From louise.renton <@t> gmail.com Fri Jul 1 07:31:43 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] formic acid decal Message-ID: Hi all, on pain of being repetitive/lazy/slovenly/slothful/unprofessional, would someone PLEASE share their recipe for formic acid decalcifying solution suitable for samples on which IHc will be performed. Many many thanks -- Louise Renton (grovelling) Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From vsailes <@t> nd.edu Fri Jul 1 09:37:13 2005 From: vsailes <@t> nd.edu (vsailes@nd.edu) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Recipe for 4%PFA w/saponin Message-ID: <1120228633.42c5551975837@webmail.nd.edu> Hi Histonetters, I'm working with mice and we want to stain the livers for albumin. I have an antibody that appears to work but we would like to perfect the technique if possible. I have done some literature searches and some of them suggested that to perfuse the animal with 4% paraformaldehyde containing saponin would give a more homogenous staining pattern but I really could not find information telling the percentage of saponin to use. If anyone out there has experience with this would you please help shed some light on this matter. Thank you in advance for your help. Have a great holiday weekend. Valerie From pruegg <@t> ihctech.net Fri Jul 1 10:20:03 2005 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Cytospin chambers In-Reply-To: <000101c57ddd$ddb4b860$0ecd080a@wsahs.nsw.gov.au> Message-ID: <200507011520.j61FK6kv091273@pro12.abac.com> Yea I heard about that too, those dirty dogs, they are recalling all the old cytospins and trying to get you to buy their new expensive ones. I cannot imagine that there is another supplier for the chambers that will fit the TE cytospin machines. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bill Sinai Sent: Thursday, June 30, 2005 7:41 PM To: histonet (E-mail) Subject: [Histonet] Cytospin chambers Dear All, I have just been informed that ThermoElectron are no longer supplying the re-usable chambers and SS clips for the Cytospin. Does anyone know of an alternate supplier for these items? Many thanks Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. 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Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mtitford <@t> aol.com Fri Jul 1 11:16:45 2005 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Formic acid decalcifying solution Message-ID: <8C74C6F68FE2369-CFC-7912@mblk-r28.sysops.aol.com> Louise Renton in South Africa requests info on formic acid based decalcifying solution for subsequent IHC. She puts "grovelling" after her name, because she probably realises we discussed this last week on the Histonet! This is Gooding & Stewarts formula. I don't have the original reference, but it is in my 1968 edition of "Handbook of Histological Technques" by C.F.A. Culling (2ed edition) I used when I trained in London: Formic acid ...5-25 ml Formaldehyde (40%)...5 ml Distilled water... to 100 ml. We use it at the 25 ml strength and it works just fine, not really fast like RDO, but fast enough! (Happy fourth of July) Mike Titford USA Pathology Mobile AL USA From gu.lang <@t> gmx.at Fri Jul 1 12:55:19 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] bone-marrow decal Message-ID: Hi, What duration of fixation is recommanded for bonemarrow trephines (3 mm diameter), if you use a formicacid-formaldehyd solution subsequently for decalcification? Please, can anyone share his/her experience in this field. (morphology, ihc) Our protocol: ca. 20 hours 9% buffered formaldehyd; then 6-8 hours decal with the combinated solution. Thank you Gudrun From mtitford <@t> aol.com Fri Jul 1 13:13:07 2005 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Formic acid decal /time Message-ID: <8C74C7FAACFBDC6-E7C-8448@mblk-r27.sysops.aol.com> Following my email about Gooding & Stewarts formula for formic acid decal, Gudron Lang asks how long we decalcify for. Well... How long you decalcify should depend on how much calcium there is! For our bone marrow biopsies (about 3 mm wide) we usually fix in B-5 for two hours, followed by decal for two hours. That is normally enough time. Sometimes there is increased calcium present which was not removed and the tissue is a little crunchy. However, with a hematopathologist breathing down your neck, we live with it! (And the IHC works). Dr Dapson will tell you there are suitable B-5 replacements and maybe there are! Happy fourth of july again! Mike Titford USA Pathology Mobile AL USA From shive003 <@t> umn.edu Fri Jul 1 14:24:01 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Nestin IHC Message-ID: <014501c57e72$6fe94fe0$41065486@auxs.umn.edu> I'm sending this out for a colleague who's not a member of Histonet. Is there a lab out there that performs anti-Nestin IHC on FFPE primate tissue? And if so, do you accept cases from outside your organization? Thanks in advance for your assistance. Jan Shivers Univ. of Minnesota Veterinary Diagnostic Lab St. Paul, MN USA From sjchtascp <@t> yahoo.com Fri Jul 1 14:24:16 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Plus slides again Message-ID: <20050701192416.86848.qmail@web90204.mail.scd.yahoo.com> I've tested 4 different brands of plus slides and they all pretty much react the same. Pick the section up, wait a few seconds and float it right off the slide. I know there not working well because I see significant tissue loosening of my sections after staining. I've used plus slides at other facilities w/o being able to float off the section. I'm using distilled, about pH 6 in my water bath, no adhesive. Temp about 46-48C. Float the ribbon out, let them flatten for a few seconds, pick them up and vertically let them air dry completely before drying them at 60-65C in a forced air dryer. Elementary problem but rather annoying. Steve --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football From Thomas.Crowell <@t> biogenidec.com Fri Jul 1 15:35:07 2005 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Thomas Crowell/Cambridge/Biogen is out of the office. Message-ID: I will be out of the office starting 07/01/2005 and will not return until 07/11/2005. I will respond to your message when I return. From mfrei <@t> sial.com Fri Jul 1 15:41:36 2005 From: mfrei <@t> sial.com (Mark Frei) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Histology Stain Contest Message-ID: Sigma-Aldrich invites histologists from around the world to participate in our stain contest. Stain categories include H&E, Special Stains and an open stain category. The entry deadline is August 26, 2005. Winners will be announced at the NSH convention this September at Fort Lauderdale, Florida. Visit sigma-aldrich.com/handh for more information. Good luck! Mark Frei MT(ASCP) Technical Specialist Sigma-Aldrich Corporation 545 South Ewing St. Louis, MO 63103 mfrei@sial.com (314) 286-8080 From denise <@t> pclv.net Fri Jul 1 17:14:25 2005 From: denise <@t> pclv.net (Denise) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Employment opportunity Seattle/Everett Area Message-ID: Phoenix Central Laboratory for Veterinarians, Inc. has a position available for a Histology Technician with ASCP certification (or eligibility). Phoenix Central Laboratory, (PCLV), is a veterinary reference laboratory that provides diagnostic services to more than 800 clinics located throughout the Pacific Northwest and Alaska. We are dedicated to providing rapid, quality service. Our laboratory is located in Everett, Washington, just thirty minutes north of downtown Seattle. Visit our web site at www.pclv.net. PCLV offers a competitive salary and benefit package including health and dental coverage, personal time, and a 401K plan. Please submit your resume to Denise DeRouchey, c/o Phoenix Central Laboratory, 11620 Airport Road, Everett, WA 98204. Alternately, you may fax your resume to 425-355-3676. From lpwenk <@t> sbcglobal.net Fri Jul 1 18:13:00 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] formic acid decal In-Reply-To: Message-ID: A milder formic acid decalcification solution is FASC = formic acid-sodium citrate. It's a buffered formic acid. Sections of vertebral bone take 5-7 days to decalcify. I don't know how long a bone biopsy takes. When my students have to submit a bone section for their certification exam, this is what we use. It gives great nuclear detail, fantastic eosinophil granules. So I expect IHC would be equally great. We use the procedure out of the Bancroft book. 65 mL of 20% aqueous trisodium citrate (13 g in 65 mL water) 35 mL of 90% formic acid (that's full strength. Don't dilute) pH is about 2.3 Change solution every 3 days. We did have a rare bone tumor, and none of the IHC worked, until we decalcified it in EDTA. Then it worked great. (Sorry, don't remember what type of tumor or what type of antibody.) But you might want to try the EDTA, if you have a lot of time. Vertebral bone takes 2-3 weeks. The little bone fragments we had from the bone tumor took 3 days. (would have been a 2-3 hours in our usual 5% hydrochloric acid) 25 g EDTA disodium salt (ethylenediaminetetracetic acid) (note: MUST be disodium salt) 175 mL d. water Solution is cloudy. Add 2.5-3.0 g of sodium hydroxide until solution becomes clear. pH should be about 7 at that point. Change solution about twice a week. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Friday, July 01, 2005 8:32 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] formic acid decal Hi all, on pain of being repetitive/lazy/slovenly/slothful/unprofessional, would someone PLEASE share their recipe for formic acid decalcifying solution suitable for samples on which IHc will be performed. Many many thanks -- Louise Renton (grovelling) Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From amosbrooks <@t> earthlink.net Sat Jul 2 15:19:45 2005 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Re: Histonet Digest, Vol 19, Issue 43 In-Reply-To: <200506301237.1dO22P9I3Nl34d0@mx-pinchot.atl.sa.earthlink.net> References: <200506301237.1dO22P9I3Nl34d0@mx-pinchot.atl.sa.earthlink.net> Message-ID: <42C6F6E1.2010501@earthlink.net> Carla, We use both depending on the antibody. Mostly we use Envision, but we tried the Impress on some of the antibodies that don't label very intensely and did see improvement. If you compare the results of a CD2, CD4, CD7 or CD8 odds are you'll see some improvement. Not all the antibodies showed marked improvement and in some cases envision was better so we use Envision on most but the success we've had with Impress made it worth switching some of the procedures. Vector might give you a sample if you ask nicely. Give it a whirl and see if you like it. Best of luck, Amos Message: 17 Date: Thu, 30 Jun 2005 07:34:30 -0700 From: Carla M Conway Subject: [Histonet] Dako EnVision G2 or Vector ImmPRESS comments wanted To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hello all, I would like any comments (pro and con) regarding Dako's EnVision G2/AP system or Vector's ImmPRESS kit. We are currently using EnVision + , but are wondering if these new systems could be even better (more sensitive). Thanks for your help, Carla Conway Western Fisheries Research Center 6505 NE 65th St Seattle, WA 98115 ph: 206-526-6282 ext. 242 fax: 206-526-6654 cmconway@usgs.gov From mpowersathome <@t> clearviewcatv.net Sun Jul 3 12:53:08 2005 From: mpowersathome <@t> clearviewcatv.net (Marian Powers) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Employment opportunity Message-ID: <000c01c57ff8$12714fc0$6bfe8543@MEGAN> We are seeking a qualified Histology Technician to work part-time in Dover, Delaware. Interested candidates may visit our website at www.dpspa.com. Alternately, you may fax your resume to 302-677-0010. From bamur <@t> alaska.net Sun Jul 3 16:58:29 2005 From: bamur <@t> alaska.net (The Murray's) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Cassette printer Message-ID: Greetings, We are in the process of purchasing a cassette label printer. I would like input on the Shur/Mark and the Sakura label printer or other brands if you have one. Thanks in advance for your replies and Happy 4th! Barbara A. Murray, Histologist, (ASCP) Pathology/Histology Dept.The Alaska Native Medical Center Anchorage, Alaska From kccatunda <@t> terra.com.br Sun Jul 3 19:55:18 2005 From: kccatunda <@t> terra.com.br (Katia Cristina Catunda) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Histology Stain Contest References: Message-ID: <001001c58033$0ce52b40$a279fea9@privatexx> Great way to motivate our employees!!! Love it, Katia ----- Original Message ----- From: "Mark Frei" To: Sent: Friday, July 01, 2005 5:41 PM Subject: [Histonet] Histology Stain Contest > Sigma-Aldrich invites histologists from around the world to participate in > our stain contest. Stain categories include H&E, Special Stains and an > open stain category. The entry deadline is August 26, 2005. Winners > will be announced at the NSH convention this September at Fort Lauderdale, > Florida. Visit sigma-aldrich.com/handh for more information. > Good luck! > > Mark Frei MT(ASCP) > Technical Specialist > Sigma-Aldrich Corporation > 545 South Ewing > St. Louis, MO 63103 > mfrei@sial.com > (314) 286-8080 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From jxw387 <@t> psu.edu Sun Jul 3 22:34:25 2005 From: jxw387 <@t> psu.edu (Jianwen Wei) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] IHC with neonatal mouse bones Message-ID: <200507040334.XAA18497@webmail7.cac.psu.edu> Hi all, When I do IHC on neonatal mouse bones, I always find that there are very strong auto-fluorescence background signals from blood cells in the bone marrow. Do you know how can I avoid these background singals? Thanks, Jianwen Wei Dept. of Biology Penn State University Jianwen Wei 208 Mueller Lab Department of Biology Pennsylvania State University University Park, PA 16802 Tel: (814) 865-1769 From louise.renton <@t> gmail.com Mon Jul 4 02:53:14 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] formic acid thanks Message-ID: Thank you to all who responded so kindly and patiently to my request for a recipe, (especially Mike Titford who obviously liked the grovelling bit)...happy belated 4th of July to y'all. best regards -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From Jacqueline.Malam <@t> rli.mbht.nhs.uk Mon Jul 4 03:17:52 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] RE: trephine decal Message-ID: We use 5% glacial acetic acid in formalin for the trephine biopsy pots so it is gently decalcifying on receipt then, when fixed (usually left overnight if received late in day), we transfer it to EDTA solution (50g disod. salt in 350ml water plus 5g sodium hydroxide) and leave about 8 hours, then process as normal. Is the recommended protocol in our pathologists' text book on lymphomas, and routine immunos work well but haven't tried the whole range so can't guarantee them all Jacqui Malam Lancaster DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From carl.hobbs <@t> kcl.ac.uk Mon Jul 4 04:06:09 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Deformol Message-ID: <002301c58077$9ead7ac0$112b5c9f@Carlos> High , would be very grateful for any info on "deformol". Only ref I found was by Elvers in 1943....can't get the paper, tho. Would like to know what's in it. Said to be able to remove formalin from tissue! Thanks Carl From pruegg <@t> ihctech.net Mon Jul 4 10:24:16 2005 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] formic acid decal In-Reply-To: <200507012317.j61NH11t047068@pro12.abac.com> Message-ID: <200507041524.j64FOGvV064693@pro12.abac.com> For I IHC I use ImmunoCal for Decal Chemicals, it is a 5% formic acid with buffers. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Friday, July 01, 2005 5:13 PM To: 'louise renton'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] formic acid decal A milder formic acid decalcification solution is FASC = formic acid-sodium citrate. It's a buffered formic acid. Sections of vertebral bone take 5-7 days to decalcify. I don't know how long a bone biopsy takes. When my students have to submit a bone section for their certification exam, this is what we use. It gives great nuclear detail, fantastic eosinophil granules. So I expect IHC would be equally great. We use the procedure out of the Bancroft book. 65 mL of 20% aqueous trisodium citrate (13 g in 65 mL water) 35 mL of 90% formic acid (that's full strength. Don't dilute) pH is about 2.3 Change solution every 3 days. We did have a rare bone tumor, and none of the IHC worked, until we decalcified it in EDTA. Then it worked great. (Sorry, don't remember what type of tumor or what type of antibody.) But you might want to try the EDTA, if you have a lot of time. Vertebral bone takes 2-3 weeks. The little bone fragments we had from the bone tumor took 3 days. (would have been a 2-3 hours in our usual 5% hydrochloric acid) 25 g EDTA disodium salt (ethylenediaminetetracetic acid) (note: MUST be disodium salt) 175 mL d. water Solution is cloudy. Add 2.5-3.0 g of sodium hydroxide until solution becomes clear. pH should be about 7 at that point. Change solution about twice a week. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Friday, July 01, 2005 8:32 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] formic acid decal Hi all, on pain of being repetitive/lazy/slovenly/slothful/unprofessional, would someone PLEASE share their recipe for formic acid decalcifying solution suitable for samples on which IHc will be performed. Many many thanks -- Louise Renton (grovelling) Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere _______________________________________________ Histonet mailing list Histonet@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 cbass <@t> bidmc.harvard.edu Mon Jul 4 12:54:08 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] PCR and ISH in fixed tissues Message-ID: <9E94278E-ECB4-11D9-8BC8-0003930771EE@bidmc.harvard.edu> Hey guys, My boss is submitting a grant and he does not believe that you can do PCR (RT) or ISH with fixed tissues. I thought you could but I don't know any details. Could someone elaborate? Is there a particular way the tissue should be fixed? In other words do I have to use FFPE sections or can I use 4% paraformaldehyde perfusion? Any advice would be helpful. Thanks, Caroline Bass From ASuarez <@t> mrl.ubc.ca Mon Jul 4 13:09:57 2005 From: ASuarez <@t> mrl.ubc.ca (Agripina Suarez) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] PCR and ISH in fixed tissues Message-ID: Hi Caroline. Most of the ISH I do are on paraformaldehyde/formalin-fixed paraffin-embedded sections and they work very well. The tissues are fixed either in 4% paraformaldehyde or in commercially available 10% neutral buffered formalin for 12 to 24 hours, depends on the size of the tissue. Hope this helps. Agripina Agripina C. Suarez McDonald Research Laboratories iCAPTURE Centre, St Paul's Hospital 1081 Burrard St, Vancouver B.C., Canada V6Z 1Y6 Tel no. 604 682 2344 local 62562 Fax no. 604 806 8351 >>> Caroline Bass 7/4/2005 10:54:08 AM >>> Hey guys, My boss is submitting a grant and he does not believe that you can do PCR (RT) or ISH with fixed tissues. I thought you could but I don't know any details. Could someone elaborate? Is there a particular way the tissue should be fixed? In other words do I have to use FFPE sections or can I use 4% paraformaldehyde perfusion? Any advice would be helpful. Thanks, Caroline Bass _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From miffordclark <@t> optonline.net Mon Jul 4 15:10:06 2005 From: miffordclark <@t> optonline.net (clifford berger) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] formic acid decal References: <200507041524.j64FOGvV064693@pro12.abac.com> Message-ID: <000e01c580d4$5f1e71e0$0200a8c0@dellovo0ll7kuk> For more information on this product, technical papers and much more information, you can visit our website, www.decal-bone.com. You can also call us at 1-800-428-5856 for free samples. Best regards and happy 4rth Cliff Berger Decal Chemical Corp The Real Decal 1-800-428-5856 ----- Original Message ----- From: "patsy ruegg" To: ; "'louise renton'" ; Sent: Monday, July 04, 2005 11:24 AM Subject: RE: [Histonet] formic acid decal > For I IHC I use ImmunoCal for Decal Chemicals, it is a 5% formic acid with > buffers. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy > Wenk > Sent: Friday, July 01, 2005 5:13 PM > To: 'louise renton'; Histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] formic acid decal > > A milder formic acid decalcification solution is FASC = formic acid-sodium > citrate. It's a buffered formic acid. Sections of vertebral bone take 5-7 > days to decalcify. I don't know how long a bone biopsy takes. When my > students have to submit a bone section for their certification exam, this is > what we use. It gives great nuclear detail, fantastic eosinophil granules. > So I expect IHC would be equally great. We use the procedure out of the > Bancroft book. > > 65 mL of 20% aqueous trisodium citrate (13 g in 65 mL water) > 35 mL of 90% formic acid (that's full strength. Don't dilute) > pH is about 2.3 > Change solution every 3 days. > > We did have a rare bone tumor, and none of the IHC worked, until we > decalcified it in EDTA. Then it worked great. (Sorry, don't remember what > type of tumor or what type of antibody.) But you might want to try the EDTA, > if you have a lot of time. Vertebral bone takes 2-3 weeks. The little bone > fragments we had from the bone tumor took 3 days. (would have been a 2-3 > hours in our usual 5% hydrochloric acid) > > 25 g EDTA disodium salt (ethylenediaminetetracetic acid) (note: MUST be > disodium salt) > 175 mL d. water > Solution is cloudy. Add 2.5-3.0 g of sodium hydroxide until solution becomes > clear. pH should be about 7 at that point. > Change solution about twice a week. > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > Lee & Peggy Wenk > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise > renton > Sent: Friday, July 01, 2005 8:32 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] formic acid decal > > Hi all, > > on pain of being repetitive/lazy/slovenly/slothful/unprofessional, > would someone PLEASE share their recipe for formic acid decalcifying > solution suitable for samples on which IHc will be performed. Many many > thanks > > -- > Louise Renton (grovelling) > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "....I know who I am. No-one else knows who I am. If I was a giraffe, and > someone said I was a snake, I'd think no, actually I'm a giraffe" > Richard Gere > > _______________________________________________ > Histonet mailing list > Histonet@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 AnthonyH <@t> chw.edu.au Mon Jul 4 18:10:29 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] PCR and ISH in fixed tissues Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E2C0@simba.kids> Caroline, We have successfully done ISH for EBERs, Adenovirus RNA, CMV RNA, Chromogranin mRNA, Estrogen Receptor mRNA, and nCAM RNA to name a few on both Frozen sections and Formalin Fixed Paraffin Embedded Tissue (FFPE). We have also successfully extracted RNA and DNA from FFPE tissues for PCR (Ewing's and Rhabdo translocation PCR studies)and do this routinely. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Caroline Bass [mailto:cbass@bidmc.harvard.edu] Sent: Tuesday, 5 July 2005 3:54 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] PCR and ISH in fixed tissues Hey guys, My boss is submitting a grant and he does not believe that you can do PCR (RT) or ISH with fixed tissues. I thought you could but I don't know any details. Could someone elaborate? Is there a particular way the tissue should be fixed? In other words do I have to use FFPE sections or can I use 4% paraformaldehyde perfusion? Any advice would be helpful. Thanks, Caroline Bass _______________________________________________ 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 wolf <@t> webhart.net Mon Jul 4 19:07:23 2005 From: wolf <@t> webhart.net (Patrick Paulusse) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] ER/PR staining of normal breast tissue Message-ID: <000701c56962$85217100$57b05bd1@wolf> Good Evening All, Should normal breast duct tissue always stain positively with estrogen and progesterone antibodies. If not, is this a natural occurrence or is there something physiologically occuring to prevent the staining. I have had several occasions when the normal breast duct tissue will not stain (the corresponding tumour tissue in the same block was also negative). An external control on the same slide stains wonderfully. Fixation is never less than 24 hours and seldom over 72 hours in 10% NBF. I have several control blocks and use the one that mimics the fixation and processing of the test specimen. Any comments and references are always welcome. Thanking you in advance, Patrick Paulusse Anatomical Pathology Pembroke Regional Hospital From ree3 <@t> leicester.ac.uk Tue Jul 5 06:32:03 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] non-muscle myosin IIA Message-ID: Looking for an antibody to the above for immunohistochemistry... Thanks Richard Edwards MRC TOX UNIT.....LEICESTER.....U.K........ From TMcNemar <@t> lmhealth.org Tue Jul 5 08:25:27 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Rotating shifts... Message-ID: <6CD94D97ED7D924BA5C2B588FA9528213967D9@nt_exchange.lmhealth.org> I was wondering if most histo labs use rotating shifts or if people mostly work the same hours. Do you have people that just work one shift and primarily do one job (gross, cut, stain, etc.)? At full staff we have 4 techs and 3 of them rotate between 2 weeks of 6-2:30 and 1 week of 7:30-4 (surgicals). I wondered if switching the hours to a straight 7:30 - 4 position might help attract someone. I was also thinking that we would have a more consistency/continuity if everyone had a certain job/area that they were primarily responsible for. All thoughts appreciated. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 From louise.renton <@t> gmail.com Tue Jul 5 08:52:42 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] antibody search for BMP Message-ID: Dear all, I am looking for antibodies to BMP (bone morphogenic protein) 3, 6 and 7 (AKA Op1) for use on FFPE embedded tissue. I would be grateful for any input, as an internet search has come up with very little information. Thank you -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From JNocito <@t> Pathreflab.com Tue Jul 5 09:01:51 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] revisiting expired immmuno reagents Message-ID: Hello histoland, okay, I've been shooting my mouth off a lot lately and I want y'all to know that I contacted the CAP about revising the question about using expired immuno reagents. The question in question (like that huh?) in the latest CAP checklist is ANP.22432. Whether they contact me or not is another question (so many questions). My rationale for changing the question are these 1. Immuno reagents are too expensive to throw away. 2. Each time a run is performed, the reagents are revalidated 3. A laboratory must have a written procedure in place detailing the revalidating process. 4. A quality control form must be used and signed by the medical director assuring that the reagents work. I am open to suggestions and other ideas that I may have over looked. With your support, maybe we can, at least, have this question revised. Of course, I expect some fallout because of this, but hey, it wouldn't be me if I didn't Enjoy!!! Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX From PIXLEYSK <@t> UCMAIL.UC.EDU Tue Jul 5 09:25:23 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] RE: Formic Acid decalcification Message-ID: Dear Louise Renton: Here is a method from a research lab. We use 10% formic acid, but add an ion exchange resin that soaks up the calcium. This eliminates the need to change solutions, but brings its own problems since the resin must be stirred. This is faster than EDTA, but I don't know if it is faster than just a 10% formic acid solution alone. The mixture is good for many decalcifications. Great ICC staining. Formic Acid Decalcification: Purchase (i.e., Fisher Scientific): Rexyn 101 H ion exchange resin formic acid (full strength, ~90%). Make 6-10 liters of 10% formic acid (in double distilled water) Add 500 g of resin (So ~50-83 g resin per liter of 10% formic acid) Need stir bar to keep resin suspended, so put samples in cloth bags (i.e., homemade cotton bags) and suspend from side. Alternatively, use lab dessicator. Samples sit above dessicator plate while stir bar goes underneath. Sincerely, Sarah Pixley, Ph.D. Dept. of Cell Biology, Neurobiology and Anatomy University of Cincinnati College of Medicine Cincinnati, OH 45267-0521 From deb.vaneyck <@t> phci.org Tue Jul 5 09:28:07 2005 From: deb.vaneyck <@t> phci.org (Van Eyck, Deb) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Impress Message-ID: Dear-histonetters-----I have also wondered about the Impress Kit----Amos and to otheres who tried it---was it pretty interchangeable- time wise with the Envision , or Envision + also what about antibody dilutions? same strength? Another immuno question for you all? is anyone doing Toxoplasmosis staining? What antibody are you using? What protocol? Thanks much-Deb Van Eyck > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [SMTP:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > histonet-request@lists.utsouthwestern.edu > Sent: Sunday, July 03, 2005 12:06 PM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 20, Issue 3 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Re: Histonet Digest, Vol 19, Issue 43 (Amos Brooks) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sat, 02 Jul 2005 16:19:45 -0400 > From: Amos Brooks > Subject: [Histonet] Re: Histonet Digest, Vol 19, Issue 43 > To: histonet@lists.utsouthwestern.edu > Message-ID: <42C6F6E1.2010501@earthlink.net> > Content-Type: text/plain; charset=ISO-8859-1; format=flowed > > Carla, > > We use both depending on the antibody. Mostly we use Envision, but > we tried the Impress on some of the antibodies that don't label very > intensely and did see improvement. If you compare the results of a CD2, > CD4, CD7 or CD8 odds are you'll see some improvement. Not all the > antibodies showed marked improvement and in some cases envision was better > so we use Envision on most but the success we've had with Impress made it > worth switching some of the procedures. Vector might give you a sample if > you ask nicely. Give it a whirl and see if you like it. > > Best of luck, > Amos > > > > Message: 17 > Date: Thu, 30 Jun 2005 07:34:30 -0700 > From: Carla M Conway > Subject: [Histonet] Dako EnVision G2 or Vector ImmPRESS comments > wanted > To: Histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=US-ASCII > > Hello all, > > I would like any comments (pro and con) regarding Dako's EnVision G2/AP > system or Vector's ImmPRESS kit. We are currently using EnVision + , but > are wondering if these new systems could be even better (more sensitive). > > Thanks for your help, > > Carla Conway > > Western Fisheries Research Center > 6505 NE 65th St > Seattle, WA 98115 > ph: 206-526-6282 ext. 242 > fax: 206-526-6654 > cmconway@usgs.gov > > > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 20, Issue 3 > *************************************** > > This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. From TillRenee <@t> uams.edu Tue Jul 5 10:39:33 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] liver cryosections Message-ID: Can anyone offer some advice on cutting liver cryosections? My every attempt produces shredded tissues. I know in paraffin embedding that means they are dry and I would soak it in ice water before I cut, but what do you do with cryosections? I'm pretty sure it's not my cutting technique, though I am fairly new at it. I've adjusted the roll plate and the vacume window. And I've tried it without the roll plate, though as we are just starting out with our cryostat I don't have any good brushes for pulling the section. Any ideas? Maybe they did not have enough time to come down from -80 to -20? I left them in for about 2 hours. Renee' From PMonfils <@t> Lifespan.org Tue Jul 5 10:45:01 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] liver cryosections Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717565@lsexch.lsmaster.lifespan.org> The block is too cold. -20 degrees is an appropriate temperature for sectioning many tissues, but some tissues require a lower temperature and others require a higher temperature. For liver, try -15 degrees. For some samples you may have to go as high as -12 degrees. