this as a better resource as it actually lists the carcinogens in alphabetically rather than a whole list of chemicals that you have to trawl through to find which ones are carcinogens and which are not.. http://ehp.niehs.nih.gov/roc/toc10.html Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of John Kiernan Sent: 18 September 2003 06:08 To: Roy Ellis Cc: Histonet Subject: [Histonet] ABC of Safe Practices (Gripe!) Dear Roy, The ABC of Safe Practices web site that you recommend is impossibly slow! After 20 minutes I had not collected all the decorated titles, and I didn't collect a single word of actual information. Why do some writers of web sites use complicated graphics instead of simple words? Simple HTML provides for all sizes, fonts and colours, and allows rapid presentation of real graphics (diagrams, photographs etc), with the option to make pictures available by clicking on a link. The www.hoslink.com site is an abomination. It should be plain text, not modern art. End of growl. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ __________________ Roy Ellis wrote: > A useful resource for members of the list is 'The ABC of Safe Practices in the Biological Sciences Laboratory' ... written by histologists ... > can be found at http://www.hoslink.com/aaindex.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nick.kirk3 <@t> btopenworld.com Thu Sep 18 01:10:58 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] More Health and Safety Message-ID: Hi all Following on from the thread about carcinogens, I thought that some of you might be interested to know that here in the UK we now have to have creatinine clearance tests done if we handle or are exposed to xylene. It's part of the Health & Safety Executive documentation for which I attach a link here http://www.hse.gov.uk/pubns/eh40sup.pdf This is an update document and not the whole one which lists all the chemicals that have exposure limits in the UK. Might make interesting reading for those of you non-UK histonetters who are interested in Health and Safety. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England From kemlo <@t> tiscali.co.uk Thu Sep 18 01:48:22 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Microscope with camera and projector In-Reply-To: Message-ID: <001c01c37db0$de003ee0$22dce150@KEMLOS> We need to purchase a microscope, camera and projector for clinical meetings. Anyone got good kit in the UK? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts:? ???????????? kemlo@tiscali.co.uk?or kemlo1@btinternet.com? Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. ? ? ? -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Amy Self Sent: 17 September 2003 14:36 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] receiving un-registered specimens Hi all Netters, I was wandering how everyone rec'd specimens in the histology lab? Are they registered into the hospital system before they are grossed in or are they brought straight to the histo lab, grossed and then registered. We have an on-going problem with specimens that are coming to the histology lab before they are getting registered in the hospital system. This causes a lot of confusion with everyone. Also does anyone have a specific policy for handling formalin spills that they could share with me? We have a general policy for lab spills but I don't think that it is detailed enough for the histology lab. Thanks for all the help that you all send out not only to me but to everyone.....Much appreciated. Amy Self Georgetown Hospital Systems 843-527-7179 (home) 843-520-7882 (fax) 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 agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Felix.Rintelen <@t> serono.com Thu Sep 18 03:18:35 2003 From: Felix.Rintelen <@t> serono.com (Felix.Rintelen@serono.com) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] how to embed/section mesentery? Message-ID: Dear all, Is there anybody who may give me advice how to make good sections of parafin embedded mesentery (mouse)? How to prepare the tissue for embedding and how to handle it to embed/orient it properly in the parafin block? My concern is especially the soft consistency of the tissue making me feel that the proper orientation in the block might be difficult. Thank you very much in advance, Felix Felix Rintelen Serono Pharmaceutical Research Institute Geneva, Switzerland ******************************************************************************************** S - This message contains confidential information and is intended only for the individual named. If you are not the named addressee, you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. 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 malware. The presence of this disclaimer is not a proof that it was originated at Serono International S.A. or one of its affiliates. Serono International S.A and its affiliates therefore do not accept liability for any errors or omissions in the content of this message, which arise as a result of e-mail transmission. If verification is required, please request a hard-copy version. Serono International SA, 15bis Chemin Des Mines, Geneva, Switzerland, www.serono.com. ********************************************************************************************* From nick.kirk3 <@t> btopenworld.com Thu Sep 18 02:40:59 2003 From: nick.kirk3 <@t> btopenworld.com (nick.kirk3@btopenworld.com) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Microscope with camera and projector Message-ID: <1511490.1063870859734.JavaMail.root@127.0.0.1> Kemlo We use a Leica DMLB microscope mounted with a Leica digital camera for our Multi-disciplinary team (MDT) meetings. It's connected to a standard ceiling mounted digital projector in our MDT suite. It works very well. We're just about to get the digital camera upgraded to a Leica 3 megapixel camera from the current one which is only about 1.5 megapixels which is OK for most low to medium magnifications but is a little lacking at high magnification. Hope that is of some use. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England > from: Kemlo Rogerson > date: Thu, 18 Sep 2003 07:48:22 > to: histonet@lists.utsouthwestern.edu > subject: Re: [Histonet] Microscope with camera and projector > > We need to purchase a microscope, camera and projector for clinical > meetings. > > Anyone got good kit in the UK? > > Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS > Tel: 01270 877625 > Mob: 07830 196072 > Mobile E-Mail kemlorogerson@3mail.com > FAX & Answer Phone 0871 242 8094 > E-mail Accounts:? > ???????????? kemlo@tiscali.co.uk?or > kemlo1@btinternet.com? > Disclaimer: The information contained in this message and/or any > attachments(s) may be of a private and confidential nature, and is > intended solely for the attention of the addressee. If you have received > this message in error or feel you should not have been the intended > recipient, please return it and any attachments to the sender > immediately. All messages relating to this communication should then be > deleted from your system. Unauthorised usage, copying, disclosure or > alteration of this message and/or attachment(s) is strictly prohibited. > Barking, Havering and Redbridge Hospitals NHS Trust will not be held > responsible for any direct or indirect damages which may arise from > alteration of this message or any attachment(s), by a third party or > resulting from the transmission of a virus. > > ? > ? > ? > > > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Amy Self > Sent: 17 September 2003 14:36 > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] receiving un-registered specimens > > > > Hi all Netters, > > I was wandering how everyone rec'd specimens in the histology > lab? > Are they registered into the hospital system before they are grossed in > or > are they brought straight to the histo lab, grossed and then registered. > We > have an on-going problem with specimens that are coming to the histology > lab > before they are getting registered in the hospital system. This causes > a > lot of confusion with everyone. > > Also does anyone have a specific policy for handling formalin > spills > that they could share with me? We have a general policy for lab spills > but > I don't think that it is detailed enough for the histology lab. > > > Thanks for all the help that you all send out not only to me but > to > everyone.....Much appreciated. > > Amy Self > Georgetown Hospital Systems > 843-527-7179 (home) > 843-520-7882 (fax) > > > 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 agent responsible for > delivering this message to the intended recipient, you are hereby > notified > that any dissemination, distribution or copying of this communication is > strictly prohibited. If you have received this communication in error, > please notify us immediately by replying to the message and deleting it > from > your computer. Thank you. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mezey <@t> ana.sote.hu Thu Sep 18 03:34:18 2003 From: mezey <@t> ana.sote.hu (Mezey Szilvia) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] many thanks Message-ID: <3F698A29.9573.3C006C@localhost> Thank you everybody for your replies. I'll try the suggested methods and let you know which worked best. Szilvi Mezey Szilvia Mezey PhD student Semmelweis University Dept. of Anatomy, Histology and Embryology Tuzolto u. 58. Budapest, 1094, Hungary T.: +36-12156920/3687 F.: +36-12155158 E-mail: mezey@ana.sote.hu From mezey <@t> ana.sote.hu Thu Sep 18 03:59:28 2003 From: mezey <@t> ana.sote.hu (Mezey Szilvia) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] BrdU and EM? In-Reply-To: Message-ID: <3F69900F.11083.530BD5@localhost> Yes, I have tried with all kinds of concentrations of HCl and different times of incubation as well. The BrdU labelling works even with 10 minutes of HCl incubation (this is not very consistent, though) but the ultrastructure is 'dead' by that time. Has anyone got any experience with breaking DNA strands using UV light? What's the wavelength that could be used and how long does the tissue need to be exposed? My first try was our fluorescent microscope (at 360-370 nm), exposure time 1 hour, but it doesn't seem to do the job . Szilvi Mezey Subject: RE: [Histonet] BrdU and EM? Date sent: Thu, 18 Sep 2003 09:40:23 +0100 From: "Edwards, R.E." To: "Mezey Szilvia" > Have you tried cutting down the HCl time to a minimum????? > Richard Edwards > MRC TOX UNIT....U.K....... > > > > -----Original Message----- > From: Mezey Szilvia [mailto:mezey@ana.sote.hu] > Sent: 17 September 2003 12:54 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] BrdU and EM? > > > Hello All, > > I'm trying to do BrdU staining (in paraformaldehyde fixed, free- > floating sections) and preserve the ultrastructure of the tissue for > EM at the same time. HCl works fine for denaturing DNA for BrdU- > ICC but doesn't leave much of the tissue for EM. DNase would be > more EM-friendly but doesn't penetrate the tissue enough. > > Does anybody know a method that could provide a fair enough > compromise between BrdU and EM? Maybe by increasing the > penetration of DNase? > > Best regards to you all, > > Szilvi > > Szilvia Mezey > PhD student > Semmelweis University > Dept. of Anatomy, Histology and Embryology > Tuzolto u. 58. Budapest, 1094, Hungary > T.: +36-12156920/3687 > F.: +36-12155158 > E-mail: mezey@ana.sote.hu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Szilvia Mezey PhD student Semmelweis University Dept. of Anatomy, Histology and Embryology Tuzolto u. 58. Budapest, 1094, Hungary T.: +36-12156920/3687 F.: +36-12155158 E-mail: mezey@ana.sote.hu From pdelvent <@t> wyoming.com Thu Sep 18 06:56:43 2003 From: pdelvent <@t> wyoming.com (Priscilla Delventhal) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] temp positions Message-ID: Hi - I'm sorry to do this but someone called me on my cell phone yesterday and I've lost the message. I think it was a Joe Vialpando concerning a temp histology position. I am so new to the cell phone that I must have erased the message instead of saving it. I apologise for using the histonet for this, but could think of no other way. Please whoever called, call me again. Thanks Priscilla Delventhal From kgrobert <@t> rci.rutgers.edu Thu Sep 18 08:57:39 2003 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] buying used lab equipment References: <61E9F2400F53D5119CFC00508B44E33B019F54D5@khmcexch.uhsi.org> Message-ID: <3F69B9D2.BD95FFBF@rci.rutgers.edu> We have bought used lab equipment on eBay, and (I think) from one of the used equipment companies,though I am not sure which one as I was not the one who made the purchase. As for the equipment lasting, all our purchases were fairly recent, so all I can say right now is "so far, so good". :o) Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Rutgers University Piscataway, NJ "Kapoor, Sue" wrote: > Can anyone tell me their experience with purchasing used lab > equipment...did the equipment last? how long? if you had any problems with > it, was it resolved to your satisfaction? did you purchase from a company > dealing in used equipment? from another lab? and anything else you care to > share. > > I'm looking for a used slide dryer, the type you open the lid and set the > rack of slides in. One source only had a very old model and another that was > recommended on histonet was similar to ebay....I just want a simple and > affordable slide drying oven. > > Thanks, > Sue Kapoor, HT (ASCP) > Histology Supervisor > Kenosha Medical Center > Kenosha, WI > sue.kapoor@uhsi.org > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hectorjito26 <@t> hotmail.com Thu Sep 18 09:18:22 2003 From: hectorjito26 <@t> hotmail.com (Brent Norris) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Finger Nail Softening Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/c8cc2a7a/attachment.htm From funderwood <@t> mcohio.org Thu Sep 18 09:16:10 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] how to embed/section mesentery? Message-ID: A trick used for placental membranes is to wrap the membrane around a wood applicator stick, then process it. When embedding slide the membrane "roll" off the stick, orient it on end in the embedding mold. Fred >>> 09/18/03 04:18AM >>> Dear all, Is there anybody who may give me advice how to make good sections of parafin embedded mesentery (mouse)? How to prepare the tissue for embedding and how to handle it to embed/orient it properly in the parafin block? My concern is especially the soft consistency of the tissue making me feel that the proper orientation in the block might be difficult. Thank you very much in advance, Felix Felix Rintelen Serono Pharmaceutical Research Institute Geneva, Switzerland ******************************************************************************************** S - This message contains confidential information and is intended only for the individual named. If you are not the named addressee, you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. 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 malware. The presence of this disclaimer is not a proof that it was originated at Serono International S.A. or one of its affiliates. Serono International S.A and its affiliates therefore do not accept liability for any errors or omissions in the content of this message, which arise as a result of e-mail transmission. If verification is required, please request a hard-copy version. Serono International SA, 15bis Chemin Des Mines, Geneva, Switzerland, www.serono.com. ********************************************************************************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ctsmolde <@t> capeheart.uct.ac.za Thu Sep 18 09:10:08 2003 From: ctsmolde <@t> capeheart.uct.ac.za (Jenny Molde) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] For sale Message-ID: Anyone out there looking to buy a Reichert Jung ultratome for cutting Tem samples. It is in good working order. Price negotiable. We have recently purchased a new one for ourselves. You can contact me at the above e-mail if you are interested. Many thanks. Jenny Molde Cardiovascular Research Unit University of Cape Town Observatory Cape Town South Africa From mcauliff <@t> umdnj.edu Thu Sep 18 09:42:03 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Details of existing protocol In-Reply-To: <3CFAA0108952D111A5BF00805FA6FB0F0464D437@PEDSNTAS.csmc.edu> References: <3CFAA0108952D111A5BF00805FA6FB0F0464D437@PEDSNTAS.csmc.edu> Message-ID: <3F69C43B.6060701@umdnj.edu> Dear Gillian: Your tissue is over-processed. Very, very over-processed. 1. Fix at room temperature, not cold. For most things cold fixation is a myth, it just slows down the reaction of the fix with the tissue. 2. Cut the heart in half and fix in a volume of fix at least 10X that of the tissue. If you are not going to cut the whole heart, why fix the whole heart? If you are going to cut the whole heart your results will be better if it is cut in half for processing. 3. Fix longer, 48 hours is the minimum for full fixation with formaldehyde. Depending on what you are looking for there may be better fixatives. 4. After fixation wash the tissue in running water for an hour or more. 5. Go directly to 70% ethanol. Any alcohol lower than 50% is a waste of time and money. 6. Dehydrate in ethanols at room temperature: 95% 1 or 2 changes, 1 hour each, 100% 2 changes one hour each, a 1:1 mix of 100%:xylene one hour, 2 changes of xylene. No hot xylene!! Also consider using cholorform as a clearing agent, it is easier to remove with paraffin. 7. Melted paraffin, 3 changes 30 minutes to one hour each. The was should be just a few degrees above its melting point, 65 degrees is probably too hot. A vacuum oven is good. 8. You don't need a "machine" to embed. Get some "peel-a-way" molds, pour the wax in, wait a 15 seconds, put the tissue in, put the mold in cold water in a dish, put the dish in the refrigerator. Make your own "molds". Get a block of wood 3X bigger than the tissue. Fold some notebook paper around the block to make a box with the top open. Pour in hot wax. Wait 15-20 seconds for the bottom to set up a bit. Put tissue in. In a minute or two, put block in cold water as above. 9. Make sure your knife is sharp. Good luck! Geoff Barlow, Gillian wrote: >Dear All > >Several of you have replied asking for details of my protocol, so I thought >I would post it and ask for comments. Please note we do not have a >processor, everything is done by hand. Many thanks for your help, this >seems to be a really nice group. > >Fixation (day 1): >Fixative is 4% paraformaldehyde, made fresh each time and pH'd to 7.3-7.4 >and chilled on ice. >Hearts are harvested whole, dropped directly into fixative on ice (25-30ml >each) and fixed overnight at 4 degrees. >Overnight is usually 16 to 20 hours. I have also tried cutting the hearts >in half before fixing, and substituting 10% phosphate-buffered formalin >(Allegiance) for 4% paraformaldehyde. > >Dehydration and embedding (day 2): >10% Ethanol, 2 hours, 4 degrees >20% Ethanol, 2 hours, 4 degrees >30% Ethanol, 2 hours, 4 degrees >50% Ethanol, 1 hour, 4 degrees >70% Ethanol, overnight or until we are ready to process the tissue, 4 >degrees > >Dehydration and embedding (day 3): >80% Ethanol, 1 hour, 4 degrees >95% Ethanol,1 hour, 4 degrees >100% Ethanol, 1 hour, 4 degrees >Xylene Bath 1, 1 hour, room temp. >Xylene Bath 2, 1 hour, 65 degrees (usually but not always in vacuum oven) >Xylene Bath 3, 1 hour, 65 degrees (usually but not always in vacuum oven) >Paraffin Bath 1, 1 hour, 65 degrees (usually but not always in vacuum oven). >Paraffin is Paraplast Plus. >Paraffin Bath 2, 1 hour, 65 degrees (usually but not always in vacuum oven) > >At this point we usually take the casettes out of the paraffin and allow >them to solidify at room temperature overnight. The following day, we put >them back in to a third paraffin bath and leave them there for 2 to three >hours (65 degrees with or without vacuum again). > >Then we embed, which for us is the difficult part. The embedding machine we >have access to is three floors away, so we take the jar of paraffin >containing our cassettes downstairs (transporting in a larger container >containing some hot water to stop the paraffin from solidifying), transfer >the casettes into the holding tank of the embedding machine, give them 10 to >20 minutes to equilibrate, and then embed. > >Gillian Barlow, PhD >Postdoctoral Fellow >Laboratory of Julie Korenberg, PhD, MD >Cedars-Sinai Medical Center >Davis Bldg, Lab 2007 >110 George Burns Rd >Los Angeles, CA 90048 > >Phone: (310) 423 7650 >Fax: (310) 423 0302 > > -- -- ********************************************** 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 jbrod <@t> tvmdl.tamu.edu Thu Sep 18 09:41:22 2003 From: jbrod <@t> tvmdl.tamu.edu (Jordan Brod) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Purpose of Oven Prior to Staining? Message-ID: Good Morning All: My lab is in a huge discussion about the reason why we put our slides in an oven before staining. I told them that the reason was to dry all of the water off of them, because xylene and water do not mix. This statement is also backed up in literature that I have read. They still do not believe me, because they think the purpose of the oven is to deparaffinize the slides. Can anyone help me out, so I can put this discussion to rest? I will print off all of your comments for my technicians to read. Is what I told them correct or not? Thanks. ******************************************************************** Jordan W. Brod Diagnostic Lab Supervisor - Pathology Texas Veterinary Medical Diagnostic Laboratory (TVMDL) 1 Sippel Road TAMU 4471 College Station, Texas 77843 (979) 845-3414 (979) 845-1794 fax jbrod@tvmdl.tamu.edu From Stanley.Lupo <@t> gsk.com Thu Sep 18 08:51:31 2003 From: Stanley.Lupo <@t> gsk.com (Stanley.Lupo@gsk.com) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] how to embed/section mesentery? Message-ID: Stanley.Lupo@GSK.Com Safety Assessment Mail Code: UE0461 Phone: 610 270-7340 Fax: 610 270-7202 Fellow Histonetter, In our lab, proper orientation begins at prosection. The mesentery is laid out flat, then rolled around the rubber plunger of a 1 mL syringe. The button that is formed when the rolled mesentery is removed from the plunger is placed in a plastic capsule and fixed in 10% Neutral Buffered Formalin. Processing involves 40 minute cycles through 95% EtOH, then 60 minute cycles through 100% EtOH, 40 minutes each through 2 changes of toluene, followed by 4 changes of paraffin at 150 minutes (total). Total processing time is 9 hours. The mesenteric roll is embedded on edge, and care is taken not to oversoak the block. Good luck. Stan Lupo Department of Safety Assessment GlaxoSmithKline Felix.Rintelen@serono.com Sent by: histonet-admin@lists.utsouthwestern.edu 18-Sep-2003 04:18 To: Histonet cc: Subject: [Histonet] how to embed/section mesentery? Dear all, Is there anybody who may give me advice how to make good sections of parafin embedded mesentery (mouse)? How to prepare the tissue for embedding and how to handle it to embed/orient it properly in the parafin block? My concern is especially the soft consistency of the tissue making me feel that the proper orientation in the block might be difficult. Thank you very much in advance, Felix Felix Rintelen Serono Pharmaceutical Research Institute Geneva, Switzerland ******************************************************************************************** S - This message contains confidential information and is intended only for the individual named. If you are not the named addressee, you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. 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 malware. The presence of this disclaimer is not a proof that it was originated at Serono International S.A. or one of its affiliates. Serono International S.A and its affiliates therefore do not accept liability for any errors or omissions in the content of this message, which arise as a result of e-mail transmission. If verification is required, please request a hard-copy version. Serono International SA, 15bis Chemin Des Mines, Geneva, Switzerland, www.serono.com. ********************************************************************************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/4983213d/attachment.htm From juan.gutierrez <@t> christushealth.org Thu Sep 18 09:44:55 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Finger Nail Softening Message-ID: I don't know if it is still available, but in the old days we used Nair to soften toe nails. We just covered the nail with it for a couple of days and then processed it. Good luck. Juan C Gutierrez, HT(ASCP) -----Original Message----- From: Brent Norris [mailto:hectorjito26@hotmail.com] Sent: Thu 9/18/2003 9:18 AM To: histonet@pathology.swmed.edu Cc: Subject: [Histonet] Finger Nail Softening Hey all! I was asked a question about softening finger nails before or during processing that would make them easier to cut and allow them to hold on to the slide. They want to do this instead of any kind of treatment to the nails after they are embedded in paraffin so that it isn't just on the surface. If anyone has a procedure for this or a reference from a manual I would really appreciate it. You can email me directly at banorris@utmb.edu thanks once again Brent A. Norris _____ Compare Cable, DSL or Satellite plans: As low as $29.95. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Linke_Noelle <@t> Allergan.com Thu Sep 18 09:45:47 2003 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] how to embed/section mesentery? Message-ID: <805C4A052A253B4A8E4948B13F54D12704D181@IRMAIL102.irvine.allergan.com> Hi Felix, I guess it depends on what you are looking for in the mesentery. Last time we did this (in rats), we trimmed by starting with the mesenteric lymph node, and trimmed all the mesentery off by following the GI tract. Then, using the lymph node as a center point, spread out the mesentery between 2 sponges and processed flat. That way it comes out of the processor ready to embed flat! Hope this helps, Noelle Linke, BS, HTL(ASCP) Allergan, Inc Irvine, CA 92612 -----Original Message----- From: Felix.Rintelen@serono.com [mailto:Felix.Rintelen@serono.com] Sent: Thursday, September 18, 2003 1:19 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] how to embed/section mesentery? Dear all, Is there anybody who may give me advice how to make good sections of parafin embedded mesentery (mouse)? How to prepare the tissue for embedding and how to handle it to embed/orient it properly in the parafin block? My concern is especially the soft consistency of the tissue making me feel that the proper orientation in the block might be difficult. Thank you very much in advance, Felix Felix Rintelen Serono Pharmaceutical Research Institute Geneva, Switzerland ************************************************************************ ******************** S - This message contains confidential information and is intended only for the individual named. If you are not the named addressee, you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. 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 malware. The presence of this disclaimer is not a proof that it was originated at Serono International S.A. or one of its affiliates. Serono International S.A and its affiliates therefore do not accept liability for any errors or omissions in the content of this message, which arise as a result of e-mail transmission. If verification is required, please request a hard-copy version. Serono International SA, 15bis Chemin Des Mines, Geneva, Switzerland, www.serono.com. ************************************************************************ ********************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Thu Sep 18 09:55:23 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] how to embed/section mesentery? In-Reply-To: References: Message-ID: <3F69C75B.6080009@umdnj.edu> Hi Felix: For mesentaries some people like a "spread" rather than a section. Take out several loops of intestine and pin them out on a sheet of cork or dental wax with a tiny bit of tension on the mesentary so it is flat. Submerge in fix. You can process the whole "block" this way throught alcohols and clearing, then cut out and mount the sheet of mesentary on a slide without embedding. Yes it will be thicker than a section but you will see a nice 3D tissue with capillaries etc. Geoff Felix.Rintelen@serono.com wrote: >Dear all, > >Is there anybody who may give me advice how to make good sections of >parafin embedded mesentery (mouse)? How to prepare the tissue for embedding >and how to handle it to embed/orient it properly in the parafin block? My >concern is especially the soft consistency of the tissue making me feel >that the proper orientation in the block might be difficult. > >Thank you very much in advance, > >Felix > >Felix Rintelen >Serono Pharmaceutical Research Institute >Geneva, Switzerland > > > > >******************************************************************************************** >S - This message contains confidential information and is intended only for the individual >named. If you are not the named addressee, you should not disseminate, distribute or copy >this e-mail. Please notify the sender immediately by e-mail if you have received this >e-mail by mistake and delete this e-mail from your system. >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 malware. The >presence of this disclaimer is not a proof that it was originated at Serono International S.A. >or one of its affiliates. Serono International S.A and its affiliates therefore do not accept >liability for any errors or omissions in the content of this message, which arise as a result >of e-mail transmission. If verification is required, please request a hard-copy version. >Serono International SA, 15bis Chemin Des Mines, Geneva, Switzerland, www.serono.com. >********************************************************************************************* > > >_______________________________________________ >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 peoshel <@t> wisc.edu Thu Sep 18 10:00:49 2003 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] ABC of Safe Practices (Gripe!) In-Reply-To: <3F693DAF.92772E77@uwo.ca> References: <001601c37d6d$2714fce0$a88a140a@imvs.sa.gov.au> <3F693DAF.92772E77@uwo.ca> Message-ID: I have to agree with John here. There is *far* too much glitz and glamor on web sites that are supposedly for information. HTML does a lot, and what it doesn't do, we don't need. Or want. The extra junk just slows down browsers and creates compatibility problems. Recall the old advertising adage "sell the sizzle, not the steak"? It doesn't work here. Give us the steak and hang the sizzle. Phil >Dear Roy, > >The ABC of Safe Practices web site that you >recommend is impossibly slow! > >After 20 minutes I had not collected all >the decorated titles, and I didn't collect >a single word of actual information. > >Why do some writers of web sites >use complicated graphics instead of simple >words? Simple HTML provides for all sizes, >fonts and colours, and allows rapid >presentation of real graphics (diagrams, >photographs etc), with the option to make >pictures available by clicking on a link. > >The www.hoslink.com site is an abomination. >It should be plain text, not modern art. >End of growl. > >-- >------------------------- >John A. Kiernan >Department of Anatomy and Cell Biology >The University of Western Ontario >London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ >__________________ >Roy Ellis wrote: >> A useful resource for members of the list is 'The ABC of Safe >>Practices in the Biological Sciences Laboratory' ... written by >>histologists ... >> can be found at http://www.hoslink.com/aaindex.html -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From giorgia.setti <@t> kcl.ac.uk Thu Sep 18 10:38:45 2003 From: giorgia.setti <@t> kcl.ac.uk (Giorgia Setti) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] (no subject) Message-ID: <000801c37dfa$f2175fe0$39515c9f@umds.ac.uk> Dear all, has anyone stained MCP-1 in the rat kidney??And if yes which is the distribution of MCP-1 in the rat kidney?? Thank you Giorgia -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/fa353fb4/attachment.htm From MGomez <@t> ameripath.com Thu Sep 18 10:48:56 2003 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] IHC control Message-ID: Good morning everyone: I'm interested in purchasing control tissue/slides for Immunohistochemistry, particularly melanoma controls. Thank you, Milton From g.lang <@t> bigfoot.de Thu Sep 18 10:51:17 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Finger Nail Softening References: Message-ID: <000f01c37dfc$b2bdf550$0d12a8c0@SERVER> 10% KOH soften nail-tissue. We let the tissue in the solution for a few hours. Burck writes: tissue should be fixed in formalin, Alk. till 70%, than treat with "Chlordioxydessigs?ure" (=Diaphenol, =Chloratren from Chroma). This solution bleach Melanin. I hope this helps. Gundi Lang general hospital, Linz, Austria ----- Original Message ----- From: Brent Norris To: histonet@pathology.swmed.edu Sent: Thursday, September 18, 2003 4:18 PM Subject: [Histonet] Finger Nail Softening Hey all! I was asked a question about softening finger nails before or during processing that would make them easier to cut and allow them to hold on to the slide. They want to do this instead of any kind of treatment to the nails after they are embedded in paraffin so that it isn't just on the surface. If anyone has a procedure for this or a reference from a manual I would really appreciate it. You can email me directly at banorris@utmb.edu thanks once again Brent A. Norris ------------------------------------------------------------------------------ Compare Cable, DSL or Satellite plans: As low as $29.95. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/10d74688/attachment.htm From mcauliff <@t> umdnj.edu Thu Sep 18 10:37:12 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Purpose of Oven Prior to Staining? In-Reply-To: References: Message-ID: <3F69D128.1080108@umdnj.edu> Hi Jordan: I'm with you on this one. If the oven will deparafinize sections it should be possible to stain sections with an aqueous stain after "deparafinization" in the oven. Have them try this while you remove wax the normal way. Compare results. Maybe then they will believe you (unelss they are very stubborn). Geoff Jordan Brod wrote: >Good Morning All: > >My lab is in a huge discussion about the reason why we put our slides in an oven before staining. I told them that the reason was to dry all of the water off of them, because xylene and water do not mix. This statement is also backed up in literature that I have read. They still do not believe me, because they think the purpose of the oven is to deparaffinize the slides. > >Can anyone help me out, so I can put this discussion to rest? I will print off all of your comments for my technicians to read. Is what I told them correct or not? > >Thanks. > > > > >******************************************************************** >Jordan W. Brod >Diagnostic Lab Supervisor - Pathology >Texas Veterinary Medical Diagnostic Laboratory (TVMDL) >1 Sippel Road >TAMU 4471 >College Station, Texas 77843 >(979) 845-3414 >(979) 845-1794 fax >jbrod@tvmdl.tamu.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From jqb7 <@t> cdc.gov Thu Sep 18 10:55:46 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Purpose of Oven Prior to Staining? Message-ID: I agree about the removal of water. It also can aid in helping sections adhere to slides, but if you are using "plus" slides that aspect is probably irrelevant. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Thursday, September 18, 2003 11:37 AM To: Jordan Brod Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Purpose of Oven Prior to Staining? Hi Jordan: I'm with you on this one. If the oven will deparafinize sections it should be possible to stain sections with an aqueous stain after "deparafinization" in the oven. Have them try this while you remove wax the normal way. Compare results. Maybe then they will believe you (unelss they are very stubborn). Geoff Jordan Brod wrote: >Good Morning All: > >My lab is in a huge discussion about the reason why we put our slides >in an oven before staining. I told them that the reason was to dry all >of the water off of them, because xylene and water do not mix. This >statement is also backed up in literature that I have read. They still >do not believe me, because they think the purpose of the oven is to >deparaffinize the slides. > >Can anyone help me out, so I can put this discussion to rest? I will >print off all of your comments for my technicians to read. Is what I >told them correct or not? > >Thanks. > > > > >******************************************************************** >Jordan W. Brod >Diagnostic Lab Supervisor - Pathology >Texas Veterinary Medical Diagnostic Laboratory (TVMDL) >1 Sippel Road >TAMU 4471 >College Station, Texas 77843 >(979) 845-3414 >(979) 845-1794 fax >jbrod@tvmdl.tamu.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JGordon <@t> cellmarque.com Thu Sep 18 10:49:50 2003 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] IHC control Message-ID: Milton, Cell Marque sells IHC controls for ALL the antibodies that we offer, including skin controls for S-100, HMB-45, MART-1 melanA, Tyrosinase, and Factor 13a. Check out our website at www.cellmarque.com . All sets of controls are $59 each, regardless of the tissue (we even have rarer tissues like mantle cell lymphoma and anaplastic large cell lymphoma). If you have any further questions, please e-mail me or call us at 1-800-665-7284. Jeff Gordon Domestic Sales and Marketing Manager Cell Marque Corp. -----Original Message----- From: MGomez@ameripath.com [mailto:MGomez@ameripath.com] Sent: Thursday, September 18, 2003 10:49 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC control Good morning everyone: I'm interested in purchasing control tissue/slides for Immunohistochemistry, particularly melanoma controls. Thank you, Milton _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From g.lang <@t> bigfoot.de Thu Sep 18 11:22:30 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:54 2005 Subject: Fw: [Histonet] Purpose of Oven Prior to Staining? Message-ID: <003901c37e01$0f2cb0c0$0d12a8c0@SERVER> ----- Original Message ----- From: "Gudrun Lang" To: "Jordan Brod" Sent: Thursday, September 18, 2003 6:00 PM Subject: Re: [Histonet] Purpose of Oven Prior to Staining? > I think the purpose is to melt the paraffin, because warm liquid paraffin is > easier to get off the tissue in xylene. > The second point is, that the tissue will be baked on the glass. So it hold > better. > When you put the slides to early out of the oven, the cuts will swim off the > slide. > Gundi Lang > general hospital, Linz, Austria > > > ----- Original Message ----- > From: "Jordan Brod" > To: > Sent: Thursday, September 18, 2003 4:41 PM > Subject: [Histonet] Purpose of Oven Prior to Staining? > > > Good Morning All: > > My lab is in a huge discussion about the reason why we put our slides in an > oven before staining. I told them that the reason was to dry all of the > water off of them, because xylene and water do not mix. This statement is > also backed up in literature that I have read. They still do not believe > me, because they think the purpose of the oven is to deparaffinize the > slides. > > Can anyone help me out, so I can put this discussion to rest? I will print > off all of your comments for my technicians to read. Is what I told them > correct or not? > > Thanks. > > > > > ******************************************************************** > Jordan W. Brod > Diagnostic Lab Supervisor - Pathology > Texas Veterinary Medical Diagnostic Laboratory (TVMDL) > 1 Sippel Road > TAMU 4471 > College Station, Texas 77843 > (979) 845-3414 > (979) 845-1794 fax > jbrod@tvmdl.tamu.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From ree3 <@t> leicester.ac.uk Thu Sep 18 11:20:03 2003 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] actinomyces antibody Message-ID: Anyone know of an antibody to actinomyces that would work on paraffin sections???? Many thanks Richard Edwards MRC TOX UNIT....LEICESTER...U.K.... From g.lang <@t> bigfoot.de Thu Sep 18 11:34:54 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Glass cleaning for silver References: <9DF8DDFE3736D3119D6D0008C79148D60A5BFD21@groexmbcr17.pfizer.com> <3F67F0ED.77A2DCF5@uwo.ca> Message-ID: <007701c37e02$caae49c0$0d12a8c0@SERVER> What happens with the silver stain, when class with traces of metallic silver is used? We don't do this cleaning in our lab, and had no problems until now. Gundi Lang ----- Original Message ----- From: "John Kiernan" To: "Behan, Rosemarie G" Cc: Sent: Wednesday, September 17, 2003 7:28 AM Subject: [Histonet] Glass cleaning for silver > Rosemary Behan > asked about cleaning glassware > for silver staining. > A reply follows. > > It's necessary to remove traces of metallic silver, > which are not necessarily visible. The only common > acid that does this is nitric. Here is what I do, > with rationale. Let's assume the vessel is a Coplin > jar. > > Rinse with one or two changes of distilled water. > [Tap water contains chloride ions and silver > chloride precipitation must be avoided.] > > Pour a few ml of concentrated nitric acid into > the vessel and carefully let it wet all the > inside surface. For a Coplin jar it's important > to dissolve silver from all corners and from the > slots for supporting slides. It takes 15-30 > seconds to make black deposits or mirrors > disappear. A minute of turning and tilting has > to be enough to remove all the visible and > invisible silver. > > Safely discard the nitric acid and fill up the > vessel with distilled or otherwise purified > water, three times. > [Pure water must be used because tap water > will precipitate traces of silver chloride > from the residual nitric acid - which contains > dissolved silver nitrate.] > > Wash in tap water, detergent etc as for any > other dirty lab glassware, and don't spare the > brush. > [The nitric acid treatment does not remove > all types of dirt. Bits of detached section, > stuck to the glass, are made yellow by nitric > acid.] > > Rinse in tap water, repeatedly, to get > rid of the detergent (no more froth with > shaking) and then in 3 generous changes of > pure water to dilute out residual chloride > from the tap water. > > Let the vessel dry by drainage and evaporation, > then keep it in a closed cupboard, with its > lid on (if it has a lid; and don't forget to > clean the lid). > > If glassware is contaminated by insoluble silver > compounds such as silver chloride, 5% sodium > thiosulphate (10 minutes) will remove the silver. > [The thiosulphate ion strongly complexes silver > ions and will remove them from solid silver > halides. This is the "fixation" of photographers.] > > In the above remarks I have not given detailed > safety and disposal instructions. Conc. nitric acid > is nasty stuff but becomes harmless when diluted > with water. > > Do not use hydrochloric acid or an HCl-alcohol > mixture for "acid washing" of glassware that will > contain silver nitrate or protargol. > [Reason is obvious from above discussion.] > > There are silver solvents less noxious than > concentrated nitric acid. The best known one is > Farmer's reducer. This is used in black & white > photography for controlled removal of darkness > (= silver) from negatives or prints. It is a > solution containing potassium ferricyanide and > sodium thiosulphate. Its actions on photo media > take several minutes, but it takes much longer > to weaken silver deposits on glassware and in > overstained sections (my unpublished anecdotal > observations). > > For what it's worth, I think concentrated > nitric acid is the best cleaner of glassware > used for silver methods. I also think that > poor glass-hygiene (dishwashers etc etc etc) > often causes failure or poor results with > many staining techniques. > -- > ------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ > -- > (Rosemary Behan: I've deleted many irrelevant > messages from the tail of your email, which > contained a megabyte of unrelated stuff. > Please be careful about what to quote!) > _____________________________________ > "Behan, Rosemarie G" wrote: > > > > I am looking for a recipe for acid cleaning glassware to do silver stains, > > can anyone help me? > __________________________________________________ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From weipingren <@t> yahoo.com Thu Sep 18 11:36:23 2003 From: weipingren <@t> yahoo.com (weiping Ren) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] immunostaining of VEGF and CD31 in mouse decalcified bone tissue Message-ID: <20030918163623.13822.qmail@web41613.mail.yahoo.com> Hi, Histonetter: Does anybody have experience of immunohistochemical staining of VEGF and CD31 in parraffin embedded and decalcified mouse bone tissue? We have tried using VEGF (santa Cruz) and CD31(Phamgen) antibody in mouse bone (formalin fixed and EDTA decalcified) with very weak staining. The antigen retrieval method we have tried is using proteinase K incubation for 10 min. I would appreciate any suggestions and advice. Thanks. Weiping Wayne State University --------------------------------- Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/1c0f6920/attachment.htm From gentras <@t> vetmed.auburn.edu Thu Sep 18 11:39:54 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] acid-cleaning glassware Message-ID: <5.2.0.9.0.20030918112303.009fd0c0@mailhost.vetmed.auburn.edu> I have used the potassium dichromate/sulfuric acid method, chromerge/ sulfuric and I'm currently using Nochromix/sulfuric acid all of which work just fine for silver staining. Both the chromerge cleaning solution (# C577-12) and Nochromix Reagent (# 04-345-20) I order from Fisher Scientific ( 800-766-7000). Best wishes. Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From la.sebree <@t> hosp.wisc.edu Thu Sep 18 11:47:49 2003 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Purpose of Oven Prior to Staining? Message-ID: I agree with you Jordan. There is still paraffin on the slides after they come out of the oven. If your cut slides are already as dry as the Sahara, you could forgo the oven all together and go straight into xylene; that's what is deparaffinizing the slides, not the oven. There are also commercial HIER buffers available that have detergents in them that will achieve deparaffinization simultaneously with the retrieval procedure, with or without the oven. Stick to your guns, Jordan; you're right! Linda A. Sebree, HT(ASCP) University of Wisconsin Hospital & Clinics IHC/ISH Laboratory A4/204-2472 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: Jordan Brod [mailto:jbrod@tvmdl.tamu.edu] Sent: Thursday, September 18, 2003 9:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Purpose of Oven Prior to Staining? Good Morning All: My lab is in a huge discussion about the reason why we put our slides in an oven before staining. I told them that the reason was to dry all of the water off of them, because xylene and water do not mix. This statement is also backed up in literature that I have read. They still do not believe me, because they think the purpose of the oven is to deparaffinize the slides. Can anyone help me out, so I can put this discussion to rest? I will print off all of your comments for my technicians to read. Is what I told them correct or not? Thanks. ******************************************************************** Jordan W. Brod Diagnostic Lab Supervisor - Pathology Texas Veterinary Medical Diagnostic Laboratory (TVMDL) 1 Sippel Road TAMU 4471 College Station, Texas 77843 (979) 845-3414 (979) 845-1794 fax jbrod@tvmdl.tamu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Thu Sep 18 11:49:12 2003 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Carcinogenic materials Message-ID: <0BE6ADFAE4E7E04496BF21ABD34662801D0C47@EXCHANGE1.huntingtonhospital.com> I've had several requests for some of the information I received on carcinogenic materials. The information listed below is from an NSH publication, "Health and Safety Committe Information Packet." You can contact NSH at histo@nsh.org or (301) 262-6221. Their website is www.nsh.org and there are numerous publications you can order from this website. Carcinogens in the Histology Lab: Arsenic compounds (cacodylate buffers) Benzene (solvent) Chromium oxide (oxidizer for GMS) Formaldehyde (fixative) Benzidine (chromagen) Benzideine-based dyes such as: biebrich scarlet, congo red, oil red O, woodstain scarlet, ponceau S, trypan blue, bordeaux. Laurie Colbert Huntington Hospital Pasadena, CA From chris <@t> ptplab.com Thu Sep 18 12:21:17 2003 From: chris <@t> ptplab.com (Christopher L. Robertson) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Used Photodyne & Printer for sale Message-ID: Anyone in histoland need a used Photodyne machine and printer? Model V701 Photo Machine with monitor Sony color video printer UP1220A Sony color print pack for video printer, 3 Boxes Original price for Photodyne $8, 200 Original Price for video printer $1,587 Asking price: $4,000 (or best offer) Chris PTPl chris@ptplab.com From dellav <@t> musc.edu Thu Sep 18 12:39:16 2003 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Carcinogenic materials Message-ID: Laurie, do you think it would be beneficial to also include "suspected carcinogens" to your list? don't know if the materials you checked listed these as well. I find it interesting that formaldehyde didn't make your list. our safety dept here claims it's no longer suspected but a confirmed carcinogen. Vinnie Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Laurie Colbert" 09/18/03 12:49PM >>> I've had several requests for some of the information I received on carcinogenic materials. The information listed below is from an NSH publication, "Health and Safety Committe Information Packet." You can contact NSH at histo@nsh.org or (301) 262-6221. Their website is www.nsh.org and there are numerous publications you can order from this website. Carcinogens in the Histology Lab: Arsenic compounds (cacodylate buffers) Benzene (solvent) Chromium oxide (oxidizer for GMS) Formaldehyde (fixative) Benzidine (chromagen) Benzideine-based dyes such as: biebrich scarlet, congo red, oil red O, woodstain scarlet, ponceau S, trypan blue, bordeaux. Laurie Colbert Huntington Hospital Pasadena, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dellav <@t> musc.edu Thu Sep 18 12:53:47 2003 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] The NSH needs your help NOW! Message-ID: The National Society for Histotechnology is interested in learning how histology practitioners utilize microwave devices in their work. In this vein, we have created a brief, anonymous survey that will enable us to obtain data that reflects the standard of practice with microwave accelerated procedures. This survey will take approximately 5 minutes to complete. This information will assist us in the creation of microwave user guidelines as part of our collaboration with the National Committee for Clinical Laboratory Standards (NCCLS). We cannot meet our goal without your help. Your input is key to enabling us to better understand how microwave devices are used in the field. You can help us by going to the NSH homepage at http://www.nsh.org and clicking on the link at the top of the page to complete the survey. We would like to hear from everyone, even if you have never used a microwave device in your work. It is open to everyone who practices in our field, not just NSH members. Please take a moment of your time to provide this important information. We must compile this information before the NSH Symposium/Convention in October so time is running short. Please complete your survey TODAY !!!! Vinnie Della Speranza NSH Vice President From Diane.Gladney <@t> se.amedd.army.mil Thu Sep 18 13:08:16 2003 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Purpose of Oven Prior to Staining? Message-ID: <9D41AB7C56F8304F98537ABD87B249F6626129@dasmthgbz001.amedd.army.mil> Jordan, The purpose of the slide dryer or oven is to dry the water off of the slides. The slides should be well drained to remove as much water as possible before placing them in the rack before it is placed in the oven. It is not and will not deparaffinize the slides. The slides must be completely free of water before they are stained either by hand or on an automatic slide stainer. Any histological technique book will tell you that the deparaffinization is accomplished by using xylene, xylene substitute or other appropriate paraffin solvent. Ideally, the slides should be dried at room temperature overnight especially if photographs are going to be made. But few labs have this much time. A drier that has a temperature in excess of 60 degrees C will cause some artifacts. Our autostainer has a built-in forced air dryer that the slides stay in for 10 - 15 minutes. If we have a rack on the stainer and have another rack ready for the stainer, we place this rack in a small forced air dryer set for 15 minutes then remove and wait on the stainer to finish before adding another rack. The slide dryer then is by-passed and the dried slides go directly to the first xylene substitute (what we use instead of xylene) for deparaffinization. If they still won't believe you, have them try to stain some "dried slides" and see the results. Good Luck, Diane Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: Jordan Brod [mailto:jbrod@tvmdl.tamu.edu] Sent: Thursday, September 18, 2003 10:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Purpose of Oven Prior to Staining? Good Morning All: My lab is in a huge discussion about the reason why we put our slides in an oven before staining. I told them that the reason was to dry all of the water off of them, because xylene and water do not mix. This statement is also backed up in literature that I have read. They still do not believe me, because they think the purpose of the oven is to deparaffinize the slides. Can anyone help me out, so I can put this discussion to rest? I will print off all of your comments for my technicians to read. Is what I told them correct or not? Thanks. ******************************************************************** Jordan W. Brod Diagnostic Lab Supervisor - Pathology Texas Veterinary Medical Diagnostic Laboratory (TVMDL) 1 Sippel Road TAMU 4471 College Station, Texas 77843 (979) 845-3414 (979) 845-1794 fax jbrod@tvmdl.tamu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Thu Sep 18 13:17:00 2003 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Carcinogenic materials Message-ID: <0BE6ADFAE4E7E04496BF21ABD34662801D0C48@EXCHANGE1.huntingtonhospital.com> Formaldehyde is on the list. I'm sure this list is not meant to be all-inclusive. I think it's just a listing of some of the most common chemicals we use in histology. What I printed was just a small portion of the publication. There is lots more info in the "expanded" version. -----Original Message----- From: Vinnie Della Speranza [mailto:dellav@musc.edu] Sent: Thursday, September 18, 2003 10:39 AM To: Laurie Colbert; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Carcinogenic materials Laurie, do you think it would be beneficial to also include "suspected carcinogens" to your list? don't know if the materials you checked listed these as well. I find it interesting that formaldehyde didn't make your list. our safety dept here claims it's no longer suspected but a confirmed carcinogen. Vinnie Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Laurie Colbert" 09/18/03 12:49PM >>> I've had several requests for some of the information I received on carcinogenic materials. The information listed below is from an NSH publication, "Health and Safety Committe Information Packet." You can contact NSH at histo@nsh.org or (301) 262-6221. Their website is www.nsh.org and there are numerous publications you can order from this website. Carcinogens in the Histology Lab: Arsenic compounds (cacodylate buffers) Benzene (solvent) Chromium oxide (oxidizer for GMS) Formaldehyde (fixative) Benzidine (chromagen) Benzideine-based dyes such as: biebrich scarlet, congo red, oil red O, woodstain scarlet, ponceau S, trypan blue, bordeaux. Laurie Colbert Huntington Hospital Pasadena, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jbrod <@t> tvmdl.tamu.edu Thu Sep 18 10:27:01 2003 From: jbrod <@t> tvmdl.tamu.edu (Jordan Brod) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Thanks For the Responses Message-ID: Thank you for responding to my question. I have figured out that my technicians are just wanting to argue with me (all in good fun)!! They know better, they are just having a good time. They have now re-named our oven "The Depariffinization Facilitator." What a Technical Staff!!!! Thanks again, Gig'em Aggies!!!! Jordan Brod From jkiernan <@t> uwo.ca Thu Sep 18 13:44:36 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Glass cleaning for silver References: <9DF8DDFE3736D3119D6D0008C79148D60A5BFD21@groexmbcr17.pfizer.com> <3F67F0ED.77A2DCF5@uwo.ca> <007701c37e02$caae49c0$0d12a8c0@SERVER> Message-ID: <3F69FD14.DA2D2441@uwo.ca> If there are traces of mmetallic silver on the glass, they act as catalytic centres in the reduction (development) stage of the silver method and more silver is deposited on them, instead of in the correct places in the sections. If you do gold toning, the silver deposits are largely changed to gold, which cannot be removed with nitric acid. Deposition of silver on glass is a problem especially with techniques in which concentrated silver nitrate is deliberately carried over into the developer. The Gros-Schultz method for axons is an example. There can also be deposition if solutions are left in Coplin jars or staining tanks after use. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ _______________________________ Gudrun Lang wrote: > > What happens with the silver stain, when class with traces of metallic > silver is used? > We don't do this cleaning in our lab, and had no problems until now. > > Gundi Lang > > ----- Original Message ----- > From: "John Kiernan" > To: "Behan, Rosemarie G" > Cc: > Sent: Wednesday, September 17, 2003 7:28 AM > Subject: [Histonet] Glass cleaning for silver > > > Rosemary Behan > > asked about cleaning glassware > > for silver staining. > > A reply follows. > > > > It's necessary to remove traces of metallic silver, > > which are not necessarily visible. The only common > > acid that does this is nitric. Here is what I do, > > with rationale. Let's assume the vessel is a Coplin > > jar. > > > > Rinse with one or two changes of distilled water. > > [Tap water contains chloride ions and silver > > chloride precipitation must be avoided.] > > > > Pour a few ml of concentrated nitric acid into > > the vessel and carefully let it wet all the > > inside surface. For a Coplin jar it's important > > to dissolve silver from all corners and from the > > slots for supporting slides. It takes 15-30 > > seconds to make black deposits or mirrors > > disappear. A minute of turning and tilting has > > to be enough to remove all the visible and > > invisible silver. > > > > Safely discard the nitric acid and fill up the > > vessel with distilled or otherwise purified > > water, three times. > > [Pure water must be used because tap water > > will precipitate traces of silver chloride > > from the residual nitric acid - which contains > > dissolved silver nitrate.] > > > > Wash in tap water, detergent etc as for any > > other dirty lab glassware, and don't spare the > > brush. > > [The nitric acid treatment does not remove > > all types of dirt. Bits of detached section, > > stuck to the glass, are made yellow by nitric > > acid.] > > > > Rinse in tap water, repeatedly, to get > > rid of the detergent (no more froth with > > shaking) and then in 3 generous changes of > > pure water to dilute out residual chloride > > from the tap water. > > > > Let the vessel dry by drainage and evaporation, > > then keep it in a closed cupboard, with its > > lid on (if it has a lid; and don't forget to > > clean the lid). > > > > If glassware is contaminated by insoluble silver > > compounds such as silver chloride, 5% sodium > > thiosulphate (10 minutes) will remove the silver. > > [The thiosulphate ion strongly complexes silver > > ions and will remove them from solid silver > > halides. This is the "fixation" of photographers.] > > > > In the above remarks I have not given detailed > > safety and disposal instructions. Conc. nitric acid > > is nasty stuff but becomes harmless when diluted > > with water. > > > > Do not use hydrochloric acid or an HCl-alcohol > > mixture for "acid washing" of glassware that will > > contain silver nitrate or protargol. > > [Reason is obvious from above discussion.] > > > > There are silver solvents less noxious than > > concentrated nitric acid. The best known one is > > Farmer's reducer. This is used in black & white > > photography for controlled removal of darkness > > (= silver) from negatives or prints. It is a > > solution containing potassium ferricyanide and > > sodium thiosulphate. Its actions on photo media > > take several minutes, but it takes much longer > > to weaken silver deposits on glassware and in > > overstained sections (my unpublished anecdotal > > observations). > > > > For what it's worth, I think concentrated > > nitric acid is the best cleaner of glassware > > used for silver methods. I also think that > > poor glass-hygiene (dishwashers etc etc etc) > > often causes failure or poor results with > > many staining techniques. > > -- > > ------------------------- > > John A. Kiernan > > Department of Anatomy and Cell Biology > > The University of Western Ontario > > London, Canada N6A 5C1 > > kiernan@uwo.ca > > http://publish.uwo.ca/~jkiernan/ > > -- > > (Rosemary Behan: I've deleted many irrelevant > > messages from the tail of your email, which > > contained a megabyte of unrelated stuff. > > Please be careful about what to quote!) > > _____________________________________ > > "Behan, Rosemarie G" wrote: > > > > > > I am looking for a recipe for acid cleaning glassware to do silver > stains, > > > can anyone help me? > > __________________________________________________ > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Thu Sep 18 13:46:19 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] oven drying slides Message-ID: <5.2.0.9.0.20030918133949.009fc960@mailhost.vetmed.auburn.edu> In my opinion the purpose is to complete the drying of water from the sections to avoid loss of valuable tissue and to aid in easier removable or the warmed paraffin during deparaffinization. Therefore, yes you are correct. Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From DDittus787 <@t> aol.com Thu Sep 18 13:51:28 2003 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] IHC control Message-ID: <687779E6.1432F4D8.0A1F969F@aol.com> zymed carries control tissue for formalin fixed antibodies 1-800-874-4494 Dana From Myri37 <@t> aol.com Thu Sep 18 13:59:15 2003 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] MMA sections Message-ID: <1ea.fe0f9b0.2c9b5a83@aol.com> hello everyone i cut bloks of methylmetacrylate, and i have sections of 5 um thikness, i need to stain tissue embedded, do you think it's necessary to deplasticise MMA do you have any protocol : how to deplasticise MMA, how to stick sections with MMA on slides, and do you know any stain to show collagen fibers, and mineralizing tissue thank you very much for anyhelp and any advice Myriam Natural Implant marseille -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/5912234f/attachment.htm From emry <@t> u.washington.edu Thu Sep 18 14:02:32 2003 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] msds Message-ID: I have not been able to print msds information from several sights. Is there a sight where this is easier? Thanks, Trisha From carl.hobbs <@t> kcl.ac.uk Thu Sep 18 14:24:30 2003 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] how to embed mesentery Message-ID: <000e01c37e1a$88348a60$89c98451@home> Fred, perfect! We did same with hamster cheek pouch...worked a treat. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/26041d9c/attachment.htm From gentras <@t> vetmed.auburn.edu Thu Sep 18 14:37:24 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] under processed tissue Message-ID: <5.2.0.9.0.20030918135516.00a0b140@mailhost.vetmed.auburn.edu> Hello, my processor recently malfunctioned rendering poorly processed sections. I placed the sections in the 60C incubator for 30 min. to aid in melting of the paraffin. Then I closely monitored the samples as I sent them backwards through the processing cycle. I actually reversed the beakers ( ie. placed #10 at station 1 and beaker #1 at station 10 and so on and so forth). If you decide to try this be sure to remember to switch them back to their proper stations and replace all solutions. Also, don't allow them to go past #10 ( avoiding the paraffin infiltration). I Washed sections for 30 min. in running tap H2O. Then placed them in a rehydrating solution: 0.6g sodium carbonate+ 42ml DHOH +18ml absolute alcohol overnight. Although, this method advises processing the following day on the standard 16-hour procedure, starting in 80% alcohol. I usually processed as usual starting in 70% ETOH. This rehydrating solution method was taken from Sheehan/Mosby Theory and Practice of Histotechnology pge 4. My most recent samples I'm currently holding in 70% ETOH until I can get my processor problem resolved. However, I've used this method before and it renders fairly decent sections with a bit of trial and error. Best wishes. Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/bb3af01e/attachment.htm From huffpw <@t> uleth.ca Thu Sep 18 14:46:07 2003 From: huffpw <@t> uleth.ca (Phillip Huff) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] msds In-Reply-To: References: Message-ID: <1690.142.66.42.15.1063914367.squirrel@webmail.uleth.ca> The following site contains a listing of everything one needs to know about MSDS. http://www.ilpi.com/msds/ Scroll down about 2 pages and you will come to a set of links for MSDSs. Hope it helps Phil > I have not been able to print msds information from several sights. Is > there a sight where this is easier? > Thanks, > > Trisha > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From RITA.ANGEL <@t> UC.EDU Thu Sep 18 14:50:30 2003 From: RITA.ANGEL <@t> UC.EDU (Rita Angel) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] HT Exam Message-ID: <5.1.0.14.2.20030918154606.00b646e8@ucmail3.uc.edu> Hi everyone, I have a co-worker that has is interested in taking the HT Exam. She has filled out the paperwork and has mailed it, but is wondering what advice everyone has in which resources to study. There seems to be a low success rate in passing the exam, about 56%, according to recent emails I've received. I took the exam 10 years ago, so I'm not sure my advice would be the best for her now. Thanks for your help, Rita Angel, HT (ASCP) University of Cincinnati From padunnje <@t> iupui.edu Thu Sep 18 14:52:19 2003 From: padunnje <@t> iupui.edu (Dunn-Jena, Patsy A) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Antibody-Anti-Lactobacillis-casei Message-ID: Does anyone use this antibody that can help me with a vendor and a protocol? I am almost 1/2 way through a study and now the advisors want the student to change the procedure. And of course it needs to be done yesterday! Rabbit or monoclonal is preferred for primary and secondary would be goat. Any help that anyone can offer would be appreciated. Patsy Dunn-Jena, LAT, RVT, HT (ASCP) Indiana University School of Dentistry Mineralized Tissue and Histology Research Laboratory (317) 274-0544 padunnje@iupui.edu -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/49224f4d/attachment.htm From tpmorken <@t> labvision.com Thu Sep 18 15:05:04 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] HT Exam Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CB8E@usca0082k03.rallansci.apogent.com> " ... taking the HT Exam ...is wondering what advice everyone has in which resources to study." Study EVERYTHING you can get your hands on. No kidding! Tim Morken HTL(ASCP) Lab Vision / NeoMarkers www.labvision.com -----Original Message----- From: Rita Angel [mailto:RITA.ANGEL@UC.EDU] Sent: Thursday, September 18, 2003 12:51 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HT Exam Hi everyone, I have a co-worker that has is interested in taking the HT Exam. She has filled out the paperwork and has mailed it, but is wondering what advice everyone has in which resources to study. There seems to be a low success rate in passing the exam, about 56%, according to recent emails I've received. I took the exam 10 years ago, so I'm not sure my advice would be the best for her now. Thanks for your help, Rita Angel, HT (ASCP) University of Cincinnati _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Thu Sep 18 15:08:11 2003 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] HT Exam Message-ID: See: http://www.ascp.org/bor/directors/stats/ht_stat.asp for current stats. Gary Gill -----Original Message----- From: Rita Angel [mailto:RITA.ANGEL@UC.EDU] Sent: Thursday, September 18, 2003 2:51 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HT Exam Hi everyone, I have a co-worker that has is interested in taking the HT Exam. She has filled out the paperwork and has mailed it, but is wondering what advice everyone has in which resources to study. There seems to be a low success rate in passing the exam, about 56%, according to recent emails I've received. I took the exam 10 years ago, so I'm not sure my advice would be the best for her now. Thanks for your help, Rita Angel, HT (ASCP) University of Cincinnati _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rfail <@t> toolkitmail.com Thu Sep 18 15:12:40 2003 From: rfail <@t> toolkitmail.com (rfail@toolkitmail.com) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] SAB for amyloid Message-ID: <01L0TE1BEHKM99CROU@SMTP00.InfoAve.Net> Does anyone know of an amyloid stain with the abbreviation SAB? Rena Fail From gareth.davis <@t> Vanderbilt.Edu Thu Sep 18 15:52:39 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Opinion on storing sections on slides Message-ID: <082C721AF78DB34983E8BA2CD085462105C676@mailbe07> Hello All, I would like to get some input on storing sections. I have cut frozen (PFA perfused and post fixated) tissue on a sliding microtome, kept the cut sections in PBS until mounting on slides. Then I applied the sections to a SuperFrost Plus slide with water and dried down on a slide warmer-overnight. So, the staining has turned out okay, but I wonder if I should keep the sections I don't use in the freezer, like typical frozen sections, or keep them at room temperature, like paraffin sections (since they are dry and already fixed). I was going to try to stain free floating sections, but it just didn't work. So, any suggestions would be greatly appreciated. Thanks, Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/538c966f/attachment.htm From JWEEMS <@t> sjha.org Thu Sep 18 15:57:08 2003 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] SAB for amyloid Message-ID: <9E75DAB5F369D84ABF84FAB7A0243B44B6D42E@exch4.sjha.org> It's sulfonated alcian blue -----Original Message----- From: rfail@toolkitmail.com [mailto:rfail@toolkitmail.com] Sent: Thursday, September 18, 2003 4:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] SAB for amyloid Does anyone know of an amyloid stain with the abbreviation SAB? Rena Fail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gareth.davis <@t> Vanderbilt.Edu Thu Sep 18 15:59:56 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Question for Colleen Forster Message-ID: <082C721AF78DB34983E8BA2CD085462105C677@mailbe07> Colleen, I have question for you on your c-fos protocol. "I do a steam retrievel using a 6.0pH buffer for 30 minutes followed by a 10 minute cool down" Exactly what buffer do you use and how do you go about the steam retrievel? I used to place my coplin jar (with slides and buffer) in a beaker of boiling water - is that what you do? Thanks, Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/87c4d110/attachment.htm From amosbrooks <@t> earthlink.net Thu Sep 18 15:57:56 2003 From: amosbrooks <@t> earthlink.net (amosbrooks@earthlink.net) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Re: Histonet digest, Vol 1 #53 - 32 msgs In-Reply-To: <20030918170001.24794.15784.Mailman@swlx167.swmed.edu> Message-ID: <3F69E414.7577.225AAE7@localhost> Jordan, You are right they are wrong. I agree with what Geoff said about having them stain an un- deparaffinized section. He makes a good point. When you pick up a section there is a tiny film of water between the section and the slide. If you don't remove the film of water by drying the slide, the water will try to pull off the slide and mix with whatever solution it is in. (Regaurdless of using charged slides or not) That would be bad because the water pulling away from the slide would pull the section off with it. Like a sandwich and pulling the jelly out thru the bread. That is the reason for drying the slide. Another experiment to try. Don't dry some extra slides. Just run them down to water. Most of the tissue will have fallen off. That is just to the water water, then try staining them, especially with any harsh chemicals (peroxide, silver etc), there will be no tissue left. It's nice being right isn't it? later, Amos Brooks > Message: 14 > Date: Thu, 18 Sep 2003 09:41:22 -0500 > From: "Jordan Brod" > To: > Subject: [Histonet] Purpose of Oven Prior to Staining? > > Good Morning All: > > My lab is in a huge discussion about the reason why we put our slides > in = an oven before staining. I told them that the reason was to dry > all of = the water off of them, because xylene and water do not mix. > This = statement is also backed up in literature that I have read. > They still do = not believe me, because they think the purpose of the > oven is to deparaffin= ize the slides. > > Can anyone help me out, so I can put this discussion to rest? I will > = print off all of your comments for my technicians to read. Is what > I told = them correct or not? > > Thanks. From rbruggeman <@t> psu.edu Thu Sep 18 13:24:33 2003 From: rbruggeman <@t> psu.edu (Richard Bruggeman) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Automated IHC machine Message-ID: We are looking to purchase a new automated stainer for our pathology research lab and would like to hear some recommendations from the group. Our needs include the ability to be highly adaptable and variable, almost on a daily basis (this is purely for research.) We do not necessarily need extremely high throughput (20 slides per run would be fine.) The ability to perform ISH and antigen retrieval would be nice as well. If anyone has a suggestion as to what unit to purchase, we would be grateful. Thanks! Richard "Trey" Bruggeman Milton S. Hershey Medical Center Penn State College of Medicine Department of Pathology, H179 500 University Drive Hershey, PA 17033 (717) 531-1044 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/009953a3/attachment.htm From koyano <@t> umn.edu Thu Sep 18 17:10:01 2003 From: koyano <@t> umn.edu (Naoko Koyano) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Microtome knife sharpener in Minneapolis area? Message-ID: Hello everyone. I recently moved to Minneapolis and am looking for a knife sharpener that I could come and use in the Twin cities area, preferably in U of M. Does anyone have a sharpener around here? If you are not in the Twin Cities but know a good place to send knives to get sharpened, I would appreciate that information, too. Thank you very much. Naoko Koyano -- ****************************** Naoko Koyano-Nakagawa E-mail: koyano@umn.edu Office: 612-625-7687 Lab: 612-624-3131 Department Fax: 612-626-5009 ****************************** From AnthonyH <@t> chw.edu.au Thu Sep 18 17:59:31 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Purpose of Oven Prior to Staining? Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E03E@simba.kids> Jordan, Reasons for drying slides above the melting point of wax: 1. Remove excess water 2. Allow sections to attach to slides (otherwise they will lift off in the dewaxing xylene). 3. Aid in final spreading out of sections so that there are fewer folds (should I say NO folds!!) 4. Remove some of the excess wax (ie it flows off the slide). Remember this requires regular cleaning of the oven to remove excess wax. 5. Can't think of any other reasons Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Jordan Brod [mailto:jbrod@tvmdl.tamu.edu] Sent: Friday, 19 September 2003 0:41 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Purpose of Oven Prior to Staining? Good Morning All: My lab is in a huge discussion about the reason why we put our slides in an oven before staining. I told them that the reason was to dry all of the water off of them, because xylene and water do not mix. This statement is also backed up in literature that I have read. They still do not believe me, because they think the purpose of the oven is to deparaffinize the slides. Can anyone help me out, so I can put this discussion to rest? I will print off all of your comments for my technicians to read. Is what I told them correct or not? Thanks. ******************************************************************** Jordan W. Brod Diagnostic Lab Supervisor - Pathology Texas Veterinary Medical Diagnostic Laboratory (TVMDL) 1 Sippel Road TAMU 4471 College Station, Texas 77843 (979) 845-3414 (979) 845-1794 fax jbrod@tvmdl.tamu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Thu Sep 18 18:11:05 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] SAB for amyloid Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E03F@simba.kids> The technique is as follows: SULPHATED ALCIAN BLUE FOR AMYLOID PRINCIPLE: Amyloid has an increased avidity for alcian blue in the presence of added salt. By fixing with an akali solution, SAB can be followed by various trichromic counterstains. FIXATION: 10% buffered formalin MICROTOMY: 7um paraffin sections SOLUTIONS: 1. Acetic/Alcohol Rinse: 95% ethanol 45ml Distilled water 45ml Glacial Acetic Acid 10ml 2. Stock Solutions: a) 1% Alcian Blue (CI 74240) in 95% ethanol. b) 1% Sodium Sulphate Hydrate in distilled water. 3. SAB Working Solution - prepare fresh: Stock solution (a) 18ml Stock solution (b) 18ml Glacial Acetic Acid 4ml Mix, stand 30 minutes before use. 4. 80% ethanol saturated with borax. 5. Van Gieson counterstain. METHOD: 1. Hydrate section and wash for 10 minutes. 2. Rinse in acetic/alcohol, 2 minutes. 3. Place in SAB, 2 hours. 4. Rinse in acetic/alcohol, 2 minutes. 5. Wash in water. 6. Alkalinise in borax/alcohol, 30 minutes. 7. Wash in water. 8. Counterstain in Van Giesons. 9. Blot dry, dehydrate, clear and mount. RESULTS: Amyloid blue-green Collagen red Muscle, cytoplasm yellow Mast cell granules and some colloids are also blue-green. REFERENCE: Lendrum, A.C., Slidders, W., Fraser, D.S., (1972). "Renal Hyalin: A Study of Amyloidosis and Diabetic Fibrinous Vasculosis with new Staining Methods". J. Clin. Path. 25, 373-396. This is from an old manual. Sorry that the Hazardous warning phrases are missing. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: rfail@toolkitmail.com [mailto:rfail@toolkitmail.com] Sent: Friday, 19 September 2003 6:13 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] SAB for amyloid Does anyone know of an amyloid stain with the abbreviation SAB? Rena Fail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From lpwenk <@t> mail.netquest.com Thu Sep 18 18:42:22 2003 From: lpwenk <@t> mail.netquest.com (Lee & Peggy Wenk) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] msds References: Message-ID: <00f501c37e3e$826783c0$8732fea9@hppav> I like SIRI = Safety Information Resources Inc. http://www.hazard.com/index.php In the middle of the page, click on SIRI MSDS Collection On the new page, type in the name of the chemical you want to look up in the first rectangle area. Also, click on "whole word", rather than "partial word". The difference is - if the chemical you want is aluminum ammonium sulfate - "whole word" gives you all the MSDS for aluminum ammonium sulfate. "Partial word" gives you all MSDS that have the word aluminum or the word ammonium or the word sulfate. I like this site, as you can then click on the company you need - Fisher, Sigma, Biochemical Sciences, Mallinckrodt , to name a few. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "P. Emry" To: Sent: Thursday, September 18, 2003 3:02 PM Subject: [Histonet] msds > I have not been able to print msds information from several sights. Is > there a sight where this is easier? > Thanks, > > Trisha > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From i_stain <@t> yahoo.com Thu Sep 18 18:35:41 2003 From: i_stain <@t> yahoo.com (Scott Mordue) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Automated IHC machine Message-ID: <20030918233541.19417.qmail@web42004.mail.yahoo.com> There is ST5050 stainer from Zymed. It's an open system for 20 slides. I think it is good for your purpose-pathology research. Processing temp from RT to 45C. You can call them to learn more 800-874-4494. Scott CSU We are looking to purchase a new automated stainer for our pathology research lab and would like to hear some recommendations from the group. Our needs include the ability to be highly adaptable and variable, almost on a daily basis (this is purely for research.) We do not necessarily need extremely high throughput (20 slides per run would be fine.) The ability to perform ISH and antigen retrieval would be nice as well. If anyone has a suggestion as to what unit to purchase, we would be grateful. Thanks! Richard "Trey" Bruggeman Milton S. Hershey Medical Center Penn State College of Medicine Department of Pathology, H179 500 University Drive Hershey, PA 17033 (717) 531-1044 __________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com From Laurie.Reilly <@t> jcu.edu.au Thu Sep 18 18:34:08 2003 From: Laurie.Reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Purpose of Oven Prior to Staining? In-Reply-To: <9D41AB7C56F8304F98537ABD87B249F6626129@dasmthgbz001.amedd. army.mil> Message-ID: <5.1.0.14.0.20030919092927.00a29e30@mail.jcu.edu.au> Dear Diane, Your statement that"Ideally, the slides should be dried at room temperature overnight especially if photographs are going to be made" fascinates me. Could you please explain the significance of room temperature drying for photomicrography. Thanks and regards, Laurie. At 02:08 PM 09/18/03 -0400, Gladney, Diane C Ms MACH wrote: >Jordan, > >The purpose of the slide dryer or oven is to dry the water off of the >slides. The slides should be well drained to remove as much water as >possible before placing them in the rack before it is placed in the oven. >It is not and will not deparaffinize the slides. The slides must be >completely free of water before they are stained either by hand or on an >automatic slide stainer. Any histological technique book will tell you that >the deparaffinization is accomplished by using xylene, xylene substitute or >other appropriate paraffin solvent. Ideally, the slides should be dried at >room temperature overnight especially if photographs are going to be made. >But few labs have this much time. A drier that has a temperature in excess >of 60 degrees C will cause some artifacts. Our autostainer has a built-in >forced air dryer that the slides stay in for 10 - 15 minutes. If we have a >rack on the stainer and have another rack ready for the stainer, we place >this rack in a small forced air dryer set for 15 minutes then remove and >wait on the stainer to finish before adding another rack. The slide dryer >then is by-passed and the dried slides go directly to the first xylene >substitute (what we use instead of xylene) for deparaffinization. If they >still won't believe you, have them try to stain some "dried slides" and see >the results. > >Good Luck, >Diane > >Diane C. Gladney, HT (ASCP) >Histology /Cytology Supervisor >Moncrief Army Community Hospital >P.O. BOX 484 >4500 Stuart Ave. >FT. Jackson, SC 29207 > >(803) 751-2530 >DSN 734-2530 > >EMAIL: diane.gladney@se.amedd.army.mil OR > dcgx1@aol.com > > >-----Original Message----- >From: Jordan Brod [mailto:jbrod@tvmdl.tamu.edu] >Sent: Thursday, September 18, 2003 10:41 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Purpose of Oven Prior to Staining? > > >Good Morning All: > >My lab is in a huge discussion about the reason why we put our slides in an >oven before staining. I told them that the reason was to dry all of the >water off of them, because xylene and water do not mix. This statement is >also backed up in literature that I have read. They still do not believe >me, because they think the purpose of the oven is to deparaffinize the >slides. > >Can anyone help me out, so I can put this discussion to rest? I will print >off all of your comments for my technicians to read. Is what I told them >correct or not? > >Thanks. > > > > >******************************************************************** >Jordan W. Brod >Diagnostic Lab Supervisor - Pathology >Texas Veterinary Medical Diagnostic Laboratory (TVMDL) >1 Sippel Road >TAMU 4471 >College Station, Texas 77843 >(979) 845-3414 >(979) 845-1794 fax >jbrod@tvmdl.tamu.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 Physiology & Pharmacology Fax 07 4779 1526 Aust.Inst.of Tropical Vet.& Animal Sc. James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From lpwenk <@t> mail.netquest.com Thu Sep 18 18:48:19 2003 From: lpwenk <@t> mail.netquest.com (Lee & Peggy Wenk) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Re: Carcinogenic Materials References: Message-ID: <010601c37e3f$56bef360$8732fea9@hppav> NSH has a packet of histology laboratory safety information, free to NSH members. There is a charge for non-NSH members. http://www.nsh.org/education/materials.html About 1/2 way down the page is the heading HEALTH and SAFETY RESOURCE MATERIAL. At the bottom of the page is a link to the order form. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Histology Royal Oak, MI ----- Original Message ----- From: JENNIFER SCHUMACHER To: laurie.colbert@huntingtonhospital.com ; histonet@lists.utsouthwestern.edu Sent: Wednesday, September 17, 2003 6:24 PM Subject: [Histonet] Re: Carcinogenic Materials Are these items that could be sent to the list as a resource for everyone? Sounds like a handy thing to have on hand. Thanks. Jen >>> "Laurie Colbert" 09/17/03 05:10PM >>> Mary North from Oregon forwarded me some information from an NSH publication, "Health and Safety Committee Information Packet." This was exactly what I was looking for. It was very informative and even listed carcinogens in the Histology Lab. Thanks again, Mary! I also have the hazourdous materials manual from Dick and Janet Dapson that Vinnie suggested. I would recommend this manual for every Histology lab. Laurie Colbert Huntington Memorial Hospital _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030918/4e5b7772/attachment.htm From lpwenk <@t> mail.netquest.com Thu Sep 18 19:04:33 2003 From: lpwenk <@t> mail.netquest.com (Lee & Peggy Wenk) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] HT Exam References: <5.1.0.14.2.20030918154606.00b646e8@ucmail3.uc.edu> Message-ID: <011801c37e41$9b94cc60$8732fea9@hppav> The NSH webpage on Education has a list of recommended study guides/textbooks. I believe it is the same list as that given out by ASCP to candidates who have signed up to take the HT or HTL exams. http://www.nsh.org/education/studyaids.html I would also like to suggest the HT/HTL Study Guide and Workbook put together by histotechs in the Michigan Society of Histotechnologists, including program directors of histotechnology programs and speakers on various topics at NSH and state symposiums. It is a comprehensive outline of what is on the exam, along with areas that the person has to fill in for themselves, such as definitions. Also, there are worksheets to copy, to be used as templates for each of the fixatives, special stains, decalcifiers, etc., in which the person again has to look up the information in order to complete the worksheets. It is a way to organize your studying, and make you look up the needed information. It was designed for those people having to study on their own. It is $18 (US funds)(check made out to "MSH", no charge cards or POs). Mail to: Michigan Society of Histotechnologists, c/o Dick Dapson, 1020 Harts Lake Road, Battle Creek, MI 49015 (Disclaimer: Yes, I am a MSH member. No, I don't get any money out of this.) Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Rita Angel" To: Sent: Thursday, September 18, 2003 3:50 PM Subject: [Histonet] HT Exam > Hi everyone, > > I have a co-worker that has is interested in taking the HT Exam. She has > filled out the paperwork and has mailed it, but is wondering what advice > everyone has in which resources to study. There seems to be a low success > rate in passing the exam, about 56%, according to recent emails I've received. > > I took the exam 10 years ago, so I'm not sure my advice would be the best > for her now. > > Thanks for your help, > > Rita Angel, HT (ASCP) > University of Cincinnati > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kacw <@t> citlink.net Thu Sep 18 19:52:04 2003 From: kacw <@t> citlink.net (Kristen&Chuck) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] (no subject) Message-ID: Hi! I have recently switched my e-mail address. How do I get this one of the subscriber list? Thanks Kristen From kemlo <@t> tiscali.co.uk Fri Sep 19 01:51:18 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:21:54 2005 Subject: [Histonet] Purpose of Oven Prior to Staining? In-Reply-To: <1CF2E2E5BB36D5119E7A0008C791F3740800E03E@simba.kids> Message-ID: <001e01c37e7a$72d530b0$0b062850@KEMLOS> 5. To create artifacts. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts:? ???????????? kemlo@tiscali.co.uk?or kemlo1@btinternet.com? Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. ? ? ? -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Tony Henwood Sent: 19 September 2003 00:00 To: histonet@lists.utsouthwestern.edu Cc: 'Jordan Brod' Subject: RE: [Histonet] Purpose of Oven Prior to Staining? Jordan, Reasons for drying slides above the melting point of wax: 1. Remove excess water 2. Allow sections to attach to slides (otherwise they will lift off in the dewaxing xylene). 3. Aid in final spreading out of sections so that there are fewer folds (should I say NO folds!!) 4. Remove some of the excess wax (ie it flows off the slide). Remember this requires regular cleaning of the oven to remove excess wax. 5. Can't think of any other reasons Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Jordan Brod [mailto:jbrod@tvmdl.tamu.edu] Sent: Friday, 19 September 2003 0:41 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Purpose of Oven Prior to Staining? Good Morning All: My lab is in a huge discussion about the reason why we put our slides in an oven before staining. I told them that the reason was to dry all of the water off of them, because xylene and water do not mix. This statement is also backed up in literature that I have read. They still do not believe me, because they think the purpose of the oven is to deparaffinize the slides. Can anyone help me out, so I can put this discussion to rest? I will print off all of your comments for my technicians to read. Is what I told them correct or not? Thanks. ******************************************************************** Jordan W. Brod Diagnostic Lab Supervisor - Pathology Texas Veterinary Medical Diagnostic Laboratory (TVMDL) 1 Sippel Road TAMU 4471 College Station, Texas 77843 (979) 845-3414 (979) 845-1794 fax jbrod@tvmdl.tamu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo <@t> tiscali.co.uk Fri Sep 19 01:32:41 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Purpose of Oven Prior to Staining? In-Reply-To: Message-ID: <001701c37e77$d47dad40$0b062850@KEMLOS> How could the oven de-wax the section? The wax is in the tissue isn't it? Does it drain out? Nah, xylene removes the wax. The oven gets rid of the water, allows the section to fully 'uncrinkle' and brings the molecules of the tissue closer to that of the glass, thus causing intermolecular forces to keep the section on the slide (my hypothesis). I mean how else do sections adhere to glass if the slide is not treated? How do the sections stay on the glass? Any ideas? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts:? ???????????? kemlo@tiscali.co.uk?or kemlo1@btinternet.com? Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. ? ? ? -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Jordan Brod Sent: 18 September 2003 15:41 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Purpose of Oven Prior to Staining? Good Morning All: My lab is in a huge discussion about the reason why we put our slides in an oven before staining. I told them that the reason was to dry all of the water off of them, because xylene and water do not mix. This statement is also backed up in literature that I have read. They still do not believe me, because they think the purpose of the oven is to deparaffinize the slides. Can anyone help me out, so I can put this discussion to rest? I will print off all of your comments for my technicians to read. Is what I told them correct or not? Thanks. ******************************************************************** Jordan W. Brod Diagnostic Lab Supervisor - Pathology Texas Veterinary Medical Diagnostic Laboratory (TVMDL) 1 Sippel Road TAMU 4471 College Station, Texas 77843 (979) 845-3414 (979) 845-1794 fax jbrod@tvmdl.tamu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ree3 <@t> leicester.ac.uk Fri Sep 19 03:20:15 2003 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] RE: actinomyces antibody Message-ID: Anyone know of an antibody to actinomyces that would work on paraffin sections???? Many thanks Richard Edwards MRC TOX UNIT....LEICESTER...U.K.... From Myri37 <@t> aol.com Fri Sep 19 05:08:25 2003 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] (no subject) Message-ID: <5F95E580.4E4194A1.0005167B@aol.com> hi i use Masson trichrome stain since a long time for paraffin sections, and i don't know why now collagen doesn't stain in blue, i make my blue anilin with methyl blue of SIGMA in water and acetic acid, do you know any reason that makes collagen does'tn stain in blue ? thank you very much for any help Myriam From DDDeltour <@t> sig.med.navy.mil Fri Sep 19 05:34:58 2003 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] (no subject) Message-ID: Any reason why you are not using "Aniline" blue stain? If it is not available you can substitute Light green for the Aniline blue. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy LPO Histology Histology Technician DSN 624-4669 FAX 624-4680 COMM (From US) 011-39-095-56-4669 -----Original Message----- From: Myri37@aol.com [mailto:Myri37@aol.com] Sent: Friday, September 19, 2003 12:08 PM To: histonet@pathology.swmed.edu Subject: [Histonet] (no subject) hi i use Masson trichrome stain since a long time for paraffin sections, and i don't know why now collagen doesn't stain in blue, i make my blue anilin with methyl blue of SIGMA in water and acetic acid, do you know any reason that makes collagen does'tn stain in blue ? thank you very much for any help Myriam _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From g.lang <@t> bigfoot.de Fri Sep 19 05:39:17 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] (no subject) References: <5F95E580.4E4194A1.0005167B@aol.com> Message-ID: <00b401c37e9a$46d0d3a0$0d12a8c0@SERVER> Hi, Look at this histologic-site, usefull information about masson trichrom. http://206.137.77.9/sakura/pdf/9n10179.pdf Gundi Lang ----- Original Message ----- From: To: Sent: Friday, September 19, 2003 12:08 PM Subject: [Histonet] (no subject) > hi > i use Masson trichrome stain since a long time for paraffin sections, and i don't know why now collagen doesn't stain in blue, i make my blue anilin with methyl blue of SIGMA in water and acetic acid, do you know any reason that makes collagen does'tn stain in blue ? > thank you very much for any help > Myriam > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From IKirbis <@t> onko-i.si Fri Sep 19 05:33:42 2003 From: IKirbis <@t> onko-i.si (=?iso-8859-2?Q?Kirbi=B9_Srebotnik_Irena?=) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] research, routine workload Message-ID: Dear Histoneters, I'm interesting to know how in others pathology/cytology labs combine routine and research work. Is it expecting that technicians participate in research projects beside routine work during normal hours for no additional money?, or they can choose if they want participate and work an extra hours? Irena Kirbi? Institute of Oncology. Ljubljana, Slovenia -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030919/52b126f6/attachment.htm From IKirbis <@t> onko-i.si Fri Sep 19 05:34:55 2003 From: IKirbis <@t> onko-i.si (=?iso-8859-2?Q?Kirbi=B9_Srebotnik_Irena?=) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] FW: research, routine workload Message-ID: > Dear Histoneters, > > I'm interesting to know how in others pathology/cytology labs combine routine and research work. Is it expecting that technicians participate in research projects beside routine work during normal hours for no additional money?, or they can choose if they want participate and work an extra hours? > > Irena Kirbi? > Institute of Oncology. Ljubljana, Slovenia > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030919/c0263069/attachment.htm From Tora.Bardal <@t> bio.ntnu.no Fri Sep 19 06:23:59 2003 From: Tora.Bardal <@t> bio.ntnu.no (Tora Bardal) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] promoting the art of histology/microscopy Message-ID: <5.1.0.14.2.20030919132341.00a1dc60@pop.nt.ntnu.no> Hello Just wanted to share with you a succesful way to show the beauty of microscopy to the public. This spring I had an exhibition in the Natural Science Library at the Norwegian University of Science and Technology, the pictures are linked from my homepage: http://www.nt.ntnu.no/users/tbardal/ I even got 5 min on national TV, in a science program. And yes, I know the homepage is in norwegian, but the pictures are international............. Tora Bardal Chief Engineer Department of Biology, Norwegian University of Science and Technology (NTNU) Bratt?ra Research Center N-7491 Trondheim Norway tlf: + (47)73 59 09 38 E-mail: Tora.Bardal@bio.ntnu.no fax:+ (47)73 59 63 11 From juan.gutierrez <@t> christushealth.org Fri Sep 19 07:24:10 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Automated IHC machine Message-ID: What you are describing is the Ventana Benchmark. In my opinion the best platform in the market. No wait, the second best platform. The Benchmark XT is the best platform, but maybe more than you need. They also have a research platform, but I'm not familiar with it, I think is called the Discovery. www.ventanamed.com. 1-800-227-2155. Juan C. Gutierrez, HT(ASCP) not affiliated with Ventana. -----Original Message----- From: Richard Bruggeman [mailto:rbruggeman@psu.edu] Sent: Thu 9/18/2003 1:24 PM To: histonet@pathology.swmed.edu Cc: Subject: [Histonet] Automated IHC machine We are looking to purchase a new automated stainer for our pathology research lab and would like to hear some recommendations from the group. Our needs include the ability to be highly adaptable and variable, almost on a daily basis (this is purely for research.) We do not necessarily need extremely high throughput (20 slides per run would be fine.) The ability to perform ISH and antigen retrieval would be nice as well. If anyone has a suggestion as to what unit to purchase, we would be grateful. Thanks! Richard "Trey" Bruggeman Milton S. Hershey Medical Center Penn State College of Medicine Department of Pathology, H179 500 University Drive Hershey, PA 17033 (717) 531-1044 From DKUMISKI <@t> mail.mcg.edu Fri Sep 19 07:25:45 2003 From: DKUMISKI <@t> mail.mcg.edu (Donna Kumiski) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] how to embed/section mesentery? Message-ID: How do you mount the tissue on the slide? Donna ---------------------------------------------- Donna Kumiski HT (ASCP) Medical College of Georgia Dept. of Cellular Biology and Anatomy CB 1202 Augusta, Georgia 30912-2000 (706) 721-6278 dkumiski@mail.mcg.edu ----------------------------------------------- >>> Geoff McAuliffe 9/18/2003 10:55:23 AM >>> Hi Felix: For mesentaries some people like a "spread" rather than a section. Take out several loops of intestine and pin them out on a sheet of cork or dental wax with a tiny bit of tension on the mesentary so it is flat. Submerge in fix. You can process the whole "block" this way throught alcohols and clearing, then cut out and mount the sheet of mesentary on a slide without embedding. Yes it will be thicker than a section but you will see a nice 3D tissue with capillaries etc. Geoff Felix.Rintelen@serono.com wrote: >Dear all, > >Is there anybody who may give me advice how to make good sections of >parafin embedded mesentery (mouse)? How to prepare the tissue for embedding >and how to handle it to embed/orient it properly in the parafin block? My >concern is especially the soft consistency of the tissue making me feel >that the proper orientation in the block might be difficult. > >Thank you very much in advance, > >Felix > >Felix Rintelen >Serono Pharmaceutical Research Institute >Geneva, Switzerland > > > > >******************************************************************************************** >S - This message contains confidential information and is intended only for the individual >named. If you are not the named addressee, you should not disseminate, distribute or copy >this e-mail. Please notify the sender immediately by e-mail if you have received this >e-mail by mistake and delete this e-mail from your system. >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 malware. The >presence of this disclaimer is not a proof that it was originated at Serono International S.A. >or one of its affiliates. Serono International S.A and its affiliates therefore do not accept >liability for any errors or omissions in the content of this message, which arise as a result >of e-mail transmission. If verification is required, please request a hard-copy version. >Serono International SA, 15bis Chemin Des Mines, Geneva, Switzerland, www.serono.com. >********************************************************************************************* > > >_______________________________________________ >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 juan.gutierrez <@t> christushealth.org Fri Sep 19 07:38:28 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] The NSH needs your help NOW! Message-ID: Where is the survey? The only survey in there is the member satisfaction one, and is not short. Juan -----Original Message----- From: Vinnie Della Speranza [mailto:dellav@musc.edu] Sent: Thu 9/18/2003 12:53 PM To: Histonet@lists.utsouthwestern.edu Cc: lenaspencer@insightbb.com; peggy@nsh.org Subject: [Histonet] The NSH needs your help NOW! The National Society for Histotechnology is interested in learning how histology practitioners utilize microwave devices in their work. In this vein, we have created a brief, anonymous survey that will enable us to obtain data that reflects the standard of practice with microwave accelerated procedures. This survey will take approximately 5 minutes to complete. This information will assist us in the creation of microwave user guidelines as part of our collaboration with the National Committee for Clinical Laboratory Standards (NCCLS). We cannot meet our goal without your help. Your input is key to enabling us to better understand how microwave devices are used in the field. You can help us by going to the NSH homepage at http://www.nsh.org and clicking on the link at the top of the page to complete the survey. We would like to hear from everyone, even if you have never used a microwave device in your work. It is open to everyone who practices in our field, not just NSH members. Please take a moment of your time to provide this important information. We must compile this information before the NSH Symposium/Convention in October so time is running short. Please complete your survey TODAY !!!! Vinnie Della Speranza NSH Vice President _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Fri Sep 19 09:36:14 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] storing sections on slides Message-ID: <5.2.0.9.0.20030919093336.00a0ac90@mailhost.vetmed.auburn.edu> Hello, I think if you return those sections to the freezer you might encounter artifacts when staining later. I would just leave them at 4C. Best wishes. Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From gentras <@t> vetmed.auburn.edu Fri Sep 19 09:36:14 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] storing sections on slides Message-ID: <5.2.0.9.0.20030919093336.00a0ac90@mailhost.vetmed.auburn.edu> Hello, I think if you return those sections to the freezer you might encounter artifacts when staining later. I would just leave them at 4C. Best wishes. Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From gcallis <@t> montana.edu Fri Sep 19 09:40:11 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Update to Study Guides for HT/HTL exams found on NSH webpage In-Reply-To: <011801c37e41$9b94cc60$8732fea9@hppav> References: <5.1.0.14.2.20030918154606.00b646e8@ucmail3.uc.edu> Message-ID: <3.0.6.32.20030919084011.00b849e8@gemini.msu.montana.edu> Please note these updates to texts on NSH webpagelist http://www.nsh.org/education/studyaids.html. *IMMUNOHISTOPATHOLOGY: A PRACTICAL APPROACH TO DIAGNOSIS, J. M. Elias, 1990 (Out of Print) ASCP Press, 800-621-4142 has a new edition put out this year. I am not sure of title, but it covers more subjects than previous edition which has been out of print for years. *THEORY & PRACTICE OF HISTOLOGICAL TECHNIQUES, J. Bancroft & A. Stevens, 4th ed., 1996, (ISBN #0443-04760-X). Churchill-Livingstone, 800-553-5426 has a new FIFTH edition, same title, with M Gamble and J Bancroft as Editors, 2001. John Kiernan also has a new editon in paperback, titled Histological and Histotechnical Methods: Theory and Practice, ISBN# 0750649364, Oxford University Press, 800 451-7566 or www.oup-usa.org. I think you can get this one through Amazon.com. I strongly suggest anyone taking these exams get a histology atlas in order to identify/familiarize themselves on tissue/organ morphology. Wheaters Functional Histology, a text and colour atlas by B Young and JW Heath, Fourth Edition, 2001 is excellent, and comes with a CD, and is an inexpensive paperback. ISBN# 0443 05612 9 from Churchill Livingstone. The color photos plus written explanations are superior and simple to follow. Hopefully, NSH will update their webpage soon with the newer titles/texts available. 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 Sep 19 10:08:07 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Please use subject lines Message-ID: <3.0.6.32.20030919090807.00b849e8@gemini.msu.montana.edu> Dear subscribers, PLEASE!!!!!! Put something in your subject line for your Histonet message. If there is nothing in subject line, I delete the messages without ever opening the messages. 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 Sep 19 10:15:56 2003 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] promoting the art of histology/microscopy Message-ID: Excellent site (NB John K.) and spectacular images. A very well done to you Tora. 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: Tora Bardal [mailto:Tora.Bardal@bio.ntnu.no] Sent: 19 September 2003 12:24 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] promoting the art of histology/microscopy Hello Just wanted to share with you a succesful way to show the beauty of microscopy to the public. This spring I had an exhibition in the Natural Science Library at the Norwegian University of Science and Technology, the pictures are linked from my homepage: http://www.nt.ntnu.no/users/tbardal/ I even got 5 min on national TV, in a science program. And yes, I know the homepage is in norwegian, but the pictures are international............. Tora Bardal Chief Engineer Department of Biology, Norwegian University of Science and Technology (NTNU) Bratt?ra Research Center N-7491 Trondheim Norway tlf: + (47)73 59 09 38 E-mail: Tora.Bardal@bio.ntnu.no fax:+ (47)73 59 63 11 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Fri Sep 19 10:34:38 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] promoting the art of histology/microscopy Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CB96@usca0082k03.rallansci.apogent.com> Fantastic - Beautiful! do I take it right that some of these images are for sale? Tim Morken -----Original Message----- From: Tora Bardal [mailto:Tora.Bardal@bio.ntnu.no] Sent: Friday, September 19, 2003 4:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] promoting the art of histology/microscopy Hello Just wanted to share with you a succesful way to show the beauty of microscopy to the public. This spring I had an exhibition in the Natural Science Library at the Norwegian University of Science and Technology, the pictures are linked from my homepage: http://www.nt.ntnu.no/users/tbardal/ I even got 5 min on national TV, in a science program. And yes, I know the homepage is in norwegian, but the pictures are international............. Tora Bardal Chief Engineer Department of Biology, Norwegian University of Science and Technology (NTNU) Bratt?ra Research Center N-7491 Trondheim Norway tlf: + (47)73 59 09 38 E-mail: Tora.Bardal@bio.ntnu.no fax:+ (47)73 59 63 11 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ander093 <@t> gold.tc.umn.edu Fri Sep 19 11:02:21 2003 From: ander093 <@t> gold.tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Microtome knife sharpener in Minneapolis area? In-Reply-To: Message-ID: <5.2.0.9.0.20030919110020.00a330e0@ander093.email.umn.edu> Naoko, Where on campus are you located? LuAnn Anderson Neuropathology Lab University of Minnesota 612-625-1120 At 05:10 PM 9/18/03 -0500, Naoko Koyano wrote: >Hello everyone. > >I recently moved to Minneapolis and am looking for a knife sharpener that I >could come and use in the Twin cities area, preferably in U of M. >Does anyone have a sharpener around here? > >If you are not in the Twin Cities but know a good place to send knives to >get sharpened, I would appreciate that information, too. > >Thank you very much. >Naoko Koyano >-- >****************************** >Naoko Koyano-Nakagawa >E-mail: koyano@umn.edu >Office: 612-625-7687 >Lab: 612-624-3131 >Department Fax: 612-626-5009 >****************************** > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Erica.Schaffer <@t> aventis.com Fri Sep 19 11:08:13 2003 From: Erica.Schaffer <@t> aventis.com (Erica.Schaffer@aventis.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Performing ISH with CryoJane Message-ID: Hello Everyone, I need to be able to cut 3 micron frozen sections that will be used in ISH and IHC. I researched the internet and continue to see the CryoJane tape transfer system. My question is Does anyone have experience doing ISH on samples that were cut using the CryoJane? If so, what were the results? Was there significant background staining? Any information or advice would help. Thank you, Erica Erica Schaffer From DRG <@t> Stowers-Institute.org Fri Sep 19 11:12:04 2003 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Alizarin red/Alcian blue double skeletal stain Message-ID: Does anyone have a protocol for Alizarin Red/Alcian Blue on chick embryo? I have several protocols for mouse. I have tried to adjust times in solution, but I have some problems with the cartilage staining in some of the chick embryos. Thank you in advance. Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org From Rcartun <@t> harthosp.org Fri Sep 19 11:21:03 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] IHC Meeting Message-ID: The Society for Applied Immunohistochemisty will be holding its annual meeting on Saturday, October 4th at New York Hospital in Queens. Topics include Neuropathology, Veterinary Oncologic Pathology, and Emerging Infectious Diseases. Meeting and membership information is available on our website "appliedimmuno.org". Please contact me if you require further information. Richard Cartun Director, Immunopathology Hartford Hospital Hartford, CT From jkiernan <@t> uwo.ca Fri Sep 19 11:44:54 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Alizarin red/Alcian blue double skeletal stain References: Message-ID: <3F6B3285.484B9A9F@uwo.ca> There's this method: Webb GN & Byrd RA 1994. simultaneous differential staining of cartilage and bone in rodent fetuses: an alcian blue and alizarin red S procedure without glacial acetic acid. Biotechnic & Histochemistry 69(4):181-185. The colour photo looks excellent. The instructions take up 2 printed pages so more is needed than a simple "protocol." The journal is available in all good medical libraries. The method could probably be simplified for chick embryos because they are smaller than rat fetuses. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ _______________________________________ "Grant, Debra" wrote: > > Does anyone have a protocol for Alizarin Red/Alcian Blue on chick embryo? I have several protocols for mouse. I have tried to adjust times in solution, but I have some problems with the cartilage staining in some of the chick embryos. > Thank you in advance. > > Debby Grant > Research Technician II > Histology Core Facility > Stowers Institute for Medical Research > 1000 E. 50th Street > Kansas City, MO 64110 > drg@stowers-institute.org From Jacqueline.Miller <@t> UTSouthwestern.edu Fri Sep 19 12:11:49 2003 From: Jacqueline.Miller <@t> UTSouthwestern.edu (Jacqueline Miller) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] background problem Message-ID: The following is information about a high background problem when using RatIgG1 control antibodies on sections of human placental tissue: The primary Ab we are using is a rat anti-human. We dilute the primary to get 4 ug/ml. We dilute the RatIgG1 control Ab to get the same working concentration. We were diluting all the antibodies in 0.01M PBS (pH 7.2-7.4), but switched to a 1%BSA in 0.01M PBS solution in an attempt to correct the background problem. The primary is on for 60' RT. We have had the background problem with two different Rat IgG1 controls from two different companies. The one we are using now is much worse. The secondary is a biotinylated donkey anti-rat IgG F(ab')2 without cross reactivity, and is on for 30'RT. However, we have done several test runs substituting PBS for the primary, and there is no staining. We also used a MsIgG1 control antibody on the same type of tissue with a Vectastain MsIgG1 ABC kit (serum, secondary, and ABC), and had no staining. We wash the slides in PBS after every step except for the serum block (before the primary). We've tried increasing the time of the endogenous block (after the secondary) from 5 min. to 30 min. and decreasing the time of the ABC (avidin and biotinylated horseradish peroxidase complex) from 45' to 30'. We stained the paraffin sections using the citrate antigen retrieval method, once with a trypsin treatment, and once with no treatment at all. In all the treatments/no treatment, all the slides with the RatIgG1 control stained dark red, and all the slides with only PBS for the primary did not stain at all. Just a couple of days ago, we stained some frozen sections of the same type of tissue, as well as some sections of an older tissue, and once again, the sections with the RatIgG1 stained and the ones with only PBS did not stain. Any advice or suggestions from anyone on how to solve this background problem would be greatly appreciated. Thanks in advance, Jacqueline Miller Research Asst I Department of Obstetrics and Gynecology UT Southwestern Medical Center at Dallas >>> Gayle Callis 09/15/03 04:20PM >>> Did you do a dilution panel at beginning to determine that 4 ug/ml is the correct optimal working concentration? 4 ug/ml could be too high a concentration, depending on what you are staining for, and some kit secondaries may also be too concentrated. You did not say exactly how you were doing the staining and what for?? A kit, Strepavidin, etc, etc. specific secondary, ??? F(ab')2 secondary??? A few more specifics help as there are too many sources of background lurking to shoot us down! 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 joycejudge259 <@t> hotmail.com Fri Sep 19 12:11:55 2003 From: joycejudge259 <@t> hotmail.com (joyce judge) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] job opening Message-ID: Enclosed is the job description for a histology position we currently have at Ardais Corp. Joyce Judge Ardais Corp Lexington, MA Histotechnologist At ARDAIS, we have embarked upon a very special mission. We want to improve healthcare by accelerating the development of safe and effective new therapies. We look forward eagerly to the day when there is a “right” therapeutic solution for every patient, and when the most intractable diseases are addressed successfully. We believe that our work is critically important to that goal, and we invite you to join our team. We currently have an opportunity for a Histotechnologist. As a member of our Histology team, this individual will prepare sections of formalin-fixed, paraffin-embedded and flash frozen OCT embedded tissue samples. The selected candidate will be working with human tissues and other biological materials producing microscopic slides and related high quality products as well as performing automated immunohistochemistry (IHC) analysis. Additionally, he/she will contribute to the training of new members of the histology team on various histology processes. Maintaining high standards of quality as it relates to both process and finished product is critical to success in this role. We require an AS/BS in Biological Sciences or equivalent and 5+ years of related experience, preferably in biotechnology. Strong preference for candidates with experience in either IHC analysis or tissue microarray construction. In addition to excellent technical skills, we are looking for a team player that can work with a diverse group of scientific and clinical professionals in a growing and rapidly evolving environment. The ideal candidate will also possess excellent interpersonal and communication skills. Strong preference for candidates with HT/HTL (ASCP) certification and demonstrated experience gained in either biotechnology or a research environment. This is an exceptional opportunity for an individual who is interested in working in a fast paced setting with a group of talented professionals. Interested applicants please send resume, referencing this position title as well as salary requirements to: deb_shapiro@ardais.com or mail to Human Resources, Ardais Corporation, 128 Spring Street, Lexington, MA 02421 781-698-0100 Ardais is an equal opportunity employer. _________________________________________________________________ Need more e-mail storage? Get 10MB with Hotmail Extra Storage. http://join.msn.com/?PAGE=features/es From Jacqueline.Miller <@t> UTSouthwestern.edu Fri Sep 19 12:13:01 2003 From: Jacqueline.Miller <@t> UTSouthwestern.edu (Jacqueline Miller) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] background problem Message-ID: Nidal, Thanks for the advice. In the past, we've used a 0.5% hydrogen peroxide solution for 5 minutes. We tried increasing the time, but it didn't help. We block with 1.5% normal donkey serum for 20 minutes, and our secondary antibody is a donkey anti-rat. I haven't had a chance to talk to my boss yet about using Tween 20 in the washes. Jacqueline Miller Research Asst I Department of Obstetrics and Gynecology UTSouthwestern Medical Center at Dallas >>> NIDAL E MUVARAK 09/15/03 03:55PM >>> Jacqueline, The background could be due to several factors. If you're doing immunoperoxidase staining, make sure you quench any endogenous peroxidase activity well before adding the substrate-DAB (if applicable). Some people recommend 0.03% hydrogen peroxide for 5-10 min; others do it @ 0.1-1% for 5-10 min. Also, blocking helps reduce background. Blocking works best when the serum used for blocking is derived from the same species in which the secondary antibody is raised. Some people use 1% blocking serum for 10 min. Others use it @ 1-2% for one hour. Last, I add Tween 20 to my washing buffer at 0.3%. Not only does it reduce the background, but it helps break the surface tension of drops when adding a drop of solution to the tissue, which makes the drop spread evenly on the tissue. Hope this helps. Goodluck. Nidal E Muvarak Associate Research Specialist Vascular Tissue Biomechanics Laboratory Department of Biomedical Engineering University of Wisconsin-Madison 1550 Engineering Dr.; Rm. 2158 Madison, WI 53706-1609 Lab: (608) 265-8921; Office: (608) 265-4205; Home: (608) 256-7934; Cell: (608) 332-6068 http://vtb.bme.wisc.edu ----- Original Message ----- From: Jacqueline Miller Date: Monday, September 15, 2003 3:02 pm Subject: [Histonet] background problem > Hi, > > We're having trouble with high background when using Rat IgG1 > antibodies (4 ?g/ml) on paraffin-embedded sections of human > tissues, with or without retrieval procedures. Does anyone have > any advice? > > Thank you, > > Jacqueline Miller > Research Asst. I > OB/GYN Dept. > University of Texas Southwestern Medical Center > Dallas, TX > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From tpmorken <@t> labvision.com Fri Sep 19 12:32:34 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] background problem Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CB99@usca0082k03.rallansci.apogent.com> Jacqueline, for your non-specific background problem I suggest one of the following: 1) Adsorb the nonspecific antibody with liver acetone powder (any mammal, available from Sigma and others) to the diluted antibody. Add the powder (about one-tenth the volume of the antisera), mix well and let sit overnight. Then spin down to remove the powder, decant and use the antibody. It should adsorb non-specific antibodies. 2) Use a primary diluent that includes human sera to trap non-specific antibodies. 3) boost the percentage of animal sera in the diluent - up to 20 percent if necessary. Tim Morken Lab Vision / NeoMarkers www.labvision.com -----Original Message----- From: Jacqueline Miller [mailto:Jacqueline.Miller@UTSouthwestern.edu] Sent: Friday, September 19, 2003 10:12 AM To: gcallis@montana.edu Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] background problem The following is information about a high background problem when using RatIgG1 control antibodies on sections of human placental tissue: The primary Ab we are using is a rat anti-human. We dilute the primary to get 4 ug/ml. We dilute the RatIgG1 control Ab to get the same working concentration. We were diluting all the antibodies in 0.01M PBS (pH 7.2-7.4), but switched to a 1%BSA in 0.01M PBS solution in an attempt to correct the background problem. The primary is on for 60' RT. We have had the background problem with two different Rat IgG1 controls from two different companies. The one we are using now is much worse. The secondary is a biotinylated donkey anti-rat IgG F(ab')2 without cross reactivity, and is on for 30'RT. However, we have done several test runs substituting PBS for the primary, and there is no staining. We also used a MsIgG1 control antibody on the same type of tissue with a Vectastain MsIgG1 ABC kit (serum, secondary, and ABC), and had no staining. We wash the slides in PBS after every step except for the serum block (before the primary). We've tried increasing the time of the endogenous block (after the secondary) from 5 min. to 30 min. and decreasing the time of the ABC (avidin and biotinylated horseradish peroxidase complex) from 45' to 30'. We stained the paraffin sections using the citrate antigen retrieval method, once with a trypsin treatment, and once with no treatment at all. In all the treatments/no treatment, all the slides with the RatIgG1 control stained dark red, and all the slides with only PBS for the primary did not stain at all. Just a couple of days ago, we stained some frozen sections of the same type of tissue, as well as some sections of an older tissue, and once again, the sections with the RatIgG1 stained and the ones with only PBS did not stain. Any advice or suggestions from anyone on how to solve this background problem would be greatly appreciated. Thanks in advance, Jacqueline Miller Research Asst I Department of Obstetrics and Gynecology UT Southwestern Medical Center at Dallas >>> Gayle Callis 09/15/03 04:20PM >>> Did you do a dilution panel at beginning to determine that 4 ug/ml is the correct optimal working concentration? 4 ug/ml could be too high a concentration, depending on what you are staining for, and some kit secondaries may also be too concentrated. You did not say exactly how you were doing the staining and what for?? A kit, Strepavidin, etc, etc. specific secondary, ??? F(ab')2 secondary??? A few more specifics help as there are too many sources of background lurking to shoot us down! 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 tora.bardal <@t> bio.ntnu.no Fri Sep 19 13:51:01 2003 From: tora.bardal <@t> bio.ntnu.no (Tora Bardal) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] promoting the art of histology/microscopy In-Reply-To: Message-ID: <5.2.0.9.2.20030919194217.00b3e968@pop.nt.ntnu.no> At 19.09.2003 07:51, you wrote: >Although I do not know Norweigan, the web page is beautiful!! I have >often thought I could hand a microscopic image on my walls for art. It >does not look like too many words, would you mind translating into >English? Just what the photos are depicting (organ and stain >type/microscopic method)? Dear Jennifer English is not at all my first language, I'll do my best :-) As you may have understood, the computer is an important tool "on the way" to the final picture. I use Adobe Photoshop, playing with colours and effects. I'll try to explain , the page http://chembio.ntnu.no/users/tbardal/mikroskopibilder/index.htm shows 35 pictures, from left to right 1) a gut feeling, semithin section of cod gut embedded in LR White, Prot.A-gold and Silver Enhancement to show the lipase labeling and Toluidin Blue as counterstain LM (ordinary light microscope) 2) algeoppblomstring - algal bloom LM 3) chive (from my garden), digital camera 4) Artemia, a small Crustacea, LM 5) also Artemia, a little older. These Crustaceans are used as marin fish feed.LM 6) otifer, also used as fish feed, LM 7) playing around with non miscible fluids, LM with DIC 8) Eye from cod larvae, semithin EPON-section, Toluidin Blue 9) midgut from turbot larvae with a lot of lipid droplets, semithin EPON-section, Toluidin Blue 10) Just an illustration of the importance of narrowing the search when looking for fish, not FISH 11) gill development, turbot. Stereomicroscope 12) Halibut larvae, newly hatched and 140 days old, Stereomicroscope 13) Scanning electrone micrograph, halibut gut 14) Snow melting in the light beam, LM with DIC 15) shell (scallop??) digital camera 16) Halibut eggs, Stereomicroscope 17) Heart muscle from Atlantic salmon, immunfluorescense ( musclespesific protein). Fluorescence microscope/paraffin 18) Halibut tail fin, Stereomicroscope 19) Golgi from cod enterocyte, EPON, TEM 20) Grain dissolving, LM with DIC 21) Bacteria in the gut of a turbot larvae, EPON, TEM 22) another rotifer, LM with DIC 23) Japanese Eel, 5 days old LM 24) Vacuole in a liver section, EPON, TEM 25) cod "trying to eat" NTNU's logo 26) moth - digital camera 27) tail, lobster 9 days, Stereomicroscope 28-30 pigment celles in cod skin, Stereomicroscope 31) cross section of 1 day old turbot larvae, just behind the eye region, semithin EPON-section, Toluidin Blue 32) Grain dissolving, LM with DIC 33) no. 32 and some "roses" from cod skin 34) vertebra from cod, originally stained with Alcian Blue and Alizarin Red, LM with DIC 35) muscle gene expression in halibut embryo, wholemount in situ hybridization, Stereomicroscope and from left to right in page http://chembio.ntnu.no/users/tbardal/nye2003/index.htm 1) turbot larvae, 25 days after hatching, originally stained with Alcian Blue and Alizarin Red, LM 2) also turbot larvae, 25 days after hatching, originally stained with Alcian Blue and Alizarin Red, LM 3) Japanese Eel, from day 0 to day 8, LM 4) tail fin, turbot larvae, 25 days after hatching, originally stained with Alcian Blue and Alizarin Red, LM From settembr <@t> umdnj.edu Fri Sep 19 14:05:59 2003 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Validation of ISH Message-ID: Hello, I would like to ask how laboratories have gone about validating their In Situ Hybridization protocols to stay within the guidelines of CAP and pass their inspections. Please respond. Thank you. Dana Settembre University Hospital - UMDNJ Newark, NJ USA From mcauliff <@t> umdnj.edu Fri Sep 19 14:36:12 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] how to embed/section mesentery? In-Reply-To: References: Message-ID: <3F6B5AAC.2050604@umdnj.edu> Assuming that the tissue is flat enough (from tension applied during fixation and processing) you can put cleared tissue on a slide and coverslip it. Of course, it was stained (which I forgot to mention, blush!) after fixation and before dehydration and clearing. We use such spreads to teach the histology of loose connective tissue to first year medical students. Probably not something a clinical lab would do. Geoff Donna Kumiski wrote: >How do you mount the tissue on the slide? > Donna > >---------------------------------------------- >Donna Kumiski HT (ASCP) >Medical College of Georgia >Dept. of Cellular Biology and Anatomy >CB 1202 >Augusta, Georgia 30912-2000 >(706) 721-6278 > dkumiski@mail.mcg.edu >----------------------------------------------- > > > > >>>>Geoff McAuliffe 9/18/2003 10:55:23 AM >>> >>>> >>>> >Hi Felix: > > For mesentaries some people like a "spread" rather than a section. >Take out several loops of intestine and pin them out on a sheet of cork >or dental wax with a tiny bit of tension on the mesentary so it is flat. >Submerge in fix. You can process the whole "block" this way throught >alcohols and clearing, then cut out and mount the sheet of mesentary on >a slide without embedding. Yes it will be thicker than a section but you >will see a nice 3D tissue with capillaries etc. > >Geoff > >Felix.Rintelen@serono.com wrote: > > > >>Dear all, >> >>Is there anybody who may give me advice how to make good sections of >>parafin embedded mesentery (mouse)? How to prepare the tissue for embedding >>and how to handle it to embed/orient it properly in the parafin block? My >>concern is especially the soft consistency of the tissue making me feel >>that the proper orientation in the block might be difficult. >> >>Thank you very much in advance, >> >>Felix >> >>Felix Rintelen >>Serono Pharmaceutical Research Institute >>Geneva, Switzerland >> >> >> >> >>******************************************************************************************** >>S - This message contains confidential information and is intended only for the individual >>named. If you are not the named addressee, you should not disseminate, distribute or copy >>this e-mail. Please notify the sender immediately by e-mail if you have received this >>e-mail by mistake and delete this e-mail from your system. >>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 malware. The >>presence of this disclaimer is not a proof that it was originated at Serono International S.A. >>or one of its affiliates. Serono International S.A and its affiliates therefore do not accept >>liability for any errors or omissions in the content of this message, which arise as a result >>of e-mail transmission. If verification is required, please request a hard-copy version. >>Serono International SA, 15bis Chemin Des Mines, Geneva, Switzerland, www.serono.com. >>********************************************************************************************* >> >> >>_______________________________________________ >>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 ********************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030919/06db97fe/attachment.htm From mcauliff <@t> umdnj.edu Fri Sep 19 14:46:14 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Opinion on storing sections on slides In-Reply-To: <082C721AF78DB34983E8BA2CD085462105C676@mailbe07> References: <082C721AF78DB34983E8BA2CD085462105C676@mailbe07> Message-ID: <3F6B5D06.40206@umdnj.edu> Hi Gareth: I would keep them in buffer in the refrigerator until I needed them. I have done immunos for tyrosine hydroylase and GFAP on sections stored at minus 80 C for several months. The GFAP was ok but the TH suffered from storage. Storage for just a few days at room temp ruined Mac1 for mouse microglia. Once sections dry out the immuno seems to go downhill. Geoff Davis, Gareth wrote: > > Hello All, > I would like to get some input on storing sections. I have cut frozen > (PFA perfused and post fixated) tissue on a sliding microtome, kept > the cut sections in PBS until mounting on slides. Then I applied the > sections to a SuperFrost Plus slide with water and dried down on a > slide warmer-overnight. So, the staining has turned out okay, but I > wonder if I should keep the sections I don't use in the freezer, like > typical frozen sections, or keep them at room temperature, like > paraffin sections (since they are dry and already fixed). I was going > to try to stain free floating sections, but it just didn't work. So, > any suggestions would be greatly appreciated. > Thanks, > Gareth > > Ms. Gareth B. Davis > Research Assistant II > Neuro-magnetics Division of > the Department of Neurology > Vanderbilt University Medical Center > 615-936-3318 > > -- -- ********************************************** 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 ********************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030919/72306292/attachment.htm From Cheryl.Clarke <@t> fraserhealth.ca Fri Sep 19 15:10:40 2003 From: Cheryl.Clarke <@t> fraserhealth.ca (Clarke, Cheryl) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] unsubscibe Message-ID: Please take me off the list From Ted_pella <@t> tedpella.com Fri Sep 19 15:25:23 2003 From: Ted_pella <@t> tedpella.com (Ted Pella) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] (no subject) Message-ID: <000901c37eec$27bbb320$5020fdcc@domain> Please take me off the list _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DOOLEEO <@t> shands.ufl.edu Fri Sep 19 15:27:40 2003 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] automated IHC and ISH machine Message-ID: For automated IHC (immunohistology) and ISH (insitu hybridization) we use Ventana Benchmarks. We really like them and insitu for EBER, HPV,and Kappa and Lambda are really nice and easy. The machine also can do antigen retrial. For research purposes you might consider Ventana's Discovery machine which is more adapted for research applications. Elaine Dooley HTL Shands Teaching Hospital 352-265-0111 ext 72117 From weipingren <@t> yahoo.com Fri Sep 19 15:31:06 2003 From: weipingren <@t> yahoo.com (weiping Ren) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] thanks Message-ID: <20030919203106.27070.qmail@web41602.mail.yahoo.com> Hi, Histonetter: Thanks for all who gave me good advice for VEGF and CD31 staining. I will try Cell Marque's antigen retrival protocol first. Weiping --------------------------------- Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030919/4e5288b2/attachment.htm From gcallis <@t> montana.edu Fri Sep 19 15:32:43 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] immunostaining of VEGF and CD31 in mouse decalcified bone tissue In-Reply-To: <20030918181311.61898.qmail@web41610.mail.yahoo.com> References: <3.0.6.32.20030918111340.00b83d60@gemini.msu.montana.edu> Message-ID: <3.0.6.32.20030919143243.00ba21b8@gemini.msu.montana.edu> Publication on bone using EDTA on fresh bone before snap freezing and fixation of a frozen section. Mori S et al. A new decalcifying technique for immunohistochenical studies of calcified tissue especially applicable to cell surface marker demonstration, J Histochem Cytochem 36:111-114, 1988. Abstract: We have developed a new decalcifying technique for use in light and electron microscopy studies utilizing immunohistochemical staining of calcified tissues. Specimens containing calcified tissues can be adequately decalcified at a pH of 7.1-7.4 and temperature of -5 degrees C, without freezing, by use of a solution containing EDTA, sodium hydroxide, and glycerol. In this study, Leu-2a, Leu-3a, Leu-4, Leu-7, Leu-12, Leu-14, Leu-M1, Leu-M2, Leu-M3, and HLA-DR-positive cells in destructive lesions of bone tissues from patients with rheumatoid arthritis and osteomyelitis were successfully detected immunohistochemically. Furthermore, the presence of HLA-DR antigen on the surface of the infiltrating cells in the same lesions could be demonstrated using the immunoelectron microscopic technique. The results reported here have not previously been obtainable using conventional decalcifying techniques. Be sure to search in this journal, there have been some excellent, more recent publications on decalcificatin of bone to do IHC on CD markers, etc. 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 Cheryl.Clarke <@t> fraserhealth.ca Fri Sep 19 17:12:14 2003 From: Cheryl.Clarke <@t> fraserhealth.ca (Clarke, Cheryl) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] RE:UNSUBSCRIBE Message-ID: Please remove my name from the mailing list From georgecole <@t> ev1.net Sat Sep 20 13:18:43 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] The DVD's in the muscle and nerve packets Message-ID: <000001c37fa3$a14f9550$0a4dbad0@hppav> MUSCLE AND NERVE PACKET RECIPIENTS: I have discovered that the 3 recordable DVD's sent to you are a bit softer than commercial audio and video DVD's. Handle them carefully. They seem to scratch easily. I hope you all find you can serve biopsy patients better with the new methods---I sure did. georgecole@ev1.net -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030920/ac4ea626/attachment.htm From pengbaowei <@t> 21cn.com Sat Sep 20 21:23:50 2003 From: pengbaowei <@t> 21cn.com (Baowei Peng) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] AEC substrate for IHC Message-ID: hi, all histoneters, We can use ethanol to dehydrate and xylene to clear slides if we used DAB as substrate in IHC. Since AEC is organically soluble, we can not use these reagents. I am wondering what reagents we should use and should we dehydrate and clear AEC substrate slides? Baowei Peng Shanghai JiaoTong University, Shanghai, China,PRC ---------------------------------------------- Óµ±§ÀËÂþ£¬³¢ÊÔ¼¤Ç飬¼ÓÈë¿¡ÄÐÃÀÅ®µÄ¼¤Çé½»ÓÑÀÖÔ° http://y.21cn.com 21CNÐÂÎÅÖÐÐÄ£ºÐÂÎÅÌìÌì¶Á£¬Ç¿µµÖð¸öÊý http://news.21cn.com ¸öÈËÍøÕ¾Á÷Á¿±ä½ðÇ®£¬»¶Ó­¼ÓÈë21CNÓʼþÁªÃË£¡ http://mail.21cn.com/alliance/ ̨ÍåµÚÒ»¸»ÆÅÉí¼ÛÓâ°ÙÒÚ µÚÒ»¸»Í¯Éí¼ÛÁùÒÚ http://news.21cn.com/social/ From histo20 <@t> hotmail.com Sat Sep 20 23:07:25 2003 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Ergonomic chairs Message-ID: Can anyone recommend ergonomic chairs for Histology? Any information will be greatly appreciated. Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 _________________________________________________________________ Help protect your PC. Get a FREE computer virus scan online from McAfee. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From C.Gorrie <@t> unsw.edu.au Sun Sep 21 19:17:43 2003 From: C.Gorrie <@t> unsw.edu.au (Cathy Gorrie) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] AEC substrate for IHC Message-ID: >Dear Baowei, You will need some sort of water based mountant (available from all the IHC companies). Then you coverslip directly from water. The slides don't need to be dehydrated through alcohol or cleared in xylene. cheers, Cath --------------------------------------------------- Catherine Gorrie School of Medical Sciences University of New South Wales Sydney, N.S.W. 2052 Phone: 61-2-9385 2462 Fax : 61-2-9385 8016 e-mail: c.gorrie@unsw.edu.au >hi, all histoneters, > >We can use ethanol to dehydrate and xylene to clear slides if we >used DAB as substrate in IHC. Since AEC is organically soluble, we >can not use these reagents. >I am wondering what reagents we should use and should we dehydrate >and clear AEC substrate slides? > >Baowei Peng >Shanghai JiaoTong University, >Shanghai, >China,PRC > > >---------------------------------------------- >??????????"??????????????????????????????????? >http://y.21cn.com >21CN???????????????????????????????? >http://news.21cn.com >???????????????????????-????21CN?????????? >http://mail.21cn.com/alliance/ >?????????????????????? ???????????????? >http://news.21cn.com/social/ > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- From jkiernan <@t> uwo.ca Mon Sep 22 00:03:04 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] AEC substrate for IHC References: Message-ID: <3F6E8288.597C1A3A@uwo.ca> The answer is WATER. You must use an aqueous mounting medium to preserve the product of oxidation of AEC. You must not dehydrate or clear the preparations. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ______________________ Baowei Peng wrote: > > hi, all histoneters, > > We can use ethanol to dehydrate and xylene to clear slides if we used DAB as substrate in IHC. Since AEC is organically soluble, we can not use these reagents. > I am wondering what reagents we should use and should we dehydrate and clear AEC substrate slides? From L.Driessen <@t> orthop.umcn.nl Mon Sep 22 00:55:16 2003 From: L.Driessen <@t> orthop.umcn.nl (Driessen, L.) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Adhesives suitable for Antigen Retrieval Message-ID: <5B31B9F59E138A409E8F7C2183FF36F00408A0@umcnet13.umcn.nl> Hello, Is someone familiar with adhesives that can be used in combination with antigen retrieval techniques? Recently I've tried to perform this AR-technique on MMA-sections that where fixed to a glass-carrier using a gelatine-adhesive. Unfortunately the slices came loose. The AR-technique I used was heating the slides for 10 minutes at 100 ?C. Needs to say that I use a mixture of ethanol and butoxy-ethanol for stretching out the slices onto the glass-carrier, which might or might not influence the adhesive capacity of certain adhesives. Leon Driessen Orthopaedic Research Lab UMCN-St. Radboud, Nijmegen The Netherlands From Myri37 <@t> aol.com Mon Sep 22 02:33:31 2003 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] anilin blue Message-ID: <3BF82E25.7F7AC0E2.0005167B@aol.com> hello i am searching anilin blue, for masson trichrome stain, i've only found methyl blue at SIGMA,it's a synonym but it does'nt give the same result i tried it, but collagen is not stained blue, does anyone know where could i find the reel blue anilin powder ? thank you for anyhelp Myriam Natural implant France From nick.kirk3 <@t> btopenworld.com Mon Sep 22 03:18:43 2003 From: nick.kirk3 <@t> btopenworld.com (nick.kirk3@btopenworld.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] anilin blue Message-ID: <6146769.1064218723749.JavaMail.root@127.0.0.1> Myriam VWR supply Aniline blue, the catalogue number is 34003 4C. You can get it in 25g amounts. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England > from: Myri37@aol.com > date: Mon, 22 Sep 2003 08:33:31 > to: histonet@pathology.swmed.edu > subject: Re: [Histonet] anilin blue > > hello > i am searching anilin blue, for masson trichrome stain, i've only found methyl blue at SIGMA,it's a synonym but it does'nt give the same result i tried it, but collagen is not stained blue, does anyone know where could i find the reel blue anilin powder ? > thank you for anyhelp > Myriam > Natural implant > France > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pengbaowei <@t> 21cn.com Mon Sep 22 06:28:10 2003 From: pengbaowei <@t> 21cn.com (Baowei Peng) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] antibody for DC cell Message-ID: Hi all, Thanks for responses of my previous question. Does anyone can tell me which campany sell anti mouse CD11c antibody? ---------------------------------------------- Óµ±§ÀËÂþ£¬³¢ÊÔ¼¤Ç飬¼ÓÈë¿¡ÄÐÃÀÅ®µÄ¼¤Çé½»ÓÑÀÖÔ° http://y.21cn.com 21CNÐÂÎÅÖÐÐÄ£ºÐÂÎÅÌìÌì¶Á£¬Ç¿µµÖð¸öÊý http://news.21cn.com ¸öÈËÍøÕ¾Á÷Á¿±ä½ðÇ®£¬»¶Ó­¼ÓÈë21CNÓʼþÁªÃË£¡ http://mail.21cn.com/alliance/ ̨ÍåµÚÒ»¸»ÆÅÉí¼ÛÓâ°ÙÒÚ µÚÒ»¸»Í¯Éí¼ÛÁùÒÚ http://news.21cn.com/social/ From mparker <@t> epl-inc.com Mon Sep 22 07:05:23 2003 From: mparker <@t> epl-inc.com (Mary Parker) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Fw: Brain Tissue in Buffer Message-ID: <004d01c38101$cd87b800$6801a8c0@93F1H11> ----- Original Message ----- From: Mary Parker To: histonet@pathology.swmed.edu.com Sent: Monday, September 22, 2003 8:03 AM Subject: Brain Tissue in Buffer If anyone has any experience about how to successfully process and microtome serial sections of rat brain tissue that was perfused, then stored in buffer solution for approx one year in the refrigerator...please help! We were given this tissue by an outside investigator and are having a miserable time of it. I need all the information I can gather about tissue stored in buffer for an extended period of time. Is it hopeless? Thanks Mary Parker, H.T., A.S.C.P. Histology Manager EPL, Inc. P.O. Box 12766 RTP, N.C. 27709 (919) 998-9407 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030922/cc01179e/attachment.htm From nick.kirk3 <@t> btopenworld.com Mon Sep 22 07:21:39 2003 From: nick.kirk3 <@t> btopenworld.com (nick.kirk3@btopenworld.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] antibody for DC cell Message-ID: <7286657.1064233299866.JavaMail.root@127.0.0.1> Dako do a mouse anti-human one that works on frozen tissue. Cat No. M0732 Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England > from: Baowei Peng > date: Mon, 22 Sep 2003 12:28:10 > to: histonet@lists.utsouthwestern.edu > subject: Re: [Histonet] antibody for DC cell > > Hi all, > Thanks for responses of my previous question. > Does anyone can tell me which campany sell anti mouse CD11c antibody? > ---------------------------------------------- > ?????????????????????????????????????????????? > http://y.21cn.com > 21CN???????????????????????????????? > http://news.21cn.com > ????????????????????????????21CN?????????? > http://mail.21cn.com/alliance/ > ?????????????????????? ???????????????? > http://news.21cn.com/social/ > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfavara <@t> niaid.nih.gov Mon Sep 22 07:50:39 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Fw: Brain Tissue in Buffer Message-ID: Mary, I had some investigators that insisted on putting mouse brain in PBS @4C to insure fixation was for a uniform time. Sometimes these were in PBS[sterile] for months. When I got them ran them through the processor with and initial NBF station. I don't recall them being any different unless the fixation was inadequate. Good Luck! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Mary Parker [mailto:mparker@epl-inc.com] Sent: Monday, September 22, 2003 6:05 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fw: Brain Tissue in Buffer ----- Original Message ----- From: Mary Parker To: histonet@pathology.swmed.edu.com Sent: Monday, September 22, 2003 8:03 AM Subject: Brain Tissue in Buffer If anyone has any experience about how to successfully process and microtome serial sections of rat brain tissue that was perfused, then stored in buffer solution for approx one year in the refrigerator...please help! We were given this tissue by an outside investigator and are having a miserable time of it. I need all the information I can gather about tissue stored in buffer for an extended period of time. Is it hopeless? Thanks Mary Parker, H.T., A.S.C.P. Histology Manager EPL, Inc. P.O. Box 12766 RTP, N.C. 27709 (919) 998-9407 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030922/f48ff084/attachment.htm From mcauliff <@t> umdnj.edu Mon Sep 22 08:30:18 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Fw: Brain Tissue in Buffer In-Reply-To: <004d01c38101$cd87b800$6801a8c0@93F1H11> References: <004d01c38101$cd87b800$6801a8c0@93F1H11> Message-ID: <3F6EF96A.5090108@umdnj.edu> What exactly is the problem? If the tissue was properly fixed it should cut fine, no matter if storage was one week or one year (assuming no mold growth). Are you trying to embed in wax or cut frozen sections? Geoff Mary Parker wrote: > > ----- Original Message ----- > From: Mary Parker > To: histonet@pathology.swmed.edu.com > > Sent: Monday, September 22, 2003 8:03 AM > Subject: Brain Tissue in Buffer > > If anyone has any experience about how to successfully process and > microtome serial sections of rat brain tissue that was perfused, then > stored in buffer solution for approx one year in the > refrigerator...please help! We were given this tissue by an outside > investigator and are having a miserable time of it. > I need all the information I can gather about tissue stored in buffer > for an extended period of time. > Is it hopeless? > Thanks > > > Mary Parker, H.T., A.S.C.P. > Histology Manager > EPL, Inc. > P.O. Box 12766 > RTP, N.C. 27709 > (919) 998-9407 -- -- ********************************************** 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 ********************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030922/7f7846f1/attachment.htm From gcallis <@t> montana.edu Mon Sep 22 09:14:32 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] AEC substrate for IHC In-Reply-To: Message-ID: <3.0.6.32.20030922081432.00b7ac48@gemini.msu.montana.edu> You cannot use any solvent to dehydrate, mount your coverslip from rinse water with an aqueous mounting media - AquaMount from VWR, or any other vendors product. Some people like liquid coverslip called Crystal Mount placed directly over the section, allowed to dry, and after it is dry, mount a permanent coverslip with regular toluene or xylene based mounting media. Other companies have equivalents to all these mounting medias, you can shop around or others will make suggestions. At 10:23 AM 9/21/2003 +0800, you wrote: >hi, all histoneters, > >We can use ethanol to dehydrate and xylene to clear slides if we used DAB as substrate in IHC. Since AEC is organically soluble, we can not use these reagents. >I am wondering what reagents we should use and should we dehydrate and clear AEC substrate slides? > >Baowei Peng >Shanghai JiaoTong University, >Shanghai, >China,PRC > > >---------------------------------------------- >?????????????????????????????????????????????? >http://y.21cn.com >21CN???????????????????????????????? >http://news.21cn.com >????????????????????????????21CN?????????? >http://mail.21cn.com/alliance/ >?????????????????????? ???????????????? >http://news.21cn.com/social/ > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Sep 22 09:27:17 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Dendritic cell CD11c antibody In-Reply-To: Message-ID: <3.0.6.32.20030922082717.00b7ac48@gemini.msu.montana.edu> This is the N418 clone, either BD PHarmingen or Serotec. Try the latter first. 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 gentras <@t> vetmed.auburn.edu Mon Sep 22 09:40:11 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] AEC substrate in IHC Message-ID: <5.2.0.9.0.20030922093735.009e8d70@mailhost.vetmed.auburn.edu> Hello, there is no need to dehydrate and clear after washing off of the substrate you just cover slip in an aqueous mounting medium. Best wishes. Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From dennijc <@t> vetmed.auburn.edu Mon Sep 22 10:31:56 2003 From: dennijc <@t> vetmed.auburn.edu (John C. Dennis) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] AEC substrate for IHC In-Reply-To: Message-ID: Dear Baowei Peng Use an aqueous mountant or, depending on your substrate, dehydrate in ethanol and air dry. Then mount without using the clearing agent. John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Sun, 21 Sep 2003, Baowei Peng wrote: > hi, all histoneters, > > We can use ethanol to dehydrate and xylene to clear slides if we used DAB= as substrate in IHC. Since AEC is organically soluble, we can not use thes= e reagents. > I am wondering what reagents we should use and should we dehydrate and cl= ear AEC substrate slides? > > Baowei Peng > Shanghai JiaoTong University, > Shanghai, > China,PRC > > > ---------------------------------------------- > =D3=B5=B1=A7=C0=CB=C2=FE=A3=AC=B3=A2=CA=D4=BC=A4=C7=E9=A3=AC=BC=D3=C8=EB= =BF=A1=C4=D0=C3=C0=C5=AE=B5=C4=BC=A4=C7=E9=BD=BB=D3=D1=C0=D6=D4=B0 > http://y.21cn.com > 21CN=D0=C2=CE=C5=D6=D0=D0=C4=A3=BA=D0=C2=CE=C5=CC=EC=CC=EC=B6=C1=A3=AC=C7= =BF=B5=B5=D6=F0=B8=F6=CA=FD > http://news.21cn.com > =B8=F6=C8=CB=CD=F8=D5=BE=C1=F7=C1=BF=B1=E4=BD=F0=C7=AE=A3=AC=BB=B6=D3=AD= =BC=D3=C8=EB21CN=D3=CA=BC=FE=C1=AA=C3=CB=A3=A1 > http://mail.21cn.com/alliance/ > =CC=A8=CD=E5=B5=DA=D2=BB=B8=BB=C6=C5=C9=ED=BC=DB=D3=E2=B0=D9=D2=DA =B5=DA= =D2=BB=B8=BB=CD=AF=C9=ED=BC=DB=C1=F9=D2=DA > http://news.21cn.com/social/ > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From g.lang <@t> bigfoot.de Mon Sep 22 10:46:44 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] anilin blue References: <3BF82E25.7F7AC0E2.0005167B@aol.com> Message-ID: <001401c38120$c27aa520$0d12a8c0@SERVER> Fa. Chroma, 1B-501 Anilinblau wasserl?slich www.chroma.de greeting from Austria Gundi Lang general hospital Linz ----- Original Message ----- From: To: Sent: Monday, September 22, 2003 9:33 AM Subject: [Histonet] anilin blue > hello > i am searching anilin blue, for masson trichrome stain, i've only found methyl blue at SIGMA,it's a synonym but it does'nt give the same result i tried it, but collagen is not stained blue, does anyone know where could i find the reel blue anilin powder ? > thank you for anyhelp > Myriam > Natural implant > France > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From dr_pooja_chopra <@t> hotmail.com Mon Sep 22 12:34:56 2003 From: dr_pooja_chopra <@t> hotmail.com (pooja chopra) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] isopropanol as dehydrating agent in microwave processing Message-ID: Dear Histonetter, I am a new user of this email group. I am a resident doctor doing my thesis research on microwave fixation and processing in a moderately funded government laboratory in India. I am using a domestic microwave oven, as a lab oven is not affordable. I have been able to use only isopropanol (IPA) in the microwave processing schedule as it is very difficult to get license for ethanol in India. I have read lots of discouraging things that IPA should not be used as dehydrator, even in microwave method. I would like to hear from someone, regarding whether there is any way of making use of IPA both as dehydrator and clearing agent. Thanks, Pooja Chopra _________________________________________________________________ Yearning for ol' pals? For ol' school days? http://www.batchmates.com/msn.asp Here's a surprise! From muololi <@t> umdnj.edu Mon Sep 22 13:46:09 2003 From: muololi <@t> umdnj.edu (Lilia Muolo) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Keeping tissues on membrane slides Message-ID: Hi, does anyone have a good way to keep tissues from falling off membrane slides. When I deparaffinize in Xylene for 2 min. I find a lot of the tissue falls off, but if I keep it less time in Xylene, then it doesn't stain well in Toluidine Blue. The membrane slides are made by a lab researcher in another lab and given to me to use. I would appreciate any help the Histonet can provide. Lil Immuno. Lab Cancer Institute of New Jersey -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030922/dc6ee268/attachment.htm From georgecole <@t> ev1.net Mon Sep 22 16:16:42 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Whereabouts of Dr. kohjie Shima, neurologist Message-ID: <000001c3814e$d3ad7ba0$074dbad0@hppav> Dr, Shima and I studied muscles and nerves together 25 years ago at Good Samaritan Hospital.. I am trying to get in touch with Dr. Shima .I would appreciate hearing from anyone who knows how to contact him. georgecole@ev1.net -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030922/695d350c/attachment.htm From jkiernan <@t> uwo.ca Tue Sep 23 00:05:38 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] isopropanol as dehydrating agent in microwave processing References: Message-ID: <3F6FD4A2.2926A25F@uwo.ca> For almost all applications in practical histotechnology you can use isopropanol instead of ethanol. Both these alcohols mix similarly with water and with clearing agents such as xylene or chloroform. Neither ethanol nor isopropanol will mix with solid or melted paraffin wax. In some microwave processing techniques, specimens are moved from isopropanol into wax. The microwave heat is said to evaporate the isopropanol and allow infiltration with wax, without the need for an intermediate solvent. This method is recommended in Kok & Boon (1992) Microwave for Microscopists, 3rd ed. There was some heated Histonet discussion about all this in the late 1990s. Some of it may still be in the archives: www.histosearch.com For a thesis on microwave processing you will need a copy of Kok & Boon's book as a starting point, and you will need to follow up their references. If your library subscribes to Web of Science you will be able to find recent papers that cite earlier papers and books. Without access to Web of Science it is impossible to find out if someone else has recently done and published your intended research. Hope this helps, -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ________________________ pooja chopra wrote: > > Dear Histonetter, > I am a new user of this email group. I am a resident doctor doing my thesis > research on microwave fixation and processing in a moderately funded > government laboratory in India. I am using a domestic microwave oven, as a > lab oven is not affordable. > I have been able to use only isopropanol (IPA) in the microwave processing > schedule as it is very difficult to get license for ethanol in India. I have > read lots of discouraging things that IPA should not be used as dehydrator, > even in microwave method. I would like to hear from someone, regarding > whether there is any way of making use of IPA both as dehydrator and > clearing agent. > Thanks, > Pooja Chopra ____________________ From kemlo <@t> tiscali.co.uk Tue Sep 23 01:34:11 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Sullivan Nicclaides Pathology In-Reply-To: <3F688EEC.7070203@rmy.emory.edu> Message-ID: <000501c3819c$b6d85ce0$06cce150@KEMLOS> Anyone in that Organisation in the Brisbane area? Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts:? ???????????? kemlo@tiscali.co.uk?or kemlo1@btinternet.com? Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. ? ? ? -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Andrew Fedanov Sent: 17 September 2003 17:42 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Clot sections Hi Histonetters, Does anybody has an experience in preparation of clot sections? Thanks -- Andrew Fedanov Ph.D. Emory Vaccine Center at Yerkes 954 Gatewood Rd. NE Atlanta, GA 30329 Phone: 404-727-3043 Fax: 404-727-8199 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From damianmole <@t> ntlworld.com Tue Sep 23 02:56:09 2003 From: damianmole <@t> ntlworld.com (damianmole@ntlworld.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] ED2 Kupffer cell marker Message-ID: <20030923075609.LZVP13360.mta05-svc.ntlworld.com@[10.137.100.72]> Dear all, Can anyone help me? I am trying to visualise Kupffer cell subpopulations using ED1 and ED2. ED1 is working fantastically well but I am getting nothing with ED2. Both primaries are from Serotec Oxford. I have tried both unmasking and not unmasking by autoclave. I am using a DAKO strepABC visualisation kit with DAB substrate. I would be indebted to anyone who could offer me some advice. Damian Mole Queen?s University Belfast Northern Ireland ----------------------------------------- Email provided by http://www.ntlhome.com/ From mezey <@t> ana.sote.hu Tue Sep 23 03:53:02 2003 From: mezey <@t> ana.sote.hu (Mezey Szilvia) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Re: Brain Tissue in Buffer In-Reply-To: <20030922170001.13196.20737.Mailman@swlx167.swmed.edu> Message-ID: <3F70260D.14280.B8188@localhost> Have you tried embedding the tissue in gelatin - Agar-agar? Szilvi Mezey Szilvia Mezey PhD student Semmelweis University Dept. of Anatomy, Histology and Embryology Tuzolto u. 58. Budapest, 1094, Hungary T.: +36-12156920/3687 F.: +36-12155158 E-mail: mezey@ana.sote.hu From Nancy.Walker <@t> sanofi-synthelabo.com Tue Sep 23 06:36:53 2003 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Re: Brain Tissue in Buffer Message-ID: Hi, I've been told that if you fix in PFA then store in PBS for long periods the PBS "unfixes" the tissue. So I would suggest redoing your fixation process. Good luck! From dr_pooja_chopra <@t> hotmail.com Tue Sep 23 08:16:02 2003 From: dr_pooja_chopra <@t> hotmail.com (pooja chopra) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Reply- isopropanol as dehydrator in microwave method Message-ID: Dear Histonetters, Thanks for your helpful notes I am sending a copy of this email on the forum as it will be good histonet etiquette. Dear Diane, Thanks for the reply. The size of block is not measured, but I try to be as close to a maximum of 1.5X1.0X0.5. Actually with the 5 types of tissues I am including in my study, lymph node, bowel, gall bladder, prostatic chips (TURP), and ovary(mostly cysts) mostly my tissue size is much less in size, at least the minimum thickness is less than 5 mm in most cases which I believe is most important. Here is my protocol. FIXATION (stabilization) immersing the specimen in normal saline and irradiating in the microwave oven for 5 to 10 minutes(maximum 15 minutes) I check after 6 minutes and then at my discretion. Tissue pieces are kept in microwavable plastic cassettes submerged in sufficient amount of saline in 250 ml beaker and subjected to irradiation. PROCESSING: Isopropyl alcohol (60 %) for 8 minutes (4+4) Isopropyl alcohol (80%) for 6 minutes (2+4) Isopropyl alcohol (100%) for 5 minutes(1+4) (generally I open the cassette to check at least at this step) Isopropyl alcohol (100%) for 5 minutes (1+4) Molten Paraffin wax (bath I) for 5 minutes Molten paraffin wax (bath II) for 5 minutes Since my oven has only powers: cook and defrost and all the working is possible only on defrost mode, (cook is very high) I have to put water load after the initial few minutes of each stage.e.g. (4+4) in 1st step of dehydration. Please review the procedure at your discretion, are you using a domestic oven too? Please do let me know the email id of Cheryl Crowder, if possible I would like to contact her. Dear Jeff, I have tried to lower the power of the oven, by using in defrost mode and also by using water load. But I don’t think I can do much about the cycle duration, it is fixed in any given model. At times I get very good results but consistent good results have only been with prostatic chips. My lymph nodes are very unpredictable, once I had actually fixed fresh lymph node and processed, all in the microwave with excellent results. But since then all my lymph nodes have been jinxed. I would like to know what a PID controller is. Can it be used in domestic oven? Dear John A. Kiernan, Can you please let me know what is the Web of Science? Is it a journal or a website? I do have the book by Drs Boon and Kok, and am trying to find the toolbook by Dr Login, but I have a problem following up all the references I find, because some of the specialized journals (especially European) are hard to find in the Post Graduate Institute library over here in Chandigarh, India. Thanks a ton, Pooja Hi Pooja, It is actually better to use isopropanol as the final step between the alcohol and paraffin, since it has a lower boiling point, it evaporates out faster than ethanol, once you have transferred the tissue into the molten paraffin. I don't know what size your tissues are, but why don't you send me your protocol with tissue size and if you're having any problems I could review it. In some routine labs, with commercial tissue processors, they only use Isopropanol and not ethanol, since they can't afford it. Best Regards, Diane If you are having a problem with your outcomes, it is because of your microwave and not the use of IPA as a dehydrant or a "clearing agent". I have used it in a Milestone Lab Oven with great success. Your microwave is probably too long in duration of the power application at too high a level (wattage). This was our problem previously too. You need a very short duty cycle and a low wattage. A PID controller comes in very handy too. Jeff Jurczak Dear Pooja: I have used isopropyl alcohol for many years in processing and staining without problems. I've never done microwave processing so I can not comment on that aspect but I've heard that adequate fixation is absolutely necessary before doing microwave processing denise@pclv.net For almost all applications in practical histotechnology you can use isopropanol instead of ethanol. Both these alcohols mix similarly with water and with clearing agents such as xylene or chloroform. Neither ethanol nor isopropanol will mix with solid or melted paraffin wax. For a thesis on microwave processing you will need a copy of Kok & Boon's book as a starting point, and you will need to follow up their references. If your library subscribes to Web of Science you will be able to find recent papers that cite earlier papers and books. Without access to Web of Science it is impossible to find out if someone else has recently done and published your intended research. Hope this helps, -- ------------------------- John A. Kiernan _________________________________________________________________ The hottest things. The coolest deals. http://www.msn.co.in/Shopping/ Get them online! From cwscouten <@t> myneurolab.com Tue Sep 23 08:46:05 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] isopropanol as dehydrating agent in microwave processing Message-ID: Are you familiar with the Pelco Neurowave? It is a quite different product than the kind of microwave written about in Kok & Boon. A circulating coolant keeps the contents from getting warm, and absorbs microwaves before a standing wave develops. Sounds like it does nothing (a microwave oven that doesn't heat) but I have seen really sharp results out of it. When an IHC sample is split, and run through a protocol that is 10% as long as standard, but half in and half out of the microwave, the difference is dramatic. The microwave sample is better than conventional, the non microwave sample is not noticeably stained. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=472001&catdesc=Histology+Equipment&CatThreeID=714&CatOneID=4&subcatdesc=Microwave+Reaction+Facilitation&idsubcategory=204 Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com www.myneurolab.com -----Original Message----- From: John Kiernan [mailto:jkiernan@uwo.ca] Sent: Tuesday, September 23, 2003 12:06 AM To: pooja chopra Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] isopropanol as dehydrating agent in microwave processing For almost all applications in practical histotechnology you can use isopropanol instead of ethanol. Both these alcohols mix similarly with water and with clearing agents such as xylene or chloroform. Neither ethanol nor isopropanol will mix with solid or melted paraffin wax. In some microwave processing techniques, specimens are moved from isopropanol into wax. The microwave heat is said to evaporate the isopropanol and allow infiltration with wax, without the need for an intermediate solvent. This method is recommended in Kok & Boon (1992) Microwave for Microscopists, 3rd ed. There was some heated Histonet discussion about all this in the late 1990s. Some of it may still be in the archives: www.histosearch.com For a thesis on microwave processing you will need a copy of Kok & Boon's book as a starting point, and you will need to follow up their references. If your library subscribes to Web of Science you will be able to find recent papers that cite earlier papers and books. Without access to Web of Science it is impossible to find out if someone else has recently done and published your intended research. Hope this helps, -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ________________________ pooja chopra wrote: > > Dear Histonetter, > I am a new user of this email group. I am a resident doctor doing my thesis > research on microwave fixation and processing in a moderately funded > government laboratory in India. I am using a domestic microwave oven, as a > lab oven is not affordable. > I have been able to use only isopropanol (IPA) in the microwave processing > schedule as it is very difficult to get license for ethanol in India. I have > read lots of discouraging things that IPA should not be used as dehydrator, > even in microwave method. I would like to hear from someone, regarding > whether there is any way of making use of IPA both as dehydrator and > clearing agent. > Thanks, > Pooja Chopra ____________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From marirose.satterfield <@t> MercyMemorial.org Tue Sep 23 09:03:58 2003 From: marirose.satterfield <@t> MercyMemorial.org (Satterfield, Marirose) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] SLN bx Message-ID: We are looking into the process of sentinel lymph node biopsies. I have been told that the levels of radiation are nothing to be concerned about for handling of the biopsies. But when I have done some research on the Web for these biopsies it talks about a lead box for transport, rings and badges for monitoring and labeling of the specimen as radioactive. Could I please hear from the labs that are doing these and how they handle them. Thanks in advance Mari Satterfield Mercy Memorial Hospital From tpmorken <@t> labvision.com Tue Sep 23 10:07:57 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] isopropanol as dehydrating agent in microwave proc essing Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CBA3@usca0082k03.rallansci.apogent.com> Pooja, One concern I have about your experiments is that a domestic mw oven is not meant to be used with volatile, flammable chemicals. The lab versions are well vented (most must be vented to a fume hood, or have a vent system over the mw oven) and if intended for processing, are designed specifically to prevent vapor ignition and explosion (for example with non-sparking electronics and very strong door locks). Your home model has none of that and may be extremely dangerous to use with a flammable chemical. Tim Morken -----Original Message----- From: pooja chopra [mailto:dr_pooja_chopra@hotmail.com] Sent: Monday, September 22, 2003 10:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] isopropanol as dehydrating agent in microwave processing Dear Histonetter, I am a new user of this email group. I am a resident doctor doing my thesis research on microwave fixation and processing in a moderately funded government laboratory in India. I am using a domestic microwave oven, as a lab oven is not affordable. I have been able to use only isopropanol (IPA) in the microwave processing schedule as it is very difficult to get license for ethanol in India. I have read lots of discouraging things that IPA should not be used as dehydrator, even in microwave method. I would like to hear from someone, regarding whether there is any way of making use of IPA both as dehydrator and clearing agent. Thanks, Pooja Chopra _________________________________________________________________ Yearning for ol' pals? For ol' school days? http://www.batchmates.com/msn.asp Here's a surprise! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pengbaowei <@t> 21cn.com Tue Sep 23 10:10:34 2003 From: pengbaowei <@t> 21cn.com (Baowei Peng) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] H&E staining on GFP slides Message-ID: Hi all, I want to do H&E staining on GFP expressing slides. So I can view both the tructure of my slide and also GFP expressing. Is that possibile? If not, what should I do? Baowei Peng Shanghai JiaoTong University, Shanghai, China,PRC ---------------------------------------------- Óµ±§ÀËÂþ£¬³¢ÊÔ¼¤Ç飬¼ÓÈë¿¡ÄÐÃÀÅ®µÄ¼¤Çé½»ÓÑÀÖÔ° http://y.21cn.com 21CNÐÂÎÅÖÐÐÄ£ºÐÂÎÅÌìÌì¶Á£¬Ç¿µµÖð¸öÊý http://news.21cn.com ¸öÈËÍøÕ¾Á÷Á¿±ä½ðÇ®£¬»¶Ó­¼ÓÈë21CNÓʼþÁªÃË£¡ http://mail.21cn.com/alliance/ ̨ÍåµÚÒ»¸»ÆÅÉí¼ÛÓâ°ÙÒÚ µÚÒ»¸»Í¯Éí¼ÛÁùÒÚ http://news.21cn.com/social/ From dellav <@t> musc.edu Tue Sep 23 10:24:13 2003 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Microwave survey - Please Help Message-ID: Shortly after I posted our request for your participation in a brief survey about the use of Microwave devices in the workplace, the NSH website experienced a power outtage resulting from Hurricane Isabel. The NSH server is back up and running. We very much need your help. information you possess will enable us to collect data on how microwave devices are used by histology practitioners in their work. The response rate thus far has been low. Please consider donating FIVE minutes of your time to complete this survey. This survey is open to EVERYONE who practices in a histology lab, even if you do not currently use microwave devices at this time.. here is the original announcement I posted last week. The National Society for Histotechnology is interested in learning how histology practitioners utilize microwave devices in their work. In this vein, we have created a brief, anonymous survey that will enable us to obtain data that reflects the standard of practice with microwave accelerated procedures. This survey will take approximately 5 minutes to complete. This information will assist us in the creation of microwave user guidelines as part of our collaboration with the National Committee for Clinical Laboratory Standards (NCCLS). We cannot meet our goal without your help. Your input is key to enabling us to better understand how microwave devices are used in the field. You can help us by going to the NSH homepage at http://www.nsh.org and clicking on the link at the top of the page to complete the survey. We would like to hear from everyone, even if you have never used a microwave device in your work. It is open to everyone who practices in our field, not just NSH members. Please take a moment of your time to provide this important information. We must compile this information before the NSH Symposium/Convention in October so time is running short. Please complete your survey TODAY !!!! Vinnie Della Speranza NSH Vice President From jkiernan <@t> uwo.ca Tue Sep 23 10:34:37 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Literature searching (was Re: isopropanol etc) References: Message-ID: <3F70680C.1A8162A1@uwo.ca> Web of Science is the website version of the Science Citation Index. The address is: http://isi10.isiknowledge.com/portal.cgi/WOS but it will only work if your institution's library subscribes to Web of Science. If you know an important early paper that is relevant to your work, Web of Science will show you all the other papers that have cited the earlier one in their bibliographies. This allows you to search forwards, rather than searching backwards by looking up references from a recent paper or review article. Searching forward is also possible with the printed Science Citation Index, if this is still published. I have not used it since our library started subscribing to Web of Science 4 or 5 years ago. You can search much of the literature using PubMed (http://www.ncbi.nlm.nih.gov/PubMed/), which is free for everyone, a gift to the world from the US Government. (Not everything they do is bad.) Pubmed covers only the biomedical field and cannot be used for forward-looking citation searches, but it's still very useful. Web of Science covers the whole gamut. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ___________________________________ pooja chopra wrote: > Can you please let me know what is the Web of Science? Is it a journal or a website? From jkiernan <@t> uwo.ca Tue Sep 23 10:43:44 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Re: Reply- isopropanol as dehydrator in microwave method References: Message-ID: <3F706A30.EBBF63C2@uwo.ca> Someone wrote: > It is actually better to use isopropanol > as the final step between the alcohol > and paraffin, since it has a lower > boiling point, it evaporates out faster > than ethanol, ... Merck Index says the boiling points are: Ethanol 78.5 Isopropanol 82.5 There has to be another reason why you can get away without using a clearing agent! -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ From tpmorken <@t> labvision.com Tue Sep 23 10:42:36 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Continuing education on the web - free Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CBA4@usca0082k03.rallansci.apogent.com> Dear Histonetters, Lab Vision/NeoMarkers has always followed the philosophy that the needs of the customer come before any concern with products. Lab Vision started out as a resource for antibody information and was known among researchers as a place to go to find out how to get an antibody even if Lab Vision did not carry the product. In other cases we have offered technical help to laboratories even if they did not use our products. Continuing with that philosophy Lab Vision recently instituted a free tutorial program for histotechnologists that can be used for personal and in-house continuing education. Our feeling is that continuing education for histotechnologists is still a difficult proposition since so many do not have the opportunity to attend meetings and conventions. The internet offers a way to bring high-quality continuing education opportunities to every histotechnologist at their convenience. We have one tutorial posted now covering "The Role of the Histology Lab in Bioterrorism" (aimed at the US audience) which was first presented at the 2002 NSH meeting. Other tutorials will be added later as we develop them. These are different than some others because tests must be passed in order to move to the successive sections of the tutorial. After finishing the tutorial the user will receive a certificate confirming completion. We hope that these tutorials will help histotechnologists and laboratories with their requirements for continuing education. We are in discussions to become a provider of programs that the ASCP and state licensing boards will accept for their requirements of continuing education. To access the tutorial please go to the Lab Vision website (www.labvision.com ) and click on the navigation button "Tutorial Program." Each user must create a user account and give some information that will be used only to send a certificate at the finish of the program. No information given will be used for sales or marketing purposes. This program is intended solely for educational purposes. Please contact me with any questions. Tim Morken Product Development Lab Vision / NeoMarkers 47790 Westinghouse Dr Fremont, CA 94539 PH: 510-991-2840 www.labvision.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030923/62c26844/attachment.htm From lcornett <@t> highlandspath.com Tue Sep 23 10:56:07 2003 From: lcornett <@t> highlandspath.com (Cornett, Lorraine D) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] unsubscribe Message-ID: Unsubscribe -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030923/0a819b9d/attachment.htm From gcallis <@t> montana.edu Tue Sep 23 11:00:40 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] H&E staining on GFP slides In-Reply-To: Message-ID: <3.0.6.32.20030923100040.00b95c38@gemini.msu.montana.edu> WE quenched out GFP, although not eGFP with hematoxylin and Eosin will fluorescse. It is best to view GFP without trying to counterstain. Maybe some people have had success, but we did not. At 11:10 PM 9/23/2003 +0800, you wrote: > > >Hi all, > >I want to do H&E staining on GFP expressing slides. So I can view both the tructure of my slide and also GFP expressing. Is that possibile? If not, what should I do? > > >Baowei Peng >Shanghai JiaoTong University, >Shanghai, >China,PRC > >---------------------------------------------- >?????????????????????????????????????????????? >http://y.21cn.com >21CN???????????????????????????????? >http://news.21cn.com >????????????????????????????21CN?????????? >http://mail.21cn.com/alliance/ >?????????????????????? ???????????????? >http://news.21cn.com/social/ > > > > > > >_______________________________________________ >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 MGomez <@t> ameripath.com Tue Sep 23 11:48:56 2003 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Breast Tissue Processing Message-ID: Hello Histonetters: I'm looking for a good working protocol for Breast tissue, including fatty tissue, lymph nodes, etc.. Thank you very much, Milton From MGomez <@t> ameripath.com Tue Sep 23 11:50:05 2003 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Special stain control Message-ID: Hello: I would like to purchase special stain control. Do you know who sells them? Thank you, Milton From MGomez <@t> ameripath.com Tue Sep 23 11:51:08 2003 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Microtome Service Message-ID: Hello: Do you know who can provide maintenance and service to my microtomes? Thank you, Milton From DDittus787 <@t> aol.com Tue Sep 23 12:04:41 2003 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] SLN bx Message-ID: <176FC1A8.76B041A9.0A1F969F@aol.com> WE HAVE THEM HAND CARRIED TO US FROM THE O.R. ,THEY ARE LABELED WITH A BIOHAZARD STICKER AND RADIATION STICKER, WE HOLD THEM FOR 24 HRS(VERY SHORT HALF LIFE),WE PERIODICALLY HAVE OUR RADIATION OFFICER READ THEM AND MY PA WEARS A DOLCIMETER AND IS MONITORED MONTHLY. DANA From Jackie.O'Connor <@t> abbott.com Tue Sep 23 13:20:22 2003 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Microtome Service Message-ID: Check out Dorn and Hart Microedge in Villa Park, Illinois #630-832-3843. I once sent him (Ken Hart) a microtome from Honolulu which needed an overhaul. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics Jackie.O'Connor@abbott.com MGomez@ameripath.com Sent by: histonet-admin@lists.utsouthwestern.edu 09/23/2003 11:51 AM To: Histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Microtome Service Hello: Do you know who can provide maintenance and service to my microtomes? Thank you, Milton _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030923/6008fbb8/attachment.htm From siksik <@t> vgernet.net Tue Sep 23 13:57:07 2003 From: siksik <@t> vgernet.net (Steven Slap) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] isopropanol as dehydrating agent in microwave processing Message-ID: Hi HistoNetters Pooja Chopra asks about the use of isopropanol in place of 100% ethanol as a dehydrating agent in microwave processing. In my experience, isopropanol is more drying than ethanol. For this reason it is essential (as always) that the specimens be well-fixed prior to dehydration. However, given proper fixation, dehydration in a microwave can successfully be carried out using ethanol alone, isopropanol alone or some combination alocohol (like reagent alcohol, which, in addition to ethanol and isopropanol, also contains methanol) alone, preferably in two changes. That being said, Pooja Chopra mentions using a domestic microwave oven, and I do not recommend the use of any flammable solvents in such a device for safety reasons. best regards, Steven Slap ********************************************** Marketing Manager/Microwave Product Specialist Hacker Instruments & Industries, Inc. http://www.hackerinstruments.com ********************************************** -- From dellav <@t> musc.edu Tue Sep 23 15:51:04 2003 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] Microtome Service Message-ID: Help us to help you Milton. where are you located? >>> 09/23/03 12:51PM >>> Hello: Do you know who can provide maintenance and service to my microtomes? Thank you, Milton _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ckiely128 <@t> aol.com Tue Sep 23 15:48:36 2003 From: Ckiely128 <@t> aol.com (Ckiely128@aol.com) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] (no subject) Message-ID: <1e5.1056e98a.2ca20ba4@aol.com> Unsubscribe -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030923/147c9701/attachment.htm From bruyntjes <@t> voeding.tno.nl Wed Sep 24 03:50:38 2003 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] chromogranine Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079C5F@ntexch1.voeding.tno.nl> Hi all Is anyone familiar with a commercial available antibody against chromogranine that cross-reacts with FFPE rat tssues? Joost Bruijntjes TNO Nutrition and Food Research PO Box 360 3700 AJ Zeist The Netherlands From SBarnes <@t> elch.org Wed Sep 24 06:03:38 2003 From: SBarnes <@t> elch.org (Sue Barnes) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] HCV antibody for IHC Message-ID: <3F74CB2B769A5843A0C746D3F13256FD1279EE@ELCH2> Hi, I have been using HCV antibody from Biogenex and have just been told that it is no longer available and will not be replaced. I now need a new source for this antibody. Anyone doing HCV please send me your source. Thanks Sue Sue Barnes East Liverpool City Hospital East Liverpool, Ohio 43968 Ph 330 386-2968 fax: 330 386-2048 From stancelb <@t> msn.com Wed Sep 24 06:54:47 2003 From: stancelb <@t> msn.com (Barbara Stancel) Date: Fri Sep 16 15:21:55 2005 Subject: [Histonet] QC/Validation of Histo Chemicals Message-ID: To all histology laboratories inspected by any agency: Are you required to validate (do they work?) chemicals such as: 1% acetic acid, acid alcohol, dilutions of hydroquinone, Scott's Tap Water, 10% NBF, Bouin's, aqueous solutions of sodium thiosulfate, periodic acid, sodium borate, picric acid solutions, etc. (get my drift?)? Do your auditors consider the QC you do for stains and dyes as a satisfactory confirmation/validation that an individual solutions within a procedure work? I am not talking about QC of stains. In this lab that is a given...after preparation, all stains and dyes are QC'ed and the slide is labeled and kept as a permenent record. Auditors are wanting us to mark chemicals with a big ole "needs approval" label. After we prove they work, we apply a big ole "validate" label on all the bottles. We already have a labels on the bottles. Each contains the batch number, solution, stain, date, QC date and the person who performed the QC. The batch number is tracable on a special sheet on which we record even more information...like all the inventory numbers for the chemcials, stains or dyes used in the preparation of that solution, date prepared, date in use, date out of use, etc. Since all our auditors are chemist or microbiologist we are trying to make them see our point of view. This needs approval/validation method is used in chemistry and microbiology. Any suggestions? (please tell me which agency audits your lab) Histologically yours, Barbara (who feels like I am recording my life on spreadsheets with batch and inventory numbers!) Barbara H. Stancel, HTL(ASCP)HT USDA, FSIS, OPHS, Eastern Laboratory, Pathology RRC, 950 College Station Road Athens, Georgia 30604 phone: (706) 546-3556 fax: (706) 546-3589 _________________________________________________________________ Add MSN 8 Internet Software to your existing Internet access and enjoy patented spam protection and more. Sign up now! http://join.msn.com/?page=dept/byoa From bill501 <@t> mindspring.com Wed Sep 24 08:27:20 2003 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] QC/Validation of Histo Chemicals In-Reply-To: References: Message-ID: At 11:54 AM +0000 9/24/03, Barbara Stancel wrote: >Any suggestions? (please tell me which agency audits your lab) I would suggest covering the auditors with all of your "Needs Approval" stickers. The proof is in the pudding. Keep Histology Laboratories free of the bureaucrats Bill -- ____________________________ "The songs of our ancestors are also the songs of our children" -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030924/24caf2dc/attachment.htm From pxs76 <@t> po.cwru.edu Wed Sep 24 08:27:38 2003 From: pxs76 <@t> po.cwru.edu (phyllis) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] DNA extraction from Tissue Blocks Message-ID: <5.1.0.14.0.20030924092422.00c19d28@pop.cwru.edu> I would like to try using a DNA extraction method on formalin fixed tissue blocks. Can anyone recommend a method or kit? we have had difficulty with consistent results. Phyllis Scalzo, HT(ASCP) Case Western Reserve University 2085 Adelbert Rd. Cleveland, Ohio 44106 Ph: 216-368-0822 Fax: 216-368-2546 From Linke_Noelle <@t> Allergan.com Wed Sep 24 09:01:39 2003 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] chromogranine Message-ID: <805C4A052A253B4A8E4948B13F54D12704D183@IRMAIL102.irvine.allergan.com> Chromogranin A/B from Progen works beautifully on rat FFPE! http://www.progen.de/data/produktkatalog/11422.pdf Noelle Linke, BS, HTL(ASCP) Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 -----Original Message----- From: Bruijntjes, J.P. [mailto:bruyntjes@voeding.tno.nl] Sent: Wednesday, September 24, 2003 1:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] chromogranine Hi all Is anyone familiar with a commercial available antibody against chromogranine that cross-reacts with FFPE rat tssues? Joost Bruijntjes TNO Nutrition and Food Research PO Box 360 3700 AJ Zeist The Netherlands _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Helene.Seegers <@t> nottingham.ac.uk Wed Sep 24 09:14:00 2003 From: Helene.Seegers <@t> nottingham.ac.uk (Helene Seegers) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Hi, Message-ID: Hi, does anybody knows if the human monocytic THP-1 cells from culture are expressing CD14? I am using the CD14 antibodies from SIGMA (ref. C7673) without any success... Thanks Helene -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030924/c78a70f7/attachment.htm From tpmorken <@t> labvision.com Wed Sep 24 10:28:51 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] QC/Validation of Histo Chemicals Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CBAA@usca0082k03.rallansci.apogent.com> Barbara, I've never heard of anything like that in histology. Every inspection I've been involved in has been more interested in freshness of chemicals, date of production, documentation of who made it up, written procedures for producing the reagents, etc. The controls run during a procedure are supposed to "validate" whether the procedure actually works - which validates whether the chemicals "work." I guess that you would need to run a procedure on a control and replace only the given chemical, leaving all others the same. If you make up all new reagents for a procedure then you would need to do the same for every other chemical in the procedure. So for a stain that requires 10 chemicals, you would need to do ten different control runs, each with only one chemical as the variable. Sound like a lot of work for little or no real value - after all, most of these reagents are very simple solutions that "work" all the time. Tim Morken Lab Vision / NeoMarkers www.labvision.com -----Original Message----- From: Barbara Stancel [mailto:stancelb@msn.com] Sent: Wednesday, September 24, 2003 4:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] QC/Validation of Histo Chemicals To all histology laboratories inspected by any agency: Are you required to validate (do they work?) chemicals such as: 1% acetic acid, acid alcohol, dilutions of hydroquinone, Scott's Tap Water, 10% NBF, Bouin's, aqueous solutions of sodium thiosulfate, periodic acid, sodium borate, picric acid solutions, etc. (get my drift?)