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Till, > Renee > Sent: Tuesday, July 5, 2005 8:39 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] liver cryosections > > Can anyone offer some advice on cutting liver cryosections? My every > attempt produces shredded tissues. I know in paraffin embedding that > means they are dry and I would soak it in ice water before I cut, but > what do you do with cryosections? I'm pretty sure it's not my cutting > technique, though I am fairly new at it. I've adjusted the roll plate > and the vacume window. And I've tried it without the roll plate, though > as we are just starting out with our cryostat I don't have any good > brushes for pulling the section. Any ideas? Maybe they did not have > enough time to come down from -80 to -20? I left them in for about 2 > hours. > > > > Renee' > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From algranth <@t> u.arizona.edu Tue Jul 5 12:10:50 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] liver cryosections In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE01717565@lsexch.lsmaster.l ifespan.org> Message-ID: <4.3.2.7.2.20050705100141.00cc1258@algranth.inbox.email.arizona.edu> Renee, I do many frozen sections on liver tissue and have found that the manner in which the tissue is frozen makes a huge difference in the way it cuts. Check to make sure that your samples are being frozen properly. There have been numerous discussions on histonet about the proper way to freeze tissue and you can find them in the histonet archives. I have found that if your tissue looks chalky and whitish then it may have been left in whatever froze the tissue too long. This makes for real fly away shattered sections and sometimes cracks the OCT which makes for another nightmare. Next, I cut my liver frozens at -18 to -16 degrees C. Before taking the section I often have to rub my thumb gently over the surface of the tissue (wearing gloves) to slightly warm the surface. This helps to produce a section that is not shattered. Sometimes a puff of warm air helps too. Breathe gently on the tissue as you tease down the section and it doesn't want to fly away. Andi At 11:45 AM 7/5/2005 -0400, you wrote: >The block is too cold. -20 degrees is an appropriate temperature for >sectioning many tissues, but some tissues require a lower temperature and >others require a higher temperature. For liver, try -15 degrees. For some >samples you may have to go as high as -12 degrees. > >Paul M. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Till, > > Renee > > Sent: Tuesday, July 5, 2005 8:39 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] liver cryosections > > > > Can anyone offer some advice on cutting liver cryosections? My every > > attempt produces shredded tissues. I know in paraffin embedding that > > means they are dry and I would soak it in ice water before I cut, but > > what do you do with cryosections? I'm pretty sure it's not my cutting > > technique, though I am fairly new at it. I've adjusted the roll plate > > and the vacume window. And I've tried it without the roll plate, though > > as we are just starting out with our cryostat I don't have any good > > brushes for pulling the section. Any ideas? Maybe they did not have > > enough time to come down from -80 to -20? I left them in for about 2 > > hours. > > > > > > > > Renee' > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From GauchV <@t> mail.amc.edu Tue Jul 5 12:32:03 2005 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Rotating shifts... Message-ID: Tom, We have our techs on a set schedule with no rotations as far as hours are concerned. We rotate job duties monthly with the flexiblity to cover techs who are off as needed. At one point in time, we DID have a person dedicated solely to our IHC area which ultimately was a big mistake. When that person decided to leave, no one had really rotated through there in quite a while and so all of the little "tweeks" that had been worked out were not known to anyone except the person leaving. Unfortunately, he was not of the mindset to show anyone before he left which left us in a bit of a jam. Having learned from that we now have tech specialists who oversee each area and the techs rotate so that several people are well trained and up to speed in each part of the lab at any given time. We do limit the number of people who rotate through IHC to a few techs and find that the consistency and quality of the staining has been fine. Hope that helps, Vicki AMCH Albany, NY >>> Tom McNemar 7/5/2005 9:25:27 AM >>> I was wondering if most histo labs use rotating shifts or if people mostly work the same hours. Do you have people that just work one shift and primarily do one job (gross, cut, stain, etc.)? At full staff we have 4 techs and 3 of them rotate between 2 weeks of 6-2:30 and 1 week of 7:30-4 (surgicals). I wondered if switching the hours to a straight 7:30 - 4 position might help attract someone. I was also thinking that we would have a more consistency/continuity if everyone had a certain job/area that they were primarily responsible for. All thoughts appreciated. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Tue Jul 5 12:41:15 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:17 2005 Subject: Tom McNemar [Histonet] Rotating shifts... In-Reply-To: <6CD94D97ED7D924BA5C2B588FA9528213967D9@nt_exchange.lmhealth.org> Message-ID: <001d01c58188$be6b0ff0$1d2a14ac@wchsys.org> I use three shifts but have different benches associated with the shifts. Embedding 6-2:30, Accessioning(non tech) 7:30-4:00, Two cutting benches 7-3:30 (these two benches are separated into different jobs) The techs say they like the rotation because it keeps them able to do all aspects of the job and prevents "same ole same ole" boredom. Histotechs are hard to come by no matter how the jobs are arranged. -----Original Message I was wondering if most histo labs use rotating shifts or if people mostly work the same hours. Do you have people that just work one shift and primarily do one job (gross, cut, stain, etc.)? At full staff we have 4 techs and 3 of them rotate between 2 weeks of 6-2:30 and 1 week of 7:30-4 (surgicals). I wondered if switching the hours to a straight 7:30 - 4 position might help attract someone. I was also thinking that we would have a more consistency/continuity if everyone had a certain job/area that they were primarily responsible for. All thoughts appreciated. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From DEllenburg2 <@t> stfrancishealth.org Tue Jul 5 12:47:53 2005 From: DEllenburg2 <@t> stfrancishealth.org (DEllenburg2@stfrancishealth.org) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Rotating Shifts Message-ID: <0A502E8156DAA4468CB8979B27177555206193@bssmsx5501.servers.intranet.stfrancishealth.com> Tom, At full staff we have 2 techs and one assistant. At the present time we do not rotate shifts. We cover from 5a to 5p. However, both of these options have advantages and disadvantages. For my lab it works better not using the rotating shifts. You just have to make sure that each employee is aware of the job expectations and is competent in each area of the various duties of each shift. Hope this helps. Please feel free to give me a call if I can be or further help. Deborah Ellenburg, HT, (ASCP) Histology Supervisor 864-255-1582 -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: None To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 5 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. PCR and ISH in fixed tissues (Caroline Bass) 2. Re: PCR and ISH in fixed tissues (Agripina Suarez) 3. Re: formic acid decal (clifford berger) 4. RE: PCR and ISH in fixed tissues (Tony Henwood) 5. ER/PR staining of normal breast tissue (Patrick Paulusse) 6. non-muscle myosin IIA (Edwards, R.E.) 7. Rotating shifts... (Tom McNemar) 8. antibody search for BMP (louise renton) 9. revisiting expired immmuno reagents (Joe Nocito) 10. RE: Formic Acid decalcification (Pixley, Sarah (pixleysk)) 11. Impress (Van Eyck, Deb) 12. liver cryosections (Till, Renee) 13. RE: liver cryosections (Monfils, Paul) ---------------------------------------------------------------------- Message: 1 Date: Mon, 04 Jul 2005 13:54:08 -0400 From: Caroline Bass Subject: [Histonet] PCR and ISH in fixed tissues To: 'histonet@lists.utsouthwestern.edu' Message-ID: <9E94278E-ECB4-11D9-8BC8-0003930771EE@bidmc.harvard.edu> Content-Type: text/plain; charset=US-ASCII; format=flowed Hey guys, My boss is submitting a grant and he does not believe that you can do PCR (RT) or ISH with fixed tissues. I thought you could but I don't know any details. Could someone elaborate? Is there a particular way the tissue should be fixed? In other words do I have to use FFPE sections or can I use 4% paraformaldehyde perfusion? Any advice would be helpful. Thanks, Caroline Bass ------------------------------ Message: 2 Date: Mon, 04 Jul 2005 11:09:57 -0700 From: "Agripina Suarez" Subject: Re: [Histonet] PCR and ISH in fixed tissues To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Caroline. Most of the ISH I do are on paraformaldehyde/formalin-fixed paraffin-embedded sections and they work very well. The tissues are fixed either in 4% paraformaldehyde or in commercially available 10% neutral buffered formalin for 12 to 24 hours, depends on the size of the tissue. Hope this helps. Agripina Agripina C. Suarez McDonald Research Laboratories iCAPTURE Centre, St Paul's Hospital 1081 Burrard St, Vancouver B.C., Canada V6Z 1Y6 Tel no. 604 682 2344 local 62562 Fax no. 604 806 8351 >>> Caroline Bass 7/4/2005 10:54:08 AM >>> Hey guys, My boss is submitting a grant and he does not believe that you can do PCR (RT) or ISH with fixed tissues. I thought you could but I don't know any details. Could someone elaborate? Is there a particular way the tissue should be fixed? In other words do I have to use FFPE sections or can I use 4% paraformaldehyde perfusion? Any advice would be helpful. Thanks, Caroline Bass _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Mon, 04 Jul 2005 16:10:06 -0400 From: clifford berger Subject: Re: [Histonet] formic acid decal To: patsy ruegg , lpwenk@sbcglobal.net, 'louise renton' , Histonet@lists.utsouthwestern.edu Message-ID: <000e01c580d4$5f1e71e0$0200a8c0@dellovo0ll7kuk> Content-Type: text/plain; charset=iso-8859-1 For more information on this product, technical papers and much more information, you can visit our website, www.decal-bone.com. You can also call us at 1-800-428-5856 for free samples. Best regards and happy 4rth Cliff Berger Decal Chemical Corp The Real Decal 1-800-428-5856 ----- Original Message ----- From: "patsy ruegg" To: ; "'louise renton'" ; Sent: Monday, July 04, 2005 11:24 AM Subject: RE: [Histonet] formic acid decal > For I IHC I use ImmunoCal for Decal Chemicals, it is a 5% formic acid with > buffers. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy > Wenk > Sent: Friday, July 01, 2005 5:13 PM > To: 'louise renton'; Histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] formic acid decal > > A milder formic acid decalcification solution is FASC = formic acid-sodium > citrate. It's a buffered formic acid. Sections of vertebral bone take 5-7 > days to decalcify. I don't know how long a bone biopsy takes. When my > students have to submit a bone section for their certification exam, this is > what we use. It gives great nuclear detail, fantastic eosinophil granules. > So I expect IHC would be equally great. We use the procedure out of the > Bancroft book. > > 65 mL of 20% aqueous trisodium citrate (13 g in 65 mL water) > 35 mL of 90% formic acid (that's full strength. Don't dilute) > pH is about 2.3 > Change solution every 3 days. > > We did have a rare bone tumor, and none of the IHC worked, until we > decalcified it in EDTA. Then it worked great. (Sorry, don't remember what > type of tumor or what type of antibody.) But you might want to try the EDTA, > if you have a lot of time. Vertebral bone takes 2-3 weeks. The little bone > fragments we had from the bone tumor took 3 days. (would have been a 2-3 > hours in our usual 5% hydrochloric acid) > > 25 g EDTA disodium salt (ethylenediaminetetracetic acid) (note: MUST be > disodium salt) > 175 mL d. water > Solution is cloudy. Add 2.5-3.0 g of sodium hydroxide until solution becomes > clear. pH should be about 7 at that point. > Change solution about twice a week. > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > Lee & Peggy Wenk > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise > renton > Sent: Friday, July 01, 2005 8:32 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] formic acid decal > > Hi all, > > on pain of being repetitive/lazy/slovenly/slothful/unprofessional, > would someone PLEASE share their recipe for formic acid decalcifying > solution suitable for samples on which IHc will be performed. Many many > thanks > > -- > Louise Renton (grovelling) > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "....I know who I am. No-one else knows who I am. If I was a giraffe, and > someone said I was a snake, I'd think no, actually I'm a giraffe" > Richard Gere > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Tue, 5 Jul 2005 09:10:29 +1000 From: Tony Henwood Subject: RE: [Histonet] PCR and ISH in fixed tissues To: "'Caroline Bass'" , "'histonet@lists.utsouthwestern.edu'" Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E2C0@simba.kids> Content-Type: text/plain Caroline, We have successfully done ISH for EBERs, Adenovirus RNA, CMV RNA, Chromogranin mRNA, Estrogen Receptor mRNA, and nCAM RNA to name a few on both Frozen sections and Formalin Fixed Paraffin Embedded Tissue (FFPE). We have also successfully extracted RNA and DNA from FFPE tissues for PCR (Ewing's and Rhabdo translocation PCR studies)and do this routinely. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Caroline Bass [mailto:cbass@bidmc.harvard.edu] Sent: Tuesday, 5 July 2005 3:54 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] PCR and ISH in fixed tissues Hey guys, My boss is submitting a grant and he does not believe that you can do PCR (RT) or ISH with fixed tissues. I thought you could but I don't know any details. Could someone elaborate? Is there a particular way the tissue should be fixed? In other words do I have to use FFPE sections or can I use 4% paraformaldehyde perfusion? Any advice would be helpful. Thanks, Caroline Bass _______________________________________________ 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. ********************************************************************** ------------------------------ Message: 5 Date: Sat, 4 Jun 2005 20:06:38 -0400 From: "Patrick Paulusse" Subject: [Histonet] ER/PR staining of normal breast tissue To: "Histonet" Message-ID: <000701c56962$85217100$57b05bd1@wolf> Content-Type: text/plain; charset="iso-8859-1" Good Evening All, Should normal breast duct tissue always stain positively with estrogen and progesterone antibodies. If not, is this a natural occurrence or is there something physiologically occuring to prevent the staining. I have had several occasions when the normal breast duct tissue will not stain (the corresponding tumour tissue in the same block was also negative). An external control on the same slide stains wonderfully. Fixation is never less than 24 hours and seldom over 72 hours in 10% NBF. I have several control blocks and use the one that mimics the fixation and processing of the test specimen. Any comments and references are always welcome. Thanking you in advance, Patrick Paulusse Anatomical Pathology Pembroke Regional Hospital ------------------------------ Message: 6 Date: Tue, 5 Jul 2005 12:32:03 +0100 From: "Edwards, R.E." Subject: [Histonet] non-muscle myosin IIA To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Looking for an antibody to the above for immunohistochemistry... Thanks Richard Edwards MRC TOX UNIT.....LEICESTER.....U.K........ ------------------------------ Message: 7 Date: Tue, 5 Jul 2005 09:25:27 -0400 From: Tom McNemar Subject: [Histonet] Rotating shifts... To: Histonet Message-ID: <6CD94D97ED7D924BA5C2B588FA9528213967D9@nt_exchange.lmhealth.org> Content-Type: text/plain; charset="iso-8859-1" I was wondering if most histo labs use rotating shifts or if people mostly work the same hours. Do you have people that just work one shift and primarily do one job (gross, cut, stain, etc.)? At full staff we have 4 techs and 3 of them rotate between 2 weeks of 6-2:30 and 1 week of 7:30-4 (surgicals). I wondered if switching the hours to a straight 7:30 - 4 position might help attract someone. I was also thinking that we would have a more consistency/continuity if everyone had a certain job/area that they were primarily responsible for. All thoughts appreciated. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 ------------------------------ Message: 8 Date: Tue, 5 Jul 2005 15:52:42 +0200 From: louise renton Subject: [Histonet] antibody search for BMP To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Dear all, I am looking for antibodies to BMP (bone morphogenic protein) 3, 6 and 7 (AKA Op1) for use on FFPE embedded tissue. I would be grateful for any input, as an internet search has come up with very little information. Thank you -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere ------------------------------ Message: 9 Date: Tue, 5 Jul 2005 09:01:51 -0500 From: "Joe Nocito" Subject: [Histonet] revisiting expired immmuno reagents To: "Histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello histoland, okay, I've been shooting my mouth off a lot lately and I want y'all to know that I contacted the CAP about revising the question about using expired immuno reagents. The question in question (like that huh?) in the latest CAP checklist is ANP.22432. Whether they contact me or not is another question (so many questions). My rationale for changing the question are these 1. Immuno reagents are too expensive to throw away. 2. Each time a run is performed, the reagents are revalidated 3. A laboratory must have a written procedure in place detailing the revalidating process. 4. A quality control form must be used and signed by the medical director assuring that the reagents work. I am open to suggestions and other ideas that I may have over looked. With your support, maybe we can, at least, have this question revised. Of course, I expect some fallout because of this, but hey, it wouldn't be me if I didn't Enjoy!!! Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX ------------------------------ Message: 10 Date: Tue, 5 Jul 2005 10:25:23 -0400 From: "Pixley, Sarah (pixleysk)" Subject: [Histonet] RE: Formic Acid decalcification To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Dear Louise Renton: Here is a method from a research lab. We use 10% formic acid, but add an ion exchange resin that soaks up the calcium. This eliminates the need to change solutions, but brings its own problems since the resin must be stirred. This is faster than EDTA, but I don't know if it is faster than just a 10% formic acid solution alone. The mixture is good for many decalcifications. Great ICC staining. Formic Acid Decalcification: Purchase (i.e., Fisher Scientific): Rexyn 101 H ion exchange resin formic acid (full strength, ~90%). Make 6-10 liters of 10% formic acid (in double distilled water) Add 500 g of resin (So ~50-83 g resin per liter of 10% formic acid) Need stir bar to keep resin suspended, so put samples in cloth bags (i.e., homemade cotton bags) and suspend from side. Alternatively, use lab dessicator. Samples sit above dessicator plate while stir bar goes underneath. Sincerely, Sarah Pixley, Ph.D. Dept. of Cell Biology, Neurobiology and Anatomy University of Cincinnati College of Medicine Cincinnati, OH 45267-0521 ------------------------------ Message: 11 Date: Tue, 5 Jul 2005 09:28:07 -0500 From: "Van Eyck, Deb" Subject: [Histonet] Impress To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Dear-histonetters-----I have also wondered about the Impress Kit----Amos and to otheres who tried it---was it pretty interchangeable- time wise with the Envision , or Envision + also what about antibody dilutions? same strength? Another immuno question for you all? is anyone doing Toxoplasmosis staining? What antibody are you using? What protocol? Thanks much-Deb Van Eyck > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [SMTP:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > histonet-request@lists.utsouthwestern.edu > Sent: Sunday, July 03, 2005 12:06 PM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 20, Issue 3 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Re: Histonet Digest, Vol 19, Issue 43 (Amos Brooks) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sat, 02 Jul 2005 16:19:45 -0400 > From: Amos Brooks > Subject: [Histonet] Re: Histonet Digest, Vol 19, Issue 43 > To: histonet@lists.utsouthwestern.edu > Message-ID: <42C6F6E1.2010501@earthlink.net> > Content-Type: text/plain; charset=ISO-8859-1; format=flowed > > Carla, > > We use both depending on the antibody. Mostly we use Envision, but > we tried the Impress on some of the antibodies that don't label very > intensely and did see improvement. If you compare the results of a CD2, > CD4, CD7 or CD8 odds are you'll see some improvement. Not all the > antibodies showed marked improvement and in some cases envision was better > so we use Envision on most but the success we've had with Impress made it > worth switching some of the procedures. Vector might give you a sample if > you ask nicely. Give it a whirl and see if you like it. > > Best of luck, > Amos > > > > Message: 17 > Date: Thu, 30 Jun 2005 07:34:30 -0700 > From: Carla M Conway > Subject: [Histonet] Dako EnVision G2 or Vector ImmPRESS comments > wanted > To: Histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=US-ASCII > > Hello all, > > I would like any comments (pro and con) regarding Dako's EnVision G2/AP > system or Vector's ImmPRESS kit. We are currently using EnVision + , but > are wondering if these new systems could be even better (more sensitive). > > Thanks for your help, > > Carla Conway > > Western Fisheries Research Center > 6505 NE 65th St > Seattle, WA 98115 > ph: 206-526-6282 ext. 242 > fax: 206-526-6654 > cmconway@usgs.gov > > > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 20, Issue 3 > *************************************** > > This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. ------------------------------ Message: 12 Date: Tue, 5 Jul 2005 10:39:33 -0500 From: "Till, Renee" Subject: [Histonet] liver cryosections To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Can anyone offer some advice on cutting liver cryosections? My every attempt produces shredded tissues. I know in paraffin embedding that means they are dry and I would soak it in ice water before I cut, but what do you do with cryosections? I'm pretty sure it's not my cutting technique, though I am fairly new at it. I've adjusted the roll plate and the vacume window. And I've tried it without the roll plate, though as we are just starting out with our cryostat I don't have any good brushes for pulling the section. Any ideas? Maybe they did not have enough time to come down from -80 to -20? I left them in for about 2 hours. Renee' ------------------------------ Message: 13 Date: Tue, 5 Jul 2005 11:45:01 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] liver cryosections To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717565@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="ISO-8859-1" The block is too cold. -20 degrees is an appropriate temperature for sectioning many tissues, but some tissues require a lower temperature and others require a higher temperature. For liver, try -15 degrees. For some samples you may have to go as high as -12 degrees. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Till, > Renee > Sent: Tuesday, July 5, 2005 8:39 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] liver cryosections > > Can anyone offer some advice on cutting liver cryosections? My every > attempt produces shredded tissues. I know in paraffin embedding that > means they are dry and I would soak it in ice water before I cut, but > what do you do with cryosections? I'm pretty sure it's not my cutting > technique, though I am fairly new at it. I've adjusted the roll plate > and the vacume window. And I've tried it without the roll plate, though > as we are just starting out with our cryostat I don't have any good > brushes for pulling the section. Any ideas? Maybe they did not have > enough time to come down from -80 to -20? I left them in for about 2 > hours. > > > > Renee' > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 5 *************************************** The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at (864) 255-1000, and permanently delete the original e-mail, attachment(s), and any copies. From GauchV <@t> mail.amc.edu Tue Jul 5 12:56:40 2005 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Zymed Kit Message-ID: Hi, I am posting this for a colleague of mine...Is anyone using and having success with Zymed's Rembrandt In Situ Hybridization and Detection- DISH and AP Detection kit for HPV typing or for the 16/18 probe? Is anyone having success with any other company's 16/18/ HPV In Situ methods? Any help would be greatly appreciated. You can send any responses to me through the Histonet and I will get them to her. Thank you in advance for your assistance, Vicki AMCH Albany, NY From dmccaig <@t> ckha.on.ca Tue Jul 5 12:59:30 2005 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] STAFFING VS WORKLOAD Message-ID: <3E5A3F039F0BD8118B4700C00D002024043605@CKHA9> Can you give some insite to how many tech's you have working and the number of cases you do/year. What is the expectation of number of blocks cut. For example, if you have someone else embedding and staining and the tech is responsible solely for cutting, how long would you expect 50 blocks of various tissues (large specimens as well as endoscopic biopsies) take to be microtomed? Diana McCaig, MLT From PIXLEYSK <@t> UCMAIL.UC.EDU Tue Jul 5 13:04:34 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] RE: frozen sections shredding Message-ID: Dear Renee: I agree that the block is too cold. Probably because you went from -80 to -20 too quickly (in other words, 2 hrs is not long enough). We have found the same problem when we fix in isopentane at dry ice temp. We have to let the block sit in a -20 freezer overnight before it is appropriate to cut. Now I freeze in the cryostat, using only some dry ice powder to speed up freezing, so I don't have to wait that long. I am actually looking hoping this list will give me some better freezing methods. The isopentane is too cold and a lot of my blocks crack. Sarah Pixley From quinntl <@t> umkc.edu Tue Jul 5 13:08:57 2005 From: quinntl <@t> umkc.edu (Quinn, Tim L.) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Image analysis Message-ID: <3563FA9377A5DC41BDDA1E8A6C06BEF802DDFDA6@KC-MAIL6.kc.umkc.edu> Dear Listies, I need to collect data from immuno stained skin tissue for stat analysis. I want to convert stained areas to pixel values for comparison to non stained areas. I have NIH Image J and PhotoShop 5.5. Has anyone worked with this software to evaluate for stats? Cheers, Timothy L. Quinn Senior Lab Techician Missouri University at Kansas City Medical School- Med Lab 2411 Holmes Kansas City, MO 64108 816-235-1904 quinntl@umkc.edu From petepath <@t> yahoo.com Tue Jul 5 13:48:57 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] liver cryosections Message-ID: <20050705184857.18715.qmail@web30414.mail.mud.yahoo.com> Hi Renee, I agree with the others who have suggested that the tissue is too cold. My frozen section tutorial has some pics of shattered sections on this page so you can see compare your results to these over-cooled sections. http://www.pathologyinnovations.com/new_page_2.htm Liver is one of the easier tissues to cut. It will cut best at the warmest temp that it will cut at all. Get your blocks down to cryostat temp and then use your thumb to warm it to a point where it cuts without shattering. Stephen From mpowersathome <@t> clearviewcatv.net Tue Jul 5 14:16:54 2005 From: mpowersathome <@t> clearviewcatv.net (Marian Powers) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Employment opportunity in Delaware Message-ID: <000b01c58196$1b4bd800$6bfe8543@MEGAN> We are seeking a qualified part-time histotech to work in Dover, DE. Interested candidates may call 302-677-0000 or visit our website at www.dpspa.com. From lu_ze <@t> sbcglobal.net Tue Jul 5 15:31:47 2005 From: lu_ze <@t> sbcglobal.net (Ze Lu) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Tissue processor (LEICA TP 1050) paraffin blockage, help needed Message-ID: <00c701c581a0$90a28e50$5b02a8c0@OPTIMUM2> Hi, histonet friends, We had a TP 1050 tissue processor runs well before. However there is a consistent problem recently. When running retort cleanning after fininsh of tissue processing, there is always error message of "paraffin blockage". We did notice that paraffin blocked the white plastic tube (transfer the solvent from the solvent station into the retort) right below the retort. We need to use blow dryer to get the cleanning cycle go through every time. It seemd that paraffin leaks into that tube and cold down there duing the last three steps (in paraffin step) in tissue process. I am wondering whether somebody else has the same problem. We do use "P/V" combined setting in the three paraffin steps. If using AMB, P or Vac alone, will that make the leakage less? Thank you very much. Ze From ploykasek <@t> phenopath.com Tue Jul 5 16:30:32 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Phospho-stat5 Message-ID: HI all. I'm hoping that someone out there can help me out with a problem antibody. We are trying to work up an antibody to phospho-stat5 - not having much luck. Mainly too much background staining. To get any staining at all have had to try all kinds of pretreatments, blocking, overnight incubation, etc.... Would appreciate any insight from anyone who has tried this antibody. Thanks so much. Patti Loykasek PhenoPath Laboratories Seattle, WA From histojock <@t> hotmail.com Tue Jul 5 20:05:58 2005 From: histojock <@t> hotmail.com (Histo Jock) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] RE: PCR and ISH in fixed tissues In-Reply-To: Message-ID: Hi Caroline, Here's what I know: The issue isn't whether or not you can "do" RNA work on fixed tissues, it's whether or not you can trust the result. At my former place of employment we ran a study comparing ISH and IHC results on FFPE and matched snap frozen specimens. The frozen results were always rock steady and matched the IHC dead on. The ISH on FFPE was all over the map - when you actually got staining that was. Results I've seen from other places follow the same pattern. Absolutely no consistancy with the fixed material and much, much tougher to optimize the signal. I know that fixation procedure is a least partly to blame for this. If the initial specimen was a big piece you'll often see less signal in the center as it took longer for fixative to infiltrate there. Where perfusion isn't an option, a lot of researchers will insist on cutting their animal specimens into tiny little pieces and dropping them in fixative as quickly as possible to prevent this. Real pain to work with those! You also might as well forget about anything that was embedded before the age of buffered formalin. Formic acid is death to nucleic acids. Old, unbuffered formalin that's gone acidic is a sure way to kill an ISH signal. I've never seen even a hint of a signal out of anything embedded pre-1980's Finally, keep in mind that different species of RNA are subject to different rates of degradation. I've seen FFPE specimens that showed great results for total RNA, but were useless when probed for a specific species. This is particularly true if you try standardizing against an rRNA which is almoist necer a good idea. All of the above do not apply if you use snap frozen material. Never had any problems at all with it. I'm not doubting that some places do ISH on fixed slides and get a signal from it. The literature is full a this and some times that's all you've got so you go with it. Given the choice, though, I'd stick with the frozen stuff every time despite the extra hassle involved. If you do use fixed stuff only trust it for localization, not quantitation. HistoJock. >Hey guys, > >My boss is submitting a grant and he does not believe that you can do >PCR (RT) or ISH with fixed tissues. I thought you could but I don't >know any details. Could someone elaborate? Is there a particular way >the tissue should be fixed? In other words do I have to use FFPE >sections or can I use 4% paraformaldehyde perfusion? > >Any advice would be helpful. > >Thanks, > >Caroline Bass _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From Eric <@t> ategra.com Tue Jul 5 18:55:26 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] HistoTech Job Openings Message-ID: Histonetters - My name is Eric Dye and I am the new Histo Tech recruiter here at Ategra. I took over all Histo Tech recruiting since Pam Barker retired. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job opening is in Central Ohio seeking a HistoTech Manager. I also have bench histo tech jobs available in: Michigan, Pennsylvania, Illinois, New York, Missouri,West Virginia, Georgia, Iowa, Rhode Island, and Florida. I also have blood bank positions available in Upstate New York, Alaska and Arizona. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------- From amosbrooks <@t> earthlink.net Tue Jul 5 21:04:45 2005 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Impress In-Reply-To: <200507051300.1dPQMUXo3Nl34k0@mx-avoceta.atl.sa.earthlink.net> References: <200507051300.1dPQMUXo3Nl34k0@mx-avoceta.atl.sa.earthlink.net> Message-ID: <42CB3C3D.2080504@earthlink.net> Deb, The contact time for the Impress is the same as Envision. We ran side by side titers of primary antibodies using each kit. There was some variation as I recall. I believe the CD4 ended up with a higher dilution with Impress than Envision. Many however remained the same but had a better signal to noise ratio. (Stronger labeling with less background) It should also be pointed out that some antibodies were better with Envision so it is something that you would have to try out to see what works best for you, but you may find it to be well worth your while. Hope this helps, Amos /////////////////\\\\\\\\\\\\\\\\\\\\\\\\ Dear-histonetters-----I have also wondered about the Impress Kit----Amos and to otheres who tried it---was it pretty interchangeable- time wise with the Envision , or Envision + also what about antibody dilutions? same strength? From christinmyhart <@t> yahoo.com Tue Jul 5 23:10:11 2005 From: christinmyhart <@t> yahoo.com (Venita Capaldo) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Job Opening in Phoenix Message-ID: <20050706041012.57895.qmail@web32608.mail.mud.yahoo.com> Hi there all you Histonetters. I have a hot proposition. This job is open to new grads as well as experienced. This position is full time, Monday thru Friday.....No weekends, Holidays or call. The position entails running a small DermPath lab. You would be the begin all and end all here. From accessioning to labeling and packaging, you would do between 20-60 specimens daily. We have all state of the art equipment and the employers are awesome. So if you are ready for a change of pace and a great work atmosphere give me a call or fax me your resume. Venita Ballard Histology Lab Manager (602) 277-7686 (602) 277-0407 Fax --------------------------------- Sell on Yahoo! Auctions - No fees. Bid on great items. From abright <@t> brightinstruments.com Wed Jul 6 04:57:22 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] liver cryosections Message-ID: Dear Renee, Is the liver shredding horizontally or vertical ? If horizontal the likely causes are that the liver is to cold, the knife, disposable blade or holder is not held firm enough, or there is wear in the microtome slideways or drive bearings. If vertical, the knife or disposable blade has small nicks along the cutting edge. 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: Till, Renee [mailto:TillRenee@uams.edu] Sent: 05 July 2005 16:40 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] liver cryosections Can anyone offer some advice on cutting liver cryosections? My every attempt produces shredded tissues. I know in paraffin embedding that means they are dry and I would soak it in ice water before I cut, but what do you do with cryosections? I'm pretty sure it's not my cutting technique, though I am fairly new at it. I've adjusted the roll plate and the vacume window. And I've tried it without the roll plate, though as we are just starting out with our cryostat I don't have any good brushes for pulling the section. Any ideas? Maybe they did not have enough time to come down from -80 to -20? I left them in for about 2 hours. Renee' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BriggsK <@t> drmc.drhsi.org Wed Jul 6 05:47:54 2005 From: BriggsK <@t> drmc.drhsi.org (Kevin Briggs) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Microtome Service Message-ID: <7C1D5BE1ADF06E488AEB7A6247214A86015FB4D2@HORNET.drmc.drhsi.org> Hi Netters! We are in search of a service and repair person for our microtomes and would like your recommendations for anyone who covers the North Carolina-Virginia area. Thanks, Kevin D. Briggs, MS, CT(ASCP) Team Leader-Cytopathology/Histopathology Services Danville Regional Medical Center Department of Pathology 142 South Main Street Danville, VA 24541 (434) 799-4470 ext.5451 (phone) (434) 773-6806 (fax) email: briggsk@drhsi.org From dsnider <@t> shrinenet.org Wed Jul 6 06:13:13 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] no nuclei in frozen sections Message-ID: Hello again everyone! I need your advice and suggestions again! I have begun to have holes in frozen sections where the nuclues should be. I am cutting clinically engineered skin specimens, at -21. NO flash freeze spray. The specimens are cut at 5-6 microns and are embedded in Shandons' M-1 embedding matrix. This problem seems to have come up in the last 3 weeks or so, but I cannot unravel why. Nothing has been changed on my end, but the problem has been in all the researchers submissions. I have done recuts and they are still there. I have wracked my brain and am coming up clueless! I have contacted the manufacturer for additional information on the properties of M-1, but I am still coming up with no answers. Thanks in advance, Deanna Shriners Hospital for Children Research Dept. Cincinnati, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From cgfields <@t> lexhealth.org Wed Jul 6 07:09:23 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] Microtome Service Message-ID: Hi Kevin, We are located in Columbia South Carolina and Southeast Pathology Instruments Service, 843-588-2559 does all our service. Michael Dietrich is factory trained on Tissue Tek, Micron, Leica and others. I have had no problems with this company and I know he covers North Carolina. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 -----Original Message----- From: Kevin Briggs [mailto:BriggsK@drmc.drhsi.org] Sent: Wednesday, July 06, 2005 6:48 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Microtome Service Hi Netters! We are in search of a service and repair person for our microtomes and would like your recommendations for anyone who covers the North Carolina-Virginia area. Thanks, Kevin D. Briggs, MS, CT(ASCP) Team Leader-Cytopathology/Histopathology Services Danville Regional Medical Center Department of Pathology 142 South Main Street Danville, VA 24541 (434) 799-4470 ext.5451 (phone) (434) 773-6806 (fax) email: briggsk@drhsi.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From petepath <@t> yahoo.com Wed Jul 6 08:56:27 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:17 2005 Subject: [Histonet] University of Pittsburgh Message-ID: <20050706135627.30566.qmail@web30406.mail.mud.yahoo.com> Are the nuclei totally missing? or are there holes in the nuclei? If there are holes in the nuclei I believe this is one of the manifestations of freeze artefact which becomes very obvious in sections in the under 3 micra range. Even though a cryostat is set for 5 micra it is very common for the first few sections to be different thickness, commonly a thick one followed by a thin one. I show pics of what I believe are nuclear ice crystals and the effect of section thickness on this page of my frozen section tutorial: http://pathologyinnovations.com/new_page_3.htm. If the nuclei are falling out, I have no answer. Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From Michael.Rice <@t> holy-cross.com Wed Jul 6 07:16:28 2005 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] RE: Histonet Digest, Vol 20, Issue 5 Message-ID: <3BC92F29BE821745AB15E04C98EE028D6D38C7@HCH2KMAIL.holy-cross.com> Joe, I could not agree with you more, in these days of declining revenue and increasing costs, it seems almost criminal to discard a reagent in histology because of an arbitrary date. Having once worked for a antibody manufacturer, we determined expiration dates by subjecting the Ab to extreme high and low temperatures to accelerate the rate at which the reagent would stop working. In routine circumstances in a lab, this would not be the case. In any event, as long as the reagent passes QC each time it is used should be the determining point. Since CAP does not pay for my reagents, I feel as you do that the question should be revisited Mike Rice patholgy supervisor holy cross hospital ft lauderdale -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, July 05, 2005 1:06 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 5 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. PCR and ISH in fixed tissues (Caroline Bass) 2. Re: PCR and ISH in fixed tissues (Agripina Suarez) 3. Re: formic acid decal (clifford berger) 4. RE: PCR and ISH in fixed tissues (Tony Henwood) 5. ER/PR staining of normal breast tissue (Patrick Paulusse) 6. non-muscle myosin IIA (Edwards, R.E.) 7. Rotating shifts... (Tom McNemar) 8. antibody search for BMP (louise renton) 9. revisiting expired immmuno reagents (Joe Nocito) 10. RE: Formic Acid decalcification (Pixley, Sarah (pixleysk)) 11. Impress (Van Eyck, Deb) 12. liver cryosections (Till, Renee) 13. RE: liver cryosections (Monfils, Paul) ---------------------------------------------------------------------- Message: 1 Date: Mon, 04 Jul 2005 13:54:08 -0400 From: Caroline Bass Subject: [Histonet] PCR and ISH in fixed tissues To: 'histonet@lists.utsouthwestern.edu' Message-ID: <9E94278E-ECB4-11D9-8BC8-0003930771EE@bidmc.harvard.edu> Content-Type: text/plain; charset=US-ASCII; format=flowed Hey guys, My boss is submitting a grant and he does not believe that you can do PCR (RT) or ISH with fixed tissues. I thought you could but I don't know any details. Could someone elaborate? Is there a particular way the tissue should be fixed? In other words do I have to use FFPE sections or can I use 4% paraformaldehyde perfusion? Any advice would be helpful. Thanks, Caroline Bass ------------------------------ Message: 2 Date: Mon, 04 Jul 2005 11:09:57 -0700 From: "Agripina Suarez" Subject: Re: [Histonet] PCR and ISH in fixed tissues To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Caroline. Most of the ISH I do are on paraformaldehyde/formalin-fixed paraffin-embedded sections and they work very well. The tissues are fixed either in 4% paraformaldehyde or in commercially available 10% neutral buffered formalin for 12 to 24 hours, depends on the size of the tissue. Hope this helps. Agripina Agripina C. Suarez McDonald Research Laboratories iCAPTURE Centre, St Paul's Hospital 1081 Burrard St, Vancouver B.C., Canada V6Z 1Y6 Tel no. 604 682 2344 local 62562 Fax no. 604 806 8351 >>> Caroline Bass 7/4/2005 10:54:08 AM >>> Hey guys, My boss is submitting a grant and he does not believe that you can do PCR (RT) or ISH with fixed tissues. I thought you could but I don't know any details. Could someone elaborate? Is there a particular way the tissue should be fixed? In other words do I have to use FFPE sections or can I use 4% paraformaldehyde perfusion? Any advice would be helpful. Thanks, Caroline Bass _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Mon, 04 Jul 2005 16:10:06 -0400 From: clifford berger Subject: Re: [Histonet] formic acid decal To: patsy ruegg , lpwenk@sbcglobal.net, 'louise renton' , Histonet@lists.utsouthwestern.edu Message-ID: <000e01c580d4$5f1e71e0$0200a8c0@dellovo0ll7kuk> Content-Type: text/plain; charset=iso-8859-1 For more information on this product, technical papers and much more information, you can visit our website, www.decal-bone.com. You can also call us at 1-800-428-5856 for free samples. Best regards and happy 4rth Cliff Berger Decal Chemical Corp The Real Decal 1-800-428-5856 ----- Original Message ----- From: "patsy ruegg" To: ; "'louise renton'" ; Sent: Monday, July 04, 2005 11:24 AM Subject: RE: [Histonet] formic acid decal > For I IHC I use ImmunoCal for Decal Chemicals, it is a 5% formic acid with > buffers. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy > Wenk > Sent: Friday, July 01, 2005 5:13 PM > To: 'louise renton'; Histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] formic acid decal > > A milder formic acid decalcification solution is FASC = formic acid-sodium > citrate. It's a buffered formic acid. Sections of vertebral bone take 5-7 > days to decalcify. I don't know how long a bone biopsy takes. When my > students have to submit a bone section for their certification exam, this is > what we use. It gives great nuclear detail, fantastic eosinophil granules. > So I expect IHC would be equally great. We use the procedure out of the > Bancroft book. > > 65 mL of 20% aqueous trisodium citrate (13 g in 65 mL water) > 35 mL of 90% formic acid (that's full strength. Don't dilute) > pH is about 2.3 > Change solution every 3 days. > > We did have a rare bone tumor, and none of the IHC worked, until we > decalcified it in EDTA. Then it worked great. (Sorry, don't remember what > type of tumor or what type of antibody.) But you might want to try the EDTA, > if you have a lot of time. Vertebral bone takes 2-3 weeks. The little bone > fragments we had from the bone tumor took 3 days. (would have been a 2-3 > hours in our usual 5% hydrochloric acid) > > 25 g EDTA disodium salt (ethylenediaminetetracetic acid) (note: MUST be > disodium salt) > 175 mL d. water > Solution is cloudy. Add 2.5-3.0 g of sodium hydroxide until solution becomes > clear. pH should be about 7 at that point. > Change solution about twice a week. > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > Lee & Peggy Wenk > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise > renton > Sent: Friday, July 01, 2005 8:32 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] formic acid decal > > Hi all, > > on pain of being repetitive/lazy/slovenly/slothful/unprofessional, > would someone PLEASE share their recipe for formic acid decalcifying > solution suitable for samples on which IHc will be performed. Many many > thanks > > -- > Louise Renton (grovelling) > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "....I know who I am. No-one else knows who I am. If I was a giraffe, and > someone said I was a snake, I'd think no, actually I'm a giraffe" > Richard Gere > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Tue, 5 Jul 2005 09:10:29 +1000 From: Tony Henwood Subject: RE: [Histonet] PCR and ISH in fixed tissues To: "'Caroline Bass'" , "'histonet@lists.utsouthwestern.edu'" Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E2C0@simba.kids> Content-Type: text/plain Caroline, We have successfully done ISH for EBERs, Adenovirus RNA, CMV RNA, Chromogranin mRNA, Estrogen Receptor mRNA, and nCAM RNA to name a few on both Frozen sections and Formalin Fixed Paraffin Embedded Tissue (FFPE). We have also successfully extracted RNA and DNA from FFPE tissues for PCR (Ewing's and Rhabdo translocation PCR studies)and do this routinely. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Caroline Bass [mailto:cbass@bidmc.harvard.edu] Sent: Tuesday, 5 July 2005 3:54 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] PCR and ISH in fixed tissues Hey guys, My boss is submitting a grant and he does not believe that you can do PCR (RT) or ISH with fixed tissues. I thought you could but I don't know any details. Could someone elaborate? Is there a particular way the tissue should be fixed? In other words do I have to use FFPE sections or can I use 4% paraformaldehyde perfusion? Any advice would be helpful. Thanks, Caroline Bass _______________________________________________ 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. ********************************************************************** ------------------------------ Message: 5 Date: Sat, 4 Jun 2005 20:06:38 -0400 From: "Patrick Paulusse" Subject: [Histonet] ER/PR staining of normal breast tissue To: "Histonet" Message-ID: <000701c56962$85217100$57b05bd1@wolf> Content-Type: text/plain; charset="iso-8859-1" Good Evening All, Should normal breast duct tissue always stain positively with estrogen and progesterone antibodies. If not, is this a natural occurrence or is there something physiologically occuring to prevent the staining. I have had several occasions when the normal breast duct tissue will not stain (the corresponding tumour tissue in the same block was also negative). An external control on the same slide stains wonderfully. Fixation is never less than 24 hours and seldom over 72 hours in 10% NBF. I have several control blocks and use the one that mimics the fixation and processing of the test specimen. Any comments and references are always welcome. Thanking you in advance, Patrick Paulusse Anatomical Pathology Pembroke Regional Hospital ------------------------------ Message: 6 Date: Tue, 5 Jul 2005 12:32:03 +0100 From: "Edwards, R.E." Subject: [Histonet] non-muscle myosin IIA To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Looking for an antibody to the above for immunohistochemistry... Thanks Richard Edwards MRC TOX UNIT.....LEICESTER.....U.K........ ------------------------------ Message: 7 Date: Tue, 5 Jul 2005 09:25:27 -0400 From: Tom McNemar Subject: [Histonet] Rotating shifts... To: Histonet Message-ID: <6CD94D97ED7D924BA5C2B588FA9528213967D9@nt_exchange.lmhealth.org> Content-Type: text/plain; charset="iso-8859-1" I was wondering if most histo labs use rotating shifts or if people mostly work the same hours. Do you have people that just work one shift and primarily do one job (gross, cut, stain, etc.)? At full staff we have 4 techs and 3 of them rotate between 2 weeks of 6-2:30 and 1 week of 7:30-4 (surgicals). I wondered if switching the hours to a straight 7:30 - 4 position might help attract someone. I was also thinking that we would have a more consistency/continuity if everyone had a certain job/area that they were primarily responsible for. All thoughts appreciated. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 ------------------------------ Message: 8 Date: Tue, 5 Jul 2005 15:52:42 +0200 From: louise renton Subject: [Histonet] antibody search for BMP To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Dear all, I am looking for antibodies to BMP (bone morphogenic protein) 3, 6 and 7 (AKA Op1) for use on FFPE embedded tissue. I would be grateful for any input, as an internet search has come up with very little information. Thank you -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere ------------------------------ Message: 9 Date: Tue, 5 Jul 2005 09:01:51 -0500 From: "Joe Nocito" Subject: [Histonet] revisiting expired immmuno reagents To: "Histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello histoland, okay, I've been shooting my mouth off a lot lately and I want y'all to know that I contacted the CAP about revising the question about using expired immuno reagents. The question in question (like that huh?) in the latest CAP checklist is ANP.22432. Whether they contact me or not is another question (so many questions). My rationale for changing the question are these 1. Immuno reagents are too expensive to throw away. 2. Each time a run is performed, the reagents are revalidated 3. A laboratory must have a written procedure in place detailing the revalidating process. 4. A quality control form must be used and signed by the medical director assuring that the reagents work. I am open to suggestions and other ideas that I may have over looked. With your support, maybe we can, at least, have this question revised. Of course, I expect some fallout because of this, but hey, it wouldn't be me if I didn't Enjoy!!! Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX ------------------------------ Message: 10 Date: Tue, 5 Jul 2005 10:25:23 -0400 From: "Pixley, Sarah (pixleysk)" Subject: [Histonet] RE: Formic Acid decalcification To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Dear Louise Renton: Here is a method from a research lab. We use 10% formic acid, but add an ion exchange resin that soaks up the calcium. This eliminates the need to change solutions, but brings its own problems since the resin must be stirred. This is faster than EDTA, but I don't know if it is faster than just a 10% formic acid solution alone. The mixture is good for many decalcifications. Great ICC staining. Formic Acid Decalcification: Purchase (i.e., Fisher Scientific): Rexyn 101 H ion exchange resin formic acid (full strength, ~90%). Make 6-10 liters of 10% formic acid (in double distilled water) Add 500 g of resin (So ~50-83 g resin per liter of 10% formic acid) Need stir bar to keep resin suspended, so put samples in cloth bags (i.e., homemade cotton bags) and suspend from side. Alternatively, use lab dessicator. Samples sit above dessicator plate while stir bar goes underneath. Sincerely, Sarah Pixley, Ph.D. Dept. of Cell Biology, Neurobiology and Anatomy University of Cincinnati College of Medicine Cincinnati, OH 45267-0521 ------------------------------ Message: 11 Date: Tue, 5 Jul 2005 09:28:07 -0500 From: "Van Eyck, Deb" Subject: [Histonet] Impress To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Dear-histonetters-----I have also wondered about the Impress Kit----Amos and to otheres who tried it---was it pretty interchangeable- time wise with the Envision , or Envision + also what about antibody dilutions? same strength? Another immuno question for you all? is anyone doing Toxoplasmosis staining? What antibody are you using? What protocol? Thanks much-Deb Van Eyck > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [SMTP:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > histonet-request@lists.utsouthwestern.edu > Sent: Sunday, July 03, 2005 12:06 PM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 20, Issue 3 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Re: Histonet Digest, Vol 19, Issue 43 (Amos Brooks) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sat, 02 Jul 2005 16:19:45 -0400 > From: Amos Brooks > Subject: [Histonet] Re: Histonet Digest, Vol 19, Issue 43 > To: histonet@lists.utsouthwestern.edu > Message-ID: <42C6F6E1.2010501@earthlink.net> > Content-Type: text/plain; charset=ISO-8859-1; format=flowed > > Carla, > > We use both depending on the antibody. Mostly we use Envision, but > we tried the Impress on some of the antibodies that don't label very > intensely and did see improvement. If you compare the results of a CD2, > CD4, CD7 or CD8 odds are you'll see some improvement. Not all the > antibodies showed marked improvement and in some cases envision was better > so we use Envision on most but the success we've had with Impress made it > worth switching some of the procedures. Vector might give you a sample if > you ask nicely. Give it a whirl and see if you like it. > > Best of luck, > Amos > > > > Message: 17 > Date: Thu, 30 Jun 2005 07:34:30 -0700 > From: Carla M Conway > Subject: [Histonet] Dako EnVision G2 or Vector ImmPRESS comments > wanted > To: Histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=US-ASCII > > Hello all, > > I would like any comments (pro and con) regarding Dako's EnVision G2/AP > system or Vector's ImmPRESS kit. We are currently using EnVision + , but > are wondering if these new systems could be even better (more sensitive). > > Thanks for your help, > > Carla Conway > > Western Fisheries Research Center > 6505 NE 65th St > Seattle, WA 98115 > ph: 206-526-6282 ext. 242 > fax: 206-526-6654 > cmconway@usgs.gov > > > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 20, Issue 3 > *************************************** > > This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. ------------------------------ Message: 12 Date: Tue, 5 Jul 2005 10:39:33 -0500 From: "Till, Renee" Subject: [Histonet] liver cryosections To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Can anyone offer some advice on cutting liver cryosections? My every attempt produces shredded tissues. I know in paraffin embedding that means they are dry and I would soak it in ice water before I cut, but what do you do with cryosections? I'm pretty sure it's not my cutting technique, though I am fairly new at it. I've adjusted the roll plate and the vacume window. And I've tried it without the roll plate, though as we are just starting out with our cryostat I don't have any good brushes for pulling the section. Any ideas? Maybe they did not have enough time to come down from -80 to -20? I left them in for about 2 hours. Renee' ------------------------------ Message: 13 Date: Tue, 5 Jul 2005 11:45:01 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] liver cryosections To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717565@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="ISO-8859-1" The block is too cold. -20 degrees is an appropriate temperature for sectioning many tissues, but some tissues require a lower temperature and others require a higher temperature. For liver, try -15 degrees. For some samples you may have to go as high as -12 degrees. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Till, > Renee > Sent: Tuesday, July 5, 2005 8:39 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] liver cryosections > > Can anyone offer some advice on cutting liver cryosections? My every > attempt produces shredded tissues. I know in paraffin embedding that > means they are dry and I would soak it in ice water before I cut, but > what do you do with cryosections? I'm pretty sure it's not my cutting > technique, though I am fairly new at it. I've adjusted the roll plate > and the vacume window. And I've tried it without the roll plate, though > as we are just starting out with our cryostat I don't have any good > brushes for pulling the section. Any ideas? Maybe they did not have > enough time to come down from -80 to -20? I left them in for about 2 > hours. > > > > Renee' > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 5 *************************************** ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From petepath <@t> yahoo.com Wed Jul 6 09:20:31 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] RE:no nuclei in frozen sections Last post wrong subject Message-ID: <20050706142031.88609.qmail@web30412.mail.mud.yahoo.com> Are the nuclei totally missing? or are there holes in the nuclei? If there are holes in the nuclei I believe this is one of the manifestations of freeze artefact which becomes very obvious in sections in the under 3 micra range. Even though a cryostat is set for 5 micra it is very common for the first few sections to be different thickness, commonly a thick one followed by a thin one. I show pics of what I believe are nuclear ice crystals and the effect of section thickness on this page of my frozen section tutorial: http://pathologyinnovations.com/new_page_3.htm. If the nuclei are falling out, I have no answer. Stephen From jmahoney <@t> alegent.org Wed Jul 6 09:40:40 2005 From: jmahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Job Opening Message-ID: Histology Technicians Full Time/Casual/On-call/Flexible shifts available Alegent Health is currently looking for a Histology Tech. Registered by American Society of Clinical Pathologists or eligible. Two years experience in a clinical laboratory preferred. Alegent (logo) Apply on-line:www.alegent.com Pre-employment physical, drug screen and background check required prior to hire. AA/EEO Jan Mahoney Omaha NE From JosefaNava <@t> texashealth.org Wed Jul 6 10:17:58 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] (no subject) Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944BF@phdex03.txhealth.org> We are looking to buy a new grossing station and are deciding between the Shandon Grosslab Senior vs. the MOPEC MB600. Has anyone used the MOPEC MB600 and what do you like/ dislike about it? I thank you for any feedback you can give us. Josie Nava Presbyterian Hospital of Dallas The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From JosefaNava <@t> texashealth.org Wed Jul 6 10:27:10 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grosslab Senior Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944C0@phdex03.txhealth.org> We are looking to buy a new grossing station and are deciding between the Shandon Grosslab Senior vs. the MOPEC MB600. Has anyone used the MOPEC MB600 and what do you like about it? I thank you for any information you can give us. Josie Nava Presbyterian Hospital of Dallas The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From GoodwinD <@t> pahosp.com Wed Jul 6 10:38:44 2005 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] microm (Richard Allan) v. Leica glass coverslipper Message-ID: <992899E9EC268548AB8DDE246AF88473055F51E0@PAHEX01.uphs.upenn.edu> Soliciting comments, pro and con, for these two instruments. Thanks! Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Philadelphia ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com From info <@t> ihcworld.com Wed Jul 6 10:50:25 2005 From: info <@t> ihcworld.com (IHC World) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] HistoWiki - The Free Histology Encyclopedia - Launched! Message-ID: Hello All, Everyone on the Histoland is invited to participate the histology project - HistoWiki - the free-content histology encyclopedia that anyone can edit. The goal of setting up this site is to build the world largest, the most comprehensive and up-to-date encyclopedia about everything related to Histology. http://www.ihcworld.com/histowiki/ If you have any questions or suggestions, please drop me a email at info@ihcworld.com Thanks. Richard IHC World Email: info@ihcworld.com From pruegg <@t> ihctech.net Wed Jul 6 10:54:38 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] goat anti-galectin 1 Message-ID: <200507061554.j66FsckM006052@chip.viawest.net> Anybody using SC goat anti-galectin 1 antibody on ffpe tissue? If so please share your protocol. Thank you, Patsy P.S. Has anyone developed a labelled polymer type detection system to use with goat primary antibodies yet? From liz <@t> premierlab.com Wed Jul 6 11:07:17 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] goat anti-galectin 1 In-Reply-To: <200507061554.j66FsckM006052@chip.viawest.net> Message-ID: <000001c58244$c7f20080$a7d48a80@AMY> Patsy I use labeled polymers with goat primaries as follows: After the goat antibody I use a rabbit anti-goat secondary. I get this from vector labs and then I use the rabbit envision + next. It works great. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Wednesday, July 06, 2005 8:55 AM To: 'Histonet' Subject: [Histonet] goat anti-galectin 1 Anybody using SC goat anti-galectin 1 antibody on ffpe tissue? If so please share your protocol. Thank you, Patsy P.S. Has anyone developed a labelled polymer type detection system to use with goat primary antibodies yet? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GoodwinD <@t> pahosp.com Wed Jul 6 12:03:40 2005 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] glass coverslipper 2 Message-ID: <992899E9EC268548AB8DDE246AF88473055F51EF@PAHEX01.uphs.upenn.edu> PS--anyone out there using Sakura's glass coverslipper? Any comments? Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com From JMacDonald <@t> mtsac.edu Wed Jul 6 12:08:23 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grosslab Senior Message-ID: I cannot comment on the MB 600, but we have the MB 100. It great job of removing formalin fumes. There is absolutely no s when disecting tissues. Mopec was great to work with, but so is T hermo. Jennifer MacDonald -----his To: Se Date: 07/06/2005 08:27A Subject: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grossla We are looking to buy between the Shandon Gross MOPEC MB600 and &nbs information you can giv Josie Nava Presbyterian Hospital of Da The information contained in this message and any attac intended only for the use of the individual or entity to which it addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL you are not the distributing, or usin immediately by return e- your system. _____ _______________________ 5F_ Histonet mailing lis Histonet@lists.utsouthwestern.edu [1]http://lists.utsouthw References 1. 3D"http://lists.utsouthwe=/ From PIXLEYSK <@t> UCMAIL.UC.EDU Wed Jul 6 12:22:26 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] How to Permeabilize thick cultures? Message-ID: Dear List: I have some thick explant-like cultures (many cell layers thick). We are having great difficulty getting antibodies to penetrate. All the staining occurs in thinned out edge areas but nothing stains in the central regions, where I know there are cells that should be positive for this antigen. We have tried: 1) using 1% Triton in the blocking and primary antibody steps and letting the antibody incubate for 2 days, while secondary and ABC reagents were left on for 4-6 hours. 2) we have permeabilized by placing the cultures (grown on filter papers) in acetone (chilled with dry ice/ethanol) for 5 mins. This gave a hint of increased staining, but not as good as for monolayer cultures. Today we are trying cultures treated with acetone for 15 minutes. Antibodies to antigens on or very close to the surface work well, but anything that needs to penetrate is not working. Does anyone have any other ideas? Should I increase staining to a week? Thanks in advance. Sarah Pixley Univ. of Cincinnati From Kirsty.Geard <@t> huttvalleydhb.org.nz Mon Jul 4 23:58:10 2005 From: Kirsty.Geard <@t> huttvalleydhb.org.nz (Kirsty Geard) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Histo choice fixatives Message-ID: Hi I am just about to start trialing some of the Histo Choice fixatives; Histochoice Tissue Fixative A, Histochoice Tissue Fixative D and HC-V Tissue Fixative. Before I start trialing though I would like to hear the opinion of others who have used it, especially in a hospital setting. Feel free to email me direct if you would rather. Thanks in advance Kirsty Kirsty Geard Histology MLSO Hutt Hospital High st Lower Hutt From histolog <@t> fcv.unl.edu.ar Wed Jul 6 12:49:34 2005 From: histolog <@t> fcv.unl.edu.ar (Lab. Invest. Histol.) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Enzymatic digestion In-Reply-To: <200507061554.j66FsckM006052@chip.viawest.net> Message-ID: I would need to know the differences in the effect (degree of digestion) of each one of these enzymes used for antigen retrieval. Proteinase K Ficin Pepsin Tripsin Thank you Hugo ------------------------------------------------------------------- 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 DayDawning <@t> wideopenwest.com Wed Jul 6 13:13:08 2005 From: DayDawning <@t> wideopenwest.com (DayDawning) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] mopec Message-ID: <006b01c58256$5d3ec400$6801a8c0@schostakvdehdv> Two of the largest hospitals in my area, William Beaumont and Henry Ford both have Mopec grossing stations. Not only that, but have been repeat customers! Mopec has also supported the NSH and the Michigan Society for decades. They are a great bunch of folks! I have not personally used the grossing station but do know that the company is in tune to the needs of the customer and very reliable. Dawn M. Truscott, HT(ASCP) We are looking to buy a new grossing station and are deciding between the Shandon Grosslab Senior vs. the MOPEC MB600. Has anyone used the MOPEC MB600 and what do you like/ dislike about it? I thank you for any feedback you can give us. Josie Nava Presbyterian Hospital of Dallas From Nancy.Temple <@t> ssfhs.org Wed Jul 6 13:17:01 2005 From: Nancy.Temple <@t> ssfhs.org (Temple Nancy) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] RE:MOPEC MB600 vs. Shandon GrossLab Message-ID: We put in 2 MOPEC gross stations about 2 years ago, 1 of which is the MB600. Our PA likes it just fine. The good thing about it was that you could have it built the way you wanted. The basic station plus whatever features needed. Ours is vented to the outside, instead of filters. Raises and lowers to the height of person using. MOPEC was very easy to work with. Nancy Temple, HT(ASCP) Supervisor Histology/Cytology St. Francis Hospital Indpls., In. From ckeyes <@t> yorkcentral.on.ca Wed Jul 6 14:07:00 2005 From: ckeyes <@t> yorkcentral.on.ca (Keyes Catherine) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] (no subject) Message-ID: I would like to ask a question. Does anyone know how to clear a specimen to be late injected with a dye to detect the calcium in a tissue? Catherine From Michael.Rice <@t> holy-cross.com Wed Jul 6 14:00:03 2005 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] RE: Histonet Digest, Vol 20, Issue 7 Message-ID: <3BC92F29BE821745AB15E04C98EE028DE3C2A0@HCH2KMAIL.holy-cross.com> I have the Shandon senior, it is a very difficult unit to clean and keep clean as well as having mechanical problems to the extent that Shanon agreed to extend the warranty for an additional year. My opinion is that is over engineered, too much electro mechanical "stuff" mike rice ft lauderdale -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, July 06, 2005 1:31 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 7 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE:no nuclei in frozen sections Last post wrong subject (Stephen Peters M.D.) 2. Job Opening (Janice A Mahoney) 3. (no subject) (Nava, Josefa) 4. Urgent feedback on MOPEC MB600 VS. Shandon Grosslab Senior (Nava, Josefa) 5. microm (Richard Allan) v. Leica glass coverslipper (Goodwin, Diana) 6. HistoWiki - The Free Histology Encyclopedia - Launched! (IHC World) 7. goat anti-galectin 1 (Patsy Ruegg) 8. RE: goat anti-galectin 1 (Elizabeth Chlipala) ---------------------------------------------------------------------- Message: 1 Date: Wed, 6 Jul 2005 07:20:31 -0700 (PDT) From: "Stephen Peters M.D." Subject: [Histonet] RE:no nuclei in frozen sections Last post wrong subject To: Histonet@lists.utsouthwestern.edu Message-ID: <20050706142031.88609.qmail@web30412.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Are the nuclei totally missing? or are there holes in the nuclei? If there are holes in the nuclei I believe this is one of the manifestations of freeze artefact which becomes very obvious in sections in the under 3 micra range. Even though a cryostat is set for 5 micra it is very common for the first few sections to be different thickness, commonly a thick one followed by a thin one. I show pics of what I believe are nuclear ice crystals and the effect of section thickness on this page of my frozen section tutorial: http://pathologyinnovations.com/new_page_3.htm. If the nuclei are falling out, I have no answer. Stephen ------------------------------ Message: 2 Date: Wed, 06 Jul 2005 09:40:40 -0500 From: "Janice A Mahoney" Subject: [Histonet] Job Opening To: Message-ID: Content-Type: text/plain; charset=US-ASCII Histology Technicians Full Time/Casual/On-call/Flexible shifts available Alegent Health is currently looking for a Histology Tech. Registered by American Society of Clinical Pathologists or eligible. Two years experience in a clinical laboratory preferred. Alegent (logo) Apply on-line:www.alegent.com Pre-employment physical, drug screen and background check required prior to hire. AA/EEO Jan Mahoney Omaha NE ------------------------------ Message: 3 Date: Wed, 6 Jul 2005 10:17:58 -0500 From: "Nava, Josefa" Subject: [Histonet] (no subject) To: Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944BF@phdex03.txhealth.org> Content-Type: text/plain; charset="us-ascii" We are looking to buy a new grossing station and are deciding between the Shandon Grosslab Senior vs. the MOPEC MB600. Has anyone used the MOPEC MB600 and what do you like/ dislike about it? I thank you for any feedback you can give us. Josie Nava Presbyterian Hospital of Dallas The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. ------------------------------ Message: 4 Date: Wed, 6 Jul 2005 10:27:10 -0500 From: "Nava, Josefa" Subject: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grosslab Senior To: Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944C0@phdex03.txhealth.org> Content-Type: text/plain; charset="us-ascii" We are looking to buy a new grossing station and are deciding between the Shandon Grosslab Senior vs. the MOPEC MB600. Has anyone used the MOPEC MB600 and what do you like about it? I thank you for any information you can give us. Josie Nava Presbyterian Hospital of Dallas The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. ------------------------------ Message: 5 Date: Wed, 6 Jul 2005 11:38:44 -0400 From: "Goodwin, Diana" Subject: [Histonet] microm (Richard Allan) v. Leica glass coverslipper To: "Histonet (E-mail)" Message-ID: <992899E9EC268548AB8DDE246AF88473055F51E0@PAHEX01.uphs.upenn.edu> Content-Type: text/plain; charset="iso-8859-1" Soliciting comments, pro and con, for these two instruments. Thanks! Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Philadelphia ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com ------------------------------ Message: 6 Date: Wed, 6 Jul 2005 11:50:25 -0400 From: "IHC World" Subject: [Histonet] HistoWiki - The Free Histology Encyclopedia - Launched! To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello All, Everyone on the Histoland is invited to participate the histology project - HistoWiki - the free-content histology encyclopedia that anyone can edit. The goal of setting up this site is to build the world largest, the most comprehensive and up-to-date encyclopedia about everything related to Histology. http://www.ihcworld.com/histowiki/ If you have any questions or suggestions, please drop me a email at info@ihcworld.com Thanks. Richard IHC World Email: info@ihcworld.com ------------------------------ Message: 7 Date: Wed, 6 Jul 2005 09:54:38 -0600 From: "Patsy Ruegg" Subject: [Histonet] goat anti-galectin 1 To: "'Histonet'" Message-ID: <200507061554.j66FsckM006052@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Anybody using SC goat anti-galectin 1 antibody on ffpe tissue? If so please share your protocol. Thank you, Patsy P.S. Has anyone developed a labelled polymer type detection system to use with goat primary antibodies yet? ------------------------------ Message: 8 Date: Wed, 6 Jul 2005 10:07:17 -0600 From: "Elizabeth Chlipala" Subject: RE: [Histonet] goat anti-galectin 1 To: "'Patsy Ruegg'" , "'Histonet'" Message-ID: <000001c58244$c7f20080$a7d48a80@AMY> Content-Type: text/plain; charset="us-ascii" Patsy I use labeled polymers with goat primaries as follows: After the goat antibody I use a rabbit anti-goat secondary. I get this from vector labs and then I use the rabbit envision + next. It works great. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Wednesday, July 06, 2005 8:55 AM To: 'Histonet' Subject: [Histonet] goat anti-galectin 1 Anybody using SC goat anti-galectin 1 antibody on ffpe tissue? If so please share your protocol. Thank you, Patsy P.S. Has anyone developed a labelled polymer type detection system to use with goat primary antibodies yet? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 7 *************************************** ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From mcauliff <@t> umdnj.edu Wed Jul 6 14:46:32 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Enzymatic digestion In-Reply-To: References: Message-ID: <42CC3518.6060507@umdnj.edu> The answer to this question will depend on the tissue, fixative used, time of fixation, antigen in question, antibody used, and probably other factors as well. A literature search is in order. Geoff Lab. Invest. Histol. wrote: >I would need to know the differences in the effect (degree of digestion) of >each one of these enzymes used for antigen retrieval. > >Proteinase K >Ficin >Pepsin >Tripsin > > >Thank you > > Hugo > >------------------------------------------------------------------- >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/ > > >_______________________________________________ >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 petepath <@t> yahoo.com Wed Jul 6 15:01:54 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Clearing specimen ( no subject) Message-ID: <20050706200154.34900.qmail@web30406.mail.mud.yahoo.com> Lewin and Ridells GI path text describes a simple method to clear dye injected bowel by passing the tissue through increasing grades of ETOH followed by clearing in oil of wintergreen (methyl salicyalate). I tried this once and it worked nicely. And your lab will smell "minty fresh" for weeks. Stephen From lu_ze <@t> sbcglobal.net Wed Jul 6 15:14:46 2005 From: lu_ze <@t> sbcglobal.net (Ze Lu) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Uneven staining in same slide, any clues? Message-ID: <006201c58267$5ae4e5c0$5b02a8c0@OPTIMUM2> Hi, histonet friends, In some Ki67 staining study we have done. I always see a trend that sections in bottom half stained better than the sections in top half in the same slide. Both tissue sections are next each other during sectioning. Expect there is similar staining but always slight difference. We used hydrophobic marker pen to draw cycle to form a reservior, so the incubation should be even. Anyone has clues on it? Ze Lu, Ph.D. Optimum Therapeutics, LLC From pruegg <@t> ihctech.net Wed Jul 6 15:23:09 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] (no subject) In-Reply-To: Message-ID: <200507062023.j66KN8kM007485@chip.viawest.net> I do not understand this question, please explain further what you want to do? Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Keyes Catherine Sent: Wednesday, July 06, 2005 12:07 PM To: (histonet@lists.utsouthwestern.edu) Subject: [Histonet] (no subject) I would like to ask a question. Does anyone know how to clear a specimen to be late injected with a dye to detect the calcium in a tissue? Catherine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Jul 6 15:25:13 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Uneven staining in same slide, any clues? In-Reply-To: <006201c58267$5ae4e5c0$5b02a8c0@OPTIMUM2> Message-ID: <200507062025.j66KPDkM008212@chip.viawest.net> Are you doing the staining manually or using an Autostainer? Maybe you need to use more reagent to cover both sections. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ze Lu Sent: Wednesday, July 06, 2005 1:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Uneven staining in same slide, any clues? Hi, histonet friends, In some Ki67 staining study we have done. I always see a trend that sections in bottom half stained better than the sections in top half in the same slide. Both tissue sections are next each other during sectioning. Expect there is similar staining but always slight difference. We used hydrophobic marker pen to draw cycle to form a reservior, so the incubation should be even. Anyone has clues on it? Ze Lu, Ph.D. Optimum Therapeutics, LLC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Wed Jul 6 15:46:49 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] revisiting expired immmuno reagents Message-ID: Hi Joe: Nice work! I agree 100% with your position on this. I was thinking about having everyone in support of this change send me an e-mail indicating their support and then forwarding those e-mails to the CAP. Maybe that would get their attention! Let me think about this some more before we move forward. Richard "Joe Nocito" 07/05/05 10:01AM >>> Hello histoland, okay, I've been shooting my mouth off a lot lately and I want y'all to know that I contacted the CAP about revising the question about using expired immuno reagents. The question in question (like that huh?) in the latest CAP checklist is ANP.22432. Whether they contact me or not is another question (so many questions). My rationale for changing the question are these 1. Immuno reagents are too expensive to throw away. 2. Each time a run is performed, the reagents are revalidated 3. A laboratory must have a written procedure in place detailing the revalidating process. 4. A quality control form must be used and signed by the medical director assuring that the reagents work. I am open to suggestions and other ideas that I may have over looked. With your support, maybe we can, at least, have this question revised. Of course, I expect some fallout because of this, but hey, it wouldn't be me if I didn't Enjoy!!! Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From Melissa.Gonzalez <@t> cellgenesys.com Wed Jul 6 17:28:02 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Liver Albumin Message-ID: Hi Valerie, I have had great results just post-fixing the livers in 4% PFA for 3 hours at room temp, followed by 30% sucrose overnight at 4 degrees before freezing down in OCT in the cryostat, or -20C. Perfusion wasn't necessary. Also, I am not familiar with perfusion using saponin in the mix. But I do always use TritonX on my actual cut slides before staining, anytime I use 4% PFA fixed tissues. Detergents help permeabilize the cells, facilitating your antibody binding. Hope this helps, Melissa Message: 10 Date: Fri, 1 Jul 2005 09:37:13 -0500 From: vsailes@nd.edu Subject: [Histonet] Recipe for 4%PFA w/saponin To: histonet@lists.utsouthwestern.edu Message-ID: <1120228633.42c5551975837@webmail.nd.edu> Content-Type: text/plain; charset=ISO-8859-1 Hi Histonetters, I'm working with mice and we want to stain the livers for albumin. I have an antibody that appears to work but we would like to perfect the technique if possible. I have done some literature searches and some of them suggested that to perfuse the animal with 4% paraformaldehyde containing saponin would give a more homogenous staining pattern but I really could not find information telling the percentage of saponin to use. If anyone out there has experience with this would you please help shed some light on this matter. Thank you in advance for your help. Have a great holiday weekend. Valerie From jnocito <@t> satx.rr.com Wed Jul 6 18:33:31 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] revisiting expired immmuno reagents References: Message-ID: <004b01c58283$1eec5aa0$b4bd0b43@yourxhtr8hvc4p> thanks for your support Joe ----- Original Message ----- From: "Richard Cartun" To: ; Sent: Wednesday, July 06, 2005 3:46 PM Subject: Re: [Histonet] revisiting expired immmuno reagents Hi Joe: Nice work! I agree 100% with your position on this. I was thinking about having everyone in support of this change send me an e-mail indicating their support and then forwarding those e-mails to the CAP. Maybe that would get their attention! Let me think about this some more before we move forward. Richard "Joe Nocito" 07/05/05 10:01AM >>> Hello histoland, okay, I've been shooting my mouth off a lot lately and I want y'all to know that I contacted the CAP about revising the question about using expired immuno reagents. The question in question (like that huh?) in the latest CAP checklist is ANP.22432. Whether they contact me or not is another question (so many questions). My rationale for changing the question are these 1. Immuno reagents are too expensive to throw away. 2. Each time a run is performed, the reagents are revalidated 3. A laboratory must have a written procedure in place detailing the revalidating process. 4. A quality control form must be used and signed by the medical director assuring that the reagents work. I am open to suggestions and other ideas that I may have over looked. With your support, maybe we can, at least, have this question revised. Of course, I expect some fallout because of this, but hey, it wouldn't be me if I didn't Enjoy!!! Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Anti-Virus. Version: 7.0.323 / Virus Database: 267.8.9/42 - Release Date: 7/6/2005 From HornHV <@t> archildrens.org Thu Jul 7 07:00:05 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grosslab Senior Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDD91@EMAIL.archildrens.org> We just bought the GrossLab Sr. It's great. The docs love it. I especially like the perimeter wash. But I think they both have that. We went with the GrossLab Sr. because one of our docs was already familiar with it. I think they are both good. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer MacDonald Sent: Wednesday, July 06, 2005 12:08 PM To: Nava, Josefa Cc: histonet@pathology.swmed.edu Subject: Re: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grosslab Senior I cannot comment on the MB 600, but we have the MB 100. It =oes a great job of removing formalin fumes. There is absolutely no s=ell when disecting tissues. Mopec was great to work with, but so is T hermo. Jennifer MacDonald -----his=onet-bounces@lists.utsouthwestern.edu wrote: ----- To: From: "Nava, Josefa" Se=t by: histonet-bounces@lists.utsouthwestern.edu Date: 07/06/2005 08:27A= Subject: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grossla= Senior We are looking to buy=a new grossing station and are deciding between the Shandon Gross=ab Senior vs. the MOPEC MB600. Has anyone used the MOPEC MB600 and &nbs=;what do you like about it? I thank you for any information you can giv= us. Josie Nava Presbyterian Hospital of Da=las The information contained in this message and any attac=ments is intended only for the use of the individual or entity to which it=s addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL=, and exempt from disclosure under applicable law. If you are not the=ntended recipient, you are prohibited from copying, distributing, or usin= the information. Please contact the sender immediately by return e-=ail and delete the original message from your system. _____ _______________________ 5F_=5F________________ Histonet mailing lis= Histonet@lists.utsouthwestern.edu [1]http://lists.utsouthw=stern.edu/mailman/listinfo/histonet References 1. 3D"http://lists.utsouthwe= _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From louise.renton <@t> gmail.com Thu Jul 7 08:45:59 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Nova Red/VIP substrate Message-ID: Dear All Could those of you in the community who have used these reagents from Vector Labs for double labeled IHC reactions please comment on their ease of use & contrast when used in conjuction with DAB substrate. TIA -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From hymclab <@t> hyhc.com Thu Jul 7 08:51:57 2005 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grossla b Senior Message-ID: We have had a Grosslab Sr. for approximately 5 years with no troubles. I had a hard time picking between the Grosslab and Mopec's but chose the Grosslab and am not sorry about it. The only thing I wish I could have pushed for was the adjustable height(couldn't convice administration that we needed it as we only had 1 pathologist at the time, now we have three with 2 tall and 1 short). Dawn -----Original Message----- From: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: Thursday, July 07, 2005 7:00 AM To: Jennifer MacDonald; Nava, Josefa Cc: histonet@pathology.swmed.edu Subject: RE: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grosslab Senior We just bought the GrossLab Sr. It's great. The docs love it. I especially like the perimeter wash. But I think they both have that. We went with the GrossLab Sr. because one of our docs was already familiar with it. I think they are both good. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer MacDonald Sent: Wednesday, July 06, 2005 12:08 PM To: Nava, Josefa Cc: histonet@pathology.swmed.edu Subject: Re: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grosslab Senior I cannot comment on the MB 600, but we have the MB 100. It =oes a great job of removing formalin fumes. There is absolutely no s=ell when disecting tissues. Mopec was great to work with, but so is T hermo. Jennifer MacDonald -----his=onet-bounces@lists.utsouthwestern.edu wrote: ----- To: From: "Nava, Josefa" Se=t by: histonet-bounces@lists.utsouthwestern.edu Date: 07/06/2005 08:27A= Subject: [Histonet] Urgent feedback on MOPEC MB600 VS. Shandon Grossla= Senior We are looking to buy=a new grossing station and are deciding between the Shandon Gross=ab Senior vs. the MOPEC MB600. Has anyone used the MOPEC MB600 and &nbs=;what do you like about it? I thank you for any information you can giv= us. Josie Nava Presbyterian Hospital of Da=las The information contained in this message and any attac=ments is intended only for the use of the individual or entity to which it=s addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL=, and exempt from disclosure under applicable law. If you are not the=ntended recipient, you are prohibited from copying, distributing, or usin= the information. Please contact the sender immediately by return e-=ail and delete the original message from your system. _____ _______________________ 5F_=5F________________ Histonet mailing lis= Histonet@lists.utsouthwestern.edu [1]http://lists.utsouthw=stern.edu/mailman/listinfo/histonet References 1. 3D"http://lists.utsouthwe= _______________________________________________ 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. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kappeler <@t> patho.unibe.ch Thu Jul 7 08:59:15 2005 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] CD13 on FFPE tissue Message-ID: <008401c582fc$21ce60f0$27955c82@patho.unibe.ch> Hi all Has anybody experience with CD13 antibodies on FFPE tissues? We have tried Novocastra's clone 38C12 after different pretreatments (5 variants of 'hot', 2 enzyme) and are not really enthusiastic about it (faint staining, very few positive cells in tonsils and bone marrow). Suggestions for other clones that work on FFPE welcome! Thanks Andi Kappeler Insitute of Pathology, University of Bern, Switzerland From ploykasek <@t> phenopath.com Thu Jul 7 09:33:23 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Job opening In-Reply-To: Message-ID: I am re-posting an opening at PhenoPath Laboratories in Seattle, WA. This position is in our contract research department. It's a very unique opportunity with a growing, innovative company. This position involves research immunohistochemistry projects at the bench level. Will need to provide oversight, training, evaluation, and prioritization of work for 3+ technologists; communicate with clients and manager in planning, executing and documenting projects. Quality control, safety, and preventive maintenance oversight also required. Bachelor?s of Science with a background in immunohistochemistry and project management; strong administrative, organizational, and interpersonal skills; laboratory supervision and experience with GLP projects desired; histology experience a plus. For more information, visit www.phenopath.com. Thank you. Patti Loykasek PhenoPath Laboratories ------ End of Forwarded Message From marco.prunotto <@t> medecine.unige.ch Thu Jul 7 10:16:29 2005 From: marco.prunotto <@t> medecine.unige.ch (Marco Prunotto) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] tetrazolium salts and IHC compatibility for infarcted areas In-Reply-To: References: Message-ID: <42CD474D.6060709@medecine.unige.ch> Dear All, I'm starting to use the tetrazolium method for setting up a model of infarction in the pig. I would like to ask you if it's possible to make that staining (for macroscopic evaluation of infarcted area and "at risk" area) and then fix it in formalin and perform also immunohistochemistry. Have you got any reference for that? This is important because if it's not possible to have the same at same time I have to duplicate the animals. Thanks a lot --------------------------- Marco Prunotto, PhD Dept. of Pathology & Immunology, CMU 1, rue michel-Servet 1211 Geneva 4, Switzerland Tel: +41 22 3795763 Fax: +41 22 3795746 From LewisS <@t> ccri.net Thu Jul 7 10:35:42 2005 From: LewisS <@t> ccri.net (Lewis, Sarah) Date: Fri Sep 16 15:25:18 2005 Subject: FW: [Histonet] pap pen Message-ID: <714B9F12B4E18C4C843B66E8E190F2AD447536@res2k3ms01.CRII.ORG> Hello histoneters, I do use the IMMEDGE pen from Vector for all of my Immunofluorescence. Works great, it has a fast drying time and does not lift or run. Give it a try I am sure you will like it! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net -----Original Message----- From: Steven Coakley [mailto:sjchtascp@yahoo.com] Sent: Thursday, June 30, 2005 1:54 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] pap pen Any good progress in Pap pens for Immunofluoresence. Steve --------------------------------- Yahoo! Sports Rekindle the Rivalries. Sign up for Fantasy Football From DOOLEEO <@t> shands.ufl.edu Thu Jul 7 11:57:49 2005 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] CK17 Message-ID: Dear Histonetters, I have been trying to get CK17(M7046) from DAKO to work on the Ventana Benchmark. I have tried several different tissues for controls. I have several dilutions and tried several types of retrieval. I get staining but I also get background that does not seem to dilute out. I have tried DAKO's background reducing diluent and it helps somewhat. If anyone uses this antibody on the Ventana Benchmark I wonder if you would share your protocol? What type of control tissue do you recommend? What dilution do you use? For those who use other stainers is there a particular pretreatment that you recommend? Thanks in advance Elaine Dooley From lrichey <@t> u.washington.edu Thu Jul 7 12:06:15 2005 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] glass coverslipper 2 In-Reply-To: <992899E9EC268548AB8DDE246AF88473055F51EF@PAHEX01.uphs.upenn.edu> References: <992899E9EC268548AB8DDE246AF88473055F51EF@PAHEX01.uphs.upenn.edu> Message-ID: <42CD6107.6090700@u.washington.edu> We just purchased the Sakura glass coverslipper. We had good luck with it for a month and then started getting error 21 continually. Goodwin, Diana wrote: >PS--anyone out there using Sakura's glass coverslipper? Any comments? > >Diana Goodwin >Supervisor, Anatomic Pathology >Pennsylvania Hospital, Preston 655-C > >ph: 215-829-6532 >fax: 215-829-7564 >e-mail: goodwind@pahosp.com > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From ajaivyas <@t> gmail.com Thu Jul 7 12:48:18 2005 From: ajaivyas <@t> gmail.com (Ajai Vyas) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Luminiscence imaging in excised rat tissue Message-ID: <934be63705070710484c56a9a@mail.gmail.com> Hi, I am trying to image luminescence (firefly luciferaase) emanating from isolated cluster of cells within rat brain. Does anybody has experience about what doses of substrate I should be using and if I should add ATP in the substrate for better signal. Thanks in advance, Ajai From rschoon <@t> email.unc.edu Thu Jul 7 12:58:35 2005 From: rschoon <@t> email.unc.edu (rschoon) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Looking for part-time work in RTP area In-Reply-To: References: Message-ID: <42CD6D4B.60802@email.unc.edu> I am looking for part-time employment in the Raleigh -Durham RTP area evenings and or weekends. HT/HTL certified, extensive histology, microscopy and image analysis experiance. Please contact me via e-mail for CV and resume. Robert Schoonhoven From rich.strauss <@t> Esoterix.com Thu Jul 7 14:33:18 2005 From: rich.strauss <@t> Esoterix.com (Strauss, Rich) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Giemsa/Wright stain for automated procedures. Message-ID: <555A57F57B53D411A1EF00D0B784644B02BEFDC1@BWD0101E> I am looking for any suggestions for automated procedures to run Giemsa/Wright on blood smears. Both instrument and reagent ideas would be appreciated. Rich Strauss HT, QIHC Esoterix Inc. From emry <@t> u.washington.edu Thu Jul 7 15:24:28 2005 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] slippery pig snouts Message-ID: Hi, We are trying to do brdu on nasal septums of pig snouts. I put them on plus coated slides and they are falling off during the process. Have any of you found a way to keep cartilage on slides? How long and at what temperature can we heat the slides to see if that will help? Thanks, Trisha Seattle From wecare <@t> qualityhistology.com Thu Jul 7 15:37:47 2005 From: wecare <@t> qualityhistology.com (wecare@qualityhistology.com) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Giemsa/Wright stain for automated procedures. References: <555A57F57B53D411A1EF00D0B784644B02BEFDC1@BWD0101E> Message-ID: <003b01c58333$bcd3e020$0600a8c0@internetconnect.net> You can use EMD Chemicals Midas III Stainer. Their operator manual specifies the procedures that have been well tested. They provide all three elements for the testing: Instrument, Reagents and Technical support for instrumentation and reagents. Preyas ----- Original Message ----- From: "Strauss, Rich" To: Sent: Thursday, July 07, 2005 3:33 PM Subject: [Histonet] Giemsa/Wright stain for automated procedures. > > I am looking for any suggestions for automated procedures to run > Giemsa/Wright on blood smears. Both instrument and reagent ideas would be > appreciated. > > Rich Strauss HT, QIHC > Esoterix Inc. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From liz <@t> premierlab.com Thu Jul 7 16:17:55 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] slippery pig snouts In-Reply-To: Message-ID: <000401c58339$57b5bd70$a7d48a80@AMY> Trisha We have stained a lot of cartilage, not pig snots, but this might be helpful. We cut our slides and let them drain a bit and then place them flat on a hot plate overnight. The hot plate is not to hot so that the paraffin melts. We do not place them in a hot oven, we find that the cartilage might flip. We deparaffinize and then check if any of the cartilage is out of place. You can then dip the slides in water and gently get the cartilage to go back in place. If I'm having a difficult time with the sections falling off I would allow them to dry flat before I begin the immunostaining procedure. I hope this helps. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Trisha Emry Sent: Thursday, July 07, 2005 1:24 PM To: histo Subject: [Histonet] slippery pig snouts Hi, We are trying to do brdu on nasal septums of pig snouts. I put them on plus coated slides and they are falling off during the process. Have any of you found a way to keep cartilage on slides? How long and at what temperature can we heat the slides to see if that will help? Thanks, Trisha Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Reuel.Cornelia <@t> tsrh.org Thu Jul 7 16:48:58 2005 From: Reuel.Cornelia <@t> tsrh.org (Reuel Cornelia) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] In Situ on Human Alpha tubulin (RNA biotinylated probe) Message-ID: Hi, Can somebody help me with ISH? I have a lots of background staining (precipitation) on my negative control. I'm using Dako TSA GEN Point. If anyone who have the best of their knowledge can tell me how to eliminate this by experience, please help me. I'm using Human Alpha tubulin (RNA biotinylated probe)(Maxim biotech) applied to goat muscle and human muscle. I did antigen retrieval,proteinase K, Antigen Retreival/pepsin pre treatment, H2O2 in methanol for 10 mins., denatured at 95 C for 5 mins, hybridized for 20 hrs/ overnight at 37C,stringent wash for 20 mins at 55 C and then proceed with Dako TSA kit.Wash with TBSTween. Both negative muscle have background stain that looks like the reaction we have seen on our positive control although it is not as intense with our positive. I also used ChromaGene from immunovision but the reaction is so weak. Question? 1. Would this be because of my tissue? Human muscle fix in STF, goat muscle fix in paraformaldehyde. Goat muscle gives lesser background and cleaner but still there is a background that looks like my positive control. I wanted to make it clean. 2. Would my probe have something to do with it. 3. How about my stringent wash? I know there are so many factors and i'm looking for anything that help me answer my questions and my help will be from your expertise. i'm looking forward for your reply. Thank you. ******************************************************************************************************************* Texas Scottish Rite Hospital for Children is one of the nation's leading pediatric centers for the treatment of orthopedic conditions and learning disorders, like dyslexia. This email transmission and/or its attachments may contain confidential health information, intended only for the use of the individual or entity named above. The authorized recipient of this information is prohibited from disclosing it to any other party unless required to do so by law and is required to delete/destroy the information after its stated need has been fulfilled. If you are not the intended recipient, any disclosure, copying, distribution or action taken in reliance on the contents of this email transmission is prohibited. If you have received this information in error, please notify the sender immediately and delete this information. We appreciate your efforts to protect the children's confidential information. ******************************************************************************************************************* From histo20 <@t> hotmail.com Thu Jul 7 17:49:35 2005 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Cytoprep Tech Trainee Message-ID: Hi everyone! Thanks again to all who sent me information about the training of a histotech (checklists, etc.) This was such a big help and very much appreciated. I am again asking for help - if anyone has timelines, checklists, etc.- this time for training a new cytoprep tech. Any help would be very much appreciated! thank you, Paula Wilder From c.m.vanderloos <@t> amc.uva.nl Fri Jul 8 02:20:57 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] RE: Nova Red/VIP substrate Message-ID: <15e367415e208c.15e208c15e3674@amc.uva.nl> Dear Louise, When it comes to double staining, Vector Nova Red chromogen would not be my choice for combining with DAB. Nova Red is not like AEC, it's more brownish-orange and doesn't contrast that well with standard DAB. You may try to shift the brown-yellow color of DAB reaction product to more blueish, by adding nickel- or chromium-salts. The combination of standard DAB with VIP gives a better contrast than Nova red. Both these chromogen kits are fully monkey-proof, don't worry about that! If you perform this sequential double staining (with two HRP activities involved), please keep in mind the following: * it's not going to work for the observation of co-localization by mixed-colors, just for two different cell populations (or different cellular compartments) only. * it's obligatory to apply DAB reaction product in the first staining sequence! DAB reaction product is the only known chromogen that effectively "cover" immunoreagents used in the first staining sequence. This sheltering effect is the key mechanism for preventing unwanted cross-reactions with reagents used in the second staining sequence. * sequential double staining works satisfactory if the first primary (and secondary) antibody is well titrated. If the first antibody (or secondary) is used too concentrated, the sheltering effect from the DAB reaction product will not be effective. I do hope I didn't scare you off performing double staining! Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Thu, 7 Jul 2005 15:45:59 +0200 From: louise renton Subject: [Histonet] Nova Red/VIP substrate To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Dear All Could those of you in the community who have used these reagents from Vector Labs for double labeled IHC reactions please comment on their ease of use & contrast when used in conjuction with DAB substrate. TIA -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa From c.m.vanderloos <@t> amc.uva.nl Fri Jul 8 02:33:47 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] RE: CD13 on FFPE tissue Message-ID: <15e98b315e500d.15e500d15e98b3@amc.uva.nl> Andy, We too have experience with Novacastra reagents that doesn't do what was promised in the data sheet (CD69 on FFPE). Our problem was finally solved when we turned to tyramide amplification (TSA or CSA II). Perhaps you can boost your faint CD13 staining in this way. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Thu, 7 Jul 2005 15:59:15 +0200 From: "Andi Kappeler" Subject: [Histonet] CD13 on FFPE tissue To: "Histonet" Message-ID: <008401c582fc$21ce60f0$27955c82@patho.unibe.ch> Content-Type: text/plain; format=flowed; charset="Windows-1252"; reply-type=original Hi all Has anybody experience with CD13 antibodies on FFPE tissues? We have tried Novocastra's clone 38C12 after different pretreatments (5 variants of 'hot', 2 enzyme) and are not really enthusiastic about it (faint staining, very few positive cells in tonsils and bone marrow). Suggestions for other clones that work on FFPE welcome! Thanks Andi Kappeler Insitute of Pathology, University of Bern, Switzerland From Jacqueline.Malam <@t> rli.mbht.nhs.uk Fri Jul 8 02:37:51 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] RE: CK 17 Message-ID: We have the Benchmark XT model and, although I haven't tried CK 17 as yet, I know that there are one or two antisera whose clones it doesn't like despite trying various protocols. Have you tried any other clones - it might be worth a go. Jacqui Malam Lancaster -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 07 July 2005 18:02 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 9 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. CK17 (Elaine Dooley) ---------------------------------------------------------------------- Message: 1 Date: Thu, 07 Jul 2005 12:57:49 -0400 From: "Elaine Dooley" Subject: [Histonet] CK17 To: Message-ID: Content-Type: text/plain; charset=US-ASCII Dear Histonetters, I have been trying to get CK17(M7046) from DAKO to work on the Ventana Benchmark. I have tried several different tissues for controls. I have several dilutions and tried several types of retrieval. I get staining but I also get background that does not seem to dilute out. I have tried DAKO's background reducing diluent and it helps somewhat. If anyone uses this antibody on the Ventana Benchmark I wonder if you would share your protocol? What type of control tissue do you recommend? What dilution do you use? For those who use other stainers is there a particular pretreatment that you recommend? Thanks in advance Elaine Dooley ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 9 *************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From peter_bannister <@t> hotmail.co.uk Fri Jul 8 03:52:46 2005 From: peter_bannister <@t> hotmail.co.uk (Peter Bannister) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] What is the best way to decontaminate a cryostat? Message-ID: Hello Histonetters, We have an old cryostat (without UV) with which we wish to produce sections for subsequent laser capture micro-dissection - to extract RNA for our functional genomics projects. Naturally we need to keep the level of cross contamination between specimens to an absolute minimum. Could anyone tell me the best way to decontaminate a cryostat for this purpose (what you use and how you do it). Many thanks for your help. Pete Bannister. Dr. Peter Bannister, Thrombosis Research Institute, Emmanuel Kaye Building, Manresa Road, London SW3 6LR. _________________________________________________________________ Want to block unwanted pop-ups? Download the free MSN Toolbar now! http://toolbar.msn.co.uk/ From n.cragg <@t> epistem.co.uk Fri Jul 8 04:41:21 2005 From: n.cragg <@t> epistem.co.uk (Nicola Cragg) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] HIER - best retrieval with minimum tissue damage? Message-ID: Hello, I'm a big fan of microwaving in citrate buffer as a method of heat inducing epitope / antigen retrieval. However, I do run into problems when microwaving skin. I've done a lot of IHC on skin and have found the microwaving is great when staining antigens in the epidermis, but I do have problems if the antigens I want to look at are in the dermis and require microwaving / heat for retrieval. The tissue is often damaged and the morphology of the dermis is disrupted, so when I do see evidence of specific staining it's difficult to be confident in it. Obviously, when optimising antibodies, I simultaneously attempt it without antigen retrieval or via other methods such as proteolytic digestion, but some antigens in the dermis appear to require heat. I have tried steaming, as I've read this is a gentler method, but I found it to be more damaging than microwaving. I have had some experience with using a pressure cooker and a waterbath for HIER but have not yet tried it specifically for skin. I use APES coated slides. Does anybody have any suggestions? Kind regards, Nicola Cragg Histology & IHC EpiStem Ltd Manchester, UK From Linresearch <@t> aol.com Fri Jul 8 05:15:15 2005 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] PCNA Message-ID: Hi, Does anyone have a protocol for PCNA on decalcified rodent bones? Could you share it and give me the source of the Ab? Thanks, Lin From izzetoguz <@t> gmail.com Fri Jul 8 07:46:56 2005 From: izzetoguz <@t> gmail.com (=?ISO-8859-9?Q?izzet_o=F0uz?=) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] FGF 3 AB Message-ID: <479ad206050708054649c9eb91@mail.gmail.com> HI HISTONETTERS, WE ARE TRYING FGF-3 AB( SANTA CRUZ CO.- SC-135) FOR BONE TISSUE, BUT HAVE SOME PROBLEMS THAT ALTHOUGH WE DOUBLE-CHECKED OUR PROTOCOL THE ANTIBODY DOES NOT WORK SOMETIMES AT SAME CONDITIONS IN FFPE BONE TISSUES. INTERESTINGLY WE HAD ALSO RARELY GOOD RESULTS IN FFPE BONE TISSUES. IS THERE ANYBODY TO DEAL WITH SAME PROBLEM? From la.sebree <@t> hosp.wisc.edu Fri Jul 8 07:48:47 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] HIER - best retrieval with minimum tissue damage? Message-ID: Nicola, Try bringing your slides in the buffer just up to a bare simmer in the MW and then transfer to a waterbath at your desired temp. (we used 92 -94 degrees C) for 20 - 40 minutes. Follow with your cool down and proceed with your staining. Before we had decloaking chambers (pressure cookers) we found this method to be the gentlest way to retrieve. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Nicola Cragg Sent: Friday, July 08, 2005 4:41 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] HIER - best retrieval with minimum tissue damage? Hello, I'm a big fan of microwaving in citrate buffer as a method of heat inducing epitope / antigen retrieval. However, I do run into problems when microwaving skin. I've done a lot of IHC on skin and have found the microwaving is great when staining antigens in the epidermis, but I do have problems if the antigens I want to look at are in the dermis and require microwaving / heat for retrieval. The tissue is often damaged and the morphology of the dermis is disrupted, so when I do see evidence of specific staining it's difficult to be confident in it. Obviously, when optimising antibodies, I simultaneously attempt it without antigen retrieval or via other methods such as proteolytic digestion, but some antigens in the dermis appear to require heat. I have tried steaming, as I've read this is a gentler method, but I found it to be more damaging than microwaving. I have had some experience with using a pressure cooker and a waterbath for HIER but have not yet tried it specifically for skin. I use APES coated slides. Does anybody have any suggestions? Kind regards, Nicola Cragg Histology & IHC EpiStem Ltd Manchester, UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From specialstainsqueen <@t> hotmail.com Fri Jul 8 08:36:53 2005 From: specialstainsqueen <@t> hotmail.com (Sherri Anderson) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Shandon Cytofunnels Message-ID: This is in response to the messages posted on Histonet that Thermo Electron Corporation is recalling all the old Cytospins® and no longer supplying the "re-usable" chambers (clipped Cytofunnels®) and stainless steel clips. We would like to clarify that the above statements are not true. Thermo Electron is the original manufacturer of the Shandon Cytofunnels. We are also the only manufacturer supplying Cytofunnels in 2 versions, the original Shandon Cytofunnels with Shandon Cytoclips® and the new, clipless Shandon EZ Cytofunnels. There is no plan to recall any Cytospin from the field as the new EZ Cytofunnels are functional on Cytospin 2, 3 or 4 without any modifications. Currently the EZ Cytofunnels are available in the United States, United Kingdom and Germany together with the conventional clipped cytofunnels. Eventually both types of cytofunnels will be available worldwide. We leave the choice and decision to our customers. Thank you for the opportunity to correct the previous postings. Sincerely, Sherri L. Anderson Product Specialist Anatomical Pathology Dept. Thermo Electron Corporation _________________________________________________________________ On the road to retirement? Check out MSN Life Events for advice on how to get there! http://lifeevents.msn.com/category.aspx?cid=Retirement From cfranci <@t> rigel.com Fri Jul 8 12:19:04 2005 From: cfranci <@t> rigel.com (Christian Franci) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] seeking mouse CD31 Ab Message-ID: <1653f15a53311314423cfc915afe8294@rigel.com> I've tried many and none seem to work with FFPE sections. Human Cd31 isn't a problem at all, even in FFPe sections but, mouse is a different story. Anyone out there has had success in staining mouse vessels in formalin sections? Any good Ab's or tricks? thanks in advance. Happy Friday! Chris From PIXLEYSK <@t> UCMAIL.UC.EDU Fri Jul 8 12:50:59 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] PCNA on bone Message-ID: Dear Lin: If you are having trouble with PCNA on RAT decalcified bone, then your problem might be the decalcification procedure. We just went through a lot of trouble to learn that BrDU staining does not work on tissue treated with our rapid decalcifying solution(unknown acid decalcification). Instead, we needed either EDTA or formic acid decalcification to get it to work. But we did previously do PCNA staining using this rapid decalcifier and got great staining with Chemicon mouse MAB424 PCNA. If you are trying to stain mouse tissues then you have a different and bigger can of worms. We could not get it to stain and heard from others that you needed a MOM (mouse on mouse) kit to get it to work. I will be interested to hear if there is a PCNA antibody that works well on mouse tissues. Sarah From jbaez <@t> interscopepath.com Fri Jul 8 12:53:00 2005 From: jbaez <@t> interscopepath.com (Baez, Janet) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] salary survey Message-ID: <9E956D8FEB06C2408B08AC16498325E90139DF@scopemx1.interscope.com> I've been asked to perform a salary survey in California, so that we may keep our Histotechs happy. You can e-mail me direct. I'm interested in knowing the salaries for the following positions. HT (ASCP) with 5+ years of experience, HT certified and not certified with 20+ years of experience, HT lead tech, HT supervisor and HT manager. The manager's positions is responsible for accreditation of 3 separate Facilities. Any information would be greatly appreciated. Thanks. From koellinr <@t> amgen.com Fri Jul 8 12:59:00 2005 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] seeking mouse CD31 and mouse CD4 on FFPE Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CC7E7@wa-mb4-sea.amgen.com> Chris, We use Pham BD anti-CD31 (MEC 13.3 clone) on FFPE mouse tissue. 5 min Dako Proteinase K Overnight in primary in refrigerator at 1 to 20,000 Biotin labeled secondary SA-px 5 min TSA amplification DAB Works very nicely and if you come to nationals in Florida, there will be a photomicrograph of the stain on our automated IHC poster. There will also be a picture of anti-mouse CD4 on FFPE mouse tissue for anyone interested in that elusive goal. Ray -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Christian Franci Sent: Friday, July 08, 2005 10:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] seeking mouse CD31 Ab I've tried many and none seem to work with FFPE sections. Human Cd31 isn't a problem at all, even in FFPe sections but, mouse is a different story. Anyone out there has had success in staining mouse vessels in formalin sections? Any good Ab's or tricks? thanks in advance. Happy Friday! Chris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Fri Jul 8 13:07:06 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] HES-1 antibody on FFPE sections Message-ID: Anybody have any success getting nuclear staining with HES-1???? Thanks, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From meryl50 <@t> hotmail.com Fri Jul 8 13:23:43 2005 From: meryl50 <@t> hotmail.com (Meryl Roberts) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] seeking emplyment in the Dallas area Message-ID: Hi there, I am a histologist with 6 years experience looking for full time emplyment in Dallas, Tx- for a copy of my resume, e-mail me at meryl50@hotmail.com sincerely, Meryl Roberts. From HornHV <@t> archildrens.org Fri Jul 8 13:42:01 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Shandon Cytofunnels Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDD94@EMAIL.archildrens.org> I wanted to comment on the new cytofunnels. At first, we had a devil of a time getting them to work, we had no one show us how. BUT, now that we have gotten used to them we really to like them. Load them up, spin and throw away. Very convenient. I wish they came in a box like the old funnels, the plastic tray is a bit cumbersome and takes up a lot of our couterspace. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sherri Anderson Sent: Friday, July 08, 2005 8:37 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Shandon Cytofunnels This is in response to the messages posted on Histonet that Thermo Electron Corporation is recalling all the old Cytospins(r) and no longer supplying the "re-usable" chambers (clipped Cytofunnels(r)) and stainless steel clips. We would like to clarify that the above statements are not true. Thermo Electron is the original manufacturer of the Shandon Cytofunnels. We are also the only manufacturer supplying Cytofunnels in 2 versions, the original Shandon Cytofunnels with Shandon Cytoclips(r) and the new, clipless Shandon EZ Cytofunnels. There is no plan to recall any Cytospin from the field as the new EZ Cytofunnels are functional on Cytospin 2, 3 or 4 without any modifications. Currently the EZ Cytofunnels are available in the United States, United Kingdom and Germany together with the conventional clipped cytofunnels. Eventually both types of cytofunnels will be available worldwide. We leave the choice and decision to our customers. Thank you for the opportunity to correct the previous postings. Sincerely, Sherri L. Anderson Product Specialist Anatomical Pathology Dept. Thermo Electron Corporation _________________________________________________________________ On the road to retirement? Check out MSN Life Events for advice on how to get there! http://lifeevents.msn.com/category.aspx?cid=Retirement _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From Eric <@t> ategra.com Fri Jul 8 20:10:15 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] HistoTech Job Openings Message-ID: Histonetters - My name is Eric Dye and I am the new Histo Tech recruiter here at Ategra. I took over all Histo Tech recruiting since Pam Barker retired. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job opening is in Central Ohio seeking a HistoTech Manager. I also have bench histo tech jobs available in: Michigan, Pennsylvania, Illinois, New Hampshire, ,West Virginia, Georgia, Colorado and Florida. I also have blood bank positions available in Upstate New York, Alaska and Arizona. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------- From koellinr <@t> amgen.com Sat Jul 9 10:47:36 2005 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] seeking mouse CD31 Ab Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CC7ED@wa-mb4-sea.amgen.com> Several people wrote privately about my post re: CD31 on FFPE mouse asking for a bit more info. * 5 min Dako proteinase K * Primary is from BD Pharmingen (think their website is bdbiosciences-go to Pharmingen tab). These are mainly flow reagents but we use a lot of them on our mouse tissues. Rat(Lew/Cr1BR) anti-mouse CD31, IgG2a, kappa (cat 553370). Overnight in refrigerator at 1 to 20,000 * Vector BA-4001, biotinylated rabbit anti-rat IgG (mouse adsorbed) 30 min. * Perkin Elmer streptavidin-horseradish peroxidase NEL750 at 1 to 2,000. 30 min * Perkin Elmer Renaissance TSA kit. 5 min. * Of course obviously go back with steptavidin peroxidase again. 30 min. * DAB/ methyl green or chromagen/counterstain your choosing and we use several.. Ray Koelling Research Scientist Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Christian Franci Sent: Friday, July 08, 2005 10:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] seeking mouse CD31 Ab I've tried many and none seem to work with FFPE sections. Human Cd31 isn't a problem at all, even in FFPe sections but, mouse is a different story. Anyone out there has had success in staining mouse vessels in formalin sections? Any good Ab's or tricks? thanks in advance. Happy Friday! Chris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From opiecurt <@t> yahoo.com Sat Jul 9 12:51:52 2005 From: opiecurt <@t> yahoo.com (curt tague) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] looking for these specific spec bottles Message-ID: <20050709175152.58856.qmail@web80103.mail.yahoo.com> i posted a couple images on the histonet, "view list server images" at the right side of the front page. pictures are named "specimen bottles" and were posted 7-8-05. if anyone know's where to get these exact bottles, i'd really appreciate your help. lg. 1 oz. bottle is 7 cm tall, 2.2 cm wide at the base and 3.4 cm wide at the lid sm. 1/4 oz bottle is 6 cm tall, 1.5 cm wide at the base and 2.0 cm wide at the lid. thanks for your help, curt From Anni-Maija.Linden <@t> helsinki.fi Sun Jul 10 04:00:21 2005 From: Anni-Maija.Linden <@t> helsinki.fi (Anni-Maija Linden) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] unsubcribe Message-ID: <1120986021.42d0e3a552b95@www3.helsinki.fi> Unsubcribe. Thanks. -- From c.m.vanderloos <@t> amc.uva.nl Mon Jul 11 02:19:08 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] RE: HIER - best retrieval with minimum tissue damage? Message-ID: <1670d0b16745a9.16745a91670d0b@amc.uva.nl> Dear Nicola, In our experiments we saw less damage by HIER when applying an overnight antigen retrieval at 60-70 C. Especially fatty tissues remain in tact nicely and do not tend to fall off. Using the ovenight 60-70C procedure we saw that some antigens stain as good as with 15 min boiling, others even more intense and others somewhat weaker. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Fri, 8 Jul 2005 10:41:21 +0100 From: "Nicola Cragg" Subject: [Histonet] HIER - best retrieval with minimum tissue damage? To: Hello, I'm a big fan of microwaving in citrate buffer as a method of heat inducing epitope / antigen retrieval. However, I do run into problems when microwaving skin. I've done a lot of IHC on skin and have found the microwaving is great when staining antigens in the epidermis, but I do have problems if the antigens I want to look at are in the dermis and require microwaving / heat for retrieval. The tissue is often damaged and the morphology of the dermis is disrupted, so when I do see evidence of specific staining it's difficult to be confident in it. Obviously, when optimising antibodies, I simultaneously attempt it without antigen retrieval or via other methods such as proteolytic digestion, but some antigens in t! he dermis From c.m.vanderloos <@t> amc.uva.nl Mon Jul 11 02:26:25 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] RE: PCNA Message-ID: <16711be167177d.167177d16711be@amc.uva.nl> Dear Lin, In stead of PCNA you can also apply Ki67 antibody. The new rabbit monoclonal antibody clone SP6 (Neomarkers) works on human, mouse, rat and on many more species. The use of a rabbit antibody also circumvents mouse-on-mouse problems. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Fri, 8 Jul 2005 06:15:15 EDT From: Linresearch@aol.com Subject: [Histonet] PCNA To: histonet@pathology.swmed.edu Hi, Does anyone have a protocol for PCNA on decalcified rodent bones? Could you share it and give me the source of the Ab? Thanks, Lin From louise.renton <@t> gmail.com Mon Jul 11 02:52:45 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Nova Red/VIP substrate In-Reply-To: <5D2189E74151CC42BEC02906BA8996322B90C6@exchsrv01.barrynet.barry.edu> References: <5D2189E74151CC42BEC02906BA8996322B90C6@exchsrv01.barrynet.barry.edu> Message-ID: Thanks for all the comments & advice regarding double labelling. I will bear all the suggestions in mind when I strat the project best regars On 7/7/05, Smith, Allen wrote: > In my hands Nova Red is red-brown and DAB is yellow-brown. I have not tried > to use them on the same slide, but I think it would be difficult to tell > them apart. > > Allen A. Smith, Ph.D. > Professor of Anatomy > Barry University School of Graduate Medical Sciences > Podiatric Medicine and Surgery > Miami Shores, Florida 33161 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise > renton > Sent: Thursday, July 07, 2005 9:46 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Nova Red/VIP substrate > > Dear All > Could those of you in the community who have used these reagents from > Vector Labs for double labeled IHC reactions please comment on their > ease of use & contrast when used in conjuction with DAB substrate. > TIA > -- > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "....I know who I am. No-one else knows who I am. If I was a giraffe, > and someone said I was a snake, I'd think no, actually I'm a giraffe" > Richard Gere > > _______________________________________________ > 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) > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From pex0220 <@t> yahoo.com.cn Mon Jul 11 08:48:01 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Proteinase K Message-ID: <20050711134801.74392.qmail@web15502.mail.cnb.yahoo.com> Hello, all, I plan to use proteinase k from Dakocytomation for antigen retrieval in bone paraffin sections, but the specification only shows proteolytic enzyme solution diluted in 0.05 mol/L Tris-Hcl, not the concentrations in details, so I have no ideas, if you have some experience about it, I hope you can do me a favor!Thank you! Guofeng --------------------------------- DO YOU YAHOO!? ÑÅ»¢Ãâ·ÑGÓÊÏ䣭No.1µÄ·À¶¾·ÀÀ¬»ø³¬´óÓÊÏä From liz <@t> premierlab.com Mon Jul 11 10:50:15 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Proteinase K In-Reply-To: <20050711134801.74392.qmail@web15502.mail.cnb.yahoo.com> Message-ID: <000e01c58630$3aed9c90$a7d48a80@AMY> Guofeng I have used Dako's proteinase K for quite a few antibodies on FFPE decalcifed bone sections. The diluent of the Dako reagent is listed on the spec sheet its 0.05mol/L Tris-HCL, 0.015mol/L sodium azide, pH 7.5. Depending upon the antibody, I use the proteinase K for 2 to 5 minutes. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com =20 Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pex Sent: Monday, July 11, 2005 6:48 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Proteinase K Hello, all, =20 I plan to use proteinase k from Dakocytomation for antigen retrieval in bone paraffin sections, but the specification only shows proteolytic enzyme solution diluted in 0.05 mol/L Tris-Hcl, not the concentrations in details, so I have no ideas, if you have some experience about it, I hope you can do me a favor!Thank you! =20 =20 Guofeng =09 --------------------------------- DO YOU YAHOO!? = =D1=C5=BB=A2=C3=E2=B7=D1G=D3=CA=CF=E4=A3=ADNo.1=B5=C4=B7=C0=B6=BE=B7=C0=C0= =AC=BB=F8=B3=AC=B4=F3=D3=CA=CF=E4 =20 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emerald_lake77 <@t> yahoo.com Mon Jul 11 11:30:13 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Macrophage marker mouse tissue Message-ID: <20050711163013.60499.qmail@web31705.mail.mud.yahoo.com> Hello, I am in need of a good macrophage marker on FFPE tissue. I will be using it on mouse tissue... mainly the heart (but other tissues may be included). So far I have looked into a number of antibodies (CD68, Mac-2) but can never be certain which one to choose. Thank you in advance for your help. Sincerely, Gustave Gustave Hebert, HT (ASCP) Scientist II Wyeth Research Cardiovascular and Metabolic Diseases Cambridge MA --------------------------------- Sell on Yahoo! Auctions - No fees. Bid on great items. From liz <@t> premierlab.com Mon Jul 11 12:25:33 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Macrophage marker mouse tissue In-Reply-To: <20050711163013.60499.qmail@web31705.mail.mud.yahoo.com> Message-ID: <000b01c5863d$8b1344b0$a7d48a80@AMY> Gustave We use F4/80 as a macrophage marker in mice. We have also tried MAC-2. Both work fine on FFPE tissue. Most of our clients ask for the F4/80 marker. We purchase ours from serotec. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of GT Hebert Sent: Monday, July 11, 2005 9:30 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Macrophage marker mouse tissue Hello, I am in need of a good macrophage marker on FFPE tissue. I will be using it on mouse tissue... mainly the heart (but other tissues may be included). So far I have looked into a number of antibodies (CD68, Mac-2) but can never be certain which one to choose. Thank you in advance for your help. Sincerely, Gustave Gustave Hebert, HT (ASCP) Scientist II Wyeth Research Cardiovascular and Metabolic Diseases Cambridge MA --------------------------------- Sell on Yahoo! Auctions - No fees. Bid on great items. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From relia1 <@t> earthlink.net Mon Jul 11 12:26:51 2005 From: relia1 <@t> earthlink.net (Pam Barker) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] RELIA job alert 7/11/05 Exciting Opportunity in South Carolina Message-ID: Hello Histonetters: I have an incredible opportunity with a new client of mine located in South Carolina. They are seeking an Anatomic Pathology Manager with a strong background in Histology. This is a full time permanent position. And the manager will be responsible for overseeing the histology and cytology departments of a growing pathology lab. Requirements: ASCP HT or HTL 7-10 years experience supervising a histology lab Strong hands-on histology experience. Experience supervising a fast paced lab My client offers excellent compensation benefits and relocation assistance. In addition due to their growth they will soon be moving into a BRAND NEW LAB!! If you are interested in hearing more about this opportunity or know of someone else who might be interested please contact me - Pam Barker at 866-607-3542 or relia1@earthlink.net Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net From Luis.Chiriboga <@t> med.nyu.edu Mon Jul 11 13:07:34 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Macrophage marker mouse tissue In-Reply-To: <20050711163013.60499.qmail@web31705.mail.mud.yahoo.com> Message-ID: A couple of years ago, I used (in frozen mouse heart) Serotec (cat# MCA1957s) rat anti-mouse cd68 which targets macrosialin, the murine CD68 homologue. It's a monoclonal (FA-11) IgG2a, detected with a biotinylated rabbit anti-rat (mouse adsorbed) followed by SA-alkaline phosphatase red. It worked very nicely but at a very high concentration. At the time they had not determined whether it was viable in FFPE so you may want to give them a call and see. Hope this helps Regards Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GT Hebert Sent: Monday, July 11, 2005 12:30 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Macrophage marker mouse tissue Hello, I am in need of a good macrophage marker on FFPE tissue. I will be using it on mouse tissue... mainly the heart (but other tissues may be included). So far I have looked into a number of antibodies (CD68, Mac-2) but can never be certain which one to choose. Thank you in advance for your help. Sincerely, Gustave Gustave Hebert, HT (ASCP) Scientist II Wyeth Research Cardiovascular and Metabolic Diseases Cambridge MA --------------------------------- Sell on Yahoo! Auctions - No fees. Bid on great items. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Mon Jul 11 13:05:29 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Apoptosis - FYI Message-ID: I know that many of you are doing work in the area of apoptosis. I see that Chemicon has a new antibody (Fractin) for detecting apoptotic cells in formalin-fixed, paraffin-embedded tissue. Has anyone used this antibody for this purpose? Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From gliuygao <@t> hotmail.com Mon Jul 11 13:41:44 2005 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] HTL blocks needed Message-ID: Dear Histonet, I am prepare my practice for HTL test. I need blcoks from Artery 0.5cm (outside diameter); Cerebellum (1.0x1.0 cm square); Ovary (1.0x1.0 cm square); Pancreas (1.0x1.0 square); Thyroid (1.0x1.0cm square); Lung (1.5x1.5cm square); Uterus (1.5x1.5cm); small intestine 2.0 cm in lenght and Helicobacter pylori. If anyone so kindly provide me the blocks. I am really appreciated. Thanks. Yan Gao Novartis From specialstainsqueen <@t> hotmail.com Mon Jul 11 13:59:12 2005 From: specialstainsqueen <@t> hotmail.com (Sherri Anderson) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] HTL blocks needed In-Reply-To: Message-ID: Dear Yan: Hello. Unless the practical portion of the HTL has changed since I've taken the examination (1999), examinees are required to prepare their own blocks (I think that you can have help with grossing so that the tissues demonstrate the required structures and are of the correct size), but everything else is supposed to be done by the examinee. This would include embedding. Most of the blocks are then submitted along with the slides for grading. Good luck with your exam! Sincerely, Sherri L. Anderson, BS, HTL(ASCP) Product Specialist Anatomical Pathology Thermo Electron Corp. >From: "yan gao" >To: Histonet@lists.utsouthwestern.edu >CC: gliuygao@hotmail.com >Subject: [Histonet] HTL blocks needed >Date: Mon, 11 Jul 2005 14:41:44 -0400 > > > Dear Histonet, > > I am prepare my practice for HTL test. I need blcoks from Artery > 0.5cm (outside diameter); Cerebellum (1.0x1.0 cm square); Ovary > (1.0x1.0 cm square); Pancreas (1.0x1.0 square); Thyroid (1.0x1.0cm > square); Lung (1.5x1.5cm square); Uterus (1.5x1.5cm); small intestine > 2.0 cm in lenght and Helicobacter pylori. If anyone so kindly provide > me the blocks. I am really appreciated. Thanks. > > > Yan Gao > Novartis >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From Kristopher.Kalleberg <@t> unilever.com Mon Jul 11 14:01:39 2005 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] HIER - best retrieval with minimum tissue damage? Message-ID: I almost exclusively perform IHC on skin and always use the pressure cooker for HIER. I rarely have any problems or encounter disruptions in the morphology of the sample. I put the slides in the pressure cooker and when the pressure reaches the appropriate level I let them cook for 6 minutes. It seems to be a fail-safe. Hope this helps. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull CT 06611 203 381 5765 From Rcartun <@t> harthosp.org Mon Jul 11 14:29:22 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] HIER - best retrieval with minimum tissue damage? Message-ID: What fixative do you use? How long do you fix? Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Kristopher Kalleberg" 07/11/05 03:01PM >>> I almost exclusively perform IHC on skin and always use the pressure cooker for HIER. I rarely have any problems or encounter disruptions in the morphology of the sample. I put the slides in the pressure cooker and when the pressure reaches the appropriate level I let them cook for 6 minutes. It seems to be a fail-safe. Hope this helps. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull CT 06611 203 381 5765 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From bme_sunysb <@t> yahoo.com Mon Jul 11 15:20:28 2005 From: bme_sunysb <@t> yahoo.com (A B) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] (no subject) Message-ID: <20050711202028.98396.qmail@web31211.mail.mud.yahoo.com> Hi, I am looking to prepare ultra thin samples (~100nm) of either calcified or decalcified bone for soft x-ray microscopy (NOT for TEM though). Any suggestions? Thanks. A.B. Biomedical Engineering Dept., SUNY Stony Brook NASA Goddard Institute for Space Studies --------------------------------- Yahoo! Mail for Mobile Take Yahoo! Mail with you! Check email on your mobile phone. From rgrow <@t> bmnet.com Mon Jul 11 17:17:27 2005 From: rgrow <@t> bmnet.com (rgrow@bmnet.com) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Proper Respirator for Formalin and Organic Solvents Message-ID: Greetings Histonetters, We are investigating different respirators for dumping tissues and changing processors. We use NBF and real xylene. What are other hospital labs using? Do you have to be fit-tested yearly? Are you following hosp. respiratory policy or your own? Sorry for all the questions, but I am on a fact-finding mission. Thanks, Renee Renee Grow, BA., HT (ASCP) rgrow@bmnet.com Histology Supervisor Blount Memorial Hospital 907 E. Lamar Alexander Pkwy. Maryville, TN 37804-5016 (865) 977-4744 (865) 977-5766 Fax ________________________________ This communication may contain protected health information (PHI) that is legally protected from inappropriate disclosure by the Privacy Standards of the Health Insurance Portability and Accountability Act (HIPAA) and relevant Tennessee Laws. If you are not the intended recipient, please note that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this message in error, you should notify the sender immediately by telephone or by return e-mail and delete this message from your computer. Direct questions to the Blount Memorial Hospital Privacy Officer at 865-977-4675. From zacharyweil <@t> yahoo.com Mon Jul 11 17:39:24 2005 From: zacharyweil <@t> yahoo.com (Zach Weil) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Microglia Stain Message-ID: <20050711223924.89011.qmail@web50602.mail.yahoo.com> Does anyone have a protocol for a microglia stain for cryostat sections of mouse brain? thanks --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour From Melissa.Gonzalez <@t> cellgenesys.com Mon Jul 11 17:59:10 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] mouse macrophage Message-ID: Gustave: Serotec's rat-anti mouse F4/80 works great on all treated tissues. Use a mouse liver as your positive control. Melissa Message: 6 Date: Mon, 11 Jul 2005 09:30:13 -0700 (PDT) From: GT Hebert Subject: [Histonet] Macrophage marker mouse tissue To: Histonet@lists.utsouthwestern.edu Message-ID: <20050711163013.60499.qmail@web31705.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello, I am in need of a good macrophage marker on FFPE tissue. I will be using it on mouse tissue... mainly the heart (but other tissues may be included). So far I have looked into a number of antibodies (CD68, Mac-2) but can never be certain which one to choose. Thank you in advance for your help. Sincerely, Gustave Gustave Hebert, HT (ASCP) Scientist II Wyeth Research Cardiovascular and Metabolic Diseases Cambridge MA From chris.goodall <@t> bristol.ac.uk Tue Jul 12 03:10:39 2005 From: chris.goodall <@t> bristol.ac.uk (anclg) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] re: slippery pig snouts Message-ID: <000001c586b9$3857ff50$8f6ede89@hist143> Dear Trisha, I also dry my + charged slides flat on a Hot hot plate, just hot enough to not melt the wax, overnight. Then dewax and take to 70% alcohol, then remove the slides and check that none of the cartilage has flipped over and swish around in the 70% to get it back if it has, I then dry the slides flat on a lukewarm hotplate....about 37C for about 10 mins before staining, this works for me. I also find that leaving some other tissue, bone or muscle on the cartilage helps to anchor the cartilage to the slide and even if it then detaches you can sometimes stain and flatten after coverslipping, I use little weights on the coverslips during drying flat. Chris From ree3 <@t> leicester.ac.uk Tue Jul 12 03:53:12 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Macrophage marker mouse tissue Message-ID: F4/80 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GT Hebert Sent: 11 July 2005 17:30 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Macrophage marker mouse tissue Hello, I am in need of a good macrophage marker on FFPE tissue. I will be using it on mouse tissue... mainly the heart (but other tissues may be included). So far I have looked into a number of antibodies (CD68, Mac-2) but can never be certain which one to choose. Thank you in advance for your help. Sincerely, Gustave Gustave Hebert, HT (ASCP) Scientist II Wyeth Research Cardiovascular and Metabolic Diseases Cambridge MA --------------------------------- Sell on Yahoo! Auctions - No fees. Bid on great items. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joycejudge259 <@t> hotmail.com Tue Jul 12 06:59:33 2005 From: joycejudge259 <@t> hotmail.com (joyce judge) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Punch set for Beecher Tissue Microarray Message-ID: I have been trying for weeks to get in touch with someone at Beecher Instruments so that I may reorder some punch sets for our Beecher Tissue Microarrayer and have had no success. I have tried emails, phone calls and faxes. Does anyone know where else I may purchase .6 mm, 1.0mm or 2.0 mm punch sets? Any help or insight you can give me would be greatly appreciated. Joyce Judge Cytomyx, LLC 781-863-9720 joyce.judge@cytomyx.com From Charlene.Henry <@t> STJUDE.ORG Tue Jul 12 08:18:43 2005 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Punch set for Beecher Tissue Microarray. . Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2099A196B@SJMEMXMB02.stjude.sjcrh.local> Good luck trying to get through to them. I ordered a tissue arrayer in November, 2004 and have not received it yet. Maybe they need to hire some help. Charlene -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of joyce judge Sent: Tuesday, July 12, 2005 7:00 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Punch set for Beecher Tissue Microarray. . I have been trying for weeks to get in touch with someone at Beecher Instruments so that I may reorder some punch sets for our Beecher Tissue Microarrayer and have had no success. I have tried emails, phone calls and faxes. Does anyone know where else I may purchase .6 mm, 1.0mm or 2.0 mm punch sets? Any help or insight you can give me would be greatly appreciated. Joyce Judge Cytomyx, LLC 781-863-9720 joyce.judge@cytomyx.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Tue Jul 12 08:26:17 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Fast myosin IHC Message-ID: Is anyone doing immunohistochemistry for fast myosin on formalin-fixed human tissue? If so, whose antibody are you using? My "Pub Med" search did not turn up much. Thanks! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From pmarcum <@t> vet.upenn.edu Tue Jul 12 08:26:21 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Punch set for Beecher Tissue Microarray. . In-Reply-To: <5CB39BCA5724F349BCB748675C6CA1A2099A196B@SJMEMXMB02.stjude .sjcrh.local> References: <5CB39BCA5724F349BCB748675C6CA1A2099A196B@SJMEMXMB02.stjude.sjcrh.local> Message-ID: <6.1.1.1.2.20050712092303.0197d298@mail.vet.upenn.edu> On and off I heard they are still in business and then they were not. Does anyone know if they are still viable or have sold to another company or out of business completely? If one can not reach them it makes it very questionable about ordering anything or whether they can meet demand. I know they were always hard to reach in the past. It is great little instrument as I have taken a couple of MTA classes with it and it was easy to use. Pam Macrum At 09:18 AM 7/12/2005, Henry, Charlene wrote: >Good luck trying to get through to them. I ordered a tissue arrayer in >November, 2004 and have not received it yet. Maybe they need to hire >some help. >Charlene > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of joyce >judge >Sent: Tuesday, July 12, 2005 7:00 AM >To: histonet@pathology.swmed.edu >Subject: [Histonet] Punch set for Beecher Tissue Microarray. . > >I have been trying for weeks to get in touch with someone at Beecher >Instruments so that I may reorder some punch sets for our Beecher Tissue >Microarrayer and have had no success. I have tried emails, phone calls >and >faxes. Does anyone know where else I may purchase .6 mm, 1.0mm or 2.0 >mm >punch sets? Any help or insight you can give me would be greatly >appreciated. > >Joyce Judge >Cytomyx, LLC >781-863-9720 >joyce.judge@cytomyx.com > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From billk1ibz <@t> yahoo.com Tue Jul 12 08:29:12 2005 From: billk1ibz <@t> yahoo.com (Bill) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Macrophage marker mouse tissue In-Reply-To: Message-ID: <20050712132912.74254.qmail@web53610.mail.yahoo.com> In my experience, Serotec's CD68 clone FA-11 is a much better tissue macrophage marker than F4/80, which is not expressed on all macs (i.e. lung). A 5 min. pretreatment w/ Dako Prot K is all that is required on FFPE mouse tissue. "Edwards, R.E." wrote:F4/80 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GT Hebert Sent: 11 July 2005 17:30 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Macrophage marker mouse tissue Hello, I am in need of a good macrophage marker on FFPE tissue. I will be using it on mouse tissue... mainly the heart (but other tissues may be included). So far I have looked into a number of antibodies (CD68, Mac-2) but can never be certain which one to choose. Thank you in advance for your help. Sincerely, Gustave Gustave Hebert, HT (ASCP) Scientist II Wyeth Research Cardiovascular and Metabolic Diseases Cambridge MA --------------------------------- Sell on Yahoo! Auctions - No fees. Bid on great items. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Luis.Chiriboga <@t> med.nyu.edu Tue Jul 12 08:47:08 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:25:18 2005 Subject: [Histonet] Macrophage marker mouse tissue In-Reply-To: <20050712132912.74254.qmail@web53610.mail.yahoo.com> Message-ID: That's great, never had the time to work it out myself.... approximate working concentration/dilution and incubation? Much appreciated regards Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bill Sent: Tuesday, July 12, 2005 9:29 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Macrophage marker mouse tissue In my experience, Serotec's CD68 clone FA-11 is a much better tissue macrophage marker than F4/80, which is not expressed on all macs (i.e. lung). A 5 min. pretreatment w/ Dako Prot K is all that is required on FFPE mouse tissue. "Edwards, R.E." wrote:F4/80 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GT Hebert Sent: 11 July 2005 17:30 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Macrophage marker mouse tissue Hello, I am in need of a good macrophage marker on FFPE tissue. I will be using it on mouse tissue... mainly the heart (but other tissues may be included). So far I have looked into a number of antibodies (CD68, Mac-2) but can never be certain which one to choose. Thank you in advance for your help. Sincerely, Gustave Gustave Hebert, HT (ASCP) Scientist II Wyeth Research Cardiovascular and Metabolic Diseases Cambridge MA --------------------------------- Sell on Yahoo! Auctions - No fees. Bid on great items. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From settembr <@t> umdnj.edu Tue Jul 12 09:05:00 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Fast myosin IHC Message-ID: Richard, I see that Novocastra has Myosin Heavy Chain (fast). Cat.# NCL-MHCf. Their antibodies, I believe, are now distributed by Vision BioSystems out of Norwell, Mass www.vision-bio.com Hope this helps. Dana Settembre University Hospital - UMDNJ Immunohistochemistry Lab Newark, NJ >>> Richard Cartun 7/12/2005 9:26:17 AM >>> Is anyone doing immunohistochemistry for fast myosin on formalin-fixed human tissue? If so, whose antibody are you using? My "Pub Med" search did not turn up much. Thanks! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kappeler <@t> patho.unibe.ch Tue Jul 12 09:37:08 2005 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Fast myosin IHC References: Message-ID: <008801c586ef$40bf45f0$27955c82@patho.unibe.ch> Hi Richard We successfully use: - mo-a-Myosin II (fast), clone MY-32, Sigma M 4276, working conc. approx 10 ug Ig/ml (corresponds to 1:400 with the lot we have), HIER in citrate pH 6.0 (pressure cooker) - mo-a-Myosin I (slow), clone NOQ7.5.4D, Sigma M 8421, working conc. approx 7.5 ug Ig/ml (corresponds to 1:1000 with the lot we have), HIER in citrate pH 6.0 (pressure cooker) Both mAbs work very well (see http://www.pathology.unibe.ch/Diagn/speztech/spez_ihc.htm for a pic (german website, sorry). Can you give me a hint in return, where you get your antibodies against Hepatitis B surface Ag (HBs) and core Ag (HBc) from? Our source is no longer able to supply them, due to some bureaucratic / eurocratic rules, now I'm looking for a reliable replacement. Many thanks in advance! Best regards Andi Kappeler Institute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "Richard Cartun" To: Sent: Tuesday, July 12, 2005 3:26 PM Subject: [Histonet] Fast myosin IHC > Is anyone doing immunohistochemistry for fast myosin on formalin-fixed > human tissue? If so, whose antibody are you using? My "Pub Med" search > did not turn up much. Thanks! > > Richard > > Richard W. Cartun, Ph.D. > Director, Immunopathology & Histology > Assistant Director, Anatomic Pathology > Hartford Hospital > 80 Seymour Street > Hartford, CT 06102 > (860) 545-1596 > (860) 545-0174 Fax > > > > > ----------------------------------------------- > Confidentiality Notice > > This e-mail message, including any attachments, is for the sole use of the > intended recipient(s) and may contain confidential or proprietary > information which is legally privileged. Any unauthorized review, use, > disclosure, or distribution is prohibited. If you are not the intended > recipient, please promptly contact the sender by reply e-mail and destroy > all copies of the original message. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From pstefanova <@t> sten.sunnybrook.utoronto.ca Tue Jul 12 10:10:55 2005 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Punch set for Beecher Tissue Microarray. . References: <5CB39BCA5724F349BCB748675C6CA1A2099A196B@SJMEMXMB02.stjude.sjcrh.local> Message-ID: <005301c586f3$e8380ed0$9d194c8e@WS21203> Hi all, just to share my experience with Beecher Instruments. I have waited 3 months to get a quotation for MTA then I ordered it Oct., 2004 and I received it April, 2005. I've found that Alpha Metrix Biotech cares Manual Tissue Arrayers I and II, and I hope spare parts as well. Tel: + 49 (0)6106 63810 Fax: + 49 (0)6106 648084 Email: info@alphametrix.de Good luck ----- Original Message ----- From: "Henry, Charlene" To: "joyce judge" ; Sent: Tuesday, July 12, 2005 9:18 AM Subject: RE: [Histonet] Punch set for Beecher Tissue Microarray. . > Good luck trying to get through to them. I ordered a tissue arrayer in > November, 2004 and have not received it yet. Maybe they need to hire > some help. > Charlene > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of joyce > judge > Sent: Tuesday, July 12, 2005 7:00 AM > To: histonet@pathology.swmed.edu > Subject: [Histonet] Punch set for Beecher Tissue Microarray. . > > I have been trying for weeks to get in touch with someone at Beecher > Instruments so that I may reorder some punch sets for our Beecher Tissue > Microarrayer and have had no success. I have tried emails, phone calls > and > faxes. Does anyone know where else I may purchase .6 mm, 1.0mm or 2.0 > mm > punch sets? Any help or insight you can give me would be greatly > appreciated. > > Joyce Judge > Cytomyx, LLC > 781-863-9720 > joyce.judge@cytomyx.com > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From cdemarinis <@t> SARATOGACARE.ORG Tue Jul 12 10:19:08 2005 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis, Carolyn) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] xylene disposal Message-ID: There has been some concern about the proper disposal of paraffin from the tissue processor due to a small amount of residual xylene in the paraffin. To satisfy EPA guidelines, how do labs in New York State dispose of their paraffin and alcohols that have small traces of xylene? *************************************************************************************** CONFIDENTIALITY NOTICE: This e-mail communication and any attachmentsmay contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. From cfavara <@t> niaid.nih.gov Tue Jul 12 11:10:50 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] whole eye help Message-ID: I have searched the archives and have found great information on processing whole eyes from larger mammals. I have a few remaining questions for wither Phil Oshel or Anne in Africa. I would appreciate one or both of you contacting me. I tried contacting both privately but e-mail has bounced back. Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From mauger <@t> email.chop.edu Tue Jul 12 11:38:41 2005 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] renin Message-ID: Hi Lee and Pegy, Do you know of a source for +renin control? Thanks, Jo Mauger >>> "Lee & Peggy Wenk" 06/30/05 6:26 AM >>> Just wondering if doing an immunohistochemical stain for renin or cathepsin D would be easier, and more reliable, than doing the Bowie. Also, JG granules will stain positive with PAS (periodic acid-Schiff) and TFT (thioflavin T), though neither of these stains are specific for JG granules. I'll cover the basics of the Bowie stain (taken from the Sheehan book), and you can let me know if you want still want me to send you the procedure. Fix for 48 hours in Helly fixative (mercuric chloride, potassium dichromate, formaldehyde)(Yes, it must be Helly. Bowie will not work with formalin fixed tissue) Wash tissue in running water overnight, then process and embed. Cut sections at 4 um. Deparafinize slide to water. Lugolize (de-Zenkerize) with iodine and sodium thiosulfate Mordant in 2.5% potassium dichromate at 40 degrees C overnight Rinse in d. water Stain in Bowie solution overnight(mixture of Biebrich scarlet, ethyl violet water, filtered, filter paper with dye allowed to dry overnight, dissolving dry dye in alcohol) Blot slides Dip in acetone Differentiate in xylene:clove oil mixture (hint from me - clove oil makes the lab smell great, but is very expensive) Xylene Why the stain isn't used any more: - Fixation and staining takes several days (4 by my count). - Tissue must be fixed in Helly. - Mercury and potassium dichromate waste must be correctly disposed - Expensive clove oil must be bought(100 mL = $60+ US) Let me know if you still would like the Bowie procedure. Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jose Luis Palazon Fernandez Sent: Wednesday, June 29, 2005 9:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bowies method Dear List-fellows Could any of you send me the protocol of Bowie s method for juxtaglomerular cells in kidney. thanks in advance Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pex0220 <@t> yahoo.com.cn Tue Jul 12 11:56:48 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Thank! Message-ID: <20050712165648.28859.qmail@web15506.mail.cnb.yahoo.com> Thank for your nice helps! Guofeng Guofeng --------------------------------- DO YOU YAHOO!? ÑÅ»¢Ãâ·ÑGÓÊÏ䣭ÖйúµÚÒ»¾øÎÞÀ¬»øÓʼþɧÈų¬´óÓÊÏä From kspencer <@t> utmem.edu Tue Jul 12 12:25:23 2005 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Microglia Stain In-Reply-To: <20050711223924.89011.qmail@web50602.mail.yahoo.com> References: <20050711223924.89011.qmail@web50602.mail.yahoo.com> Message-ID: Has anyone done fluorescence IHC for GABA antibody? Thanks On Jul 11, 2005, at 5:39 PM, Zach Weil wrote: > Does anyone have a protocol for a microglia stain for cryostat > sections of mouse brain? > > thanks > > > --------------------------------- > Yahoo! Mail > Stay connected, organized, and protected. Take the tour > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LewisS <@t> ccri.net Tue Jul 12 12:48:17 2005 From: LewisS <@t> ccri.net (Lewis, Sarah) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] ADG & Dysferlin troubles Message-ID: <714B9F12B4E18C4C843B66E8E190F2AD44753D@res2k3ms01.CRII.ORG> Hello histoneters, I could really use any procedural information anyone has on Alpha-dystroglycan (antibody only available thru Upstate) and Dysferlin (antibody from VectorLabs). I have been attempting the stains for 2months now with no satisfying results. I have tried several different procedures/fixatives/incubation times/dilutions/. I perform a battery of 14 neuromuscular Immunoflourescence, they are all BEAUTIFUL fitc green with no background staining (picture perfect) .....except these two boogers. PLEASE HELP me make my pathologist proud!!! ALL THE CREDIT TO HISTONET!!