? Do your auditors consider the QC you do for stains and dyes as a satisfactory confirmation/validation that an individual solutions within a procedure work? I am not talking about QC of stains. In this lab that is a given...after preparation, all stains and dyes are QC'ed and the slide is labeled and kept as a permenent record. Auditors are wanting us to mark chemicals with a big ole "needs approval" label. After we prove they work, we apply a big ole "validate" label on all the bottles. We already have a labels on the bottles. Each contains the batch number, solution, stain, date, QC date and the person who performed the QC. The batch number is tracable on a special sheet on which we record even more information...like all the inventory numbers for the chemcials, stains or dyes used in the preparation of that solution, date prepared, date in use, date out of use, etc. Since all our auditors are chemist or microbiologist we are trying to make them see our point of view. This needs approval/validation method is used in chemistry and microbiology. Any suggestions? (please tell me which agency audits your lab) Histologically yours, Barbara (who feels like I am recording my life on spreadsheets with batch and inventory numbers!) Barbara H. Stancel, HTL(ASCP)HT USDA, FSIS, OPHS, Eastern Laboratory, Pathology RRC, 950 College Station Road Athens, Georgia 30604 phone: (706) 546-3556 fax: (706) 546-3589 _________________________________________________________________ Add MSN 8 Internet Software to your existing Internet access and enjoy patented spam protection and more. Sign up now! http://join.msn.com/?page=dept/byoa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tvedilago <@t> system1.net Wed Sep 24 10:35:16 2003 From: tvedilago <@t> system1.net (Tommy Vedilago) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Histology Supervisors Message-ID: Hello again Histonetters, I am happy, as always, to pass on jobs to as many of you as possible. I have a couple of new searches going on. One is in Wisconsin and the other is in Philadelphia. The one in Philadelphia is very exciting because it represents an outstanding opportunity to manage a high volume, highly productive laboratory. It will speak volumes on any manager's resume and offers many benefits and opens many doors for the right person. I have spoken with many of you about this and other searches but, if I have not been able to get in touch with you, please give me a call. Even if you are not particularly interested in changing jobs, I would like to be a good resource for you in the future. My number is 866-SYSTEM1 or 866-797-8361. I will look forward to being of service to you and Histology in general in the future. Tommy Tommy Vedilago System 1 Search (864) 627-0012 (864) 627-0013 Fax From gareth.davis <@t> Vanderbilt.Edu Wed Sep 24 10:37:49 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Histonet] Special stain control Message-ID: <082C721AF78DB34983E8BA2CD085462105C688@mailbe07> Milton, I know Zymed has slides for special stain controls, if that's what you are looking for. Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030924/7960de2a/attachment.htm From gcallis <@t> montana.edu Wed Sep 24 11:10:47 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Request for special staining controls Message-ID: <3.0.6.32.20030924101047.00b70670@gemini.msu.montana.edu> It would be in your best interest to access some of these controls from tissues you are working on your own rather than just purchase them. There are lists of special staining controls avialable and also immunohistochemistry controls. I know that Peggy Wenk has a list, the J of Histotechnology published a list some time ago, Sheehan and Hrapchak Theory and Practice of Histotechnology textbook has a list of controls for a given specific special stain, and HistoLogic from Sakura Finetek has published a list of controls in the past. Many normal tissues contain the specific tissue componenent you are looking for with a special stain, and you can keep a set of control blocks for Massons trichrome (connective tissue), etc, etc. and cut your own controls. In some ways, this is better as you have good quality assurance over a long period of time. It is certainly cheaper, plus one block provides many slides at far less expense. Good luck - Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Bonnie.P.Whitaker <@t> uth.tmc.edu Wed Sep 24 12:09:07 2003 From: Bonnie.P.Whitaker <@t> uth.tmc.edu (Bonnie P Whitaker) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] QC/Validation of Histo Chemicals In-Reply-To: <8CD0CDC791DCC343ABB1F15298E4F3C417CBAA@usca0082k03.rallansci.apogent.com> Message-ID: Tim, Don't you think what she means is more like checking a new detection system in immuno? You replace the whole shebang, and run a known (hopefully multi-tissue) control and compare it to the same control stained with the old lot. If you changed many variables, and there was a problem, it could be a nightmare, but most of the time everything works like a charm. In the clinical labs that I've worked in you run validations by doing the same patient tests and controls (the number can vary) using the reagent in current usage, and also using the new reagent and then check the comparison statistics to make sure the tests run with the new reagent are in an acceptable range before the new reagent is "released" for use in the lab. In histology, the multi-tissue control could be used to run one slide, but producing multiple "tests" for comparison. Bonnie Whitaker Technical Director, Histology Laboratory Department of Pathology and Laboratory Medicine UT Med School 6431 Fannin MSB 2.231 Houston, TX 77030 713-500-6792 -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Morken, Tim - Labvision Sent: Wednesday, September 24, 2003 10:29 AM To: 'Barbara Stancel'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] QC/Validation of Histo Chemicals Barbara, I've never heard of anything like that in histology. Every inspection I've been involved in has been more interested in freshness of chemicals, date of production, documentation of who made it up, written procedures for producing the reagents, etc. The controls run during a procedure are supposed to "validate" whether the procedure actually works - which validates whether the chemicals "work." I guess that you would need to run a procedure on a control and replace only the given chemical, leaving all others the same. If you make up all new reagents for a procedure then you would need to do the same for every other chemical in the procedure. So for a stain that requires 10 chemicals, you would need to do ten different control runs, each with only one chemical as the variable. Sound like a lot of work for little or no real value - after all, most of these reagents are very simple solutions that "work" all the time. Tim Morken Lab Vision / NeoMarkers www.labvision.com -----Original Message----- From: Barbara Stancel [mailto:stancelb@msn.com] Sent: Wednesday, September 24, 2003 4:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] QC/Validation of Histo Chemicals To all histology laboratories inspected by any agency: Are you required to validate (do they work?) chemicals such as: 1% acetic acid, acid alcohol, dilutions of hydroquinone, Scott's Tap Water, 10% NBF, Bouin's, aqueous solutions of sodium thiosulfate, periodic acid, sodium borate, picric acid solutions, etc. (get my drift?)? Do your auditors consider the QC you do for stains and dyes as a satisfactory confirmation/validation that an individual solutions within a procedure work? I am not talking about QC of stains. In this lab that is a given...after preparation, all stains and dyes are QC'ed and the slide is labeled and kept as a permenent record. Auditors are wanting us to mark chemicals with a big ole "needs approval" label. After we prove they work, we apply a big ole "validate" label on all the bottles. We already have a labels on the bottles. Each contains the batch number, solution, stain, date, QC date and the person who performed the QC. The batch number is tracable on a special sheet on which we record even more information...like all the inventory numbers for the chemcials, stains or dyes used in the preparation of that solution, date prepared, date in use, date out of use, etc. Since all our auditors are chemist or microbiologist we are trying to make them see our point of view. This needs approval/validation method is used in chemistry and microbiology. Any suggestions? (please tell me which agency audits your lab) Histologically yours, Barbara (who feels like I am recording my life on spreadsheets with batch and inventory numbers!) Barbara H. Stancel, HTL(ASCP)HT USDA, FSIS, OPHS, Eastern Laboratory, Pathology RRC, 950 College Station Road Athens, Georgia 30604 phone: (706) 546-3556 fax: (706) 546-3589 _________________________________________________________________ Add MSN 8 Internet Software to your existing Internet access and enjoy patented spam protection and more. Sign up now! http://join.msn.com/?page=dept/byoa _______________________________________________ Histonet mailing list Histonet@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 dellav <@t> musc.edu Wed Sep 24 12:29:02 2003 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:21:56 2005 Subject: Fwd: RE: [Histonet] Microtome Service Message-ID: >>> 09/24/03 11:40AM >>> Hello, I'm in Sandy, Utah. Thank you very much in advance. Milton -----Original Message----- From: Vinnie Della Speranza [mailto:dellav@musc.edu] Sent: Tuesday, September 23, 2003 2:51 PM To: MGomez@ameripath.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Microtome Service Help us to help you Milton. where are you located? >>> 09/23/03 12:51PM >>> Hello: Do you know who can provide maintenance and service to my microtomes? Thank you, Milton _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dellav <@t> musc.edu Wed Sep 24 12:30:02 2003 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Microtome Service Message-ID: Milton, I'm sorry I can't be of help(I'm in the southeast) but hopefully someone out your way will see this and recommend a service technician for you. Vinnie >>> 09/24/03 11:40AM >>> Hello, I'm in Sandy, Utah. Thank you very much in advance. Milton -----Original Message----- From: Vinnie Della Speranza [mailto:dellav@musc.edu] Sent: Tuesday, September 23, 2003 2:51 PM To: MGomez@ameripath.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Microtome Service Help us to help you Milton. where are you located? >>> 09/23/03 12:51PM >>> Hello: Do you know who can provide maintenance and service to my microtomes? Thank you, Milton _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cdemarinis <@t> SARATOGACARE.ORG Wed Sep 24 12:35:33 2003 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis,Carolyn) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Gimenez stain Message-ID: <2FCF6059F121BA41A672C5012BB113AA2A52A3@mercury.sarahosp.org> We use the Gimenez stain for H. Pylori and until now it has worked beautifully. Unfortunately, the bugs are not staining red, they are staining a very weak blue-green. Can anyone help? CONFIDENTIALITY NOTICE: 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. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. From jlinda <@t> ces.clemson.edu Wed Sep 24 13:02:12 2003 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Cat Scratch Disease Message-ID: <5.1.0.14.2.20030924140052.04fe8c50@mailhost.ces.clemson.edu> Is there a specific special stain or IHC marker for Cat Scratch Disease? TIA, (Thanks in Advance) 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 fmonson <@t> wcupa.edu Wed Sep 24 13:26:19 2003 From: fmonson <@t> wcupa.edu (Monson, Frederick ) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] DNA extraction from Tissue Blocks Message-ID: Afternoon Phyllis, Your problem may be that almost all methods are hydrolytic and the 'force' needed for a specimen will vary with the degree of crosslinking of the DNP (deoxyribonucleoprotein). In methods for in situ hybridization, the DNA must be exposed in order to be 'melted'. This often requires, for HCHO-fixed material, that the DNP protein be at least disrupted. Pronase is most often suggested. As it is a very active peptidase, each batch/lot will behave differently AS will each specimen. Thus, it is almost always necessary to perform an empiric test that establishes the concentration of pronase and time of action. i.e., I have discovered no easier way, though I have not done ISH for 4 years. The problem is to remove the RNP with pronase and then the DNA with dnase without removing most of the histology in the process. If I were looking for more selective methods to expose DNA, I would look to enzymes that modify histones. Look here: http://web.wi.mit.edu/young/pub/histone_modifiers.html Hope this helps, Fred Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson@wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/ -----Original Message----- From: phyllis [mailto:pxs76@po.cwru.edu] Sent: Wednesday, September 24, 2003 9:28 AM To: Histonet@pathology.swmed.edu Subject: [Histonet] DNA extraction from Tissue Blocks I would like to try using a DNA extraction method on formalin fixed tissue blocks. Can anyone recommend a method or kit? we have had difficulty with consistent results. Phyllis Scalzo, HT(ASCP) Case Western Reserve University 2085 Adelbert Rd. Cleveland, Ohio 44106 Ph: 216-368-0822 Fax: 216-368-2546 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Sep 24 13:26:59 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Control list references Message-ID: <3.0.6.32.20030924122659.00b7a828@gemini.msu.montana.edu> Here is access for staining tissue control list formerly posted on Histonet by Tim Morken (Now with Lab Vision) and Sue Barnes. Here is the Histologic newsletter web site at Sakura (all past issues): http://www.sakuraus.com/ASPages/Histo-Logic.asp And here is the pdf file for the article on normal controls for IHC: http://206.137.77.9/sakura/pdf/28n10598.pdf Tim Morken CDC, Atlanta ************************************************************************ Sue Barnes [mailto:SBarnes@elch.org] Sent: Wednesday, July 24, 2002 10:56 AM Subject: RE: Looking for a control reference list... Histo-Logic had a great list. It was from Vol xxviii, No. 1, May 1998. It has a large list of controls and antibodies. 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 Ronnie_Houston <@t> bshsi.com Wed Sep 24 13:41:53 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Cat Scratch Disease Message-ID: <530361BF03351B4CAE5270A05D3037B5FE042C@bsrexms01.BSHSIR.COM> Linda, Biocare Medical has an antibody to Cat Scratch (conc CM144, predilute PM144AA). We had previously used a modified Steiner, but the bugs are so small (0.2-0.5 micron) that some of the pathologists have had trouble seeing them clearly with the steiner. Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com -----Original Message----- From: Linda Jenkins [mailto:jlinda@ces.clemson.edu] Sent: Wednesday, September 24, 2003 2:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cat Scratch Disease Is there a specific special stain or IHC marker for Cat Scratch Disease? TIA, (Thanks in Advance) 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at 410-442-3250, and permanently delete the original e-mail, attachment(s), and any copies. From funderwood <@t> mcohio.org Wed Sep 24 14:55:50 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Purpose of Oven Prior to Staining? Message-ID: Hi Laurie. I have found that with B5 fixed lymph nodes it is mandatory to allow all water to evaporate from the section before heating. It seeemed that if the section was heated with water present, nuclear detail was destroyed. Almost like it became nuclear "soup" and the morphology was terrible. Fred >>> Laurie Reilly 09/18/03 07:34PM >>> Dear Diane, Your statement that"Ideally, the slides should be dried at room temperature overnight especially if photographs are going to be made" fascinates me. Could you please explain the significance of room temperature drying for photomicrography. Thanks and regards, Laurie. At 02:08 PM 09/18/03 -0400, Gladney, Diane C Ms MACH wrote: >Jordan, > >The purpose of the slide dryer or oven is to dry the water off of the >slides. The slides should be well drained to remove as much water as >possible before placing them in the rack before it is placed in the oven. >It is not and will not deparaffinize the slides. The slides must be >completely free of water before they are stained either by hand or on an >automatic slide stainer. Any histological technique book will tell you that >the deparaffinization is accomplished by using xylene, xylene substitute or >other appropriate paraffin solvent. Ideally, the slides should be dried at >room temperature overnight especially if photographs are going to be made. >But few labs have this much time. A drier that has a temperature in excess >of 60 degrees C will cause some artifacts. Our autostainer has a built-in >forced air dryer that the slides stay in for 10 - 15 minutes. If we have a >rack on the stainer and have another rack ready for the stainer, we place >this rack in a small forced air dryer set for 15 minutes then remove and >wait on the stainer to finish before adding another rack. The slide dryer >then is by-passed and the dried slides go directly to the first xylene >substitute (what we use instead of xylene) for deparaffinization. If they >still won't believe you, have them try to stain some "dried slides" and see >the results. > >Good Luck, >Diane > >Diane C. Gladney, HT (ASCP) >Histology /Cytology Supervisor >Moncrief Army Community Hospital >P.O. BOX 484 >4500 Stuart Ave. >FT. Jackson, SC 29207 > >(803) 751-2530 >DSN 734-2530 > >EMAIL: diane.gladney@se.amedd.army.mil OR > dcgx1@aol.com > > >-----Original Message----- >From: Jordan Brod [mailto:jbrod@tvmdl.tamu.edu] >Sent: Thursday, September 18, 2003 10:41 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Purpose of Oven Prior to Staining? > > >Good Morning All: > >My lab is in a huge discussion about the reason why we put our slides in an >oven before staining. I told them that the reason was to dry all of the >water off of them, because xylene and water do not mix. This statement is >also backed up in literature that I have read. They still do not believe >me, because they think the purpose of the oven is to deparaffinize the >slides. > >Can anyone help me out, so I can put this discussion to rest? I will print >off all of your comments for my technicians to read. Is what I told them >correct or not? > >Thanks. > > > > >******************************************************************** >Jordan W. Brod >Diagnostic Lab Supervisor - Pathology >Texas Veterinary Medical Diagnostic Laboratory (TVMDL) >1 Sippel Road >TAMU 4471 >College Station, Texas 77843 >(979) 845-3414 >(979) 845-1794 fax >jbrod@tvmdl.tamu.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 Physiology & Pharmacology Fax 07 4779 1526 Aust.Inst.of Tropical Vet.& Animal Sc. James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From adford <@t> compuserve.com Wed Sep 24 17:24:28 2003 From: adford <@t> compuserve.com (John Difford) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Re.Isopropanol in processing Message-ID: <200309241824_MC3-1-4FCE-13C2@compuserve.com> Dear All Despite some statements to the contrary that have recently been posted on Histonet, I have to inform you that isopropanol is miscible with melted paraffin wax and this is why one can use if for processing without the use of a clearing agent. John Difford Histopathology Royal Free Hospital London England, UK From ploykasek <@t> phenopath.com Wed Sep 24 18:22:04 2003 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] DNA extraction from Tissue Blocks In-Reply-To: <5.1.0.14.0.20030924092422.00c19d28@pop.cwru.edu> Message-ID: What are you wanting to do with the DNA? There is a reagent called Gene Releaser from BioVentures, Inc that can be used with FFPE sections. The DNA can then be used for pcr. Hope this helps. Patti Loykasek PhenoPath Laboratories Seattle, Wa > > > I would like to try using a DNA extraction method on formalin fixed tissue > blocks. Can anyone recommend a method or kit? we have had difficulty with > consistent results. > > > Phyllis Scalzo, HT(ASCP) > Case Western Reserve University > 2085 Adelbert Rd. > Cleveland, Ohio 44106 > Ph: 216-368-0822 > Fax: 216-368-2546 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Rcartun <@t> harthosp.org Wed Sep 24 13:40:08 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] IHC for renin Message-ID: Is anyone doing immunohistochemistry for "renin"? Thank you. Richard Cartun From ranahay <@t> fastmail.fm Wed Sep 24 21:08:19 2003 From: ranahay <@t> fastmail.fm (Rana Hay) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Fwd: cassette printer etc Message-ID: <20030925020819.76C2C3CAB0@www.fastmail.fm> On Thu, 25 Sep 2003 15:06:29 +1300, "Rana Hay" said: > Hello Histoneters, > The lab I work for in NZ is concidering buying a cassette printer.Any > advice? Who are the suppliers? > Also.I am interested in what medical use for the H&E stain .I have used > it in Virology, Histopathology,Andrology and seen it used in Cytology.Any > more ideas? > > Many Thanks,Rana > -- > Rana Hay > ranahay@fastmail.fm > > -- > http://www.fastmail.fm - Send your email first class -- Rana Hay ranahay@fastmail.fm -- http://www.fastmail.fm - The professional email service From jkiernan <@t> uwo.ca Wed Sep 24 23:43:14 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Re.Isopropanol in processing References: <200309241824_MC3-1-4FCE-13C2@compuserve.com> Message-ID: <3F727262.71CFF87B@uwo.ca> Dear All including John D., Isopropanol is not miscible with melted paraffin wax. If you don't believe the usual published references you can easily check for yourself. Add a drop of melted wax to a small beaker containing warm isopropyl alcohol. It won't dissolve, even if you heat to higher than the melting point of the wax. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ _________________________ John Difford wrote: > > Dear All > > Despite some statements to the contrary that have recently been posted on > Histonet, I have to inform you that isopropanol is miscible with melted > paraffin wax and this is why one can use if for processing without the use > of a clearing agent. > > John Difford > Histopathology > Royal Free Hospital > London > England, UK ___________________________ From bruyntjes <@t> voeding.tno.nl Thu Sep 25 02:25:22 2003 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Chromogranine Message-ID: <3B070848E7C2204F9DEB8BCFD76772800210D44E@ntexch1.voeding.tno.nl> Thanks to all who responded to my guestion about a chromogranine antibody Joost Bruijntjes TNO Nutrition and Food Research PO Box 360 3700 AJ Zeist The Netherlands From frouwke <@t> sci.kun.nl Thu Sep 25 03:39:11 2003 From: frouwke <@t> sci.kun.nl (Frouwke Kuijpers) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Re: Brain Tissue in Buffer References: Message-ID: <007301c38340$806eda80$4a8aae83@sci.kun.nl> My experience is that when fixed brain stays long in PBS, it becomes very soft. Than I don't use these brains anymore. Reason? I don't know. Somebody else maybe? Frouwke Kuijpers ----- Original Message ----- From: To: Sent: Tuesday, September 23, 2003 1:36 PM Subject: [Histonet] Re: Brain Tissue in Buffer > > Hi, > I've been told that if you fix in PFA then store in PBS for long periods > the PBS "unfixes" the tissue. So I would suggest redoing your fixation > process. > > Good luck! > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From cdemarinis <@t> SARATOGACARE.ORG Thu Sep 25 06:27:50 2003 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis,Carolyn) Date: Fri Sep 16 15:21:56 2005 Subject: FW: [Histonet] Gimenez stain Message-ID: <2FCF6059F121BA41A672C5012BB113AA2A52A5@mercury.saratogacare.org> We were using the Alcian Yellow Toluidine blue method for years, but unfortunately Alcian Yellow is not available in this country. (I cannot remember the problem). Therefore I have to find an alternative stain for H. Pylori. I have reordered a Gimenez kit hoping that the kit I am using is a bad lot. If Alcian Yellow is available for purchase, please let me know. Thanks for your help. Carolyn DeMarinis Saratoga Hospital Saratoga Springs New York Have you tried the Alcian Yellow Toluidine blue method. I've tried most methods over the years and this one, to me at least, is the best. I can send you a protocol if you like. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Demarinis,Carolyn Sent: 24 September 2003 18:36 To: HISTONET (E-mail) Subject: [Histonet] Gimenez stain We use the Gimenez stain for H. Pylori and until now it has worked beautifully. Unfortunately, the bugs are not staining red, they are staining a very weak blue-green. Can anyone help? CONFIDENTIALITY NOTICE: 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. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: 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. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. From Marjorie.Lehman <@t> unilever.com Thu Sep 25 06:58:28 2003 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] MiTF Message-ID: Hi, Has anyone stained for microthalmia transcription factor in frozen skin? I have tried Abs from 2 vendors, diluted down to 1:10, changed diluting fluids, used a vector kit, made my own solutions, incubated 1 hr, 2hrs and overnight and I'm still getting nothing. I'm using pigmented human skin which is the suggested control. Any help would be appreciated. Thanks, Marge From Tony.Fearn <@t> cd.burnleyhc-tr.nwest.nhs.uk Thu Sep 25 07:15:07 2003 From: Tony.Fearn <@t> cd.burnleyhc-tr.nwest.nhs.uk (Fearn Tony) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] FLI-1 Message-ID: <8C3DE9069A06D611B8950002A550C15943BCB1@BHC_MAIL02> > One of our pathologists has just returned from a conference in Milan and > is asking if we can get FLI-1, as an endothelial marker, that is meant to > be better than CD34. I've looked in the DakoCytomation and Novocastra > catalogues but there is no mention of it. Does anyone know where I can get > it, and is it worth buying? > > Anthony J Fearn > Chief Biomedical Scientist > Histopathology > > From Catherine.Goeden <@t> med.va.gov Thu Sep 25 07:28:13 2003 From: Catherine.Goeden <@t> med.va.gov (Goeden, Catherine) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] COX-2 antibody Message-ID: <6A6686E52FDAD5118CC30000F8034A25931819@VHASUXEXC1> I am working on a research project and wondering if anyone is using this antibody and could give me some information on it. I purchased the kit and antibody from Biocare Medical. They have told me it is sometimes tricky to use so if someone has some hints I would greatly appreciate it. Thanks in advance! From cdemarinis <@t> SARATOGACARE.ORG Thu Sep 25 07:24:16 2003 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis,Carolyn) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] ALCIAN YELLOW Message-ID: <2FCF6059F121BA41A672C5012BB113AA2A52A7@mercury.saratogacare.org> In reference to the email from Inga for alcian yellow, please give me the address and phone number for VWR International. Thank you. CONFIDENTIALITY NOTICE: 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. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. From ian.montgomery <@t> bio.gla.ac.uk Thu Sep 25 07:25:24 2003 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Dako Ark. Message-ID: <5.2.1.1.2.20030925132127.00a4fd20@udcf.gla.ac.uk> Although the brown reaction product with Dako Ark looks lovely I'd like a black product. Has anyone used intensifying techniques with Dako Ark and can they offer any hints and tips. 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 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/3ec5b0dc/attachment.htm From richard.rodriguez <@t> spcorp.com Thu Sep 25 08:08:01 2003 From: richard.rodriguez <@t> spcorp.com (Rodriguez, Richard) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] 3% Glutaraldehyde Message-ID: <4508920F80C0D411B90200508BF9A9F404932244@LAFMSG30.us.schp.com> We are currently using 3% Glutaraldehyde for fixation of eyes. The solution is phosphate buffered and stored at room temp. Recently, the glut is going bad (turning yellow, cloudy, glut coming out of solution) before the expiration time of 4 months. I've investigated other sources and discovered that most suppliers refrigerate their glut and store it in amber containers. I would like to know what other people are using for eye fixation in rodents and other larger animals. Thanks for all your help. Richard Rodriguez Schering-Plough Research Institute Lafayette, NJ 07848 973-940-4282 fax: 973-940-4120 ********************************************************************* 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/d2ba07aa/attachment.htm From cdemarinis <@t> SARATOGACARE.ORG Thu Sep 25 08:10:43 2003 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis,Carolyn) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] ALCIAN YELLOW Message-ID: <2FCF6059F121BA41A672C5012BB113AA2A52A8@mercury.saratogacare.org> I called VWR international and they do not have a listing for Alcian Yellow. Inga, do you have a product # or any other information. Thank you. CONFIDENTIALITY NOTICE: 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. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. From halsteaj <@t> ohsu.edu Thu Sep 25 08:37:10 2003 From: halsteaj <@t> ohsu.edu (Jeff Halstead) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] bielschowsky silver with lfb Message-ID: hello all just wondering if anybody has done or seen a bielschowsky silver stain for cns and then stained with the luxol fast blue. a researcher here has been told to submit slides for review with this combo of stains. i personally have never done these together. am going to try , but was wondering if anyone else has had any experience with these two stains . thankyou. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/0cdbc379/attachment.htm From cwscouten <@t> myneurolab.com Thu Sep 25 08:44:18 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Re: Brain Tissue in Buffer Message-ID: I have always heard you cannot unboil an egg, or unfix tissue. My guess is the brains were not fully fixed in the first place. Fixation takes a minimum of four hours and continues to progress for up to 7 days in perfused brains. Partial fixation may be best for immunological use. But taking out of fixative before fully fixed and storing in PBS for a long tim may have an obvious result. Partial detiorioration. Were the brains in question fully fixed in the first place? Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com www.myneurolab.com -----Original Message----- From: Frouwke Kuijpers [mailto:frouwke@sci.kun.nl] Sent: Thursday, September 25, 2003 3:39 AM To: histonet@lists.utsouthwestern.edu; Nancy.Walker@sanofi-synthelabo.com Subject: Re: [Histonet] Re: Brain Tissue in Buffer My experience is that when fixed brain stays long in PBS, it becomes very soft. Than I don't use these brains anymore. Reason? I don't know. Somebody else maybe? Frouwke Kuijpers ----- Original Message ----- From: To: Sent: Tuesday, September 23, 2003 1:36 PM Subject: [Histonet] Re: Brain Tissue in Buffer > > Hi, > I've been told that if you fix in PFA then store in PBS for long periods > the PBS "unfixes" the tissue. So I would suggest redoing your fixation > process. > > Good luck! > > > > _______________________________________________ > Histonet mailing list > Histonet@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 Ronnie_Houston <@t> bshsi.com Thu Sep 25 08:48:04 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] FLI-1 Message-ID: <530361BF03351B4CAE5270A05D3037B5FE0430@bsrexms01.BSHSIR.COM> Tony, Fli -1 is a nuclear endothelial marker, and appears to be involved in proliferation and tumorgeneisis. You can get the polyclonal antibody from Santa Cruz Biotechnology, cat sc356 (the UK # is 800 4573 8000). We have used it for quite some time now, and the pathologists think it is great. You may want to look at the following article: Expression of Fli-1, a nuclear transcription factor, distinguishes vascular neoplasms from potential mimics. Folpe AL, Chand EM, Goldblum JR, Weiss SW, Am J Surg Pathol 2001; 25(8): 1061-1066 Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com -----Original Message----- From: Fearn Tony [ mailto:Tony.Fearn@cd.burnleyhc-tr.nwest.nhs.uk ] Sent: Thursday, September 25, 2003 8:15 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] FLI-1 > One of our pathologists has just returned from a conference in Milan and > is asking if we can get FLI-1, as an endothelial marker, that is meant to > be better than CD34. I've looked in the DakoCytomation and Novocastra > catalogues but there is no mention of it. Does anyone know where I can get > it, and is it worth buying? > > Anthony J Fearn > Chief Biomedical Scientist > Histopathology > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at 410-442-3250, and permanently delete the original e-mail, attachment(s), and any copies. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/db38ce4a/attachment.htm From Ronnie_Houston <@t> bshsi.com Thu Sep 25 08:54:31 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] bielschowsky silver with lfb Message-ID: <530361BF03351B4CAE5270A05D3037B5FE0431@bsrexms01.BSHSIR.COM> Jeff, I don't have experience with the Bielschowsky and LFB, but combined LFB with a Palmgren's silver impreganation technique (far superior and easier to perform than Bielschowsky in my humble opinion). You need to do the LFB technique first before carrying out the silver. Let me know if you want any more details, and I'll try to search through my files. Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com -----Original Message----- From: Jeff Halstead [mailto:halsteaj@ohsu.edu] Sent: Thursday, September 25, 2003 9:37 AM To: montana.edu Thu Sep 25 09:09:22 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Making DAB+ in Dako Ark darker In-Reply-To: <5.2.1.1.2.20030925132127.00a4fd20@udcf.gla.ac.uk> Message-ID: <3.0.6.32.20030925080922.00b89a50@gemini.msu.montana.edu> You can make the brown endproduct a very dark, black brown (almost black!) by using the DAKO DAB enhancer. This works beautifully with their DAB+ that is in ARK kit or you can always do another enhancer to turn the product black. This has been discussed at great lengths in Histonet archives. Other companies have enhancers ready to use, Zymed and Vector, but I am not sure if they are black. Since the kit is a kit, we prefer to use the DAKO enhancing product, pretty much optimized for their DAB+ which (with their enhancer), allowed us to get a murine CD4 diluted out 1:15,000 (0.5mg/ml)and more. This was not an ARK kit method, but we were impressed with the overall chromogen/enhancer. We had to be careful about any residual peroxidase or pseudoperoxidases in tissues when using DAKO DAB enhancer. We used the glucose oxidase endogenous peroxidase method for quenching with very clean results. At 01:25 PM 9/25/2003 +0100, you wrote: > Although the brown reaction product with Dako Ark looks lovely >I'd like a black product. Has anyone used intensifying techniques with Dako >Ark and can they offer any hints and tips. > Ian. > Dr. Ian Montgomery, > Histotechnology, > Graham Kerr Building, >& 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 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 dobbin <@t> upei.ca Thu Sep 25 08:56:48 2003 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] COX-2 antibody In-Reply-To: <6A6686E52FDAD5118CC30000F8034A25931819@VHASUXEXC1> Message-ID: <3F72C9F1.12594.7634B4@localhost> Hi Catherine, I am no expert but I have a little experience. In human tissues I had to use heat retreival on my formalin-fixed, paraffin-embedded tissues as well as leaving the antibody on overnight at 4 C. In canine tissues I couldn't get it to work at all (although someone else at this institution could, but I don't know the details), and in rats it worked like a charm with ag retreival not required and incubation for only 1 hr at room temperature! There are many different COX-2 ab's out there and as many or even more different sources. I think the trick might be to find the one that suits your application best. Good luck. Greg Date sent: Thu, 25 Sep 2003 07:28:13 -0500 From: "Goeden, Catherine" Subject: [Histonet] COX-2 antibody To: "'histonet@pathology.swmed.edu'" > > I am working on a research project and wondering if anyone is using this > antibody and could give me some information on it. I purchased the kit and > antibody from Biocare Medical. They have told me it is sometimes tricky to > use so if someone has some hints I would greatly appreciate it. Thanks in > advance! > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Pathology Lab Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Making a living is getting; making a life is giving. From Linke_Noelle <@t> Allergan.com Thu Sep 25 09:37:52 2003 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] cleaved caspase-3 Message-ID: <805C4A052A253B4A8E4948B13F54D12704D184@IRMAIL102.irvine.allergan.com> Hi everyone! Is anyone using cleaved caspase-3 with a protocol other than overnight at 4 degrees? I'm having problems with my staining and I don't have weeks to get it sorted out, so I am looking for a room temp or heated protocol to try. Thanks in advance Noelle Linke, BS, HTL(ASCP) Allergan, Inc Irvine, CA 92612 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/dfbf3b38/attachment.htm From lizchlipala <@t> premierhistology.com Thu Sep 25 09:58:08 2003 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] FLI-1 In-Reply-To: <8C3DE9069A06D611B8950002A550C15943BCB1@BHC_MAIL02> Message-ID: <000001c38375$74a10db0$74d48a80@LIZ> Tony I believe the antibody you are talking about is FLK-1 or FLT-1, another name for it is VEGF-R2. From what I understand it does not stain the same as CD34, I don't believe it is specific for endothelial cells, it will stain other things also. You can get this antibody from Santa Cruz. We have worked on this in the past and in our hands it was a tough one. It does play a role in angiogenesis. Liz Elizabeth A. Chlipala, BS, HTL(ASCP) Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Fearn Tony Sent: Thursday, September 25, 2003 5:15 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] FLI-1 > One of our pathologists has just returned from a conference in Milan and > is asking if we can get FLI-1, as an endothelial marker, that is meant to > be better than CD34. I've looked in the DakoCytomation and Novocastra > catalogues but there is no mention of it. Does anyone know where I can get > it, and is it worth buying? > > Anthony J Fearn > Chief Biomedical Scientist > Histopathology > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From HornHV <@t> archildrens.org Thu Sep 25 10:21:33 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] FLI-1 Message-ID: There is an antibody Fli-1 (C-19): sc-356. We got ours from Santa Cruz. On the antibody sheets the background says: Ets-1 is the prototype member of a family of genes identified on the basis of homology to the v-Ets oncogene isolated from the E26 erythroblastosis virus...........Members of the Ets gene family exhibit varied patterns of tissue expression and share a highly conserved conboxy terminal domain containing a sequence related to the SV20 large T antigen nuclear localization signal sequence. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Fearn Tony [mailto:Tony.Fearn@cd.burnleyhc-tr.nwest.nhs.uk] Sent: Thursday, September 25, 2003 7:15 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] FLI-1 > One of our pathologists has just returned from a conference in Milan > and is asking if we can get FLI-1, as an endothelial marker, that is > meant to be better than CD34. I've looked in the DakoCytomation and > Novocastra catalogues but there is no mention of it. Does anyone know > where I can get it, and is it worth buying? > > Anthony J Fearn > Chief Biomedical Scientist > Histopathology > > _______________________________________________ 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. Arkansas Children's Hospital From MGomez <@t> ameripath.com Thu Sep 25 10:51:22 2003 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Microtome Service Message-ID: Thanks Vinnie, Milton -----Original Message----- From: Vinnie Della Speranza [mailto:dellav@musc.edu] Sent: Wednesday, September 24, 2003 11:30 AM To: MGomez@ameripath.com Cc: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microtome Service Milton, I'm sorry I can't be of help(I'm in the southeast) but hopefully someone out your way will see this and recommend a service technician for you. Vinnie >>> 09/24/03 11:40AM >>> Hello, I'm in Sandy, Utah. Thank you very much in advance. Milton -----Original Message----- From: Vinnie Della Speranza [mailto:dellav@musc.edu] Sent: Tuesday, September 23, 2003 2:51 PM To: MGomez@ameripath.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Microtome Service Help us to help you Milton. where are you located? >>> 09/23/03 12:51PM >>> Hello: Do you know who can provide maintenance and service to my microtomes? Thank you, Milton _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu Sep 25 11:17:25 2003 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] cleaved caspase-3 Message-ID: I perform caspase-3 routinely using a one hour RT incubation on human xenografts. I use the Pharmingen primary, and an SABC method. Let me know if you'd like me to send you the protocol. However, I am not using formalin fixed tissue - My tissues are zinc fixed paraffin sections. I haven't been able to get consistent results with caspase-3 on ffpe. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 Fax 847.938.3266 "Linke_Noelle" Sent by: histonet-admin@lists.utsouthwestern.edu 09/25/2003 09:37 AM To: cc: Subject: [Histonet] cleaved caspase-3 Hi everyone! Is anyone using cleaved caspase-3 with a protocol other than overnight at 4 degrees? I'm having problems with my staining and I don't have weeks to get it sorted out, so I am looking for a room temp or heated protocol to try. Thanks in advance Noelle Linke, BS, HTL(ASCP) Allergan, Inc Irvine, CA 92612 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/c00aa1a5/attachment.htm From g.lang <@t> bigfoot.de Thu Sep 25 11:48:48 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] ALCIAN YELLOW References: <2FCF6059F121BA41A672C5012BB113AA2A52A8@mercury.saratogacare.org> Message-ID: <004b01c38384$e44c1920$0d12a8c0@SERVER> Fa. Chroma offers Alcian yellow. Fa. Chroma 1F-597 Alciangelb 25 G 480,20 EUR CI 12840 www.chroma.de Gundi Lang ----- Original Message ----- From: "Demarinis,Carolyn" To: "HISTONET (E-mail)" Sent: Thursday, September 25, 2003 3:10 PM Subject: [Histonet] ALCIAN YELLOW > I called VWR international and they do not have a listing for Alcian Yellow. > Inga, do you have a product # > or any other information. Thank you. > CONFIDENTIALITY NOTICE: 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. If you have received this communication in > error, please notify Saratoga Hospital immediately by e-mail at > privacy@saratogacare.org and destroy all copies of this communication and > any attachments. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Ronnie_Houston <@t> bshsi.com Thu Sep 25 11:47:45 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] FLI-1 Message-ID: <530361BF03351B4CAE5270A05D3037B5FE0432@bsrexms01.BSHSIR.COM> Hi Liz, FLK-1 and FLT-1 are cell membrane receptors for VEGF; Fli-1 is a transcription factor/regulator. As it is a nuclear protein, yes, in a strict sense, it doesn't stain the same as CD31 or CD34 or Factor VIII, but it is specific for endothelial cells. Ronnie Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com Hi Liz, -----Original Message----- From: Elizabeth Chlipala [ mailto:lizchlipala@premierhistology.com ] Sent: Thursday, September 25, 2003 10:58 AM To: 'Fearn Tony'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] FLI-1 Tony I believe the antibody you are talking about is FLK-1 or FLT-1, another name for it is VEGF-R2. From what I understand it does not stain the same as CD34, I don't believe it is specific for endothelial cells, it will stain other things also. You can get this antibody from Santa Cruz. We have worked on this in the past and in our hands it was a tough one. It does play a role in angiogenesis. Liz Elizabeth A. Chlipala, BS, HTL(ASCP) Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [ mailto:histonet-admin@lists.utsouthwestern.edu ] On Behalf Of Fearn Tony Sent: Thursday, September 25, 2003 5:15 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] FLI-1 > One of our pathologists has just returned from a conference in Milan and > is asking if we can get FLI-1, as an endothelial marker, that is meant to > be better than CD34. I've looked in the DakoCytomation and Novocastra > catalogues but there is no mention of it. Does anyone know where I can get > it, and is it worth buying? > > Anthony J Fearn > Chief Biomedical Scientist > Histopathology > > _______________________________________________ Histonet mailing list Histonet@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 in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at 410-442-3250, and permanently delete the original e-mail, attachment(s), and any copies. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/45852db9/attachment.htm From RCHIOVETTI <@t> aol.com Thu Sep 25 12:09:07 2003 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Fwd: cassette printer etc Message-ID: In a message dated 09/24/2003 7:10:02 PM US Mountain Standard Time, ranahay@fastmail.fm writes: << On Thu, 25 Sep 2003 15:06:29 +1300, "Rana Hay" said: > Hello Histoneters, > The lab I work for in NZ is concidering buying a cassette printer.Any > advice? Who are the suppliers? > Also.I am interested in what medical use for the H&E stain .I have used > it in Virology, Histopathology,Andrology and seen it used in Cytology.Any > more ideas? >> Hi Rana, Go to Leica Microsystems' histology website and take a look at their cassette and slide printers. The address is: . You can also contact Leica from this site or their homepage for details on your local Leica rep agency. Best regards, Bob Chiovetti GTI Microsystems Leica Exclusive Regional Dealer Desert Southwest Region USA From la.sebree <@t> hosp.wisc.edu Thu Sep 25 12:12:59 2003 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] IHC stains for ABO/Rh antigens Message-ID: Hello histonetters, One of our hematopathologists would like to know where we can get a couple of immunohistochemical stains done: RBC antigen B of the ABO system and RBC antigen RH positive (D antigen). Thanks, Linda A. Sebree, HT(ASCP) University of Wisconsin Hospital & Clinics IHC/ISH Laboratory A4/204-2472 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 From northma <@t> ohsu.edu Thu Sep 25 12:18:29 2003 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:21:56 2005 Subject: FW: [Histonet] Gimenez stain Message-ID: Sigma has Alcian Yellow listed in the catalog # A 7394. American Master*Tech Scientific also has it listed. Mary North, HT(ASCP,HTL Oregon Health & Science University Portland, OR >>> "Demarinis,Carolyn" 09/25/03 04:27AM >>> We were using the Alcian Yellow Toluidine blue method for years, but unfortunately Alcian Yellow is not available in this country. (I cannot remember the problem). Therefore I have to find an alternative stain for H. Pylori. I have reordered a Gimenez kit hoping that the kit I am using is a bad lot. If Alcian Yellow is available for purchase, please let me know. Thanks for your help. Carolyn DeMarinis Saratoga Hospital Saratoga Springs New York Have you tried the Alcian Yellow Toluidine blue method. I've tried most methods over the years and this one, to me at least, is the best. I can send you a protocol if you like. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Demarinis,Carolyn Sent: 24 September 2003 18:36 To: HISTONET (E-mail) Subject: [Histonet] Gimenez stain We use the Gimenez stain for H. Pylori and until now it has worked beautifully. Unfortunately, the bugs are not staining red, they are staining a very weak blue-green. Can anyone help? CONFIDENTIALITY NOTICE: 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. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: 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. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/47c0de0d/attachment.htm From nmuvarak <@t> facstaff.wisc.edu Thu Sep 25 12:29:23 2003 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL E MUVARAK) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] DAKO ARK and Ki-67 Message-ID: <215feb2122ae.2122ae215feb@wiscmail.wisc.edu> Hello. I've been using the DAKO Ark kit to stain for Ki-67 in mouse lungs and arteries. My positive control is mouse spleen, which gives me nice dark brown nuclei. However, I've been getting intense, specific staining in the airways of the lung, even for the negative control, where I replace the primary antibody mixture with just diluent buffer. This shows up also in the experimental tissue as well. I know it's not background since the vasculature in the lung shows no staining. The alveolar space has quite a bit of "brown spots" that lead me to think that I had positive staining in the lung nuclei, but since the negative control showed that, I'm guessing it's false positive. Anybody had this problem before with lungs or other tissue? If someone has any suggestions on how to treat this problem, I'd be very grateful if they can help. Thank you for your time. Regards, Nidal E Muvarak Associate Research Specialist Vascular Tissue Biomechanics Laboratory Department of Biomedical Engineering University of Wisconsin-Madison 1550 Engineering Dr.; Rm. 2158 Madison, WI 53706-1609 Lab: (608) 265-8921; Office: (608) 265-4205; Home: (608) 256-7934; Cell: (608) 332-6068 http://vtb.bme.wisc.edu From RCHIOVETTI <@t> aol.com Thu Sep 25 12:33:57 2003 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Re: Leica Surgical Microscope Parts Message-ID: <116.2904356a.2ca48105@aol.com> In a message dated 09/24/2003 7:10:39 PM US Mountain Standard Time, bozzola@siu.edu writes: << We would like to have an alternate stand so that the optics could be used on a table top. I believe such a stand is used in the Leica M651 MSD. I would appreciate hearing from anyone who has information on this topic (vendors, users, etc.) >> John, If necessary, you can always contact Leica Customer Service if you need details on the current mounting systems and stands that are available: 1-800-248-0123 (in the USA only) Best, Bob Chiovetti GTI Microsystems Leica Exclusive Regional Dealer Desert Southwest Region USA From Gillian.Barlow <@t> cshs.org Thu Sep 25 13:00:22 2003 From: Gillian.Barlow <@t> cshs.org (Barlow, Gillian) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Excessively wrinkled sections Message-ID: <3CFAA0108952D111A5BF00805FA6FB0F0464D454@PEDSNTAS.csmc.edu> Dear Hisonetters First, many thanks to those of you who replied to help me with my cardiac fixations - I now have nice, well-fixed tissues. However, one problem till remains - the sections are very wrinkled and do not flatten out well on the waterbath, even when left for long periods of time. I store my blocks at room temperature, but put them on ice for at least two hours before sectioning. The waterbath is at 40 degrees. A colleague suggested increasing the waterbath temperature, but the sections are for IHC and I dont want to damage my antigen. Many thanks Gillian PS For anyone out there with questions on cardiac fixation and embedding, this is the protocol I have now found to work well and wanted to share: Fixation: Hearts were isolated from anaesthetized animals, cut in half and fixed overnight in 10% buffered formalin at room temperature. The following day, hearts were transferred to 70% ethanol and stored at 4 degrees prior to embedding. Embedding: 80% EtOH, 45 mins room temperature 95% EtOH, 45 mins room temperature, two changes 100% EtOH, 45 mins room temperature, two changes Xylene, 45 mins room temperature, two changes Paraffin, 45 mins, 60 degrees under vacuum, two changes Embed immediately Gillian Barlow, PhD Postdoctoral Fellow Laboratory of Julie Korenberg, PhD, MD Cedars-Sinai Medical Center Davis Bldg, Lab 2007 110 George Burns Rd Los Angeles, CA 90048 Phone: (310) 423 7650 Fax: (310) 423 0302 -------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: application/ms-tnef Size: 2394 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/81e9d7bc/attachment.bin From asmith <@t> mail.barry.edu Thu Sep 25 13:18:25 2003 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Isopropyl alcohol and paraffin wax Message-ID: <494304423C63E246A5CF87A3AEEB577011B590@bumail01.barrynet.barry.edu> Miscible means mutually soluble in ALL proportions. Isopropyl alcohol and paraffin was are definitely not miscible. However, they are mutually soluble to a limited degree. I just did a few crude solubility estimates by eyeball in my lab: I got about half a gram of "Paraplast" to dissolve in 100 ml of gently boiling isopropyl alcohol. I was able to get 30 ml of isopropyl alcohol to dissolve in 100 ml of molten "Paraplast". Both solutions become cloudy suspensions on cooling. I have not tried transferring tissue directly from isopropyl alcohol to molten paraffin, but it seems that the experiment would be worth trying. Allen A. Smith, Ph.D. Professor of Anatomy School of Graduate Medical Sciences Barry University Miami Shores, FL 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 gentras <@t> vetmed.auburn.edu Thu Sep 25 13:35:48 2003 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Nick Kirk Message-ID: <5.2.0.9.0.20030925133209.00a014f0@mailhost.vetmed.auburn.edu> Mr. Kirk will you please share with me your Alcian Yellow Toluidine blue protocol for H. pylori? Thanks, Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From lcardenas <@t> utmem.edu Thu Sep 25 14:17:01 2003 From: lcardenas <@t> utmem.edu (Luz Cardenas) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] caspase-3 Message-ID: <816d348118f5.8118f5816d34@utnet2.utmem.edu> Hi all, I would also like to have the protocol...I am just starting to try in 4% Paraformaldehyde fixed mouse brain sections... Also I read is difficult to get the staining. some kind of special conditions? Thanks! Luz C?rdenas, MD Post-doctoral Research Trainee Department of Anatomy and Neurobiology University of Tennessee Health Science Center 855 Monroe Avenue, Room 309/302 Memphis, TN 38163 Lab: (901) 448-6002 / 7318 Fax: (901) 448-7193 (Dept) > Hi everyone! > > Is anyone using cleaved caspase-3 with a protocol other than > overnight at > 4 degrees? I'm having problems with my staining and I don't have > weeks to > get it sorted out, so I am looking for a room temp or heated > protocol to > try. > > Thanks in advance > > Noelle Linke, BS, HTL(ASCP) > Allergan, Inc > Irvine, CA 92612 > > > From subratab <@t> bdonline.com Thu Sep 25 14:35:29 2003 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] how tissue perfusion affects IHC results Message-ID: <200309251942.h8PJgwPU006074@korotoa.bdonline.com> Dear All, I am staining renal tissue sections for detection of macrophages (ED1 positive cells). I am getting high number (about 10 folds) of positive cells in perfused tissue sections (perfused with saline followed by methacarn) than non-perfused tissue. My question is what happens with perfusion that makes such a great difference. Is it only the better preservation of tissue structure/antigens or there is something associated with the procedure of perfusion that brings more macrophages during the procedure? Can the tissue handling (like dissection for identification and canulation of vessels), intrarenal pressure fluctuations or relative ischemia be responsible for migration of macrophages into the tissue during perfusion procedure? I am looking for some expert opinion. Please let me know if there is any related articles on how tissue perfusion affects IHC results. Subrata Biswas PhD Student UNICAMP, SP, Brazil. From Jenny-Oblander <@t> omrf.ouhsc.edu Thu Sep 25 14:44:50 2003 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Isopropyl alcohol and paraffin wax Message-ID: When microwaving is used in histoprocessing, these procedures are not only accelerated, but fundamentally changed: Completion of fixation is achieved prior to histoprocessing in the microwave oven. Dehydration is achieved in one step, instead of the 2-6 steps used in conventional procedures. The use of a graded series of alcohols is not necessary in the microwave method. Isopropanol can be substituted for xylene as a clearing agent, and one bath is sufficient. Higher temperatures are required for the impregnation of the paraffin wax. Therefore, microwave histoprocessing becomes a three-step process: One dehydration step One clearing step One paraffin wax step (at two temperatures) This is from Microwave technique- Energy Beam Sciences -----Original Message----- From: Smith, Allen [mailto:asmith@mail.barry.edu] Sent: Thursday, September 25, 2003 1:18 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Isopropyl alcohol and paraffin wax Miscible means mutually soluble in ALL proportions. Isopropyl alcohol and paraffin was are definitely not miscible. However, they are mutually soluble to a limited degree. I just did a few crude solubility estimates by eyeball in my lab: I got about half a gram of "Paraplast" to dissolve in 100 ml of gently boiling isopropyl alcohol. I was able to get 30 ml of isopropyl alcohol to dissolve in 100 ml of molten "Paraplast". Both solutions become cloudy suspensions on cooling. I have not tried transferring tissue directly from isopropyl alcohol to molten paraffin, but it seems that the experiment would be worth trying. Allen A. Smith, Ph.D. Professor of Anatomy School of Graduate Medical Sciences Barry University Miami Shores, FL The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From adford <@t> compuserve.com Thu Sep 25 15:22:51 2003 From: adford <@t> compuserve.com (John Difford) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Re: Isopropanol in paraffin processing Message-ID: <200309251623_MC3-1-5004-E4FA@compuserve.com> Dear All Thank you Professor Smith for coming to my rescue and agreeing that isopropanol and melted paraffin wax are mutually soluble at least to some extend. i have not tested the exact proportions of each that will dissolve together but I have used isopropanol to process small biopsies without the use of a clearing agent. The advantage of this is vastly improved staining. The downside of the technique is that isopropanol is poorer at removing fat than ethanol followed by xylene and therefore tissue blocks with a higher fat content may not infiltrate with wax properly, resulting in poor sectioning. John Difford Histopathology Royal Free Hospital London England, UK From Histolady710 <@t> aol.com Thu Sep 25 16:06:40 2003 From: Histolady710 <@t> aol.com (Histolady710@aol.com) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] HT (ASCP) Exam Message-ID: Question ! What chemical is nicknamed "Microcurie"? This question is being asked on the HT (ASCP) Exam. Thanks for your help ! -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/b82e9809/attachment.htm From Jenny-Oblander <@t> omrf.ouhsc.edu Thu Sep 25 16:20:16 2003 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] HT (ASCP) Exam Message-ID: Curium -----Original Message----- From: Histolady710@aol.com [mailto:Histolady710@aol.com] Sent: Thursday, September 25, 2003 4:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HT (ASCP) Exam Question ! What chemical is nicknamed "Microcurie"? This question is being asked on the HT (ASCP) Exam. Thanks for your help ! -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/1521be38/attachment.htm From funderwood <@t> mcohio.org Thu Sep 25 14:48:26 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Excessively wrinkled sections Message-ID: I am dealing with human hearts, but I'll throw in my 2 cents. You could safely raise your water bath temp. to a range of 43 to 45 degrees C. Much higher than that and the paraffin begins to melt on the water. An aid that I use constantly is to have some kind of hot plate ( one used in making stains or solutions will work fine) and adjust the temperature to 55 degrees C, or so. Quickly touch the slide ( with section mounted) to the hotplate, this will help pull out wrinkles. It will take you a few times of trial and error to get the correct temp. and technique, but it works great. Also, I limit the amount of contact with water while the blocks are cooling, that is if you face your blocks prior to cooling. Fred >>> "Barlow, Gillian" 09/25/03 02:00PM >>> Dear Hisonetters First, many thanks to those of you who replied to help me with my cardiac fixations - I now have nice, well-fixed tissues. However, one problem till remains - the sections are very wrinkled and do not flatten out well on the waterbath, even when left for long periods of time. I store my blocks at room temperature, but put them on ice for at least two hours before sectioning. The waterbath is at 40 degrees. A colleague suggested increasing the waterbath temperature, but the sections are for IHC and I dont want to damage my antigen. Many thanks Gillian PS For anyone out there with questions on cardiac fixation and embedding, this is the protocol I have now found to work well and wanted to share: Fixation: Hearts were isolated from anaesthetized animals, cut in half and fixed overnight in 10% buffered formalin at room temperature. The following day, hearts were transferred to 70% ethanol and stored at 4 degrees prior to embedding. Embedding: 80% EtOH, 45 mins room temperature 95% EtOH, 45 mins room temperature, two changes 100% EtOH, 45 mins room temperature, two changes Xylene, 45 mins room temperature, two changes Paraffin, 45 mins, 60 degrees under vacuum, two changes Embed immediately Gillian Barlow, PhD Postdoctoral Fellow Laboratory of Julie Korenberg, PhD, MD Cedars-Sinai Medical Center Davis Bldg, Lab 2007 110 George Burns Rd Los Angeles, CA 90048 Phone: (310) 423 7650 Fax: (310) 423 0302 From weneng <@t> hotmail.com Thu Sep 25 16:52:54 2003 From: weneng <@t> hotmail.com (Wendy England) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] PAS staining Message-ID: Hi, I used Sigma PAS kits but the staining is not satisfied. If you ever done this stain before and had successful result could you share your experience, like where you purchase your kit and what I should be more concerned? Thanks in advance. Wendy _________________________________________________________________ High-speed Internet access as low as $29.95/month (depending on the local service providers in your area). Click here. https://broadband.msn.com From algranth <@t> u.arizona.edu Thu Sep 25 16:56:43 2003 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] HT (ASCP) Exam In-Reply-To: Message-ID: <4.3.2.7.2.20030925145347.00c97710@algranth.inbox.email.arizona.edu> A microcurie is a unit of radioactivity. At 05:06 PM 9/25/2003 -0400, you wrote: >Question ! What chemical is nicknamed "Microcurie"? This question is >being asked on the HT (ASCP) Exam. Thanks for your help ! ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From gcallis <@t> montana.edu Thu Sep 25 17:22:28 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Microcurium In-Reply-To: Message-ID: <3.0.6.32.20030925162228.00b959c8@gemini.msu.montana.edu> Curium is actually an element, not a chemical. Microcurie is a rate of decay - this can be found on web, but did not refer to a chemical. See description of curium, obviously named for Madame Curie and her husband. Most compounds of Cm(III) are faintly yellow. If curium enters the body it accumulates in the bones, and is therefore very toxic as its radiation destroys the red-cell forming mechanism. Curium is a radioactive rare earth metal. The most stable isotope is 247Cm which has a half-life of 16 million years. Curium is probably present in uranium ores. It has a few specialised uses but only a few of its compounds are known. Were you given choices on what "Microcurium" could be, as a nickname for a chemical? The only chemicals we spend a great deal of time with in staining procedures that are radioactive are uranyl nitrate and uranyl acetate, salt of uranium. However, curium IS found in uranium ores so it was taking a maybe (or not so) educated guess here. I'll bet John Kiernan will know this one. Strange question, I have never seen this nickname in over 30+ years in histotechnology! Guess I am missing something of historical significance??? 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 georgecole <@t> ev1.net Thu Sep 25 23:43:23 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Missing procedure for silver preparations Message-ID: <000001c383e8$b8a7ec90$044dbad0@hppav> PARDON PARDON PARDON I left out a page of the Nerve and Muscle procedures when I sent them out. It is a method that keeps the nerve tissue sections on the slide during silver impregnations. Silver nitrate is notorious for loosening sections on a slide. The preparation uses Claoue's Cement--- adapted from a process in The Microtomist's Formulary and Guide by Peter Gray, from the Blakiston Co.inc It's main ingredient: Collodion---Mallinkrodt order number UN 1993 FORMULA: Claoue's Cement: Collodion 8 ml 100% Ethyl Alcohol 92 ml Camphor 2 gm Castor Oil 3 ml PROCEDURE: I usually mixed 2 or 3 times the above amounts to make 200 or 300 ml of Claoue's. Kept tightly closed, it keeps for a long time. I know this looks like snake oil, but it works. Regular pyroxylin or celloidin solutions in alcohol and ether alone, can form a kind of sheet which can peel off taking the tissues with it. This prep forms an emulsion which keeps the tissues on the slides very nicely. Paraffin Sections: Remove paraffin as usual. After the last 95% alcohol, remove one or two slides from the alcohol to put on a clean paper towel. Place two or three sheets of bibulous paper very carefully on the slides. Blot gently with two or three fingers for two or three strokes from one end of the slide to the other. Carefully turn the bibulous paper over and repeat the blotting as before. This smoothes wrinkles and fixes the tissue to the slide. With an eye dropper, apply enough drops of Claoue's to cover the sections and to the edge of the slide where the sections are. Drain any surplus Claoue's off by touching both edges of the slides to the bibulous paper. Blow gently on the slides until the Claoue's turns opaque. Rinse in two changes of 95% ethyl alcohol and proceed with what ever stain you are doing. This preparation is compatible with silver impregnations and the Kluver-Berrera beautiful luxol fast blue and cresyl fast violet---and many others. When the silver impregnation is done, the slides are run up the alcohols. In the first two 100% alcohols, the Claoue's coating will come off with some silver giving it color. After two more 100% alcohols, clear and cover as usual. Be careful in these final stages. With the Claoue's covering removed, the sections may loosen if handled roughly. I say this, although I have never lost a section at this time in the procedure. But I'm always careful there anyway. Good luck with your silver preps. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030925/10fce419/attachment.htm From jkiernan <@t> uwo.ca Fri Sep 26 00:02:25 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Microcurium References: <3.0.6.32.20030925162228.00b959c8@gemini.msu.montana.edu> Message-ID: <3F73C861.B40CE01@uwo.ca> Congrats, Gayle on a superb injection of chemistry and lexicography into the Histonet Continuum. You're 100% on the spot as usual. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ___________________ Gayle Callis wrote: > > Curium is actually an element, not a chemical. Microcurie is a rate of > decay - this can be found on web, but did not refer to a chemical. > > See description of curium, obviously named for Madame Curie and her husband. > > Most compounds of Cm(III) are faintly yellow. If curium enters the body it > accumulates in the bones, and is > therefore very toxic as its radiation destroys the red-cell forming > mechanism. Curium is a radioactive rare earth metal. The most stable > isotope is 247Cm which has a half-life of 16 million years. Curium is > probably present in uranium ores. It has a few specialised uses but only a > few of its compounds are known. > > Were you given choices on what "Microcurium" could be, as a nickname for a > chemical? The only chemicals we spend a great deal of time with in > staining procedures that are radioactive are uranyl nitrate and uranyl > acetate, salt of uranium. However, curium IS found in uranium ores so it > was taking a maybe (or not so) educated guess here. I'll bet John Kiernan > will know this one. > > Strange question, I have never seen this nickname in over 30+ years in > histotechnology! Guess I am missing something of historical significance??? > > 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 c.m.vanderloos <@t> amc.uva.nl Fri Sep 26 02:25:50 2003 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] RE: Making DAB+ in Dako ARKdarker Message-ID: <26826e263574.26357426826e@amc.uva.nl> Hi Ian and Gayle, There are also non-kit and much cheaper ways to stain DAB any darker than the usual brown/yellow. You may add 20 ul of 1% Ni(II) chloride.6H2O (in dist. water) to 1 ml of a ready-to-use DAB chromogen solution. The result is a blue/black reaction product. Or, 20 ul of 1% Co(II)chloride.6H2O to obtain a brown/black precipitate. Both procedures are described by Hsu and Soban, JHC 30:1079-1082, 1982. There is also a possibility to post-stain a weakly DAB stained section with a diluted osmium solution, turning the weakly yellow DAB into black. Unfortunately I have lost the exact conditions and reference for this one. Have a happy (dark) staining day. Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands >----- Original Message ----- >From Gayle Callis >Date Thu, 25 Sep 2003 08:09:22 -0600 >To Ian Montgomery , >Histonet@lists.utsouthwestern.edu >Subject [Histonet] Making DAB+ in Dako Ark darker > >You can make the brown endproduct a very dark, black brown (almost >black!) by using the DAKO DAB enhancer. This works beautifully with >their DAB+ that is in ARK kit or you can always do another enhancer to >turn the product black. This has been discussed at great lengths in >Histonet archives. Other companies have enhancers ready to use, Zymed >and Vector, but I am not sure if they are black. >Since the kit is a kit, we prefer to use the DAKO enhancing product, >pretty much optimized for their DAB+ which (with their enhancer), >allowed us to get a murine CD4 diluted out 1:15,000 (0.5mg/ml) and >more. This was not an ARK kit method, but we were impressed with the >overall chromogen/enhancer. We had to be careful about any residual >peroxidase or pseudoperoxidases in tissues when using DAKO DAB >enhancer. We used the glucose oxidase endogenous peroxidase method for >quenching with very clean results. > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 > >>At 01:25 PM 9/25/2003 +0100, you wrote: >>Although the brown reaction product with Dako Ark looks lovely >>I'd like a black product. Has anyone used intensifying techniques >>with Dako Ark and can they offer any hints and tips. >>Ian. >>Dr. Ian Montgomery, >>Histotechnology, >>Graham Kerr Building, >>Institute of Biomedical& Life Sciences, >>University of Glasgow, >>Glasgow G12 8QQ. >> e-mail: ian.montgomery@bio.gla.ac.uk From docmichel <@t> netbulmail.com Fri Sep 26 03:25:01 2003 From: docmichel <@t> netbulmail.com (izzet oguz) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] apoptosis Message-ID: <3805.193.255.128.130.1064564701.webmail@mail1.netbulmail.com> Hi everyone, Is immunohistochemistry (K?-67, p53 etc.) enough for detecting the apoptotic cells or does it need to be support by another method for precise detection? Can you share your experiences? By the way I will work with kidney tissue... Thank you, ?zzet From g.lang <@t> bigfoot.de Fri Sep 26 05:43:00 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Excessively wrinkled sections References: <3CFAA0108952D111A5BF00805FA6FB0F0464D454@PEDSNTAS.csmc.edu> Message-ID: <002201c3841a$f4a84370$0d12a8c0@SERVER> It would help to put the sections first in a room temperature waterbath. (small glass). There you can flatten it carefully with a brush. Mount it on a slide and let it slowly into the warm water. It is a easy method for people that learn cutting paraffinsections. We let the slides for a few minutes on the border of the waterbath to dry. The temperature of the waterbath is 40 degrees. greetings from Austria Gudrun Lang ----- Original Message ----- From: "Barlow, Gillian" To: "'Histonet'" Sent: Thursday, September 25, 2003 8:00 PM Subject: [Histonet] Excessively wrinkled sections > Dear Hisonetters > > First, many thanks to those of you who replied to help me with my cardiac > fixations - I now have nice, well-fixed tissues. However, one problem till > remains - the sections are very wrinkled and do not flatten out well on the > waterbath, even when left for long periods of time. I store my blocks at > room temperature, but put them on ice for at least two hours before > sectioning. The waterbath is at 40 degrees. A colleague suggested > increasing the waterbath temperature, but the sections are for IHC and I > dont want to damage my antigen. > > Many thanks > Gillian > > PS For anyone out there with questions on cardiac fixation and embedding, > this is the protocol I have now found to work well and wanted to share: > > Fixation: > Hearts were isolated from anaesthetized animals, cut in half and fixed > overnight in 10% buffered formalin at room temperature. The following day, > hearts were transferred to 70% ethanol and stored at 4 degrees prior to > embedding. > > Embedding: > 80% EtOH, 45 mins room temperature > 95% EtOH, 45 mins room temperature, two changes > 100% EtOH, 45 mins room temperature, two changes > Xylene, 45 mins room temperature, two changes > Paraffin, 45 mins, 60 degrees under vacuum, two changes > Embed immediately > > > > Gillian Barlow, PhD > Postdoctoral Fellow > Laboratory of Julie Korenberg, PhD, MD > Cedars-Sinai Medical Center > Davis Bldg, Lab 2007 > 110 George Burns Rd > Los Angeles, CA 90048 > > Phone: (310) 423 7650 > Fax: (310) 423 0302 > From histo <@t> skm.org.pk Fri Sep 26 05:59:06 2003 From: histo <@t> skm.org.pk (Histology) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] semiquantitative scoring system Message-ID: Does anyone out in histoland know semiquantitative scoring system (ER & PR)? If so could you please share with me this procedure. Thanks for the help.... Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk From vanlochemstraat <@t> yahoo.com Fri Sep 26 06:46:22 2003 From: vanlochemstraat <@t> yahoo.com (damien desmond) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] unsubscribe please Message-ID: <20030926114622.81929.qmail@web60203.mail.yahoo.com> Thanks for everything but I am going to the core lab Please unsubscribe me --------------------------------- Do you Yahoo!? The New Yahoo! Shopping - with improved product search -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030926/195a8aea/attachment.htm From Mauricio.Morais <@t> tufts.edu Fri Sep 26 06:56:00 2003 From: Mauricio.Morais <@t> tufts.edu (Mauricio S. Morais) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Thank you for reply: Manipulation time of human muscle biopsies and formation of freezing artifacts Message-ID: <3F742950.3EE43B8A@tufts.edu> Hello Histonet community. I want to thank you all for sending suggestions to help me walking the pathways of the "Histo World". Special thanks to George Cole for sending the DVDs. We look forward to watch and learn. My apologies for delaying this Thanks mail. Below I add all replies I had to help others in future search through the Histonet Archives. Thank you very much. Mauricio S. Morais Senior Research Technician Nutrition, Exercise, Physiology, and Sarcopenia Laboratory (NEPS) TUFTS University Jean Mayer USDA Human Nutrition Research Center on Aging 711 Washington Street, Rm 436 Boston, MA 02111 (617) 556-3226 Replies received: 1- Mauricio; Of the thousands of muscle biopsies performed in my lab, I never saw ice crystals formed by quick searches through their containers in the freezer. They were kept about 70 0r 80 per box in a -70C freezer. I kept an empty box in the freezer. When searching for a case, I would look at the biopsies one at a time in their small individual plastic zip lock bags, ID'd by heavy lead pencil on 1.5 inch pieces of tongue blade----any paper or cardboard will curl and become hard to read---I would read the tag on the biopsy while in the freezer, putting them in to the cold empty box in the freezer until the one I was looking for showed up. I would then return the bags I had just looked at to their storage box. Put the box back where it was, and carry the bag with the biopsy I had just found 30 paces, holding it in the air, to my lab and put it in the cryostat. I would do all I had to do to the biopsy in the cryostat. The Leitz cryostat had a metal kind of shield in the bottom which I promptly took out so I could keep a 100 slide box in the botom. I cut sections onto slides and put them directly into slide boxes in the bottom of the cryostat. When I had to do anything with a block, I would put it in the cold box in the bottom of the cryostat. Never, ever did I see ice crystals added to a biopsy handled this way. Freezing artifact is the province of initial freezing of the muscle. My packet of 3 DVD's and 18 pages of procedures shows the entire orientation, freezing and cutting of muscle, I will be glad to send you one, if you wish me to. There is no charge for the packet. The DVD's are crude, but they do manage to show how it's done. georgecole@ev1.net 2- I can give you some advice on how to keep things from thawing but my experience comes from a frozen brain bank. I was given the job of cataloging every piece on frozen tissue from dissected human AD brains and normal brains as well. It was a very very large job and I am not sure about muscle but brain thaws pretty fast. You are not even supposed to handle them with gloved hands for more than a few seconds. What I used to do was get a styrofoam box of dry ice and anything that I took out of the freezer for moving around or cataloging purposes I put in the dry ice. That way it never thawed. I used a large box and lots of dry ice. I hope this helps you a little. Good luck. I had four large -80 freezers packed to the brim. It took me almost a year. Lori "Gehan, Loralee" 3- Hi! We have found it helpful, and best for specimen preservation to do the organization with the specimens setting in an open cooler on a bed of dry ice, only removing from the freezer the amount you can work with in this space. Another alternative is to work inside a cryostat. Good luck! It sounds like you have quite a challenge ahead of you! Bonnie Whitaker Technical Director, Histology Laboratory Department of Pathology and Laboratory Medicine UT Med School 6431 Fannin MSB 2.231 Houston, TX 77030 713-500-6792 4- My advice would be to work with the boxes in the cryostat that is usually set at -20C. Maybe a couple of boxes of tissue at one time until you can catalog their contents and then begin rearranging your frozen tissues to the appropriate boxes. Again a couple of boxes at a time. I would only remove the whole rack just long enough to remove the needed boxes. I work with volumes of muscle samples held at -70C. When I need to pull a particular case out I will remove a box from the -70C and work very quickly, maybe for 1-2 minutes maximum. It all begins to thaw very quickly. I don't know if there is a proven study that deals with this type of problem - but if anyone responds to you quoting a reference I would appreciate learning that information also. Hope some of this helps Jean Mitchell University of Wisconsin Hospital & Clinics Madison, WI "Mitchell (Jean A.)" 5- Mauricio: If you have access to a walk-in freezer as we do, you may be able to transfer the racks/boxes to the walk-in using styrofoam shipping containers or a large plastic cooler (like those used for camping) containing dry ice. Once in the walk-in freezer you would be able to work with the specimens for some time without having them thaw out (handle with gloves to insulate your hands from the cold and the specimens from your body heat. However, most walk-in freezers (-20 C) are not as cold as the cryofreezers commonly used here to store muscles (-70 C). But the closet-sized sized freezer might offer enough working time to allow you to sort and label without much loss or damage. Good luck, Joe Galbraith "Galbraith, Joe" 6- To sustain a lengthy sort of biopsies, buy a big block(s) of dry ice and pulverize it so you can lay things on top of ice and/or surround boxes with mutiple smaller dry ice chunks and still be able to handle the vials while sorting. Use good insulation of hands and forceps (I have some LONG Russian forceps with large rounded serrated tips - shoved into dry ice to keep cold or long, grippy hemostats to pick up vials. Vials can be laid on or shoved into pulverized dry ice and certainly stay out of a freezer for more than 5 minutes as long as they are surrounded by dry ice (hmmm approx -70 to -90C). BE sure to use a styrofoam container to hold the dry ice, open boxes can be worked with at RT for as long as you need. You could still work in a cold room with dry ice, however close quarters could be a problem - see very last comment. If possible, try to recycle some dry ice pellets (Sigma et al use to ship biologicals, antibodies, etc in styrofoam containers and if possible stockpile pellets in -80C freezer in a styrofoam box, some means for this purpose (hope you have some space!) We do this for snap freezing purposes and rarely buy dry ice blocks. We sort large 50 ml tubes w/frozen tissue blocks using dry ice method frequently and the biggest problem is cold hands, take care to insulate your fingers from snap freezing! I have seen people use thinner silk glove/liners from Winter Silks inside latex gloves and forceps to handle (juggle?) tubes. We find the dry ice easier to work with than colder, fog creating liquid nitrogen where I end up going on a frustrating fishing for tubes somewhere in bottom of a dewar. Be sure you have good ventilation, you don't want to be found passed out on the floor due to oxygen depletion by CO2 or N2 fumes. Good luck Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 7- After removal of boxes from the cryo freezer, take it to your cryostat and work there, Make sure you maintain your cryostat temp at -24 or higher. You can work with your regular gloves but try not to hold the tissue ,hold it on the cork or use forceps that have been freeze from the liquid nitrogen. After each tissue checked or organized. freeze it to liquid nitrogen to prevent ice crsytal formation and to keep the integrity of you tissue. I hope this can help you with your problem on organizing your freezer works. I'm working on muscle tissue and i don't have problems. i may have few problems only when the tissue was not properly freezed. Good luck. reuel cornelia tsrh Reuel Cornelia -------------- next part -------------- A non-text attachment was scrubbed... Name: Mauricio.Morais.vcf Type: text/x-vcard Size: 1220 bytes Desc: Card for Mauricio S. Morais Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030926/2a12abdf/Mauricio.Morais.vcf From mari.ann.mailhiot <@t> leica-microsystems.com Fri Sep 26 08:38:29 2003 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] PAS staining Message-ID: Wendy Try using Schiff's purchased from Surgipath. I had great success using it. 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 From Rcartun <@t> harthosp.org Fri Sep 26 08:21:04 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] semiquantitative scoring system Message-ID: See: D. Craig Allred, et al.: "Prognostic and predictive factors in breast cancer by immunohistochemical analysis". Modern Pathology 1998;11(2): 155-168. R. Cartun >>> "Histology" 09/26/03 06:59AM >>> Does anyone out in histoland know semiquantitative scoring system (ER & PR)? If so could you please share with me this procedure. Thanks for the help.... Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From siksik <@t> vgernet.net Fri Sep 26 09:46:34 2003 From: siksik <@t> vgernet.net (Steven Slap) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] isopropanol in processing Message-ID: Hi HistoNetters As John Kiernan pointed out, isopropanol is not miscible with molten paraffin. In microwave processing, the paraffin is heated above the boiling point of the isopropanol (in some instruments, under vacuum, allowing lower temperatures to be employed), and the remaining isopropanol is "flash evaporated" out of the tissue. Steven Slap From funderwood <@t> mcohio.org Fri Sep 26 09:55:35 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] RE: Making DAB+ in Dako ARKdarker Message-ID: A few years and brain cells ago I remember incubating slides (after DAB development) in a dilute silver nitrate solution at 60 degrees C. The DAB would blacken nicely, but incubating too long could cause background staining. The concentration and times escapes me however. Fred >>> "C.M. van der Loos" 09/26/03 03:25AM >>> Hi Ian and Gayle, There are also non-kit and much cheaper ways to stain DAB any darker than the usual brown/yellow. You may add 20 ul of 1% Ni(II) chloride.6H2O (in dist. water) to 1 ml of a ready-to-use DAB chromogen solution. The result is a blue/black reaction product. Or, 20 ul of 1% Co(II)chloride.6H2O to obtain a brown/black precipitate. Both procedures are described by Hsu and Soban, JHC 30:1079-1082, 1982. There is also a possibility to post-stain a weakly DAB stained section with a diluted osmium solution, turning the weakly yellow DAB into black. Unfortunately I have lost the exact conditions and reference for this one. Have a happy (dark) staining day. Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands >----- Original Message ----- >From Gayle Callis >Date Thu, 25 Sep 2003 08:09:22 -0600 >To Ian Montgomery , >Histonet@lists.utsouthwestern.edu >Subject [Histonet] Making DAB+ in Dako Ark darker > >You can make the brown endproduct a very dark, black brown (almost >black!) by using the DAKO DAB enhancer. This works beautifully with >their DAB+ that is in ARK kit or you can always do another enhancer to >turn the product black. This has been discussed at great lengths in >Histonet archives. Other companies have enhancers ready to use, Zymed >and Vector, but I am not sure if they are black. >Since the kit is a kit, we prefer to use the DAKO enhancing product, >pretty much optimized for their DAB+ which (with their enhancer), >allowed us to get a murine CD4 diluted out 1:15,000 (0.5mg/ml) and >more. This was not an ARK kit method, but we were impressed with the >overall chromogen/enhancer. We had to be careful about any residual >peroxidase or pseudoperoxidases in tissues when using DAKO DAB >enhancer. We used the glucose oxidase endogenous peroxidase method for >quenching with very clean results. > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 > >>At 01:25 PM 9/25/2003 +0100, you wrote: >>Although the brown reaction product with Dako Ark looks lovely >>I'd like a black product. Has anyone used intensifying techniques >>with Dako Ark and can they offer any hints and tips. >>Ian. >>Dr. Ian Montgomery, >>Histotechnology, >>Graham Kerr Building, >>Institute of Biomedical& Life Sciences, >>University of Glasgow, >>Glasgow G12 8QQ. >> e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ronnie_Houston <@t> bshsi.com Fri Sep 26 11:32:52 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Ret IHC Message-ID: <530361BF03351B4CAE5270A05D3037B5FE043A@bsrexms01.BSHSIR.COM> Richard, We were going to set this up a few months ago, and then had a new pathologist, Lisa Cerilli, join us from UVA. She had studied and compared Ret to CK19 for follicular thyroid lesions and came out in favor of CK19. They did use the monolconal from Novocastra though. Here is the abstract of her paper: Am J Clin Pathol. 2002 Aug;118(2):186-93. Interpretation of RET immunostaining in follicular lesions of the thyroid. Cerilli LA, Mills SE, Rumpel CA, Dudley TH, Moskaluk CA. Robert E Fechner Laboratory of Surgical Pathology, University of Virginia Health Sciences Center, Charlottesville, USA. We applied monoclonal antibodies against RET and cytokeratin 19 (CK19) to the following tumor sections: classic papillary carcinoma (PC), 16; Hurthle-type PC (HPC), 1; sclerosing PC with nodular fasciitis-like stroma (SPC), 1; PC, follicular variant (FVPC), 12; follicular adenoma (FA), 9; Hurthle cell adenoma (HA), 4; Hurthle cell carcinoma (HC), 3; and follicular carcinoma (FC), 7. CK19+ tumors included 16 PCs, 1HPC, 1SPC, 11 FVPCs, 7 FAs, 4 FCs, and 1HC. RET+ tumors included 4 HAs, 3 HCs, 1HPC, 12 PCs, 7 FVPCs, and 2 FAs. Reverse transcriptase-polymerase chain reaction (RT-PCR) revealed a RET transcript in 6 Hurthle cell lesions. RET immunoreactivity is less sensitive and specific for PC than CK19. CK19 is useful for identifying PC, although only lesions with diffuse, intense staining should be considered positive. The detection of RET protein by immunohistochemical analysis was corroborated by the presence of the RET transcript by RT-PCR. Further study is warranted to determine whether this represents activation by gene fusion or some other mechanism in this subset of thyroid neoplasms. I would be interested in your results though. Best wishes Ronnie Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Wednesday, September 17, 2003 12:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Ret IHC We are do "Ret" immunoperoxidase staining on thryoid tissue using a rabbit polyclonal antibody from Santa Cruz (sc-167). Does anyone have experience with this antibody on formalin-fixed, paraffin-embedded tissue? Thank you. Richard Cartun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at 410-442-3250, and permanently delete the original e-mail, attachment(s), and any copies. From weneng <@t> hotmail.com Fri Sep 26 11:56:53 2003 From: weneng <@t> hotmail.com (Wendy England) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] Thanks! (PAS staining) Message-ID: Many thanks to all who answered my PAS staining question! Your guide, suggestions and comments are really helpful. Histonet is the best! Wendy _________________________________________________________________ Get McAfee virus scanning and cleaning of incoming attachments. Get Hotmail Extra Storage! http://join.msn.com/?PAGE=features/es From funderwood <@t> mcohio.org Fri Sep 26 11:50:22 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] PAS staining Message-ID: I also use surgipath's schiff's and get good results. fred >>> 09/26/03 09:38AM >>> Wendy Try using Schiff's purchased from Surgipath. I had great success using it. 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkarazub <@t> ucsd.edu Fri Sep 26 12:24:12 2003 From: jkarazub <@t> ucsd.edu (Jeni Karazuba) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] apical positive control Message-ID: <002601c38453$00fe1770$036aef84@histology> Does anyone know of a good reliable positive control antibody that stains only the apical membrane of gut epithelium? I haven't been able to find any good information on this. Thanks! ~Jeni Karazuba UCSD Medicine -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030926/67b33cad/attachment.htm From jlambrey <@t> hotmail.com Fri Sep 26 13:43:01 2003 From: jlambrey <@t> hotmail.com (Julien Lambrey de Souza) Date: Fri Sep 16 15:21:56 2005 Subject: [Histonet] automated immunostainers Message-ID: Hi All, As a new developmental biology lab, we are looking to acquire an automated immunostainer to stain all types of collagen and probably bone precursor. Do you have a particular machine to recommend? Although we have looked at different stainers (Zymed, Genex, Optimax, Dako, Labvision), each compagny seems to have the best machine for our particular needs. We will be processing small amounts (compared to hospitals for instance) but need the automatisation because of time consuming other histological techniques we use. The brochures are all very interesting but hardly reflect the reality of laboratory routine work. Any help is welcome. Thanks a lot, Julien. _________________________________________________________________ MSN 8 with e-mail virus protection service: 2 months FREE* http://join.msn.com/?page=features/virus From settembr <@t> umdnj.edu Fri Sep 26 15:14:04 2003 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] automated immunostainers Message-ID: I had a Dako and it worked well and the training went well and their technical support is fantastic, so we got another. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> Julien Lambrey de Souza 9/26/2003 11:43:01 AM >>> Hi All, As a new developmental biology lab, we are looking to acquire an automated immunostainer to stain all types of collagen and probably bone precursor. Do you have a particular machine to recommend? Although we have looked at different stainers (Zymed, Genex, Optimax, Dako, Labvision), each compagny seems to have the best machine for our particular needs. We will be processing small amounts (compared to hospitals for instance) but need the automatisation because of time consuming other histological techniques we use. The brochures are all very interesting but hardly reflect the reality of laboratory routine work. Any help is welcome. Thanks a lot, Julien. _________________________________________________________________ MSN 8 with e-mail virus protection service: 2 months FREE* http://join.msn.com/?page=features/virus _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tflore <@t> lsuhsc.edu Fri Sep 26 15:30:32 2003 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Histotechnology Registry Study Group New Orleans, LA Message-ID: FYI There will be a monthly study group, for the Histotechnology ASCP Registry. Classes will be held at LSUHSC, MEB Building, 3 rd floor, Seminar Room 8, beginning tomorrow (Saturday) from 8am-Noon. Rosemary Velasquez, Histotechnologist from Ochsner Foundation Hospital, will head the study group. This will be Ms Velasquez second study group. There were five technologist in Ms Velasquez first study group and all five passed the HT(ASCP) registry the first time they took it. Please bring pencil and paper for notes. There is parking around the MEB building, as meters are free on Saturday. Entrance is in front of the LSU Research Building (the other door of the MEBuilding, on the side of I-10 is locked on weekends. Please sign in at the front desk with the security guard and come up the 3rd floor. Signs will be posted. Vending machines are available, bring change. I will also be here to see if Rosemary needs my help with anything. Teresa Flores -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030926/b9f80128/attachment.htm From GrobeA <@t> saintpatrick.org Fri Sep 26 16:40:50 2003 From: GrobeA <@t> saintpatrick.org (Albert Grobe) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Re:Excessively Wrinkled sections Message-ID: Gillian, Since your samples have been through the 60C paraffin, raising the waterbath temperature a few degrees should not affect you epitopes. Ideally, the waterbath should be a couple degrees below the melting temp of the wax but some fine adjustment is sometimes necessary. I have had the same problem in the past, and raising the temp has helped. Albert Albert C. Grobe, MS Tissue Engineering Lab International Heart Institute Missoula, MT 59802 Office: (406) 329-5634 E-Mail: GrobeA@saintpatrick.org From GrobeA <@t> saintpatrick.org Fri Sep 26 16:40:50 2003 From: GrobeA <@t> saintpatrick.org (Albert Grobe) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Re:Excessively Wrinkled sections Message-ID: Gillian, Since your samples have been through the 60C paraffin, raising the waterbath temperature a few degrees should not affect you epitopes. Ideally, the waterbath should be a couple degrees below the melting temp of the wax but some fine adjustment is sometimes necessary. I have had the same problem in the past, and raising the temp has helped. Albert Albert C. Grobe, MS Tissue Engineering Lab International Heart Institute Missoula, MT 59802 Office: (406) 329-5634 E-Mail: GrobeA@saintpatrick.org From jkiernan <@t> uwo.ca Fri Sep 26 23:30:59 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Excessively Wrinkled sections References: Message-ID: <3F751283.D2167356@uwo.ca> Is it really OK to float out sections on water that is "a couple degrees below the melting temp of the wax"? Ribbons of 58C wax extend nicely on water at 40-45C. I've never dared to float ribbons on a water bath in the 50s because I've seen the bad effects of high temperature when doing on-the-slide flotation on a 55C hotplate. That's the way I was taught, by technicians who were skilled enough to do it well. Hot water under a paraffin ribbon yields up its dissolved air in the form of many tiny bubbles. These form under the sections and cause lousy staining for obvious reasons. When wax melts in the presence of even a thin film of water under a ribbon of sections you have a recipe for disaster. If you can cut, float and mount "ideal" sections from water that's only 2C below the nominal melting point of the wax, please post detailed instructions. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ____________________________________ Albert Grobe wrote: Ideally, the waterbath should be a couple degrees below the melting temp of the wax but some fine adjustment is sometimes necessary. I have had the same problem in the past, and raising the temp has helped. ____________________________________ From jkiernan <@t> uwo.ca Fri Sep 26 23:30:59 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Excessively Wrinkled sections References: Message-ID: <3F751283.D2167356@uwo.ca> Is it really OK to float out sections on water that is "a couple degrees below the melting temp of the wax"? Ribbons of 58C wax extend nicely on water at 40-45C. I've never dared to float ribbons on a water bath in the 50s because I've seen the bad effects of high temperature when doing on-the-slide flotation on a 55C hotplate. That's the way I was taught, by technicians who were skilled enough to do it well. Hot water under a paraffin ribbon yields up its dissolved air in the form of many tiny bubbles. These form under the sections and cause lousy staining for obvious reasons. When wax melts in the presence of even a thin film of water under a ribbon of sections you have a recipe for disaster. If you can cut, float and mount "ideal" sections from water that's only 2C below the nominal melting point of the wax, please post detailed instructions. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ____________________________________ Albert Grobe wrote: Ideally, the waterbath should be a couple degrees below the melting temp of the wax but some fine adjustment is sometimes necessary. I have had the same problem in the past, and raising the temp has helped. ____________________________________ From nick.kirk3 <@t> btopenworld.com Sat Sep 27 00:34:20 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Excessively Wrinkled sections In-Reply-To: <3F751283.D2167356@uwo.ca> Message-ID: I've always floated sections out on a water bath in the low to mid 50s with hardly ever any problems. The key is to use de-ionised water which will reduce bubble formation and to give the water bath a tap (or thump depending on your mood) every now and again to release any bubbles that may have formed on the base or sides to allow them to float to the surface and pop harmlessly. Floating out at that temperature is also a good indicator of how well processed your tissue is, as poorly processed tissue will "explode" across the water bath and in my book, if it does that, it needs to be re-processed. Floating out at a lower temperature will still remove the wrinkles but it takes a lot longer and you have to weigh up the risk of bubbles against the extra time needed to complete the task. If your department isn't that busy, then the lower temperature is an option. If, however, you work in a busy department, you probably can't afford to sit around twiddling your thumbs all day waiting for sections to flatten out. It's a case of Risk Assessment and in the labs I've worked in over the years, we've always opted for the hotter option with no problem. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of John Kiernan Sent: 27 September 2003 05:31 To: Albert Grobe Cc: histonet@lists.utsouthwestern.edu; histonet@pathology.swmed.edu Subject: [Histonet] Excessively Wrinkled sections Is it really OK to float out sections on water that is "a couple degrees below the melting temp of the wax"? Ribbons of 58C wax extend nicely on water at 40-45C. I've never dared to float ribbons on a water bath in the 50s because I've seen the bad effects of high temperature when doing on-the-slide flotation on a 55C hotplate. That's the way I was taught, by technicians who were skilled enough to do it well. Hot water under a paraffin ribbon yields up its dissolved air in the form of many tiny bubbles. These form under the sections and cause lousy staining for obvious reasons. When wax melts in the presence of even a thin film of water under a ribbon of sections you have a recipe for disaster. If you can cut, float and mount "ideal" sections from water that's only 2C below the nominal melting point of the wax, please post detailed instructions. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ____________________________________ Albert Grobe wrote: Ideally, the waterbath should be a couple degrees below the melting temp of the wax but some fine adjustment is sometimes necessary. I have had the same problem in the past, and raising the temp has helped. ____________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From georgecole <@t> ev1.net Sat Sep 27 01:50:35 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] replacing two notices with a corrected noitice Message-ID: <000001c384c3$a86b9da0$0b4dbad0@hppav> Two notices placed 9/25 and 9/26 need to be removed. They are both titled "Missing procedure for silver preps." They tell of procedures that should be done under the hood. I gave the procedures but did not include the warning. I will replace them with the safety warning added. Thank You georgecole@ev1.net -------------- next part -------------- A non-text attachment was scrubbed... Name: winmail.dat Type: application/ms-tnef Size: 3464 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030926/487f4ed0/winmail.bin From tflore <@t> lsuhsc.edu Sat Sep 27 10:47:04 2003 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Histotechnology Registry Study Group New Orleans, LA Message-ID: Atoska, this morning we covered fixation and getting ready to go into embedding; 7 students showed up and two are from Lafayette, LA; they have had a test on fixation; visited my EM Lab and plan I plan on having a future EM workshop hands on; also planned is a cytology lecturer (as one of the techs that took the HT Registry ASCP said that there were a lot of cytology questions). So I am comfortable in saying its for both HT and HTL. The first study group was done by Rosemary Velasquez and they would meet at her house. And I was wrong about the first study group. Of 15 that would meet weekly, 6 took the HT registry and 1 took the HTL and all 7 passed! This is the first study group at LSUHSC in New Orleans, LA Teresa -----Original Message----- From: Atoska S. Gentry [mailto:gentras@vetmed.auburn.edu] Sent: Friday, September 26, 2003 4:47 PM To: Flores, Teresa Subject: Re: [Histonet] Histotechnology Registry Study Group New Orleans, LA Hi Teresa, is this only for the HT or the HTL as well? Sounds like a great plan. I don't recall hearing or reading about the first session. Thanks, Atoska At 03:30 PM 9/26/03 -0500, you wrote: FYI There will be a monthly study group, for the Histotechnology ASCP Registry. Classes will be held at LSUHSC, MEB Building, 3 rd floor, Seminar Room 8, beginning tomorrow (Saturday) from 8am-Noon. Rosemary Velasquez, Histotechnologist from Ochsner Foundation Hospital, will head the study group. This will be Ms Velasquez second study group. There were five technologist in Ms Velasquez first study group and all five passed the HT(ASCP) registry the first time they took it. Please bring pencil and paper for notes. There is parking around the MEB building, as meters are free on Saturday. Entrance is in front of the LSU Research Building (the other door of the MEBuilding, on the side of I-10 is locked on weekends. Please sign in at the front desk with the security guard and come up the 3rd floor. Signs will be posted. Vending machines are available, bring change. I will also be here to see if Rosemary needs my help with anything. Teresa Flores Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030927/c8d8713e/attachment.htm From tflore <@t> lsuhsc.edu Sat Sep 27 10:49:14 2003 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Histotechnology Registry Study Group New Orleans, LA Message-ID: Gee, I will check into that as Rosemary Velasquez material is excellent. Do you mean like a telephone linkline or computer on line? I will investigate. Where are you located? Teresa -----Original Message----- From: Dndsomi@aol.com [mailto:Dndsomi@aol.com] Sent: Friday, September 26, 2003 4:35 PM To: tflore@lsuhsc.edu Subject: Re: [Histonet] Histotechnology Registry Study Group New Orleans, LA Since I am not in the state of Louisiana, would you be available for an on-line study group of sorts? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030927/a50b4316/attachment.htm From lpwenk <@t> mail.netquest.com Sun Sep 28 07:00:03 2003 From: lpwenk <@t> mail.netquest.com (Lee & Peggy Wenk) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Re:Excessively Wrinkled sections References: Message-ID: <00d301c385b8$11ec95a0$8732fea9@hppav> Agree with it being OK to raise the temperature of the water. The tissue has already been through hot paraffin, and, for some labs, will be in a slide dryer/warmer that is set higher than your current 40 degrees C. flotation bath. Set the flotation bath about 10 degrees C. BELOW the melting point (mp) of the paraffin. (Look on side of paraffin container.) Example, if the mp of the paraffin is 55-58 degrees C., set the flotation bath at 45 - 47 degrees C. That should take care of the wrinkles. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Albert Grobe" To: ; Sent: Friday, September 26, 2003 5:40 PM Subject: [Histonet] Re:Excessively Wrinkled sections Gillian, Since your samples have been through the 60C paraffin, raising the waterbath temperature a few degrees should not affect you epitopes. Ideally, the waterbath should be a couple degrees below the melting temp of the wax but some fine adjustment is sometimes necessary. I have had the same problem in the past, and raising the temp has helped. Albert Albert C. Grobe, MS Tissue Engineering Lab International Heart Institute Missoula, MT 59802 Office: (406) 329-5634 E-Mail: GrobeA@saintpatrick.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> mail.netquest.com Sun Sep 28 07:00:03 2003 From: lpwenk <@t> mail.netquest.com (Lee & Peggy Wenk) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Re:Excessively Wrinkled sections References: Message-ID: <00d301c385b8$11ec95a0$8732fea9@hppav> Agree with it being OK to raise the temperature of the water. The tissue has already been through hot paraffin, and, for some labs, will be in a slide dryer/warmer that is set higher than your current 40 degrees C. flotation bath. Set the flotation bath about 10 degrees C. BELOW the melting point (mp) of the paraffin. (Look on side of paraffin container.) Example, if the mp of the paraffin is 55-58 degrees C., set the flotation bath at 45 - 47 degrees C. That should take care of the wrinkles. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Albert Grobe" To: ; Sent: Friday, September 26, 2003 5:40 PM Subject: [Histonet] Re:Excessively Wrinkled sections Gillian, Since your samples have been through the 60C paraffin, raising the waterbath temperature a few degrees should not affect you epitopes. Ideally, the waterbath should be a couple degrees below the melting temp of the wax but some fine adjustment is sometimes necessary. I have had the same problem in the past, and raising the temp has helped. Albert Albert C. Grobe, MS Tissue Engineering Lab International Heart Institute Missoula, MT 59802 Office: (406) 329-5634 E-Mail: GrobeA@saintpatrick.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> mail.netquest.com Sun Sep 28 07:12:29 2003 From: lpwenk <@t> mail.netquest.com (Lee & Peggy Wenk) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] HT (ASCP) Exam References: Message-ID: <00f301c385b9$ca60eea0$8732fea9@hppav> Microcurie is not a nickname. Radiation is measured in "curies", or, if the radiation is in a low dose, in "microcuries". (Similar to how distance is measured in "meters", or if in very small distances, "micrometers".) A curie is equal to 3.70 x 10(10) (That's 10 to the 10th power = 10 billion) disintegrations per second. If you remember, radioactive chemicals decay/disintegrate, by emitting particles/radiation from their atomic nucleus (proton/neutrons), and eventually turn into a non-radioactive new chemical. The curies unit of measure therefore applies to any radioactive chemical, such as uranyl acetate, which is used in EM, or could be used with measuring the amount of radiation given off by the radioactive "dyes" used in sentinel lymph nodes, or the radioactive seeds found in prostates that have had brachytherapy. (In case you are wondering, it was named after Madame Curie, who discovered radium.) Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: Histolady710@aol.com To: histonet@lists.utsouthwestern.edu Sent: Thursday, September 25, 2003 5:06 PM Subject: [Histonet] HT (ASCP) Exam Question ! What chemical is nicknamed "Microcurie"? This question is being asked on the HT (ASCP) Exam. Thanks for your help ! -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030928/e0e40c00/attachment.htm From hodges420 <@t> msn.com Sun Sep 28 08:11:45 2003 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] (no subject) Message-ID: Good Morning all, Has anyone out there used a head hunter service to find people to work at a hospital or lab. Any names would be appreciated. Thanks Tere Hodges -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030928/1b9bc41b/attachment.htm From bhewlett <@t> cogeco.ca Sun Sep 28 17:59:04 2003 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] apical positive control References: <002601c38453$00fe1770$036aef84@histology> Message-ID: <00dd01c38614$1db8bce0$b6833918@bryaniwx13voft> Jeni, CD10 antibody will stain brush borders. Bryan ----- Original Message ----- From: Jeni Karazuba To: histonet@lists.utsouthwestern.edu Sent: Friday, September 26, 2003 1:24 PM Subject: [Histonet] apical positive control Does anyone know of a good reliable positive control antibody that stains only the apical membrane of gut epithelium? I haven't been able to find any good information on this. Thanks! ~Jeni Karazuba UCSD Medicine -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030928/eea67492/attachment.htm From ranahay <@t> fastmail.fm Sun Sep 28 19:38:48 2003 From: ranahay <@t> fastmail.fm (Rana Hay) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] H&E Message-ID: <20030929003848.599DC3C9DD@www.fastmail.fm> On Thu, 25 Sep 2003 15:08:19 +1300, "Rana Hay" said: > On Thu, 25 Sep 2003 15:06:29 +1300, "Rana Hay" > said: > > Hello Histoneters, > > > >I am interested in what medical use for the H&E stain .I have used > > it in Virology, Histopathology,Andrology and seen it used in Cytology.Any > > more ideas? > > > > Many Thanks,Rana > > -- > > Rana Hay > > ranahay@fastmail.fm > > > > -- > > http://www.fastmail.fm - Send your email first class > -- > Rana Hay > ranahay@fastmail.fm > > -- > http://www.fastmail.fm - The professional email service > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Rana Hay ranahay@fastmail.fm -- http://www.fastmail.fm - I mean, what is it about a decent email service? From gakabani <@t> nc.rr.com Sun Sep 28 23:02:13 2003 From: gakabani <@t> nc.rr.com (gamal akabani hneide) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Microtome for autoradiography Message-ID: Hi All - I need some advice regarding the best microtome to use for whole body autoradiography for rats. I am planning to buy a sledge microtome SM2400 from Leica. I am going to inject a radiolabeled MAB and sacrifice the animal with few minutes to hours, what next? What about the bone sections? Etc. Any references will be appreciated. Thanks in advanced Gamal Akabani, Ph.D. Duke University Medical Center Department of Radiology Division of Nuclear Medicine From Stanley.Stylli <@t> mh.org.au Mon Sep 29 00:50:25 2003 From: Stanley.Stylli <@t> mh.org.au (Stylli, Stanley) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] antigen retrieval buffers Message-ID: <00519F9F41D2844D9C75BEE1F6AC09AF68BA18@rmhmail1.ssg.org.au> Hi Histonetters I have a question about the longevity of antigen retrieval buffers. If you make up your own buffers such as the following : 1 mM EDTA (pH 8.0) 10 mM Citrate (pH 6.0) 10 mM tri-sodium citrate (pH 6.0) 100 mM Tris / 1 mM EDTA (pH 9.0) 10 mM Tris (pH 10.0) How long can these solutions be kept at 4C without affecting the efficiency of the buffer. Thanks Stan Stan Stylli Department of Surgery, 5th Floor Clinical Sciences Building Royal Melbourne Hospital University of Melbourne Parkville, Australia, 3052. Tel: 61-3-93427616 Fax:61-3-9347-7695 -------------- next part -------------- A non-text attachment was scrubbed... Name: Stylli, Stanley.vcf Type: text/x-vcard Size: 230 bytes Desc: Stylli, Stanley.vcf Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030929/a003f5c3/StylliStanley.vcf From tissuearray <@t> hotmail.com Mon Sep 29 01:34:31 2003 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Tissue Arrays and the Block Warmer Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030929/eb32d58d/attachment.htm From Stanley.Stylli <@t> mh.org.au Mon Sep 29 02:00:22 2003 From: Stanley.Stylli <@t> mh.org.au (Stylli, Stanley) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] apoptosis marker profile of gliomas Message-ID: <00519F9F41D2844D9C75BEE1F6AC09AF68BA19@rmhmail1.ssg.org.au> Hello again Histonetters, I will be looking at the apoptotic profile of FFPE human brain tumours (various grades - low to high). I wish to correlate the expression of various apoptotic markers to the survival of the patients after certain treatments they have been subjected to against a control group that have had surgery alone. I have read a couple of papers that have used a few antibodies but I would like to hear from people who have had success with antibodies (with and without antigen retrieval). As you can imagine, you can trial antibodies from different companies until the cows come home and the bank is empty thankyou for your help Stan Stylli Stan Stylli Department of Surgery, 5th Floor Clinical Sciences Building Royal Melbourne Hospital University of Melbourne Parkville, Australia, 3052. Tel: 61-3-93427616 Fax:61-3-9347-7695 -------------- next part -------------- A non-text attachment was scrubbed... Name: Stylli, Stanley.vcf Type: text/x-vcard Size: 230 bytes Desc: Stylli, Stanley.vcf Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030929/1dfa01a8/StylliStanley.vcf From louise_renton <@t> hotmail.com Mon Sep 29 04:26:04 2003 From: louise_renton <@t> hotmail.com (louise renton) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Excessively Wrinkled sections Message-ID: Dear Nick, John et al, Further to this discussion, increasing the temp of the waterbath does help up to a point in flattening sections, especially if, as you point out, distilled or deionised water is used. However, it was my expereience that certain tissues, most notably hypertrophic thyroid, remained wrinked even at higher temps. These sections I would pick up and then "flash melt" on the hotplate for a few seconds and then allow to drain normally. This had no deleterious effect on either morphology or staining , and produced a flatter more aesthetically pleasing section. As far as high water bath temperatures are concerned, we had debates raging for many months in our lab as to the possible destruction of antigenic epitopes by excessive incubation to melt & dry sections. Then Heat assisted epitope retrieval came on the scene and knocked that argument flat, and it was determined that excessive fixation was the real culprit.. Thus does the world around us change. Best regards, Louise Renton Johannesburg South Africa _________________________________________________________________ Unsatisfied with being single? Try MSN Personals http://www.msn.co.za/personals/ From g.lang <@t> bigfoot.de Mon Sep 29 08:22:25 2003 From: g.lang <@t> bigfoot.de (Gudrun Lang) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Excessively wrinkled sections Message-ID: <001001c3868c$b91bb4d0$0d12a8c0@SERVER> ----- Original Message ----- From: "Gudrun Lang" To: "Barlow, Gillian" Sent: Monday, September 29, 2003 11:20 AM Subject: Re: [Histonet] Excessively wrinkled sections > Hi Gillian, > I think the discussion about waterbath temperature is interesting, but the > real problem is, why are the sections very wrinkled. > In my opinion heart tissue is not so difficult to cut. We use sliding > microtoms, 3 um, Aqua dest. waterbath at 40 degrees. And if there are > wrinkles, where paraffin adhere to paraffin, my experience showed me, that > in a warm waterbath it never can be seperated and flattend ( only in cold > water with a brush). > I don't know if you are a beginner on cutting or have long experience. > So my advice is to check your cutting-angle, the sharpness of the knife, the > roughness of the tissue and don't try too long on one block (temperature). > good luck > Gudrun Lang > > ----- Original Message ----- > From: "Barlow, Gillian" > To: "'Histonet'" > Sent: Thursday, September 25, 2003 8:00 PM > Subject: [Histonet] Excessively wrinkled sections > > > > Dear Hisonetters > > > > First, many thanks to those of you who replied to help me with my cardiac > > fixations - I now have nice, well-fixed tissues. However, one problem > till > > remains - the sections are very wrinkled and do not flatten out well on > the > > waterbath, even when left for long periods of time. I store my blocks at > > room temperature, but put them on ice for at least two hours before > > sectioning. The waterbath is at 40 degrees. A colleague suggested > > increasing the waterbath temperature, but the sections are for IHC and I > > dont want to damage my antigen. > > > > Many thanks > > Gillian > > > > PS For anyone out there with questions on cardiac fixation and embedding, > > this is the protocol I have now found to work well and wanted to share: > > > > Fixation: > > Hearts were isolated from anaesthetized animals, cut in half and fixed > > overnight in 10% buffered formalin at room temperature. The following > day, > > hearts were transferred to 70% ethanol and stored at 4 degrees prior to > > embedding. > > > > Embedding: > > 80% EtOH, 45 mins room temperature > > 95% EtOH, 45 mins room temperature, two changes > > 100% EtOH, 45 mins room temperature, two changes > > Xylene, 45 mins room temperature, two changes > > Paraffin, 45 mins, 60 degrees under vacuum, two changes > > Embed immediately > > > > > > > > Gillian Barlow, PhD > > Postdoctoral Fellow > > Laboratory of Julie Korenberg, PhD, MD > > Cedars-Sinai Medical Center > > Davis Bldg, Lab 2007 > > 110 George Burns Rd > > Los Angeles, CA 90048 > > > > Phone: (310) 423 7650 > > Fax: (310) 423 0302 > > > From Marion.Hiles <@t> north-bristol.swest.nhs.uk Mon Sep 29 08:52:03 2003 From: Marion.Hiles <@t> north-bristol.swest.nhs.uk (Marion Hiles) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] automated immunostainers Message-ID: <533E4A1C3061FC4EB388119D2FD7B5D402431809@nbfexch04.north-bristol.nhs> We are a Neuropath lab and stain small runs of immuno slides (varies from 5 to 30) on the Optimax Plus machine. It is an excellent machine and very easy to use and maintain. Bob Quilty Dept of Neuropathology Frenchay Hospital Bristol, UK -----Original Message----- From: Julien Lambrey de Souza [mailto:jlambrey@hotmail.com] Sent: 26 September 2003 19:43 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] automated immunostainers Hi All, As a new developmental biology lab, we are looking to acquire an automated immunostainer to stain all types of collagen and probably bone precursor. Do you have a particular machine to recommend? Although we have looked at different stainers (Zymed, Genex, Optimax, Dako, Labvision), each compagny seems to have the best machine for our particular needs. We will be processing small amounts (compared to hospitals for instance) but need the automatisation because of time consuming other histological techniques we use. The brochures are all very interesting but hardly reflect the reality of laboratory routine work. Any help is welcome. Thanks a lot, Julien. _________________________________________________________________ MSN 8 with e-mail virus protection service: 2 months FREE* http://join.msn.com/?page=features/virus _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Nancy.Walker <@t> sanofi-synthelabo.com Mon Sep 29 08:55:31 2003 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] perfusing rodent embryos Message-ID: Dear histonetters, Can anyone explain how to perfuse rodent embryos (E18) with PFA? I've tried perfusing the mothers, this works partially, but still have a lot of red blood cells in my sections. I've heard it is possible to perfuse embryos by the umbelical cord. Does anyone have an article, pictures or schema showing how and where (finding the right artery ?) what's the appropriate needle guage, pump speed, and perfusion time, etc. thanks Nancy Walker Molecular Biology Scientist Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 From Marion.Hiles <@t> north-bristol.swest.nhs.uk Mon Sep 29 08:59:29 2003 From: Marion.Hiles <@t> north-bristol.swest.nhs.uk (Marion Hiles) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] antigen retrieval buffers Message-ID: <533E4A1C3061FC4EB388119D2FD7B5D40243180A@nbfexch04.north-bristol.nhs> We make up 5 litres of 0.01M tri-sodium citrate buffer at a time and keep it at room temperature. We decant what we need and pH it. The rest seems to keep for a few weeks without any problem. Bob Quilty Dept of Neuropathology Frenchay Hospital Bristol, UK -----Original Message----- From: Stylli, Stanley [mailto:Stanley.Stylli@mh.org.au] Sent: 29 September 2003 06:50 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] antigen retrieval buffers Importance: High Hi Histonetters I have a question about the longevity of antigen retrieval buffers. If you make up your own buffers such as the following : 1 mM EDTA (pH 8.0) 10 mM Citrate (pH 6.0) 10 mM tri-sodium citrate (pH 6.0) 100 mM Tris / 1 mM EDTA (pH 9.0) 10 mM Tris (pH 10.0) How long can these solutions be kept at 4C without affecting the efficiency of the buffer. Thanks Stan Stan Stylli Department of Surgery, 5th Floor Clinical Sciences Building Royal Melbourne Hospital University of Melbourne Parkville, Australia, 3052. Tel: 61-3-93427616 Fax:61-3-9347-7695 From KSalceies <@t> salud.unm.edu Mon Sep 29 09:01:18 2003 From: KSalceies <@t> salud.unm.edu (Kelly Salceies) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Paraffin Embedding of Mice Tissue Message-ID: Hi Everyone, I am a brand new Tech and am having trouble preping some mouse tissue for paraffin embedding. I have all different tissues (liver, heart, quads, brain, lung, and gastroc) and would like to prep all tissues (if possible) under the same conditions. I have been using the Shandon Hypercenter XP or the Shandon Histowave (microwave tissue processor...). Does anybody have a good protocol for mouse tissues on either of these instruments?? I have tried a number of protocols, but all my tissue has been really dry in cutting... Any suggestions would be greatly appreciated!! Thanks, Kelly Salceies University of New Mexico Health Sciences Center Dept. of Pathology From KSalceies <@t> salud.unm.edu Mon Sep 29 11:29:30 2003 From: KSalceies <@t> salud.unm.edu (Kelly Salceies) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Paraffin Embedding of Mice Tissue - My protocols Message-ID: Hi -- I guess I should have mentioned the protocols I have already tried - sorry...I'm new to this Histonet thing :) On the Shandon Hypercenter XP I've tried: 80% ETOH 5 min 95% ETOH 15 min 2X 100% ETOH 15 min 3X Xylene 10 min 2X Xylene 5 min 2X Paraffin 15 min 2X 50% ETOH 5 min 80% ETOH 5 min 95% ETOH 15 min 2X 100% ETOH 15 min 3X Xylene 10 min 2X Xylene 5 min Paraffin 15 min 2X 80% ETOH 5 min 95% ETOH 5 min 2X 100% ETOH 5 min 2X 1:1 ETOH: Cedarwood Oil 5 min 1:1 Cedarwood Oil:Methyl Salicylate 5 min 100% Methyl Salicylate 5 min 1:1 Methyl Salicylate:Xylene 5 min Xylene 5min Paraffin 15 min 2X --The last protocol left the tissue soft - so I might just need to use longer times...I'm not sure if anybody uses this protocol.... On the Microwave I've tried: 100% ETOH 10 min 2X 1:1 ETOH:Xylene 10 min Paraffin 10 min -All steps: Max temp 65 degrees - Watt=750 100% ETOH 5 min IPA 5 min Paraffin 5 min -All steps: Max temp 65 degrees - Watt=750 100% ETOH 5 min 1:1 IPA:Xylene 5 min Paraffin 5 min -All steps: Max temp 65 degrees - Watt=750 There are the protocols I've tried....Please let me know what suggestions you have - thank you!! Kelly Salceies University of New Mexico Health Sciences Center Dept. of Pathology From Luis.Chiriboga <@t> med.nyu.edu Mon Sep 29 12:37:19 2003 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] antigen retrieval buffers In-Reply-To: <533E4A1C3061FC4EB388119D2FD7B5D40243180A@nbfexch04.north-bristol.nhs> Message-ID: We make up 20 liter carboy of citrate, pH it let it sit at RT. Most of the time it doesn't sit around long enough to have any problems. If is a slow time, add a crystal of Thymol. Same with EDTA. Luis -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Marion Hiles Sent: Monday, September 29, 2003 9:59 AM To: 'Stylli, Stanley' Cc: Histonet (E-mail) Subject: RE: [Histonet] antigen retrieval buffers We make up 5 litres of 0.01M tri-sodium citrate buffer at a time and keep it at room temperature. We decant what we need and pH it. The rest seems to keep for a few weeks without any problem. Bob Quilty Dept of Neuropathology Frenchay Hospital Bristol, UK -----Original Message----- From: Stylli, Stanley [mailto:Stanley.Stylli@mh.org.au] Sent: 29 September 2003 06:50 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] antigen retrieval buffers Importance: High Hi Histonetters I have a question about the longevity of antigen retrieval buffers. If you make up your own buffers such as the following : 1 mM EDTA (pH 8.0) 10 mM Citrate (pH 6.0) 10 mM tri-sodium citrate (pH 6.0) 100 mM Tris / 1 mM EDTA (pH 9.0) 10 mM Tris (pH 10.0) How long can these solutions be kept at 4C without affecting the efficiency of the buffer. Thanks Stan Stan Stylli Department of Surgery, 5th Floor Clinical Sciences Building Royal Melbourne Hospital University of Melbourne Parkville, Australia, 3052. Tel: 61-3-93427616 Fax:61-3-9347-7695 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Nick_Madary <@t> hgsi.com Mon Sep 29 13:02:55 2003 From: Nick_Madary <@t> hgsi.com (Nick_Madary@hgsi.com) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Jack Bart Wenger Stories needed Message-ID: Bart Wenger Special stain GURU for over 40 years at the AFIP and and THE right hand person to Lee Luna for decades is retiring. If you have any little story you would like to share with me, I am writing a little blurb about him in the Maryland Histo news letter. It would be nice to see how many comments, and how far reaching folks are that have learned from him. Please respond to this email address Nick_Madary@hgsi.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030929/3ff646ac/attachment.htm From siksik <@t> vgernet.net Mon Sep 29 14:29:45 2003 From: siksik <@t> vgernet.net (Steven Slap) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] paraffin embedding of mouse tissues Message-ID: Kelly You don't say anything about the size (especially thickness) of your tissues or the fixation time prior to processing. If they are coming out dry on such a short microwave schedule, I suspect they may be underfixed. Steven Slap From SJones <@t> cvm.tamu.edu Mon Sep 29 16:36:11 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Paraffin Embedding of Mice Tissue Message-ID: Dear Kelly, Without having a clue as to what you are doing, I will bet that you are overprocessing them. Try a 15-20 minute per station program on the XP and see how they cut. You will see what is over processed and what is underprocessed and then trim accordingly if you want to run under the same program. (Mouse spleen and liver can be trimmed in a bit bigger than others) Sarah Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Kelly Salceies" 09/29/03 09:01AM >>> Hi Everyone, I am a brand new Tech and am having trouble preping some mouse tissue for paraffin embedding. I have all different tissues (liver, heart, quads, brain, lung, and gastroc) and would like to prep all tissues (if possible) under the same conditions. I have been using the Shandon Hypercenter XP or the Shandon Histowave (microwave tissue processor...). Does anybody have a good protocol for mouse tissues on either of these instruments?? I have tried a number of protocols, but all my tissue has been really dry in cutting... Any suggestions would be greatly appreciated!! Thanks, Kelly Salceies University of New Mexico Health Sciences Center Dept. of Pathology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SJones <@t> cvm.tamu.edu Mon Sep 29 16:45:53 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Paraffin Embedding of Mice Tissue - My protocols Message-ID: Thanks for the info Kelly, I would use Pro-par or Shandon's xylene substitute instead of xylene. Your times look ok, although I don't know what size of your samples. Two absolutes should be enough. Check the temp of your paraffin, I use the lowest melting point paraffin possible on the processor and make sure the processor is programmed to be just 2-4 degrees over the melting point. Also, check the temperature with a thermometer once just to make sure. Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Kelly Salceies" 09/29/03 11:29AM >>> Hi -- I guess I should have mentioned the protocols I have already tried - sorry...I'm new to this Histonet thing :) On the Shandon Hypercenter XP I've tried: 80% ETOH 5 min 95% ETOH 15 min 2X 100% ETOH 15 min 3X Xylene 10 min 2X Xylene 5 min 2X Paraffin 15 min 2X 50% ETOH 5 min 80% ETOH 5 min 95% ETOH 15 min 2X 100% ETOH 15 min 3X Xylene 10 min 2X Xylene 5 min Paraffin 15 min 2X 80% ETOH 5 min 95% ETOH 5 min 2X 100% ETOH 5 min 2X 1:1 ETOH: Cedarwood Oil 5 min 1:1 Cedarwood Oil:Methyl Salicylate 5 min 100% Methyl Salicylate 5 min 1:1 Methyl Salicylate:Xylene 5 min Xylene 5min Paraffin 15 min 2X --The last protocol left the tissue soft - so I might just need to use longer times...I'm not sure if anybody uses this protocol.... On the Microwave I've tried: 100% ETOH 10 min 2X 1:1 ETOH:Xylene 10 min Paraffin 10 min -All steps: Max temp 65 degrees - Watt=750 100% ETOH 5 min IPA 5 min Paraffin 5 min -All steps: Max temp 65 degrees - Watt=750 100% ETOH 5 min 1:1 IPA:Xylene 5 min Paraffin 5 min -All steps: Max temp 65 degrees - Watt=750 There are the protocols I've tried....Please let me know what suggestions you have - thank you!! Kelly Salceies University of New Mexico Health Sciences Center Dept. of Pathology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From katri <@t> cogeco.ca Mon Sep 29 18:46:08 2003 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Immunocytofluorescence on smooth muscle cells References: Message-ID: <006801c386e3$dbb24f20$ce989618@hala4.on.cogeco.ca> Hi Bertrand, Two things come to mind: 1. Your blocking serum should be from the same species as what your biotinylated antibody was raised in (ie.goat in your case). 2. Rabbit antibodies sometimes can non- specifically bind to smooth muscle cells and I don't know if there is a way to prevent it. Hope this will help a bit..... Katri Katri Tuomala Anatomic Pathology St Joseph's Health Care Hamilton, Ontario, Canada ----- Original Message ----- From: "Bertrand Lefort" To: Sent: Sunday, September 29, 2002 10:01 AM Subject: [Histonet] Immunocytofluorescence on smooth muscle cells > Hello, > > > We are a new lab and we try to develop Immuno-cyto-fluorescence techniques > in the lab. > We are working with human bronchial smooth muscle cells. > I have currently a very big problem with all rabbit antibodies. All rabbit > antibodies (including IgG as isotype) give a non-specific signal, signal in > the nucleus and cytoplasm with very high intensity. There is no signal > between cells. > > This problem does not exist with Rat and mouse antibodies I have used. > > - I have tried different fixation methods (PFA 4%, acetone-methanol (1/1), > and kit like permeafix). > > - I have tried different blocking solution (Rabbit serum 2%, FBS 2%, Horse > serum and Universal blocker solution from Dako) without any results. > > - I have tried different diluents for my antibody (PBS 1X, PBS 1x-BSA 3%, > Dako diluents) > > - I have tried different permeabilization methods (saponin, ptwx) (Not with > acetone-methanol fixation). > > All I have tried gives always the same signal with a very high intensity > signal. > > More details : I use a biotin-Goat-anti-rabbit for secondary antibody that > give not a signal if first antibody is omitted. I use PBS to dilute my > secondary antibody. I use Streptavidin-alexa to detect the signal. > > I have also worked on endogen biotin on this cells and it does not appear to > be the problem. > > Has someone reported this kind of problems on human smooth muscle cells ? > Has someone suggestions or solutions to this problem ? > > Thanks in advance > > Bertrand > > > Bertrand Lefort > > Research Assistant > Laboratory of Neuro-immunology of Asthma > Research Center, Room J3155 > 5400, Blv. Gouin West > Montreal, QC > H4J 1C5 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From katri <@t> cogeco.ca Mon Sep 29 19:03:35 2003 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Paraffin Embedding of Mice Tissue References: Message-ID: <007501c386e6$4b9ebc40$ce989618@hala4.on.cogeco.ca> Hi Kelly, What is your fixative for these mice tissues and how long? How big (read thick) are your sections to be processed? These two things will determine your routine processing protocol. I'm not familiar with microwave processing, so I won't comment on that. If the tissue is well fixed, there really isn't a chance to "over process" it. The problems arise with inadequate fixation, which then leads to alcohols and xylene drying out the tissue, and hot paraffin then causes further damage. Just something to think about.... Katri Katri Tuomala Anatomic Pathology St.Joseph's Health Care Hamilton, Ontaorio, Canada ----- Original Message ----- From: "Kelly Salceies" To: Sent: Monday, September 29, 2003 10:01 AM Subject: [Histonet] Paraffin Embedding of Mice Tissue > Hi Everyone, > > I am a brand new Tech and am having trouble preping some mouse tissue > for paraffin embedding. I have all different tissues (liver, heart, > quads, brain, lung, and gastroc) and would like to prep all tissues (if > possible) under the same conditions. I have been using the Shandon > Hypercenter XP or the Shandon Histowave (microwave tissue processor...). > Does anybody have a good protocol for mouse tissues on either of these > instruments?? I have tried a number of protocols, but all my tissue has > been really dry in cutting... > > Any suggestions would be greatly appreciated!! > > > Thanks, > > Kelly Salceies > University of New Mexico > Health Sciences Center > Dept. of Pathology > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bmcmahill <@t> incytepathology.com Mon Sep 29 20:12:53 2003 From: bmcmahill <@t> incytepathology.com (Bonnie J. McMahill) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Yet Stain Message-ID: <3F7537D9A2DE6D40BA884DF1A32A08CF1F6AF1@PATH2K-SRV2.PAI.E-PATHOLOGY.COM> [Bonnie J. McMahill] Has anyone heard of the "Yet Stain" for cytology smears. Is it available commericially? Bonnie McMahill Histology Supervisor InCyte Pathology Spokane, WA > From bmcmahill <@t> incytepathology.com Mon Sep 29 20:14:39 2003 From: bmcmahill <@t> incytepathology.com (Bonnie J. McMahill) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Yet Stain Message-ID: <3F7537D9A2DE6D40BA884DF1A32A08CF1F6AF2@PATH2K-SRV2.PAI.E-PATHOLOGY.COM> Has anyone heard of the "Yet Stain" for cytology specimens? Is it available commercially? Bonnie McMahill Histology Supervisor InCyte Pathology Spokane, WA From DDDeltour <@t> sig.med.navy.mil Mon Sep 29 23:45:11 2003 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Jack Bart Wenger Stories needed Message-ID: He taught me a great deal about special stains when I went through school at AFIP. I know that he will be missed. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy LPO Histology Histology Technician DSN 624-4669 FAX 624-4680 COMM (From US) 011-39-095-56-4669 -----Original Message----- From: Nick_Madary@hgsi.com [mailto:Nick_Madary@hgsi.com] Sent: Monday, September 29, 2003 8:03 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Jack Bart Wenger Stories needed Bart Wenger Special stain GURU for over 40 years at the AFIP and and THE right hand person to Lee Luna for decades is retiring. If you have any little story you would like to share with me, I am writing a little blurb about him in the Maryland Histo news letter. It would be nice to see how many comments, and how far reaching folks are that have learned from him. Please respond to this email address Nick_Madary@hgsi.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030930/5b92f980/attachment.htm From Myri37 <@t> aol.com Tue Sep 30 00:17:40 2003 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Haupt's solution Message-ID: <1ca.119f4121.2caa6bf4@aol.com> hello everyone could you give me more information about Haupt's solution, how is it exactly used to stuck sections of MMA on glass slides do you have a protocol for acidify slides before coating with Haupt's solution ? thank you so much Myriam -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030930/b51b287b/attachment.htm From philip.bergin <@t> microbio.gu.se Tue Sep 30 03:21:14 2003 From: philip.bergin <@t> microbio.gu.se (Phil Bergin) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Immunocytofluorescence on smooth muscle cells In-Reply-To: <006801c386e3$dbb24f20$ce989618@hala4.on.cogeco.ca> Message-ID: <001201c3872b$d09d5060$a660f182@philb> Hi, We get a lot of unspecific binding in gastric smooth muscle cells and throughout the whole stomach (both human and murine) when we use rabbit antibodies, which is really difficult to block. Basically we try and stay away from rabbit antibodies as they seem to 'stick to everything' as far as our lab goes............ Best of luck to you though, Phil ----------------------------------------------------------------- Philip Bergin G?teborg University Department of Medical Microbiology and Immunology Box 435, SE-405 30 G?teborg, Sweden Phone: +46-31-3424471 Fax: +46-31-826976 -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Katri Tuomala Sent: den 30 september 2003 01:46 To: Bertrand Lefort; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Immunocytofluorescence on smooth muscle cells Hi Bertrand, Two things come to mind: 1. Your blocking serum should be from the same species as what your biotinylated antibody was raised in (ie.goat in your case). 2. Rabbit antibodies sometimes can non- specifically bind to smooth muscle cells and I don't know if there is a way to prevent it. Hope this will help a bit..... Katri Katri Tuomala Anatomic Pathology St Joseph's Health Care Hamilton, Ontario, Canada ----- Original Message ----- From: "Bertrand Lefort" To: Sent: Sunday, September 29, 2002 10:01 AM Subject: [Histonet] Immunocytofluorescence on smooth muscle cells > Hello, > > > We are a new lab and we try to develop Immuno-cyto-fluorescence techniques > in the lab. > We are working with human bronchial smooth muscle cells. > I have currently a very big problem with all rabbit antibodies. All rabbit > antibodies (including IgG as isotype) give a non-specific signal, signal in > the nucleus and cytoplasm with very high intensity. There is no signal > between cells. > > This problem does not exist with Rat and mouse antibodies I have used. > > - I have tried different fixation methods (PFA 4%, acetone-methanol (1/1), > and kit like permeafix). > > - I have tried different blocking solution (Rabbit serum 2%, FBS 2%, Horse > serum and Universal blocker solution from Dako) without any results. > > - I have tried different diluents for my antibody (PBS 1X, PBS 1x-BSA 3%, > Dako diluents) > > - I have tried different permeabilization methods (saponin, ptwx) (Not with > acetone-methanol fixation). > > All I have tried gives always the same signal with a very high intensity > signal. > > More details : I use a biotin-Goat-anti-rabbit for secondary antibody that > give not a signal if first antibody is omitted. I use PBS to dilute my > secondary antibody. I use Streptavidin-alexa to detect the signal. > > I have also worked on endogen biotin on this cells and it does not appear to > be the problem. > > Has someone reported this kind of problems on human smooth muscle cells ? > Has someone suggestions or solutions to this problem ? > > Thanks in advance > > Bertrand > > > Bertrand Lefort > > Research Assistant > Laboratory of Neuro-immunology of Asthma > Research Center, Room J3155 > 5400, Blv. Gouin West > Montreal, QC > H4J 1C5 > > > _______________________________________________ > Histonet mailing list > Histonet@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 louise_renton <@t> hotmail.com Tue Sep 30 03:49:36 2003 From: louise_renton <@t> hotmail.com (louise renton) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] over vs underprocessing Message-ID: Dear All, There have been numerous posts and replies over time regarding the quality of processed tissues, with under or over dehydration, fixation or processing being cited as the cause. I would lke some clarification and correction on my personal viewpoints on this issue as the case may be. To my understanding: a) Fixation and dehydration have an absolute end result, ie one cannot "over" dehydrate, as once the water is gone, it's gone. However, residual water is immiscible with clearing agents or wax, and would thus produce a soft poorly processed tissue. Thus "under" dehydration is a real entity. However, anecdotal evidence has stated that these tissues may become 'alcohol-fixed" thus making them difficult to cut. What is the rationale behind this? b) "Over" processing, is the result of excessive or prolonged heat coagulating proteins rendering the tissue into hard nasty uncuttable little nodules. I seem to recall this demonstrated in cookery class where eggs were fried to extinction and thus became an indigestible mass (there are many proceses and regents used in histology that bear resemblance to cooking school) I realise that this might be an oversimplified version of the real processes, but I am interested in what the experts have to say. Best regards Louise Renton Bone Research Unit MRC Johannesburg South Africa Tel & fax +27 11 717 2298 "Time flies like an arrow, fruit flies like a banana" ----Original Message Follows---- From: " Katri Tuomala" To: "Kelly Salceies" , Subject: Re: [Histonet] Paraffin Embedding of Mice Tissue Date: Mon, 29 Sep 2003 20:03:35 -0400 Hi Kelly, What is your fixative for these mice tissues and how long? How big (read thick) are your sections to be processed? These two things will determine your routine processing protocol. I'm not familiar with microwave processing, so I won't comment on that. If the tissue is well fixed, there really isn't a chance to "over process" it. The problems arise with inadequate fixation, which then leads to alcohols and xylene drying out the tissue, and hot paraffin then causes further damage. Just something to think about.... Katri Katri Tuomala Anatomic Pathology St.Joseph's Health Care Hamilton, Ontaorio, Canada ----- Original Message ----- From: "Kelly Salceies" To: Sent: Monday, September 29, 2003 10:01 AM Subject: [Histonet] Paraffin Embedding of Mice Tissue > Hi Everyone, > > I am a brand new Tech and am having trouble preping some mouse tissue > for paraffin embedding. I have all different tissues (liver, heart, > quads, brain, lung, and gastroc) and would like to prep all tissues (if > possible) under the same conditions. I have been using the Shandon > Hypercenter XP or the Shandon Histowave (microwave tissue processor...). > Does anybody have a good protocol for mouse tissues on either of these > instruments?? I have tried a number of protocols, but all my tissue has > been really dry in cutting... > > Any suggestions would be greatly appreciated!! > > > Thanks, > > Kelly Salceies > University of New Mexico > Health Sciences Center > Dept. of Pathology > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Unsatisfied with being single? Try MSN Personals http://www.msn.co.za/personals/ From Juan.Solon <@t> lshtm.ac.uk Tue Sep 30 05:52:26 2003 From: Juan.Solon <@t> lshtm.ac.uk (Juan Solon) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Reichert Jung Frigocut 2700 and disposable blades Message-ID: dear all, We are setting up a modest histochemistry lab in The Gambia, W. Africa and we will be inheriting an unused Reichert Jung Frigocut 2700. I wanted to ask if disposable blade holders can be used for this model and where I could source them. I have asked Leica at Milton Keynes (UK), and they said they do not have them in stock anymore? Thanks John Dr. Juan Antonio Solon MRC Keneba Field Station MRC Laboratories Gambia Atlantic Road, Fajara PO Box 273, Banjul The Gambia Email : juan.solon@lshtm.ac.uk Tel No: (++220) 541021 Fax No: (++220) 541022 / (++220) 496513 From abright <@t> brightinstruments.com Tue Sep 30 06:40:21 2003 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Reichert Jung Frigocut 2700 and disposable blades Message-ID: Dear John, That was most helpful of them!!!!! Please inform me the size of the knife that is used on your cryostat to enable me to inform you of the correct Disposible knife holder. It's a pity the cryostat was not one of ours as we can supply parts Ex Stock on our old instruments. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Juan Solon [mailto:Juan.Solon@lshtm.ac.uk] Sent: 30 September 2003 11:52 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Reichert Jung Frigocut 2700 and disposable blades dear all, We are setting up a modest histochemistry lab in The Gambia, W. Africa and we will be inheriting an unused Reichert Jung Frigocut 2700. I wanted to ask if disposable blade holders can be used for this model and where I could source them. I have asked Leica at Milton Keynes (UK), and they said they do not have them in stock anymore? Thanks John Dr. Juan Antonio Solon MRC Keneba Field Station MRC Laboratories Gambia Atlantic Road, Fajara PO Box 273, Banjul The Gambia Email : juan.solon@lshtm.ac.uk Tel No: (++220) 541021 Fax No: (++220) 541022 / (++220) 496513 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From van <@t> uhnres.utoronto.ca Tue Sep 30 08:14:49 2003 From: van <@t> uhnres.utoronto.ca (Rita van Bendegem) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Reichert Jung Frigocut 2700 and disposable blades References: Message-ID: <3F7981C9.3291BF7C@uhnres.utoronto.ca> Dear All, We have a Reichert Jung Frigocut 2800 and a few years back I wanted to convert to a disposable blade holder. At the time I call Leica (Canada) and they told me it was not in stock anymore . So then I called some used lab equipment places, but they couldn't find a used holder. What I did in the end was I ordered the knife holder from Leica that fit the Reichert Jung Frigocut 2800 locking system the best. Then I had our machine shop mill off a strip of metal from the bottom of the knife holder that didn't line up with the cryostat holding base. I have had no problems with the knife blade holder it works beautifully. Juan Solon wrote: > dear all, > We are setting up a modest histochemistry lab in The Gambia, W. Africa > and we will be inheriting an unused Reichert Jung Frigocut 2700. I > wanted to ask if disposable blade holders can be used for this model and > where I could source them. I have asked Leica at Milton Keynes (UK), > and they said they do not have them in stock anymore? > Thanks > > John > > Dr. Juan Antonio Solon > MRC Keneba Field Station > MRC Laboratories Gambia > Atlantic Road, Fajara > PO Box 273, Banjul > The Gambia > Email : juan.solon@lshtm.ac.uk > Tel No: (++220) 541021 > Fax No: (++220) 541022 / (++220) 496513 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Rita van Bendegem, MLT Laboratory Manager, Canadian Paraplegic Association, Spinal Cord Injury Research Centre, Mc Laughlin Pavilion Lab 12-405, 399 Bathurst St., Toronto Western Hospital, University Health Network, Toronto, Ontario M5T 2S8 Telephone 416-603-5800 ext. 2792 Fax 416-603-5745 From mari.ann.mailhiot <@t> leica-microsystems.com Tue Sep 30 08:43:40 2003 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Paraffin embedding of mice tissue Message-ID: Kelly There are several xylene substitutes on the market you can use for clearing. This will help with the dryness of the tissue. The substitutes that I am most familiar with are Propar, Clearite, Sub X and Histosolv. There is also a great book titled Animal Processing Manual that was published by NSH and edited by Gayle Callis and Diane Sterchi. This book has a wealth of information, including many processing schedules for animal tissue. You can order the book through NSH. 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 From cdemarinis <@t> SARATOGACARE.ORG Tue Sep 30 08:52:32 2003 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis,Carolyn) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Gimenez technique for helicobacter pylori Message-ID: <2FCF6059F121BA41A672C5012BB113AA2A52AA@mercury.saratogacare.org> We use the Gimenez stain for helicobacter pylori from Polyscientific. The bugs should stain bright red from the carbol fuchsin, but they always stain blue. If anyone uses this stain and gets good results, please help. Thanks. CONFIDENTIALITY NOTICE: 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. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. From djcarter <@t> dallastx.net Tue Sep 30 09:12:47 2003 From: djcarter <@t> dallastx.net (Dallas Nippert) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Paraffin Embedding of Mice Tissue - My protocols References: Message-ID: <003e01c3875c$f2166200$2ffcf3d0@djcarter> I worked with mouse tissue for 6 years, and it will almost always be dry. The only thing I did was soak my blocks in my waterbath for about 45 seconds after facing, and then put them on my ice. The protocol for processing was: 70% Etoh 20 min 80% Etoh 20 min 95% Etoh x2 for 20 min Absolute x2 for 20 min Xylene x2 for 20 min Paraffin x2 for 30 min I also used an infiltrating wax in the processor and an embedding wax in the embedder. The tissues would soak in the wax for about 15 min before I started and while I was embedding them. If they are still dry, try cutting down an alcohol time. Dallas ----- Original Message ----- From: "Kelly Salceies" To: Sent: Monday, September 29, 2003 11:29 AM Subject: [Histonet] Paraffin Embedding of Mice Tissue - My protocols > Hi -- > > I guess I should have mentioned the protocols I have already tried - > sorry...I'm new to this Histonet thing :) > > On the Shandon Hypercenter XP I've tried: > > 80% ETOH 5 min > 95% ETOH 15 min 2X > 100% ETOH 15 min 3X > Xylene 10 min 2X > Xylene 5 min 2X > Paraffin 15 min 2X > > 50% ETOH 5 min > 80% ETOH 5 min > 95% ETOH 15 min 2X > 100% ETOH 15 min 3X > Xylene 10 min 2X > Xylene 5 min > Paraffin 15 min 2X > > 80% ETOH 5 min > 95% ETOH 5 min 2X > 100% ETOH 5 min 2X > 1:1 ETOH: Cedarwood Oil 5 min > 1:1 Cedarwood Oil:Methyl Salicylate 5 min > 100% Methyl Salicylate 5 min > 1:1 Methyl Salicylate:Xylene 5 min > Xylene 5min > Paraffin 15 min 2X > > --The last protocol left the tissue soft - so I might just need to use > longer times...I'm not sure if anybody uses this protocol.... > > On the Microwave I've tried: > > 100% ETOH 10 min 2X > 1:1 ETOH:Xylene 10 min > Paraffin 10 min > -All steps: Max temp 65 degrees - Watt=750 > > 100% ETOH 5 min > IPA 5 min > Paraffin 5 min > -All steps: Max temp 65 degrees - Watt=750 > > 100% ETOH 5 min > 1:1 IPA:Xylene 5 min > Paraffin 5 min > -All steps: Max temp 65 degrees - Watt=750 > > > There are the protocols I've tried....Please let me know what > suggestions you have - thank you!! > Kelly Salceies > University of New Mexico > Health Sciences Center > Dept. of Pathology > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From m.vermehren <@t> anat.vetmed.uni-muenchen.de Tue Sep 30 08:57:11 2003 From: m.vermehren <@t> anat.vetmed.uni-muenchen.de (Margarete Vermehren) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] ISH on fresh frozen tissue Message-ID: <000001c3875a$bf6ffde0$64ae548d@HistoPC3> I am trying to do ISH on fresh frozen mouse testis. The morphology was o.k. but my protocol doesn?t work. This material should not need pretreatment, I suppose, although I must be wrong somewhere ? it doesn?t work that way. Pretreatment with protease E is very delicate. Has anybody got experience he/she could share with me? Thank you very much!! Margaret Margarete Vermehren LMU M?nchen / Tieranatomie II Tel: +49 89 21805857 Fax: +49 89 21802569 m.vermehren@anat.vetmed.uni-muenchen.de -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030930/3da03f1e/attachment.htm From Mary_Georger <@t> URMC.Rochester.edu Tue Sep 30 09:42:17 2003 From: Mary_Georger <@t> URMC.Rochester.edu (Georger, Mary) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] OT : NSH Seminar Liaisons Message-ID: Good Morning All, I have a few Seminar Liaison spots at NSH that I need to fill. They are : Monday PM Tuesday AM Wednesday PM If you would be willing to Liaison for one of these please let me know. Thank you to everyone who has so graciously offered to help out at the meeting, we couldn't do it without you! Mary Georger Center for Cardiovascular Research University of Rochester Medical Center KMRB Room 2-9816 601 Elmwood Avenue. PO Box 679 Rochester, New York 14642 585-273-1548 From tbergeron <@t> criver.com Tue Sep 30 09:44:09 2003 From: tbergeron <@t> criver.com (tbergeron@criver.com) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Paraffin Embedding of Mice Tissue - My protocols Message-ID: Another trick you can try is float the block in your waterbath for 20 or so seconds just enough to warm it a little Then I put the blocks on cold plates (don't have ice) that have a small amount of dilute fabric softener. Tracy E. Bergeron Histologist Charles River Laboratories Wilmington, MA 978-658-6000 x-1229 |---------+---------------------------------------> | | "Dallas Nippert" | | | | | | Sent by: | | | histonet-admin@lists.utsouth| | | western.edu | | | | | | | | | 09/30/2003 10:12 AM | | | | |---------+---------------------------------------> >---------------------------------------------------------------------------------------------------------------| | | | To: "Kelly Salceies" , | | cc: | | Subject: Re: [Histonet] Paraffin Embedding of Mice Tissue - My protocols | >---------------------------------------------------------------------------------------------------------------| I worked with mouse tissue for 6 years, and it will almost always be dry. The only thing I did was soak my blocks in my waterbath for about 45 seconds after facing, and then put them on my ice. The protocol for processing was: 70% Etoh 20 min 80% Etoh 20 min 95% Etoh x2 for 20 min Absolute x2 for 20 min Xylene x2 for 20 min Paraffin x2 for 30 min I also used an infiltrating wax in the processor and an embedding wax in the embedder. The tissues would soak in the wax for about 15 min before I started and while I was embedding them. If they are still dry, try cutting down an alcohol time. Dallas ----- Original Message ----- From: "Kelly Salceies" To: Sent: Monday, September 29, 2003 11:29 AM Subject: [Histonet] Paraffin Embedding of Mice Tissue - My protocols > Hi -- > > I guess I should have mentioned the protocols I have already tried - > sorry...I'm new to this Histonet thing :) > > On the Shandon Hypercenter XP I've tried: > > 80% ETOH 5 min > 95% ETOH 15 min 2X > 100% ETOH 15 min 3X > Xylene 10 min 2X > Xylene 5 min 2X > Paraffin 15 min 2X > > 50% ETOH 5 min > 80% ETOH 5 min > 95% ETOH 15 min 2X > 100% ETOH 15 min 3X > Xylene 10 min 2X > Xylene 5 min > Paraffin 15 min 2X > > 80% ETOH 5 min > 95% ETOH 5 min 2X > 100% ETOH 5 min 2X > 1:1 ETOH: Cedarwood Oil 5 min > 1:1 Cedarwood Oil:Methyl Salicylate 5 min > 100% Methyl Salicylate 5 min > 1:1 Methyl Salicylate:Xylene 5 min > Xylene 5min > Paraffin 15 min 2X > > --The last protocol left the tissue soft - so I might just need to use > longer times...I'm not sure if anybody uses this protocol.... > > On the Microwave I've tried: > > 100% ETOH 10 min 2X > 1:1 ETOH:Xylene 10 min > Paraffin 10 min > -All steps: Max temp 65 degrees - Watt=750 > > 100% ETOH 5 min > IPA 5 min > Paraffin 5 min > -All steps: Max temp 65 degrees - Watt=750 > > 100% ETOH 5 min > 1:1 IPA:Xylene 5 min > Paraffin 5 min > -All steps: Max temp 65 degrees - Watt=750 > > > There are the protocols I've tried....Please let me know what > suggestions you have - thank you!! > Kelly Salceies > University of New Mexico > Health Sciences Center > Dept. of Pathology > > > _______________________________________________ > Histonet mailing list > Histonet@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 csr <@t> meyerinst.com Tue Sep 30 10:18:47 2003 From: csr <@t> meyerinst.com (Cheryl Rehfeld - Meyer Instruments, Inc) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Reichert Jung Frigocut 2700 and disposable blades References: <3F7981C9.3291BF7C@uhnres.utoronto.ca> Message-ID: <001601c38766$26161060$7800a8c0@CSRLaptop> The Leica Knife Holder CE for disposable blades fits the Reichert Jung Frigocut. The knife base CN, CE, or CS, part number 14041918647, has a flat bottom so you won't have to mill off the bottom like Rita van Bendegem. This is an older part number but it is still available. I have put these parts on some of my customers Frigocut 2800 and they work great. Cheryl Rehfeld Meyer Instruments, Inc. Leica Exclusive Regional Dealer ----- Original Message ----- From: Rita van Bendegem To: Juan Solon Cc: Sent: Tuesday, September 30, 2003 8:14 AM Subject: Re: [Histonet] Reichert Jung Frigocut 2700 and disposable blades > Dear All, > > We have a Reichert Jung Frigocut 2800 and a few years back I wanted to > convert to a disposable blade holder. At the time I call Leica (Canada) > and they told me it was not in stock anymore . So then I called some used > lab equipment places, but they couldn't find a used holder. > > What I did in the end was I ordered the knife holder from Leica that fit > the Reichert Jung Frigocut 2800 locking system the best. Then I had our > machine shop mill off a strip of metal from the bottom of the knife holder > that didn't line up with the cryostat holding base. I have had no problems > with the knife blade holder it works beautifully. > > > Juan Solon wrote: > > > dear all, > > We are setting up a modest histochemistry lab in The Gambia, W. Africa > > and we will be inheriting an unused Reichert Jung Frigocut 2700. I > > wanted to ask if disposable blade holders can be used for this model and > > where I could source them. I have asked Leica at Milton Keynes (UK), > > and they said they do not have them in stock anymore? > > Thanks > > > > John > > > > Dr. Juan Antonio Solon > > MRC Keneba Field Station > > MRC Laboratories Gambia > > Atlantic Road, Fajara > > PO Box 273, Banjul > > The Gambia > > Email : juan.solon@lshtm.ac.uk > > Tel No: (++220) 541021 > > Fax No: (++220) 541022 / (++220) 496513 > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- > Rita van Bendegem, MLT > Laboratory Manager, > Canadian Paraplegic Association, > Spinal Cord Injury Research Centre, > Mc Laughlin Pavilion Lab 12-405, > 399 Bathurst St., > Toronto Western Hospital, > University Health Network, > Toronto, Ontario M5T 2S8 > > Telephone 416-603-5800 ext. 2792 > Fax 416-603-5745 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Juan.Solon <@t> lshtm.ac.uk Tue Sep 30 10:19:31 2003 From: Juan.Solon <@t> lshtm.ac.uk (Juan Solon) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Reichert Jung Frigocut 2700 and disposable blades Message-ID: Would this be true for the older Frigocut (2700) as well? Dr. Juan Antonio Solon MRC Keneba Field Station MRC Laboratories Gambia Atlantic Road, Fajara PO Box 273, Banjul The Gambia Email : juan.solon@lshtm.ac.uk Tel No: (++220) 541021 Fax No: (++220) 541022 / (++220) 496513 >>> "Cheryl Rehfeld - Meyer Instruments, Inc" 09/30/03 4:18 PM >>> The Leica Knife Holder CE for disposable blades fits the Reichert Jung Frigocut. The knife base CN, CE, or CS, part number 14041918647, has a flat bottom so you won't have to mill off the bottom like Rita van Bendegem. This is an older part number but it is still available. I have put these parts on some of my customers Frigocut 2800 and they work great. Cheryl Rehfeld Meyer Instruments, Inc. Leica Exclusive Regional Dealer ----- Original Message ----- From: Rita van Bendegem To: Juan Solon Cc: Sent: Tuesday, September 30, 2003 8:14 AM Subject: Re: [Histonet] Reichert Jung Frigocut 2700 and disposable blades > Dear All, > > We have a Reichert Jung Frigocut 2800 and a few years back I wanted to > convert to a disposable blade holder. At the time I call Leica (Canada) > and they told me it was not in stock anymore . So then I called some used > lab equipment places, but they couldn't find a used holder. > > What I did in the end was I ordered the knife holder from Leica that fit > the Reichert Jung Frigocut 2800 locking system the best. Then I had our > machine shop mill off a strip of metal from the bottom of the knife holder > that didn't line up with the cryostat holding base. I have had no problems > with the knife blade holder it works beautifully. > > > Juan Solon wrote: > > > dear all, > > We are setting up a modest histochemistry lab in The Gambia, W. Africa > > and we will be inheriting an unused Reichert Jung Frigocut 2700. I > > wanted to ask if disposable blade holders can be used for this model and > > where I could source them. I have asked Leica at Milton Keynes (UK), > > and they said they do not have them in stock anymore? > > Thanks > > > > John > > > > Dr. Juan Antonio Solon > > MRC Keneba Field Station > > MRC Laboratories Gambia > > Atlantic Road, Fajara > > PO Box 273, Banjul > > The Gambia > > Email : juan.solon@lshtm.ac.uk > > Tel No: (++220) 541021 > > Fax No: (++220) 541022 / (++220) 496513 > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- > Rita van Bendegem, MLT > Laboratory Manager, > Canadian Paraplegic Association, > Spinal Cord Injury Research Centre, > Mc Laughlin Pavilion Lab 12-405, > 399 Bathurst St., > Toronto Western Hospital, > University Health Network, > Toronto, Ontario M5T 2S8 > > Telephone 416-603-5800 ext. 2792 > Fax 416-603-5745 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From abright <@t> brightinstruments.com Tue Sep 30 11:02:13 2003 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Microtome for autoradiography Message-ID: Dear Gamal, You had best look into this a bit deeper. The microtome you are planning to purchase is not large enough for sectioning whole rats unless they are only 130mm in length. For WBA section you require a cryostat not a bench microtome unless you would like to place the microtome in a -20?C cold room. I am not joking this is why we started to manufacture this type of cryostat, as Bob McCulloch at Glaxo, Fulmer Hall, England, 40 years+ ago was doing so, as he could only stay at this temperature for 1/2 hour max. he asked us to produce a large sectioning cryostat for him, we did and many 100's were sold for WBA. A Tungsten Carbide Knife will have no problems sectioning through the undecalcified bone. We manufacture a range of cryostats for Whole Body Autoradiography This is an outline of the process for WBA sectioning. The appropriate safety equipment must be used e.g. eye protection and gloves. 1) Pin the specimen by the limbs onto a flat 3mm or 1/8" sheet of hardboard. 2) Immerse the above into a slurry of Petroleum Ether and Solid C02 solution, for at least 4 hours.. 3) Remove from hardboard and remove limbs and tail. 4) Wash fur with a 0.07% solution of Trinton X. 5) Immerse specimen into a mould and fill with a 2% solution of Carboxy Methyl Cellulose. 6) Finally immerse the above into a slurry of Petroleum Ether and Solid C02 solution, for at least 2 hours. 7) Remove from mould and mount the moulded specimen onto a object holder for mounting onto the microtome using a 2% solution of Carboxy Methyl Cellulose. 8) Using a Tungsten tipped knife trim specimen until the required level is reached, adhere 3m's or equivalent section mounting tape to the face of the sample and cut the required section @ 20-30? depending on requirements. I hope you find this useful. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: gamal akabani hneide [mailto:gakabani@nc.rr.com] Sent: 29 September 2003 05:02 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microtome for autoradiography Hi All - I need some advice regarding the best microtome to use for whole body autoradiography for rats. I am planning to buy a sledge microtome SM2400 from Leica. I am going to inject a radiolabeled MAB and sacrifice the animal with few minutes to hours, what next? What about the bone sections? Etc. Any references will be appreciated. Thanks in advanced Gamal Akabani, Ph.D. Duke University Medical Center Department of Radiology Division of Nuclear Medicine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From STEGTM <@t> samcstl.org Tue Sep 30 11:40:04 2003 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] imprinted slides? Message-ID: Hi! I have a question. Is there a regulation/requirement that slides, as in routine pathology cases from a hospital, have the name of the institution on them? Everywhere I've been they have had the name either imprinted, or on the paper label (back in the day...). We're facing buying several hundred gross of slides, and need to know whether it is necessary to spend the $$$$ to have them imprinted. I appreciate your help. Peace, Terre From Vickroy.Jim <@t> mhsil.com Tue Sep 30 12:23:41 2003 From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] autopsy saw Message-ID: <584AE8968B1EE14998210A0D6B97D854144647@mmcmail2.mhsil.com> Has anyone ever compared the Strker or Mopec Saw with the Mopec Quiet Cut saw? Is the Quiet Cut worth the extra dollars? James R. Vickroy BS, HT (ASCP) Technical Supervisor, Surgical Pathology 788-4046 vickroy.jim@mhsil.com This E-Mail and any attachments are intended solely for the addressee(s) and may be legally privileged and /or confidential. If you believe you received this E-Mail in error, please forward it to mhsil@mhsil.com to notify us of the error, then delete it; do not divulge, copy, forward or use the contents in any way. Please be aware that any unauthorized use or disclosure of the contents of this E-Mail may be unlawful. Memorial Health System 701 N. First Street Springfield, IL 62781-0001 217-757-7753 http://www.mhsil.com From REEVEML <@t> shands.ufl.edu Tue Sep 30 13:18:01 2003 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Drying Slides Message-ID: Currently we incubate our slides after the case is final, before we file them. What are other places doing? Air drying? How long do you air dry them? If you incubate them, how long? and at what temperature? Thanks in advance for the information. Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 From Mauricio.Morais <@t> tufts.edu Tue Sep 30 14:25:02 2003 From: Mauricio.Morais <@t> tufts.edu (Mauricio S. Morais) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] automated immunostainers References: <533E4A1C3061FC4EB388119D2FD7B5D402431809@nbfexch04.north-bristol.nhs> Message-ID: <3F79D88E.CA2938A1@tufts.edu> Hello Histonetters. Can anyone give me an idea about the costs of the machines (system) plus monthly maintenance? I wish to know if worth to have one to process 6-10 slides/day. Thank you. Mauricio S. Morais Senior Research Technician Nutrition, Exercise, Physiology, and Sarcopenia Laboratory (NEPS) TUFTS University Jean Mayer USDA Human Nutrition Research Center on Aging 711 Washington Street, Rm 436 Boston, MA 02111 (617) 556-3226 Marion Hiles wrote: > > We are a Neuropath lab and stain small runs of immuno slides (varies from 5 > to 30) on the Optimax Plus machine. It is an excellent machine and very easy > to use and maintain. > > Bob Quilty > Dept of Neuropathology > Frenchay Hospital > Bristol, UK > > -----Original Message----- > From: Julien Lambrey de Souza [mailto:jlambrey@hotmail.com] > Sent: 26 September 2003 19:43 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] automated immunostainers > > Hi All, > > As a new developmental biology lab, we are looking to acquire an automated > immunostainer to stain all types of collagen and probably bone precursor. > > Do you have a particular machine to recommend? Although we have looked at > different stainers (Zymed, Genex, Optimax, Dako, Labvision), each compagny > seems to have the best machine for our particular needs. We will be > processing small amounts (compared to hospitals for instance) but need the > automatisation because of time consuming other histological techniques we > use. The brochures are all very interesting but hardly reflect the reality > of laboratory routine work. > > Any help is welcome. > > Thanks a lot, > > Julien. > > _________________________________________________________________ > MSN 8 with e-mail virus protection service: 2 months FREE* > http://join.msn.com/?page=features/virus > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- A non-text attachment was scrubbed... Name: Mauricio.Morais.vcf Type: text/x-vcard Size: 1220 bytes Desc: Card for Mauricio S. Morais Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030930/c49760a0/Mauricio.Morais.vcf From lelliott <@t> cvh.on.ca Tue Sep 30 15:16:44 2003 From: lelliott <@t> cvh.on.ca (Lynda Elliott) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] unsubscribe Message-ID: -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030930/98ece0ab/attachment.htm From jeannie_heck <@t> yahoo.com Tue Sep 30 15:56:30 2003 From: jeannie_heck <@t> yahoo.com (Jeannie Heck) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Modified Mowry Message-ID: <20030930205630.89222.qmail@web41611.mail.yahoo.com> I would deeply appreciate a procedure for a modified Mowry Colloidal Iron using 3% as a mordant before the colloidal iron stain. I am currently using the Muller Mowry and I am getting inconsistent results. All help is appreciated. Please post to this list or e-mail me directly at heckj@hhosp.com. Thanks in advance. Jeannie Heck HT (ASCP) --------------------------------- Do you Yahoo!? The New Yahoo! Shopping - with improved product search -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030930/d9d4edce/attachment.htm From conniegrubaugh <@t> hotmail.com Sun Sep 21 15:07:16 2003 From: conniegrubaugh <@t> hotmail.com (connie grubaugh) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Histotechnologist Job in Las Vegas Message-ID: Position available in outpatient pathology laboratory to perform routine histology procedures and gross examination of Dermatopatholgy specimens. Qualifications include BA/BS degree in a biological science, and HTL (ASCP) certification. Competitive compensation and benefits, no state income tax. Please fax, mail or email resume to: Janet Kliethermes, COO 3059 South Maryland Pkwy. #100 Las Vegas Nv 89109-2201 Ph- 702-732-3441 Fax 702-732-2310 Email Kliethermes@lasvegaspathologist.com _________________________________________________________________ Help protect your PC. Get a FREE computer virus scan online from McAfee. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From jim.manavis <@t> imvs.sa.gov.au Tue Sep 30 17:39:08 2003 From: jim.manavis <@t> imvs.sa.gov.au (jim) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] apoptosis marker profile of gliomas In-Reply-To: <00519F9F41D2844D9C75BEE1F6AC09AF68BA19@rmhmail1.ssg.org.au> Message-ID: <002601c387a3$a9174780$7a6c140a@imvs.sa.gov.au> Stan We have used with great success both an active caspase-3 and -9 from Biovision. Our controls have been high grade gliomas. We retrieve with EDTA and use at a dilution of 1/500 with an overnight incubation. If you require details please let me know. Cheers Jim Manavis Senior Hospital Scientist Department of Neuropathology Institute of Medical & Veterinary Science Adelaide, SA, 5000 Australia Phone: 61-08-8222-3668 FAX: 61-08-8222 3204 e.mail: jim.manavis@imvs.sa.gov.au Disclaimer: Not this little black duck! -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Stylli, Stanley Sent: Monday, 29 September 2003 04:30 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] apoptosis marker profile of gliomas Importance: High Hello again Histonetters, I will be looking at the apoptotic profile of FFPE human brain tumours (various grades - low to high). I wish to correlate the expression of various apoptotic markers to the survival of the patients after certain treatments they have been subjected to against a control group that have had surgery alone. I have read a couple of papers that have used a few antibodies but I would like to hear from people who have had success with antibodies (with and without antigen retrieval). As you can imagine, you can trial antibodies from different companies until the cows come home and the bank is empty thankyou for your help Stan Stylli Stan Stylli Department of Surgery, 5th Floor Clinical Sciences Building Royal Melbourne Hospital University of Melbourne Parkville, Australia, 3052. Tel: 61-3-93427616 Fax:61-3-9347-7695 From AnthonyH <@t> chw.edu.au Tue Sep 30 18:06:40 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] over vs underprocessing Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E04E@simba.kids> My thoughts below Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: louise renton [ mailto:louise_renton@hotmail.com ] Sent: Tuesday, 30 September 2003 18:50 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] over vs underprocessing Dear All, There have been numerous posts and replies over time regarding the quality of processed tissues, with under or over dehydration, fixation or processing being cited as the cause. I would lke some clarification and correction on my personal viewpoints on this issue as the case may be. To my understanding: a) Fixation and dehydration have an absolute end result, ie one cannot "over" dehydrate, as once the water is gone, it's gone. However, residual water is immiscible with clearing agents or wax, and would thus produce a soft poorly processed tissue. Thus "under" dehydration is a real entity. However, anecdotal evidence has stated that these tissues may become 'alcohol-fixed" thus making them difficult to cut. What is the rationale behind this? Alcohol is a fixative as well, so inadequately formalin fixed tissues will fix in alcohols b) "Over" processing, is the result of excessive or prolonged heat coagulating proteins rendering the tissue into hard nasty uncuttable little nodules. I seem to recall this demonstrated in cookery class where eggs were fried to extinction and thus became an indigestible mass (there are many proceses and regents used in histology that bear resemblance to cooking school) Over processing (a cooked look to tissues) seems to be more pronounced in under-fixed tissue. Tissues fixed in formalin for longer than 24 hours rarely show this change. This is based on microwave processing experience where tissues are regularly under-fixed. I realise that this might be an oversimplified version of the real processes, but I am interested in what the experts have to say. Best regards Louise Renton Bone Research Unit MRC Johannesburg South Africa Tel & fax +27 11 717 2298 "Time flies like an arrow, fruit flies like a banana" ----Original Message Follows---- From: " Katri Tuomala" To: "Kelly Salceies" , Subject: Re: [Histonet] Paraffin Embedding of Mice Tissue Date: Mon, 29 Sep 2003 20:03:35 -0400 Hi Kelly, What is your fixative for these mice tissues and how long? How big (read thick) are your sections to be processed? These two things will determine your routine processing protocol. I'm not familiar with microwave processing, so I won't comment on that. If the tissue is well fixed, there really isn't a chance to "over process" it. The problems arise with inadequate fixation, which then leads to alcohols and xylene drying out the tissue, and hot paraffin then causes further damage. Just something to think about.... Katri Katri Tuomala Anatomic Pathology St.Joseph's Health Care Hamilton, Ontaorio, Canada ----- Original Message ----- From: "Kelly Salceies" To: Sent: Monday, September 29, 2003 10:01 AM Subject: [Histonet] Paraffin Embedding of Mice Tissue > Hi Everyone, > > I am a brand new Tech and am having trouble preping some mouse tissue > for paraffin embedding. I have all different tissues (liver, heart, > quads, brain, lung, and gastroc) and would like to prep all tissues (if > possible) under the same conditions. I have been using the Shandon > Hypercenter XP or the Shandon Histowave (microwave tissue processor...). > Does anybody have a good protocol for mouse tissues on either of these > instruments?? I have tried a number of protocols, but all my tissue has > been really dry in cutting... > > Any suggestions would be greatly appreciated!! > > > Thanks, > > Kelly Salceies > University of New Mexico > Health Sciences Center > Dept. of Pathology > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Unsatisfied with being single? Try MSN Personals http://www.msn.co.za/personals/ _______________________________________________ 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. ********************************************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/9374985c/attachment.htm From AnthonyH <@t> chw.edu.au Tue Sep 30 18:19:47 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] ISH on fresh frozen tissue Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E050@simba.kids> Margarete, ISH on frozen sections is possible. I have had success with the following to demonstrate mRNA. The nucleolus stains up beautifully: SOLUTIONS: 1. Acetic Alcohol Absolute Alcohol 15ml Acetic Acid 5ml 2. Absolute Alcohol 3. Cold Acetone METHOD: 1. Air dry frozen sections (6-8um) minimum 1 hour. 2. Fix in Acetic Alcohol 15 minutes. 3. Wash in absolute alcohol. 4. Place in cold acetone, allow to warm to room temperature. 5. Remove slides and air dry. 6. Circle sections with PAP pen, add probe and continue with ISH. NB do not treat with enzyme or acid. I would suggest sealing the coverslip with nail polish (after adding th probe) and heating on a hot plate to denature the DNA if searching for DNA sequences. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Margarete Vermehren [mailto:m.vermehren@anat.vetmed.uni-muenchen.de] Sent: Tuesday, 30 September 2003 23:57 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] ISH on fresh frozen tissue I am trying to do ISH on fresh frozen mouse testis. The morphology was o.k. but my protocol doesn't work. This material should not need pretreatment, I suppose, although I must be wrong somewhere - it doesn't work that way. Pretreatment with protease E is very delicate. Has anybody got experience he/she could share with me? Thank you very much!! Margaret Margarete Vermehren LMU M?nchen / Tieranatomie II Tel: +49 89 21805857 Fax: +49 89 21802569 m.vermehren@anat.vetmed.uni-muenchen.de ********************************************************************** 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. ********************************************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/4782a2d2/attachment.htm From Ellin13 <@t> adelphia.net Tue Sep 30 19:00:18 2003 From: Ellin13 <@t> adelphia.net (Jesus Ellin) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Kappa, Lambda staining??? References: <1CF2E2E5BB36D5119E7A0008C791F3740800E050@simba.kids> Message-ID: <001501c387af$00bb3040$be284344@yumaaz.adelphia.net.losaca.adelphia.net> Hello everyone ,, Need some help here, currently our lab is trying to create a IHC protocol for kappa and lambda on the Venntana benchmark,, we are having some real problems with the background staining in our control and patients,, we are using a blocker and or hydrogen peroxide, to try and minimize the peroxidase but we are still getting alot of background, if any one out there that has a benchmark and has a protocol for these stains some help would be appreciated,, forgot to mention that we are using Tonsil and Bone Marrow for control, One other thing how many out there are using the Tamtron System,, we are currently in the building phase and would like to ask some people out there questions concerning proccess, codes and end user training Jesus Ellin HTL ASCP Yuma Regional Medical Center jellin@yumaregional.org -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030930/fd82f74d/attachment.htm From Linresearch <@t> aol.com Tue Sep 30 20:00:20 2003 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] (no subject) Message-ID: Hi, Does anyone have a good processing schedule for FF rabbit tissues for PE? Would you be kind enough to post it? Lin -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030930/b6a93de0/attachment.htm From Linresearch <@t> aol.com Tue Sep 30 20:07:23 2003 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Rabbit Tissues Message-ID: <15a.255010a0.2cab82cb@aol.com> Hi, Does anyone have a good processing schedule for FF rabbit tissues for PE? Would you be kind enough to post it? Lin -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030930/5fe1428f/attachment.htm From la.sebree <@t> hosp.wisc.edu Tue Sep 30 20:12:48 2003 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Kappa, Lambda staining??? Message-ID: Hello Jesus, You don't say whether you are using polyclonal or monoclonal antibodies. It makes a difference. We use monoclonal kappa and lambda from Biocare on our Benchmark with nice clean results. Let me know if you'd like the protocols. Linda Sebree, HT(ASCP) University of Wisconsin Hospital and Clinics IHC/ISH Laboratory 600 Highland Ave. Madison, WI 53792 (608)265-6596 la.sebree@hosp.wisc.edu -----Original Message----- From: Jesus Ellin [mailto:Ellin13@adelphia.net] Sent: Tue 9/30/2003 7:00 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Kappa, Lambda staining??? Hello everyone ,, Need some help here, currently our lab is trying to create a IHC protocol for kappa and lambda on the Venntana benchmark,, we are having some real problems with the background staining in our control and patients,, we are using a blocker and or hydrogen peroxide, to try and minimize the peroxidase but we are still getting alot of background, if any one out there that has a benchmark and has a protocol for these stains some help would be appreciated,, forgot to mention that we are using Tonsil and Bone Marrow for control, One other thing how many out there are using the Tamtron System,, we are currently in the building phase and would like to ask some people out there questions concerning proccess, codes and end user training Jesus Ellin HTL ASCP Yuma Regional Medical Center jellin@yumaregional.org From lenaspencer <@t> insightbb.com Tue Sep 30 20:26:41 2003 From: lenaspencer <@t> insightbb.com (lena spencer) Date: Fri Sep 16 15:21:57 2005 Subject: [Histonet] Fw: ER Clones Message-ID: <004901c387bb$119c5cc0$21b6dc0c@insightbb.com> ----- Original Message ----- From: "lena spencer" To: Sent: Tuesday, September 30, 2003 7:45 PM Subject: ER Clones > Hi All: > I have another question from my Pathologist. By the way he was so impressed > with your response to my last question on his behalf that he sees this > forum as being an important link within the histology community. > His question: > Do you prefer one ER clone, if so which clone do you prefer and why? For > use on an automated platform. Also, do you see significant changes in your > ER staining from the core biopsy to the excisional specimen. How does > fixation change the staining results of your preferred ER clone? > Thanks in advance for your response. > Lena > From georgecole <@t> ev1.net Tue Sep 30 20:26:26 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] CLAOUE'S CEMENT MUST ALWAYS BE PUT ON UNDER THE HOOD Message-ID: <000001c387bb$093b6f30$0a4dbad0@hppav> CLAOUE'S CEMENT CONTAINS ETHER AND SHOULD ALWAYS BE USED UNDER THE HOOD. I PUT MESSAGES ON THE HISTONET ABOUT A FINE SILVER PREP FOR NERVES IN WHICH THE TISSUES ARE HELD ON THE SLIDES WITH CLAOUE'S CEMENT IT IS SECOND NATURE TO ME TO USE CLAOUE'S CEMENT UNDER THE HOOD. THE ETHER IN CLAOUE'S CEMENT IS TOXIC AND SHOULD NOT BE INHALED AND THE HIGHLY VOLATILE FUMES ARE FIRE HAZARDS. THIS OLD TECH WOKE UP THE NIGHT AFTER I PUT THAT SILVER TECHNIQUE ON THE HISTONET SQUIRMING WITH THE THOUGHT THAT I HADN'T STRESSED THIS HABITUAL CAUTIONARY STEP. MY COMPUTER WOULD NOT LET ME ADD THE CAUTION TO THE MESSAGES ABOUT THE SILVER PREP. I COULDNT GET THOSE MESSAGES DELETED, SO I COULD SEND NEW MESSAGES WITH HOOD USE INCLUDED. PLEASE FOLKS, DON'T ANYBODY PUT CLAOUE'S ON SLIDES IN THE OPEN ROOM. IT'S A GREAT CEMENT, BUT MY WIFE USED ONCE TOLD ME THAT I SMELLED LIKE ETHER 8 HOURS AFFTER I HAD USED CLAOUE'S. I HAVE USED THE HOOD EVER AFTER!! PLEASE FOLKS, DO THE SAME georgecole@ev1.net -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030930/6722180d/attachment.htm From lenaspencer <@t> insightbb.com Tue Sep 30 20:38:46 2003 From: lenaspencer <@t> insightbb.com (lena spencer) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Clones (ER) Message-ID: <002601c387bc$c1c1a280$21b6dc0c@insightbb.com> Hi All: I have another question from my Pathologist. By the way, the informative response from my last question, sold him on the value of the histonet. Question: Do you prefer on ER Clone? If so, which clone do you prefer and why? For use on an automated platform which ER clone is best? Also, do you see significant changes in ER staining from the core breast biopsy to the excisional breast specimen. How does fixation change the staining results of your preferred ER clone? Thanks in advance for your response. Lena From lenaspencer <@t> insightbb.com Tue Sep 30 20:42:40 2003 From: lenaspencer <@t> insightbb.com (lena spencer) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Fw: Clones (ER) Message-ID: <002d01c387bd$4d2efe80$21b6dc0c@insightbb.com> ----- Original Message ----- From: "lena spencer" To: Sent: Tuesday, September 30, 2003 9:38 PM Subject: Clones ER > Hi All: > I have another question from my Pathologist. By the way, the informative > response from my last question, sold him on the value of the histonet. > Question: > Do you prefer on ER Clone? If so, which clone do you prefer and why? For > use on an automated platform which ER clone is best? Also, do you see > significant changes in ER staining from the core breast biopsy to the > excisional breast specimen. How does fixation change the staining results > of your preferred ER clone? Thanks in advance for your response. > Lena > From bhewlett <@t> cogeco.ca Tue Sep 30 21:22:19 2003 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Fw: ER Clones References: <004901c387bb$119c5cc0$21b6dc0c@insightbb.com> Message-ID: <005901c387c2$d752aa30$b6833918@bryaniwx13voft> Lena, I suspect that ER clone 6F11 will prove to be the most popular. Why ? Check out the following paper; Bevitt D J et al. J Pathol. 183: 228-232 (1997). Yes, there may be significant changes in ER staining between core biopsy and excisional specimen. Particularly if you subscribe to the myth that small biopsies do not need to fix as long as larger specimens!! If your core biopsies are fixed in NBF for less than 8-12 hours and the later excisional specimens are breadloafed and properly fixed for a minimum of 24 hours. When the ER stains are compared, the core biopsies will show diminished or even negative ER staining as compared to a positive excisional specimen. We have consistently seen this effect since we started to perform ER IHC over 15 years ago. Experiments with tissues and cell lines expressing ER, using timed fixation in NBF for periods ranging from 1 hour to 3 weeks confirmed for us that 24 hours should be the minimum fixation time. We consider that ER and PgR staining seems to stabilize at a fixation time of about 12 hours, with small gains up to 24 hours. For fixation times between 24 hours and 3 weeks, no fall of in immunoreactivity was noticed with either clone 1D5 or 6F11. However, under similar experimental conditions, HER2 IHC required a minimum of 16-24 hours fixation to stabilize. Shorter times always produced diminished staining intensity, with times less than 6-8 hours resulting in a preponderance of negative results. Checking by FISH, those HER2 IHC negative core biopsies that had later excision specimens positive for HER2 IHC, confirmed the false negativity! There was a recent paper which confirmed our findings for ER. Tell your pathologist to check the following; Neal S. Goldstein, et al. Minimum formalin fixation time for consistent estrogen receptor immunohistochemical staining of invasive breast carcinoma. Am J Clin Pathol. 120(1), 86-92, July 2003. Regards, Bryan Bryan Hewlett Technical Consultant, Quality Management Program- Laboratory Services Ontario, Canada ----- Original Message ----- From: "lena spencer" To: "HistoNet Server" Sent: Tuesday, September 30, 2003 9:26 PM Subject: [Histonet] Fw: ER Clones > > ----- Original Message ----- > From: "lena spencer" > To: > Sent: Tuesday, September 30, 2003 7:45 PM > Subject: ER Clones > > > > Hi All: > > I have another question from my Pathologist. By the way he was so > impressed > > with your response to my last question on his behalf that he sees this > > forum as being an important link within the histology community. > > His question: > > Do you prefer one ER clone, if so which clone do you prefer and why? For > > use on an automated platform. Also, do you see significant changes in > your > > ER staining from the core biopsy to the excisional specimen. How does > > fixation change the staining results of your preferred ER clone? > > Thanks in advance for your response. > > Lena > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jkiernan <@t> uwo.ca Tue Sep 30 23:32:55 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Re: Modified Mowry (Long discussion) References: <20030930205630.89222.qmail@web41611.mail.yahoo.com> Message-ID: <3F7A58F7.94B4C961@uwo.ca> gJeannie Heck wrote: > > I would deeply appreciate a procedure for a modified > Mowry Colloidal Iron using 3% as a mordant before the > colloidal iron stain. I am currently using the Muller > Mowry and I am getting inconsistent results. All help > is appreciated. Please post to this list or e-mail me > directly at heckj@hhosp.com. Thanks in advance. > Jeannie Heck HT (ASCP) ___________________________________________________ 3% what? Nothing in any colloidal iron method involves mordanting, because no dye is involved (except as a counterstain, which might include a mordant as in the case of aluminium-nuclear fast red or brazalum). Mowry's technique is a modification of Hale's original, which wasn't much good. Pearse's Histochemistry reviews the various colloidal iron methods. It is the best reference, and has been translated into several languages. Is there any reason for using colloidal iron instead of an equivalent alcian blue method? Alcian blue techniques are more reproducible and have greater histochemical validity. Certified dye is available (tested in the Biological Stain Commission's lab, which is a non-profit outfit that serves both vendors and users of stains). The solid dye powder can deteriorate on the shelf, very unpredictably (I've had this happen 3 times). The deteriorated dye is insoluble; I have suspicions about why. Solutions of alcian blue at pH1 or pH3 keep working for a long time (a few years) even with repeated use. Alcian blue has its faults, but it's a better histochemical reagent than colloidal iron. There are several clever PAS variants developed by Scott et al in Britain and by Reid, Culling et al in Canada, from the late 1960s to the early 1990s. These methods have numerous steps but they stain macromolecular carbohydrates with well defined chemical specificity. These splendid methods are respectfully explained in textbooks but I suspect that they are seldom used in labs. There are also dozens of labeled lectins, which have high specificity and have been commercially available for 20+ years. There may be a lectin that binds specifically to the macromolecular carbohydrate that you are trying to stain with colloidal iron. Lectin histochemistry is EASY++ but you must do all the needed controls or your spectacularly specific pictures might be worthless artifacts. Lectins are the plant kingdoms's equivalent of antibodies. They recognize sugars and oligosaccharide sequences rather than the amino acid sequences (epitopes) that adhere to antibody molecules. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/