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net From jlinda <@t> ces.clemson.edu Tue Jul 12 12:54:06 2005 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] South Carolina Meeting Message-ID: <5.2.1.1.2.20050712134146.03257760@mailhost.ces.clemson.edu> The South Carolina Society of Histotechnology (SCSHT) is having a Summer Meeting at the Columbiana Hotel and Conference Center in Columbia, SC August 19 - 20. The program is as follows: Friday, August 19, 2005 - 5:30 - 7:00 Technical and Financial Considerations in the Selection of an Automated IHC Slide Staining System Joseph D. (Joe) Myers, MS, CT(ASCP) Saturday, August 20, 2005 - 8:00 - 11:30 Joy of Histology or What We Can Learn from the Kitchen Ada Feldman, MS, HT (ASCP) HTL Grossing Procedures for the Histotech - 1:00 - 4:30 Connie Wavrin, MT(ASCP) For more information and a program, please contact Debbie Ellenburg at : 463 Country Club Road, Liberty, SC 29657 Fax:864-255-1664 Email: DEllenburg2@stfrancishealth.org Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo.htm From sagitalcuts <@t> yahoo.com Tue Jul 12 12:59:58 2005 From: sagitalcuts <@t> yahoo.com (Yolanda Maldonado) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] PGP 9.5 Message-ID: <20050712175959.74502.qmail@web53507.mail.yahoo.com> Hello histonetters: Does anyone know of a good substitute for PGP 9.5 polyclonal Ab? Chemicon has it on back order until August and I need to find a substitute ASAP!!! Thanks in advance! Patricia Maldonado --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour From peoshel <@t> wisc.edu Tue Jul 12 12:58:12 2005 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] whole eye help In-Reply-To: References: Message-ID: Cynthia, Email bounced back? Now you've got me wondering. What can I help you with? Phil P.S. I'm copying this to the list, in case it's something weird ... >I have searched the archives and have found great information on processing >whole eyes from larger mammals. I have a few remaining questions for wither >Phil Oshel or Anne in Africa. I would appreciate one or both of you >contacting me. I tried contacting both privately but e-mail has bounced >back. > > > >Thanks, > >c > > > >Cynthia Favara >NIAID/NIH/RML/LPVD >903 South 4th Street >Hamilton, MT 59840 >406-363-9317 > >Disclaimer: >The information in this e-mail and any of its attachments is confidential >and may contain sensitive information. It should not be used by anyone who >is not the original intended recipient. If you have received this e-mail in >error please inform the sender and delete it from your mailbox or any other >storage devices. National Institute of Allergy and Infectious Diseases shall >not accept liability for any statements made that are sender's own and not >expressly made on behalf of the NIAID by one of its representatives > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm From carl.hobbs <@t> kcl.ac.uk Tue Jul 12 13:37:01 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] re macrophage marker Message-ID: <002301c58710$b1925170$0201a8c0@lynne> Bill....thanks for insight. On the look-out for just a mac marker for a PhD student's project on FFPWs of mouse lungs. Have you tried Ag retrieval other than by proteolytic digestion? ( eg HIER) Carl -------------- next part -------------- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.323 / Virus Database: 267.8.13/47 - Release Date: 12/07/2005 From MAUGER <@t> email.chop.edu Tue Jul 12 13:44:10 2005 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] ADG & Dysferlin troubles Message-ID: Hi, I use dysferlin from novocastra at 1:50 for 1 hour, followed by anti mouse FITC at 1:100 for 1 hour. You should fix slide in acetone or methanol for 20 secs prior to staining. Jo Mauger >>> "Lewis, Sarah" 07/12/05 1:48 PM >>> Hello histoneters, I could really use any procedural information anyone has on Alpha-dystroglycan (antibody only available thru Upstate) and Dysferlin (antibody from VectorLabs). I have been attempting the stains for 2months now with no satisfying results. I have tried several different procedures/fixatives/incubation times/dilutions/. I perform a battery of 14 neuromuscular Immunoflourescence, they are all BEAUTIFUL fitc green with no background staining (picture perfect) .....except these two boogers. PLEASE HELP me make my pathologist proud!!! ALL THE CREDIT TO HISTONET!!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From fladanielski <@t> yahoo.com.br Tue Jul 12 13:47:00 2005 From: fladanielski <@t> yahoo.com.br (Flavia Danielski) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] DNA extraction protocol for fixed tissues Message-ID: <20050712184701.65949.qmail@web30605.mail.mud.yahoo.com> Hi histonet people, I'd love to have some help... I need to extract DNA from paraf. 4% fixed tissues... what protocols do you use? Thank you very much greetings from Brazil Flavia Fl?via Helena da Silva Lab. de Imunogen?tica, Depto. de Gen?tica, UFRGS Av. Bento Gon?alves, 9500 pr?dio 43323/lab 212C Bairro Agronomia. POA-RS-BRAZIL phone: 0055-(51)33166737 __________________________________________________ Converse com seus amigos em tempo real com o Yahoo! Messenger http://br.download.yahoo.com/messenger/ From lrichey <@t> u.washington.edu Tue Jul 12 14:28:26 2005 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] PGP 9.5 In-Reply-To: <20050712175959.74502.qmail@web53507.mail.yahoo.com> References: <20050712175959.74502.qmail@web53507.mail.yahoo.com> Message-ID: <42D419DA.6050108@u.washington.edu> We order out PGP9.5 from biogenisis Yolanda Maldonado wrote: >Hello histonetters: >Does anyone know of a good substitute for PGP 9.5 polyclonal Ab? >Chemicon has it on back order until August and I need to find a substitute ASAP!!! >Thanks in advance! >Patricia Maldonado > > >--------------------------------- >Yahoo! Mail > Stay connected, organized, and protected. Take the tour >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From billk1ibz <@t> yahoo.com Tue Jul 12 15:11:21 2005 From: billk1ibz <@t> yahoo.com (Bill) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] re macrophage marker In-Reply-To: <002301c58710$b1925170$0201a8c0@lynne> Message-ID: <20050712201121.76339.qmail@web53605.mail.yahoo.com> Heating methods seem to diminish or abolish the staining- I've tried HIER as well as other enzymes and Proteinase K works the best. I use it at about 1 ug/ml using the following method- 30 min in primary 30 min Rabbit anti-rat (adsorbed against mouse) 30 min labeled polymer (Envision+ HRP or Mach-2 ALP) DAB or Permanent Fast Red This method works great for me. Carl Hobbs wrote: Bill....thanks for insight. On the look-out for just a mac marker for a PhD student's project on FFPWs of mouse lungs. Have you tried Ag retrieval other than by proteolytic digestion? ( eg HIER) CarlNo virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.323 / Virus Database: 267.8.13/47 - Release Date: 12/07/2005 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From eshields <@t> bhset.org Tue Jul 12 15:25:30 2005 From: eshields <@t> bhset.org (E Sharon Shields) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Need help from Ventana BenmchMark users Message-ID: Hi all, I have been asked to develop stain protocols for TdT on bone marrow aspirates and Hepatocyte Specific Antigen on FFPE tissues. Does anyone out there in Histoland doing these two IHC stains on the BenchMark? Would you be willing to share your antibody source and stain protocol? Thank you, Sharon Shields Bapitist Hospital of East Tennessee Knoxville, TN phone 865 549-4351 fax 865 632-5316 From cthouron <@t> ochsner.org Tue Jul 12 15:34:25 2005 From: cthouron <@t> ochsner.org (Carol Thouron) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Humidity chamber Message-ID: I am looking for a plexiglass incubation chamber that holds 20 slides on a raised metal tray. Can anyone suggest a vendor? C Thouron Ochsner Clinic Foundation New Orleans, LA From carl.hobbs <@t> kcl.ac.uk Wed Jul 13 02:22:47 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] re: macrophage marker in mouse tissue Message-ID: <001501c5877b$abb4b0c0$112b5c9f@Carlos> Bill....thanks for insight. " In my experience, Serotec's CD68 clone FA-11 is a much better tissue macrophage marker than F4/80" On the look-out for just a mac marker for a > PhD student's project on FFPWs of mouse lungs. Have you tried Ag > retrieval other than by proteolytic digestion ( eg HIER) for FA-11? > Carl From n.cragg <@t> epistem.co.uk Wed Jul 13 03:20:38 2005 From: n.cragg <@t> epistem.co.uk (Nicola Cragg) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Result : HIER - best retrieval with minimum tissue damage Message-ID: Hello, Thanks for everyone who has suggested gentler heat methods - waterbath heating was successful in retrieving the antigen of interest (PGP9.5 as a marker for cutaneous nerve fibres) in skin, while maintaining dermal integrity. Tried 2 buffers - citrate pH6.0 and EDTA pH8.0, preferred the citrate this time as a little cleaner, but not much between them. Will definitely use this method again as part of the optimisation process for other dermal antigens which require heat retrieval. Thanks again, Nicola Cragg EpiStem Ltd. Manchester, UK From juan.gutierrez <@t> christushealth.org Wed Jul 13 05:15:31 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Need help from Ventana BenmchMark users Message-ID: Hello Sharon, We use Ventana's TdT (SEN28)and Dakocytomation's Hepatocyte antibodies. For the TdT we have found that using the Enhanced V-Red chromogen works much better than the i-View DAB (less background). The protocols are: TdT Standard CC1 Option 1 for 8 minutes (Fc receptor block from INNOVEX Biosciences 1-800-622-7808) Antibody for 16 min (32 min if using i-View) Amplify A/B block Counterstain Hepatocyte Standard CC1 Antibody for 32 min at a 1:25-1:50 dilution A/B block Counterstain These are done on a regular Benchmark, not the LT or XT. Good luck with it and I hope this helps. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of E Sharon Shields Sent: Tuesday, July 12, 2005 3:26 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need help from Ventana BenmchMark users Hi all, I have been asked to develop stain protocols for TdT on bone marrow aspirates and Hepatocyte Specific Antigen on FFPE tissues. Does anyone out there in Histoland doing these two IHC stains on the BenchMark? Would you be willing to share your antibody source and stain protocol? Thank you, Sharon Shields Bapitist Hospital of East Tennessee Knoxville, TN phone 865 549-4351 fax 865 632-5316 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Wed Jul 13 06:06:42 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] xylene disposal Message-ID: I pour mine into a biohazard bag which is enclosed in a biohazard bin. Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: Molinari, Betsy Sent: Wednesday, July 13, 2005 6:04 AM To: 'Demarinis, Carolyn' Subject: RE: [Histonet] xylene disposal I put mine into a biohazard bag which goes into a biohazard box. Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave Houston TX, 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Demarinis, Carolyn Sent: Tuesday, July 12, 2005 10:19 AM To: HISTONET (E-mail) Subject: [Histonet] xylene disposal There has been some concern about the proper disposal of paraffin from the tissue processor due to a small amount of residual xylene in the paraffin. To satisfy EPA guidelines, how do labs in New York State dispose of their paraffin and alcohols that have small traces of xylene? ************************************************************************ *************** CONFIDENTIALITY NOTICE: This e-mail communication and any attachmentsmay contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From settembr <@t> umdnj.edu Wed Jul 13 06:43:36 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] ADG & Dysferlin troubles Message-ID: Sarah, I use Dysferlin from Novocastra on frozen sections, but not FITC, just regular IHC. If you're desperate we can talk. I use it with a detection kit for mouse, any vendor will usually do and DAB as the chromogen. No special scope is needed. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> "Lewis, Sarah" 7/12/2005 1:48:17 PM >>> Hello histoneters, I could really use any procedural information anyone has on Alpha-dystroglycan (antibody only available thru Upstate) and Dysferlin (antibody from VectorLabs). I have been attempting the stains for 2months now with no satisfying results. I have tried several different procedures/fixatives/incubation times/dilutions/. I perform a battery of 14 neuromuscular Immunoflourescence, they are all BEAUTIFUL fitc green with no background staining (picture perfect) .....except these two boogers. PLEASE HELP me make my pathologist proud!!! ALL THE CREDIT TO HISTONET!!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From n.cragg <@t> epistem.co.uk Wed Jul 13 07:59:47 2005 From: n.cragg <@t> epistem.co.uk (Nicola Cragg) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] FAO: Kris Kalleberg RE: HIER on skin Message-ID: Sorry Kris - only just spotted your reply. Thanks for your advice - I'm going to try your method. Nicola From kweidenh <@t> montefiore.org Wed Jul 13 08:10:34 2005 From: kweidenh <@t> montefiore.org (Karen Weidenheim) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] ADG & Dysferlin troubles Message-ID: Dear All, we have never been able to get the Novocastra dysferlin Ab to work on frozen sections using DAB on the Ventana. We have tried multiple permutations. Would someone be kind enough to share their exact method for this antibody on Frozen sections of Muscle? This is very important for diagnosis of muscular dystrophies. Thanks so much. Karen Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> Dana Settembre 7/13/2005 7:43:36 AM >>> Sarah, I use Dysferlin from Novocastra on frozen sections, but not FITC, just regular IHC. If you're desperate we can talk. I use it with a detection kit for mouse, any vendor will usually do and DAB as the chromogen. No special scope is needed. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> "Lewis, Sarah" 7/12/2005 1:48:17 PM >>> Hello histoneters, I could really use any procedural information anyone has on Alpha-dystroglycan (antibody only available thru Upstate) and Dysferlin (antibody from VectorLabs). I have been attempting the stains for 2months now with no satisfying results. I have tried several different procedures/fixatives/incubation times/dilutions/. I perform a battery of 14 neuromuscular Immunoflourescence, they are all BEAUTIFUL fitc green with no background staining (picture perfect) .....except these two boogers. PLEASE HELP me make my pathologist proud!!! ALL THE CREDIT TO HISTONET!!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Jul 13 10:11:36 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Re: Humidity chamber In-Reply-To: References: Message-ID: <6.0.0.22.1.20050713090409.01b51280@gemini.msu.montana.edu> Carol, There is a superb manual staining incubation chamber called Slidemaster from Scytek (www.scytek.com) out of Logan Utah. They are wonderful for rinsing, collecting waste, developing chromogen against a white background without having to pick up a slide throughout the whole procedure. The slides do NOT fall off the slide rack, a tremendous help during long, precious IHC protocols. You can put them in refrigerator, 37C incubator, or rotating platform, etc. These save time and wasted motion handling slides - we have 5 of them often with all five in use at one time. At 02:34 PM 7/12/2005, you wrote: >I am looking for a plexiglass incubation chamber that holds 20 slides on a >raised metal tray. Can anyone suggest a vendor? > >C Thouron >Ochsner Clinic Foundation >New Orleans, LA 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 conniemoss <@t> relia.net Wed Jul 13 10:16:09 2005 From: conniemoss <@t> relia.net (Connie McManus) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Humidity chamber In-Reply-To: References: Message-ID: <49479.208.186.240.165.1121267769.squirrel@email.relia.net> For many years I used a vegetable crisper for a humid chamber. I put water in the bottom, placed a metal slide tray on top of inverted staining dishes, then covered the top of the crisper with Saran Wrap. If the Saran Wrap got so it wouldn't stick, I coated the rim of the crisper with a thin coat of petroleum jelly . It worked great. Eventually, however, I bought a nifty little humid chamber tha holds about 30 slides. It is made in my very own little town of Logan, UT, but I forget the name of the company!! Gayle Callis recommended it to me. Gayle?? -- Connie McManus Mt Ogden Scientific Services -Providing full service electron microscopy for you 950 W Kershaw, Suite E Ogden UT 84401 toll-free: 877/311-6677 direct: 801/745-2583 cell: 435/757/2975 fax: 435/514-1781 conniemoss@relia.net www.mtogdensci.com ====== Carol Thouron said: > I am looking for a plexiglass incubation chamber that holds 20 slides on a raised metal > tray. Can anyone suggest a vendor? > > C Thouron > Ochsner Clinic Foundation > New Orleans, LA > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From settembr <@t> umdnj.edu Wed Jul 13 10:15:36 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] ADG & Dysferlin troubles Message-ID: Karen, I use Novocastra's Dysferlin cat.# NCL-Hamlet on human frozen muscle sections. After I let the sections air dry from 1 hour to overnight. I use it at a 1:160 dilution. The detection system that I use is DakoCytomation's LSAB2 kit and their DAB kit. Works well. Dana Settembre University Hospital-UMSNJ Newark, NJ >>> Karen Weidenheim 7/13/2005 9:10:34 AM >>> Dear All, we have never been able to get the Novocastra dysferlin Ab to work on frozen sections using DAB on the Ventana. We have tried multiple permutations. Would someone be kind enough to share their exact method for this antibody on Frozen sections of Muscle? This is very important for diagnosis of muscular dystrophies. Thanks so much. Karen Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> Dana Settembre 7/13/2005 7:43:36 AM >>> Sarah, I use Dysferlin from Novocastra on frozen sections, but not FITC, just regular IHC. If you're desperate we can talk. I use it with a detection kit for mouse, any vendor will usually do and DAB as the chromogen. No special scope is needed. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> "Lewis, Sarah" 7/12/2005 1:48:17 PM >>> Hello histoneters, I could really use any procedural information anyone has on Alpha-dystroglycan (antibody only available thru Upstate) and Dysferlin (antibody from VectorLabs). I have been attempting the stains for 2months now with no satisfying results. I have tried several different procedures/fixatives/incubation times/dilutions/. I perform a battery of 14 neuromuscular Immunoflourescence, they are all BEAUTIFUL fitc green with no background staining (picture perfect) .....except these two boogers. PLEASE HELP me make my pathologist proud!!! ALL THE CREDIT TO HISTONET!!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfavara <@t> niaid.nih.gov Wed Jul 13 10:23:00 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Re: Humidity chamber Message-ID: Ditto to this - one of the best investments for a lab - I have automated IHC and still use these on numerous occasions! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Wednesday, July 13, 2005 8:12 AM To: Carol Thouron; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Humidity chamber Carol, There is a superb manual staining incubation chamber called Slidemaster from Scytek (www.scytek.com) out of Logan Utah. They are wonderful for rinsing, collecting waste, developing chromogen against a white background without having to pick up a slide throughout the whole procedure. The slides do NOT fall off the slide rack, a tremendous help during long, precious IHC protocols. You can put them in refrigerator, 37C incubator, or rotating platform, etc. These save time and wasted motion handling slides - we have 5 of them often with all five in use at one time. At 02:34 PM 7/12/2005, you wrote: >I am looking for a plexiglass incubation chamber that holds 20 slides on a >raised metal tray. Can anyone suggest a vendor? > >C Thouron >Ochsner Clinic Foundation >New Orleans, LA 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 gentras <@t> vetmed.auburn.edu Wed Jul 13 10:41:08 2005 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] mouse brain Message-ID: <6.0.1.1.0.20050713102532.01b7d838@mailhost.vetmed.auburn.edu> Hello, please have any of you who process & section paraformaldehyde fixed, paraffin embedded mouse brain experienced asymmetry of the right & left hemispheres? Recently our coronal mouse brain sections which appear symmetrical upon gross trim and after initial facing of paraffin are asymmetrical after staining. I've rotated the block holder to the limit and sectioned the most rostral hemisphere at higher micron thickness to accommodate. However, to my dismay and disappointment none of this has helped very much. Any pointers in remedying this situation ASAP will be greatly appreciated. Thanks, Atoska From cfavara <@t> niaid.nih.gov Wed Jul 13 11:23:49 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] mouse brain Message-ID: Atoska, Questions, can you be more specific as to what you mean by asymmetrical - do you mean they do not exhibit the same anatomical structures from one side to the other or do they differ in size? I take it this is something new and the processing, etc are the same protocols that have given good results. How are you doing the gross cuts? Do you use a brain mold? How do you put them in the cassettes - lens paper or free floating? Sounds strange, would love to assist if possible. I do coronal mouse sections with a mold and place in lens paper through the processor but have had some people through the lab that don't seem to have the manual dexterity to do this consistently. Let me know! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Atoska S. Gentry [mailto:gentras@vetmed.auburn.edu] Sent: Wednesday, July 13, 2005 8:41 AM To: Histonet Subject: [Histonet] mouse brain Hello, please have any of you who process & section paraformaldehyde fixed, paraffin embedded mouse brain experienced asymmetry of the right & left hemispheres? Recently our coronal mouse brain sections which appear symmetrical upon gross trim and after initial facing of paraffin are asymmetrical after staining. I've rotated the block holder to the limit and sectioned the most rostral hemisphere at higher micron thickness to accommodate. However, to my dismay and disappointment none of this has helped very much. Any pointers in remedying this situation ASAP will be greatly appreciated. Thanks, Atoska _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GoodwinD <@t> pahosp.com Wed Jul 13 12:12:17 2005 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] overdecalcified bone Message-ID: <992899E9EC268548AB8DDE246AF88473055F5230@PAHEX01.uphs.upenn.edu> Hi, Vicki. Is there a procedure to restore the nuclear staining uptake in over-decalcified human bone? Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com From hborgeri <@t> wfubmc.edu Wed Jul 13 12:39:57 2005 From: hborgeri <@t> wfubmc.edu (Hermina Borgerink) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] overdecalcified bone Message-ID: <9AEEF1FB6254224AA355ED285F8491650999CD21@EXCHVS2.medctr.ad.wfubmc.edu> >From Theory and practice of Histotechnology, Sheehan and Hrapchak, Appendix B, p. 445. To restore basophilic properties to tissues as the result of overexposure to decalcifying solution: Method 1 1. Deparaffinize and hydrate slides 2. Place slides in 5% aqueous sodium bicarbonate solution from 4 hours to overnight 3. Wash in tap water for 5 minutes 4. Stain as desired Method 2 In step 2, use 5% aqueous solution of ammonium sulfide overnight Method 3 In step 2, use 5% aqueous solution of periodic acid overnight Hopefully one of these methods will work for you. Hermina Hermina M. Borgerink, BA, HTL(ASCP)QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax. (336) 716-1515 e-mail hborgeri@wfubmc.edu "Treat a man as he appears to be, and you make him worse. But treat a man as if he already were what he potentially could be, and you make him what he should be." (Goethe) This electronic message, including any attachments, is confidential and proprietary and is solely for the intended recipient. If you are not the intended recipient, this message was sent to you in error and you are hereby advised that any review, disclosure, copying, distribution or use of this message, or any of the information included therein, is unauthorized and strictly prohibited. If you have received this electronic transmission in error, please immediately notify the sender by reply and permanently delete all copies of this message and its attachments. Thank you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Goodwin, Diana Sent: Wednesday, July 13, 2005 1:12 PM To: 'kalschev@svm.vetmed.wisc.edu' Cc: Histonet (E-mail) Subject: [Histonet] overdecalcified bone Hi, Vicki. Is there a procedure to restore the nuclear staining uptake in over-decalcified human bone? Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jluis.palazon <@t> icman.csic.es Wed Jul 13 13:06:01 2005 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Re: renin Message-ID: <20050713180601.859E48620FB@perceval.uca.es> Hi all is renin antibody an expensive one?. O just want to know if juxtaglmerular cells are present in an aglomerular fish species I am studying. I only would use it for some 4 or 5 slides and I dont know if it is worth while to buy the antibody As my lab is not specialized in inmunostaining. thanks in advance for replys Jos? Luis El dia 12/07/2005 18:38 usted envio el siguiente mensaje: >Date: 12 de Julio de 2005 18:38:41 >From: "Joanne Mauger" >Subject: renin >To: jluis.palazon@icman.csic.es, histonet@lists.utsouthwestern.edu, lpwenk@sbcglobal.net > > Hi Lee and Pegy, > > Do you know of a source for +renin control? > > Thanks, > Jo Mauger > > >>> "Lee & Peggy Wenk" 06/30/05 6:26 AM >>> > Just wondering if doing an immunohistochemical stain for renin or cathepsin > D would be easier, and more reliable, than doing the Bowie. > > Also, JG granules will stain positive with PAS (periodic acid-Schiff) and > TFT (thioflavin T), though neither of these stains are specific for JG > granules. > > I'll cover the basics of the Bowie stain (taken from the Sheehan book), and > you can let me know if you want still want me to send you the procedure. > > Fix for 48 hours in Helly fixative (mercuric chloride, potassium dichromate, > formaldehyde)(Yes, it must be Helly. Bowie will not work with formalin fixed > tissue) > Wash tissue in running water overnight, then process and embed. Cut sections > at 4 um. > > Deparafinize slide to water. > Lugolize (de-Zenkerize) with iodine and sodium thiosulfate > Mordant in 2.5% potassium dichromate at 40 degrees C overnight > Rinse in d. water > Stain in Bowie solution overnight(mixture of Biebrich scarlet, ethyl violet > water, filtered, filter paper with dye allowed to dry overnight, dissolving > dry dye in alcohol) > Blot slides > Dip in acetone > Differentiate in xylene:clove oil mixture (hint from me - clove oil makes > the lab smell great, but is very expensive) > Xylene > > Why the stain isn't used any more: > - Fixation and staining takes several days (4 by my count). > - Tissue must be fixed in Helly. > - Mercury and potassium dichromate waste must be correctly disposed > - Expensive clove oil must be bought(100 mL = $60+ US) > > Let me know if you still would like the Bowie procedure. > > Peggy Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jose Luis > Palazon Fernandez > Sent: Wednesday, June 29, 2005 9:01 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Bowies method > > Dear List-fellows Could any of you send me the protocol of Bowie s method > for juxtaglomerular cells in kidney. thanks in advance Jos? Luis > Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de > Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: > jluis.palazon@icman.csic.es > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From weneng2004 <@t> yahoo.com Wed Jul 13 13:13:47 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] H&E after IHC Message-ID: <20050713181347.90811.qmail@web53410.mail.yahoo.com> Hi histonetters, I have to ask a basic question. I am going to do IHC using DAB. I used to do hematoxylin counterstaining. Can I do H&E after IHC? Will it cover brown color of DAB? Thanks, Wen --------------------------------- Start your day with Yahoo! - make it your home page From PMonfils <@t> Lifespan.org Wed Jul 13 13:40:33 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] H&E after IHC Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717571@lsexch.lsmaster.lifespan.org> Hematoxylin is an acceptable counterstain for immunoperoxidase techniques (though I personally prefer methyl green). I would avoid eosin however. The brown color that develops in such techniques has a definite reddish cast, and using a background stain in the red range could interfere with interpretation of results, especially in weakly stained areas. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of wen > eng > Sent: Wednesday, July 13, 2005 11:13 AM > To: histonet > Subject: [Histonet] H&E after IHC > > Hi histonetters, > > I have to ask a basic question. I am going to do IHC using DAB. I used to > do hematoxylin counterstaining. Can I do H&E after IHC? Will it cover > brown color of DAB? > > Thanks, > Wen > > > --------------------------------- > Start your day with Yahoo! - make it your home page > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gcallis <@t> montana.edu Wed Jul 13 14:07:11 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Restoring nuclear staining to bone overexposed to acids, i.e. overdecalcified bone In-Reply-To: <992899E9EC268548AB8DDE246AF88473055F5230@PAHEX01.uphs.upen n.edu> References: <992899E9EC268548AB8DDE246AF88473055F5230@PAHEX01.uphs.upenn.edu> Message-ID: <6.0.0.22.1.20050713125332.01b1b418@gemini.msu.montana.edu> Diana, You can try placing the sections into 5% sodium bicarbonate for 10 minutes after deparaffinizing and rehydration then rinse well with running tap water, distilled then onto hematoxylin. Also, stain longer in hematoxylin, even up to 10 min in Gill 2 or 3, Richard Allan hematoxylin 1, rinse with water and IF you have to clarify, do only 5 dips to get rid of any hematoxylin on the glass slide, and blue. If you use Harris, don't even use acid alcohol to differentiate, just stain 10 min, rinse and blue, rinse. If you differentiate, you will take the minimal amount of staining away with the acid alcohol, been there, done that!!! Some people have even stained overnight in hematoxylin to restore, but generally 10 minutes or so will work, hopefully. See next comments. Another possibility, and a long shot is expose the hydrated section to 1% periodic acid for 10 min, rinse and stain. This is an old trick for tissue samples stored for years in formalin that is degraded to a very acid pH. Nuclear staining may be irretrievable as the acid, even formic acid, will have performed the little protein hydrolysis chemical number on the DNA, and you may never get good nuclear staining, and can't be restored. If this is ongoing and using an HCl decalcifier, change to buffered formic acid, gentler, easier to control, and perform decalcification endpoint testing. Never leave the bones in decalcifier over a weekend, and if small, not overnight either. 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 gsennello <@t> osip.com Wed Jul 13 14:28:42 2005 From: gsennello <@t> osip.com (Sennello, Gina) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] automated microtomes Message-ID: I am in the market for fully automated microtome and have looked at Leica's RM2255, Thermo-Electron's Finesse ME and Microm's 355S. Does anyone have any strong feels for against any of these instruments? Thanks in advance for you opinions and help. Gina Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 From meryl50 <@t> hotmail.com Wed Jul 13 14:48:30 2005 From: meryl50 <@t> hotmail.com (Meryl Roberts) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] ASCP registration- urgent help needed. Message-ID: Hi there- I'm a registered histologist in the UK with 6 years experience, but I'm currently job-hunting in the USA; can anyone tell me how I go about becoming registered? I know there is a computer exam and a practical test, but is there any way I can take the exam in the UK? or is my registration in the UK transferrable? The way the visa system works- I can't enter the USA until I have an offer of emplyment, but it seems that most labs are unwilling to employ me unless I have ASCP registration- it's a no win situation!! I'm also going to contact the ASCP directly, but I thought it couldn't hurt to ask on here too. sincerely Meryl Roberts. From amosbrooks <@t> earthlink.net Wed Jul 13 17:19:57 2005 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Re: Histonet Digest, Vol 20, Issue 14 In-Reply-To: <200507121300.1dSo7r2eX3Nl34f0@mx-roseate.atl.sa.earthlink.net> References: <200507121300.1dSo7r2eX3Nl34f0@mx-roseate.atl.sa.earthlink.net> Message-ID: <42D5938D.7010308@earthlink.net> Rich & Andi, Rich, I'd like to second Andi's recommendation for Sigma as a source for Fast Myosin although we don't bother with HIER. Give it a whirl both ways, you never know her's might be better :-) Andi, while you asked Rich I figured I'd chime in for Dako as a source for Hepatitis (core & surface). Unfortunately I'm out of the lab right now so I don't have the cat #s. If you need them just drop me a line and I'll rummage them up. Good luck guys, Amos ////////////////////////////////////////////////////////////////////////////////////// Message: 22 Date: Tue, 12 Jul 2005 16:37:08 +0200 From: "Andi Kappeler" Subject: Re: [Histonet] Fast myosin IHC To: "Richard Cartun" , "Histonet" Message-ID: <008801c586ef$40bf45f0$27955c82@patho.unibe.ch> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Hi Richard We successfully use: - mo-a-Myosin II (fast), clone MY-32, Sigma M 4276, working conc. approx 10 ug Ig/ml (corresponds to 1:400 with the lot we have), HIER in citrate pH 6.0 (pressure cooker) - mo-a-Myosin I (slow), clone NOQ7.5.4D, Sigma M 8421, working conc. approx 7.5 ug Ig/ml (corresponds to 1:1000 with the lot we have), HIER in citrate pH 6.0 (pressure cooker) Both mAbs work very well (see http://www.pathology.unibe.ch/Diagn/speztech/spez_ihc.htm for a pic (german website, sorry). Can you give me a hint in return, where you get your antibodies against Hepatitis B surface Ag (HBs) and core Ag (HBc) from? Our source is no longer able to supply them, due to some bureaucratic / eurocratic rules, now I'm looking for a reliable replacement. Many thanks in advance! Best regards Andi Kappeler Institute of Pathology, University of Bern, Switzerland From billions <@t> public1.sz.js.cn Thu Jul 14 03:32:28 2005 From: billions <@t> public1.sz.js.cn (Sinoera Tech) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Re: BIOLOGICAL STAINS, DYES AND INDICATORS, DIAGNOTIC REAGENTS References: <200507121300.1dSo7r2eX3Nl34f0@mx-roseate.atl.sa.earthlink.net> <42D5938D.7010308@earthlink.net> Message-ID: <001201c5884e$9507a520$7f01a8c0@sinoerae248da4> Dear Histonetters, We manufacture following products. BIOLOGICAL STAINS, DYES AND INDICATORS, DIAGNOTIC REAGENTS sulfobromophthalein sodium hydrate thymolphthalein monophosphate magnesium salt thymolphthalein monophosphate disodium salt o-cresolphthalein monophosphate disodium salt o-cresolphthalein monophosphate N-methylglucammonium salt phenolphthalein monophosphate bis(cyclohexylamine)salt other phthalein monophosphoric acid and their salts phenolphthalein disulfate tripotassium salt alcian Blue 8GX alcian Yellow alcian Green 3',3'',5',5''-tetrachlorophenol-3,4,5,6-tetrabromosulfophthalein tri(4-morpholino)phosphine oxide aminomethanesulfonic acid Kind Regards. - Minggeng Wang, Ph.D / President SUZHOU SINOERA CHEM CO., LTD. 125 Binhe Road Suzhou New & Hi-Tech District 215011 China Fax: +86 512 68258994 Tel: +86 512 68246939 http://www.sinoeratech.com http://www.sinoerachem.com From carl.hobbs <@t> kcl.ac.uk Thu Jul 14 04:05:42 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] re macrophage marker Message-ID: <002101c58853$3676b6e0$112b5c9f@Carlos> Thanks, Bill. C From failm <@t> musc.edu Thu Jul 14 07:11:25 2005 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Ceruloplasmin Message-ID: Has anyone tried this Ab on paraffin section? Rena Fail From Susan.Ferrigon <@t> sanofi-aventis.com Thu Jul 14 08:20:35 2005 From: Susan.Ferrigon <@t> sanofi-aventis.com (Susan.Ferrigon@sanofi-aventis.com) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Double staining Message-ID: Hi I would like to perform a double staining technique with GFAP and F4/80, has anyone any advice regarding which chromogens to use. I've not done a double staining method before, so any advice would be great. Thanks Susan ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- From sjchtascp <@t> yahoo.com Thu Jul 14 11:07:15 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] low temp thermometer Message-ID: <20050714160715.47142.qmail@web90205.mail.scd.yahoo.com> We're doing allot of liquid nitrogen/isopentane snap freezing and I'd like to purchase a good low temp thermometer. Any suggestions or advise out there? Steve --------------------------------- Start your day with Yahoo! - make it your home page From tresemoles <@t> neo.rr.com Thu Jul 14 11:56:27 2005 From: tresemoles <@t> neo.rr.com (TreseMoles) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Cryostats Message-ID: <15016608.1121360187125.JavaMail.Administrator@atp> Hello fellow histonetters I need advice as to which of these 2 cryostats you find to be more "user-friendly"... the Leica CM1850 UV or the Thermo-Electron FSE or FE We are looking to purchase a new cryostat- and these two are what I have narrowed it down to. any input will be appreciated, Thanks in advance for your replies. Theresa Dobersztyn Akron Children's Hospital From Eric <@t> ategra.com Wed Jul 13 14:23:25 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] HistoTech Job Openings Message-ID: Histonetters - My name is Eric Dye and I am the new Histo Tech recruiter here at Ategra. I took over all Histo Tech recruiting since Pam Barker retired. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job opening is in Central Ohio seeking a HistoTech Manager. I also have bench histo tech jobs available in: California,Michigan, Pennsylvania, Illinois, New Hampshire, ,West Virginia, Georgia, Colorado and Florida. I also have blood bank positions available in Upstate New York, Alaska and Arizona. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------- From Michele_Marggi <@t> ssmhc.com Thu Jul 14 11:47:37 2005 From: Michele_Marggi <@t> ssmhc.com (Michele_Marggi@ssmhc.com) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Paraffin in cassette holding chamber? Message-ID: I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. From Laurie.Pereira <@t> sdcounty.ca.gov Thu Jul 14 12:18:42 2005 From: Laurie.Pereira <@t> sdcounty.ca.gov (Pereira, Laurie ) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] microwave tissue processors Message-ID: Hello Everyone, We are a small Veterinary Pathology laboratory and our Pathologist would like to look into a Microwave Tissue Processor. He would like to know the pros and cons of such a processor. We would only need a 30 specimen processor. How long do the tissues need to fix in formalin before using the microwave? Does the specimen need to be fresh or can it be 3 days up to 2 weeks old possibly autolyzed? Does the processor use more alcohols? We would appreciate any feedback from techs that have had experience with using the microwave processors. Thank you in advance. Laurie L. Pereira, RVT SDCADDL (County Veterinarian) 5555 Overland Avenue Suite 4103 San Diego, CA. 92123 Phone: 858-694-3384 Fax: 858-571-4268 From RCHIOVETTI <@t> aol.com Thu Jul 14 12:34:56 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] low temp thermometer Message-ID: In a message dated 7/14/2005 9:09:29 AM US Mountain Standard Time, sjchtascp@yahoo.com writes: > We're doing allot of liquid nitrogen/isopentane snap freezing and I'd like > to purchase a good low temp thermometer. Any suggestions or advise out > there? > Steve, I've worked in several low-temp "bio"- type labs, and they all used Fluke digital thermometers. You can match the thermocouple to the temperature range you need to measure. The website can steer you to a dealer or someone for tech questions, but I know the J- and K-type thermocouples can read temps down near -200 degrees C. That's around the temp of liquid nitrogen (-196) so it should do you well. Go to Fluke's homepage here. Then follow the links: Products -> thermometers -> Fluke 50 Series II Thermometers -> Specifications Cheers, Bob Robert (Bob) Chiovetti, Ph.D. The Microscope Works 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From RCHIOVETTI <@t> aol.com Thu Jul 14 12:39:08 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] low temp thermometer Message-ID: In a message dated 7/14/2005 10:36:00 AM US Mountain Standard Time, RCHIOVETTI@aol.com writes: > Go to Fluke's homepage here. Then follow the links: > Products -> thermometers -> Fluke 50 Series II Thermometers -> > Specifications > > Oops, sorry, can't post an active link...so...Fluke's homepage for the USA is: Bob Robert (Bob) Chiovetti, Ph.D. The Microscope Works 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From kweidenh <@t> montefiore.org Thu Jul 14 12:39:26 2005 From: kweidenh <@t> montefiore.org (Karen Weidenheim) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] low temp thermometer Message-ID: Dear Steve, I presume you are freezing muscle biopsies, or possibly research material. This isopentane/liquid nitrogen method is more an art than a science. Altitude of your lab and humidity of your ambient air, are the 2 major limiting factors. The most important thing in the freezing is not measuring the temperature but rather visual assessment of the state of the isopentane. We put the isopentane in a 250 ml stainless steel beaker and lower it slowly into the liquid nitrogen. We wait to freeze, until the isopentane has formed ice all around the walls of the beaker and all the way up the wall of the beaker, and completely across the bottom of the beaker. It will be syrupy. We use 25 seconds of freezing (I know the books say 5-10 seconds). Our lab is at sea level. It is also often humid here. We discard the isopentane every 3 months when there is no major humidity and more often (every 6 weeks) when we have a run of humid weather, or when we have a problem with freezing artifact, or when I think my assistant has been sloppy with its storage. We keep the beaker very clean i.e. we scour with steel wool. We make sure there is no residual water in the beaker (I keep 2 beakers so one is freshly clean and dry at all times). I have not used the thermometer since I was a young attending a long time ago!!! Best Karen Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> Steven Coakley 7/14/2005 12:07:15 PM >>> We're doing allot of liquid nitrogen/isopentane snap freezing and I'd like to purchase a good low temp thermometer. Any suggestions or advise out there? Steve --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu Jul 14 12:58:14 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Paraffin in cassette holding chamber? Message-ID: I've done this both ways, with and without. I once thought that the cassettes had to be held in molten paraffin to ensure a homogenous block - I've since changed my mind. It doesn't matter. I prefer a hot dry holding chamber - less messy. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist Discovery Cancer R4N2 AP3 Jackie.O'Connor@abbott.com Michele_Marggi@ssmhc.com Sent by: histonet-bounces@lists.utsouthwestern.edu 07/14/2005 11:47 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BlazekL <@t> childrensdayton.org Thu Jul 14 13:05:01 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Paraffin in cassette holding chamber? Message-ID: I've done this both with and without melted paraffin. I prefer having a dry holding tank. Less mess. Less waste of paraffin also. As long as the holding tank keeps the cassettes from solidifying I think they can sit for quite awhile. Or at least as long as it take to embed a tank full. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> 07/14/2005 12:47 PM >>> I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peoshel <@t> wisc.edu Thu Jul 14 13:29:52 2005 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] low temp thermometer In-Reply-To: References: Message-ID: You discard your isopentane every 3 months? Is this stuff you've been using for freezing, and you keep using the same isopentane? And it stays cold the whole time? I hope not. The cold isopentane is getting enriched in oxygen as it's being used, and this heightens the fire hazard. Why not freeze into (all right, everyone who's read my rants before can ignore me now) slush nitrogen? Colder, faster freezing, better freezing, and no flammable/explosive liquids or gases involved. Phil >Dear Steve, I presume you are freezing muscle biopsies, or possibly >research material. >This isopentane/liquid nitrogen method is more an art than a science. >Altitude of your lab and humidity of your ambient air, are the 2 major >limiting factors. The most important thing in the freezing is not >measuring the temperature but rather visual assessment of the state of >the isopentane. We put the isopentane in a 250 ml stainless steel >beaker and lower it slowly into the liquid nitrogen. We wait to freeze, >until the isopentane has formed ice all around the walls of the beaker >and all the way up the wall of the beaker, and completely across the >bottom of the beaker. It will be syrupy. We use 25 seconds of freezing >(I know the books say 5-10 seconds). >Our lab is at sea level. It is also often humid here. We discard the >isopentane every 3 months when there is no major humidity and more often >(every 6 weeks) when we have a run of humid weather, or when we have a >problem with freezing artifact, or when I think my assistant has been >sloppy with its storage. >We keep the beaker very clean i.e. we scour with steel wool. We make >sure there is no residual water in the beaker (I keep 2 beakers so one >is freshly clean and dry at all times). >I have not used the thermometer since I was a young attending a long >time ago!!! >Best >Karen > >Karen M. Weidenheim, M.D. >Professor of Pathology, Clinical Neurology and Clinical Neurosurgery >Albert Einstein College of Medicine >Chief, Division of Neuropathology >Montefiore Medical Center >111 East 210th Street >Bronx, NY 10467 >(718) 920-4446 >FAX (718) 653-3409 >Beeper (917) 556 3696 >CONFIDENTIALITY NOTICE: This email, including any attachments, is for >the sole use of the intended recipient(s). The information contained in >this message may be private and confidential, and may also be subject to >the work product doctrine. Any unauthorized review, use, disclosure or >distribution is prohibited. If you are not the intended recipient, >please contact the sender by reply e-mail and destroy all copies of the >original message. Thank you. > >>>> Steven Coakley 7/14/2005 12:07:15 PM >>> >We're doing allot of liquid nitrogen/isopentane snap freezing and I'd >like to purchase a good low temp thermometer. Any suggestions or advise >out there? > >Steve > > >--------------------------------- > Start your day with Yahoo! - make it your home page >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm From weneng2004 <@t> yahoo.com Thu Jul 14 13:30:37 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Thanks! Message-ID: <20050714183037.83786.qmail@web53403.mail.yahoo.com> Thanks to all who answered my question about H&E after IHC. I really appreciate the help I got from you guys. Have a nice day! Wen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From GoodwinD <@t> pahosp.com Thu Jul 14 13:50:36 2005 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] overdecalcified bone Message-ID: <992899E9EC268548AB8DDE246AF88473055F5251@PAHEX01.uphs.upenn.edu> Hermina: We used the 5% periodic acid overnight method with remarkable results. I am writing a procedure...do you have any idea why these methods work, ie, the chemistry behind the method? I found the original method in the AFIP manual, but no theory was given. Thanks, Diana G. Goodwin Department of Pathology Pennsylvania Hospital 800 Spruce St., Preston 655-C Philadelphia, PA 19107 ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com ----Original Message----- From: Hermina Borgerink [mailto:hborgeri@wfubmc.edu] Sent: Wednesday, July 13, 2005 1:40 PM To: Goodwin, Diana; kalschev@svm.vetmed.wisc.edu Cc: Histonet (E-mail) Subject: RE: [Histonet] overdecalcified bone >From Theory and practice of Histotechnology, Sheehan and Hrapchak, Appendix B, p. 445. To restore basophilic properties to tissues as the result of overexposure to decalcifying solution: Method 1 1. Deparaffinize and hydrate slides 2. Place slides in 5% aqueous sodium bicarbonate solution from 4 hours to overnight 3. Wash in tap water for 5 minutes 4. Stain as desired Method 2 In step 2, use 5% aqueous solution of ammonium sulfide overnight Method 3 In step 2, use 5% aqueous solution of periodic acid overnight Hopefully one of these methods will work for you. Hermina Hermina M. Borgerink, BA, HTL(ASCP)QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax. (336) 716-1515 e-mail hborgeri@wfubmc.edu "Treat a man as he appears to be, and you make him worse. But treat a man as if he already were what he potentially could be, and you make him what he should be." (Goethe) This electronic message, including any attachments, is confidential and proprietary and is solely for the intended recipient. If you are not the intended recipient, this message was sent to you in error and you are hereby advised that any review, disclosure, copying, distribution or use of this message, or any of the information included therein, is unauthorized and strictly prohibited. If you have received this electronic transmission in error, please immediately notify the sender by reply and permanently delete all copies of this message and its attachments. Thank you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Goodwin, Diana Sent: Wednesday, July 13, 2005 1:12 PM To: 'kalschev@svm.vetmed.wisc.edu' Cc: Histonet (E-mail) Subject: [Histonet] overdecalcified bone Hi, Vicki. Is there a procedure to restore the nuclear staining uptake in over-decalcified human bone? Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From HornHV <@t> archildrens.org Thu Jul 14 14:32:44 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Paraffin in cassette holding chamber? Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDDA0@EMAIL.archildrens.org> We do not add paraffin to our holding chamber. But they do not sit there very long, as we start embedding right away. Is there a reason you want them to sit for a time before embedding? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele_Marggi@ssmhc.com Sent: Thursday, July 14, 2005 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From bwhitaker <@t> brownpathology.com Thu Jul 14 14:42:53 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Paraffin in cassette holding chamber? In-Reply-To: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDDA0@EMAIL.archildrens.org> Message-ID: <000001c588ac$3b8fcf00$3601a8c0@brownpathology.net> If it's going to be a really long time (or even to hold extra tissue, unembedded for a great length of time) just pull the cassettes out and then put them back in to heat up right before embedding. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Thursday, July 14, 2005 2:33 PM To: Michele_Marggi@ssmhc.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin in cassette holding chamber? We do not add paraffin to our holding chamber. But they do not sit there very long, as we start embedding right away. Is there a reason you want them to sit for a time before embedding? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele_Marggi@ssmhc.com Sent: Thursday, July 14, 2005 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ 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. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Megan.Clarke <@t> hnehealth.nsw.gov.au Thu Jul 14 21:01:38 2005 From: Megan.Clarke <@t> hnehealth.nsw.gov.au (Megan Clarke) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Bcl10 Message-ID: Hello Histonetters, We would like to see if anyone can help us with an antibody optimisation for Bcl10? We have the Zymed Bcl 10 ,clone 151. We have tried HIER using citrate ph 6.0,citrate EDTA ph 8.0,and Dako TUR both on the Ventana and the Bond Max.Our dilution ranges have been from a 1 in 50 up to 1 in 2000. Our slides look "muddy" with distorted morphology.With no specific staining.We are using a reactive tonsil as our control. If anyone is familiar using this and can offer us some help we would really appreciate it. Thank you very much Megan Clarke and Zenobia Haffajee Immunohistochemisty Lab HAPS>JHH Newcastle AUSTRALIA From Megan.Clarke <@t> hnehealth.nsw.gov.au Thu Jul 14 21:11:54 2005 From: Megan.Clarke <@t> hnehealth.nsw.gov.au (Megan Clarke) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Fwd: CAP Message-ID: >>> Megan Clarke 07/15/05 12:10 PM >>> Hello Hitonetters, Could anyone please send me some information regarding you Quality Control Program on Immunohistochemisrty? I think it is called CAP?? Any information would be greatly appreciated. Thanks Megan Clarke Immunohistochemistry Lab HAPS.JHH Newcastle AUSTRALIA From Jacqueline.Malam <@t> rli.mbht.nhs.uk Fri Jul 15 03:20:43 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Ventana Benchmark XT and ALK 1 Message-ID: Does anyone out there do ALK 1 on the Ventana Benchmark? If so, I would be grateful if you could let me know what clone you use, what the protocol and dilution are, and if you could spare me a positive control block. I have tried 2 clones - Dako's ALK 1 and Lab Vision's SP8 but to no avail; I have had experience of certain clones of some antisera (Vector's CD 138 and Dako's calretinin for example) not working on the Benchmark and it's possible that these clones are not suitable. Having said that, we only had 2 cases and possibly they may have been negative for ALK 1 anyway. I used the lowest recommended dilution, both the mild and standard heat protocols, and the 3 primary incubation temperatures (room, 37 and 42) and even tried protease and no retrieval but got nowhere. I have also tried to get a known positive control block but no one I've tried has one! All assistance appreciated! Jacqui Malam Royal Infirmary Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From BMolinari <@t> heart.thi.tmc.edu Fri Jul 15 05:48:48 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Paraffin in cassette holding chamber? Message-ID: I do not put any extra paraffin into the holding chamber. If I am not going to embed right away I take the cassettes out of the processor and let them cool off with the paraffin hardening around it. I have not had any complaints with tissue handled in this way. Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Avenue Houston, TX 77030 832-355-6524 832-355-6812 ( fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele_Marggi@ssmhc.com Sent: Thursday, July 14, 2005 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nena.dimaano <@t> stryker.com Fri Jul 15 07:34:16 2005 From: nena.dimaano <@t> stryker.com (Dimaano, Nena) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Archiving of Histology Samples in Research Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD12A4F837@HOS2KEXCHCL.howost.strykercorp.com> Hi All, I was wondering if you guys can help me answer the following questions about archiving histology samples. Our Histology Lab archive the samples(including wet samples stored in 10% formalin, blocks both paraffin and plastics) indefinitely and since we are running out of space it is becoming a great deal of problem. The following are my questions: * How long do you keep the samples stored in 10% formalin? * How long do you store the blocks (paraffin and plastics) in research? * Is there any agencies like the FDA, CDC or CLIA outlining this task? thanks, Nena Nena Dimaano Orthobiologics, R&D Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com From sa.drew <@t> hosp.wisc.edu Fri Jul 15 08:14:34 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Ventana Benchmark XT and ALK 1 Message-ID: My co-worker just successfully ran a predilute polyclonal ALK from BioCare Medical on our Benchmark with an MCC1, 32"ttn and no block. We had previously only run it on the Nexes and haven't run it on our XT yet. Unfortunately, we are not flush w/ control tissue (we only have 1 positive case from 1999) so I can't send you a block. However, would a few cut slides help you get started? I know that when I first worked up this antibody I didn't have much luck w/ the rabbit monoclonals I tried. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malam Jacqueline Sent: Friday, July 15, 2005 3:21 AM To: Histonet submissions (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Ventana Benchmark XT and ALK 1 Does anyone out there do ALK 1 on the Ventana Benchmark? If so, I would be grateful if you could let me know what clone you use, what the protocol and dilution are, and if you could spare me a positive control block. I have tried 2 clones - Dako's ALK 1 and Lab Vision's SP8 but to no avail; I have had experience of certain clones of some antisera (Vector's CD 138 and Dako's calretinin for example) not working on the Benchmark and it's possible that these clones are not suitable. Having said that, we only had 2 cases and possibly they may have been negative for ALK 1 anyway. I used the lowest recommended dilution, both the mild and standard heat protocols, and the 3 primary incubation temperatures (room, 37 and 42) and even tried protease and no retrieval but got nowhere. I have also tried to get a known positive control block but no one I've tried has one! All assistance appreciated! Jacqui Malam Royal Infirmary Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jhofecker <@t> yahoo.com Fri Jul 15 08:32:51 2005 From: jhofecker <@t> yahoo.com (Jennifer Hofecker) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Re: ADG & Dysferlin troubles Message-ID: <20050715133251.93873.qmail@web33506.mail.mud.yahoo.com> Hi Everyone, sorry for the delay in responding, my histonet digest was "lost in space" for a few days. We don't do ADG here, but I also use the Dysferlin antibody from Novacastra (purchased from vector) with regular IHC (no FITC) for frozen sections. I use Clone Ham1/7B6(new vector number VP-D503). Air dry frozen sections (usually 20 min or more). Dilution: 1:20 at rm temp 1 hr. I use with Vector R.T.U. vectastain kit (use secondary 30 min@37degrees C. ABC reagent 1hr. rm temp. I use DAB from Vector for 15 min. I usually do a very pale counterstain or none at all depending on pathologist. I hope this is of some help to you. Have a good weekend, Jennifer Hofecker Nashville,TN Date: Wed, 13 Jul 2005 07:43:36 -0400 From: Dana Settembre Subject: Re: [Histonet] ADG & Dysferlin troubles To: LewisS@ccri.net, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Sarah, I use Dysferlin from Novocastra on frozen sections, but not FITC, just regular IHC. If you're desperate we can talk. I use it with a detection kit for mouse, any vendor will usually do and DAB as the chromogen. No special scope is needed. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> "Lewis, Sarah" 7/12/2005 1:48:17 PM >>> Hello histoneters, I could really use any procedural information anyone has on Alpha-dystroglycan (antibody only available thru Upstate) and Dysferlin (antibody from VectorLabs). I have been attempting the stains for 2months now with no satisfying results. I have tried several different procedures/fixatives/incubation times/dilutions/. I perform a battery of 14 neuromuscular Immunoflourescence, they are all BEAUTIFUL fitc green with no background staining (picture perfect) .....except these two boogers. PLEASE HELP me make my pathologist proud!!! ALL THE CREDIT TO HISTONET!!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From cfockler <@t> mail1.vcu.edu Fri Jul 15 08:38:00 2005 From: cfockler <@t> mail1.vcu.edu (cfockler@mail1.vcu.edu) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Xyless? In-Reply-To: <42D5938D.7010308@earthlink.net> Message-ID: <200507151338.JAA30123@despina.vcu.edu> ------------------- > Rich & Andi, > Rich, I'd like to second Andi's recommendation for Sigma as a source > for Fast Myosin although we don't bother with HIER. Give it a whirl both > ways, you never know her's might be better :-) > Andi, while you asked Rich I figured I'd chime in for Dako as a > source for Hepatitis (core & surface). Unfortunately I'm out of the lab > right now so I don't have the cat #s. If you need them just drop me a > line and I'll rummage them up. > Good luck guys, > Amos > > ////////////////////////////////////////////////////////////////////// //////////////// > > Message: 22 > Date: Tue, 12 Jul 2005 16:37:08 +0200 > From: "Andi Kappeler" > Subject: Re: [Histonet] Fast myosin IHC > To: "Richard Cartun" , "Histonet" > > Message-ID: <008801c586ef$40bf45f0$27955c82@patho.unibe.ch> > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=original > > Hi Richard > > We successfully use: > - mo-a-Myosin II (fast), clone MY-32, Sigma M 4276, working conc. approx 10 > ug Ig/ml (corresponds to 1:400 with the lot we have), HIER in citrate pH 6.0 > (pressure cooker) > - mo-a-Myosin I (slow), clone NOQ7.5.4D, Sigma M 8421, working conc. approx > 7.5 ug Ig/ml (corresponds to 1:1000 with the lot we have), HIER in citrate > pH 6.0 (pressure cooker) > Both mAbs work very well (see > http://www.pathology.unibe.ch/Diagn/speztech/spez_ihc.htm for a pic (german > website, sorry). > > Can you give me a hint in return, where you get your antibodies against > Hepatitis B surface Ag (HBs) and core Ag (HBc) from? Our source is no longer > able to supply them, due to some bureaucratic / eurocratic rules, now I'm > looking for a reliable replacement. > Many thanks in advance! > > Best regards > Andi Kappeler > Institute of Pathology, University of Bern, Switzerland > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Hello All . .. I just recently moved to a lab that uses a few changes of Xyless, (xylene substitute) in their stainer and processor. .can anyone give me any information about this product? I can't find anything useful on the web. Thanks in advance! Candyce LeTellier Histotechnologist From phillipst <@t> missouri.edu Fri Jul 8 13:43:57 2005 From: phillipst <@t> missouri.edu (Tom Phillips) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] paraffin microtome recommendation? Message-ID: <6.0.0.22.2.20050708134252.04bab6e0@pop.missouri.edu> I am considering buying a paraffin microtome for our core facility. I don't personally use one much so if any knowledgeable users have recommendations on brands or models to buy (or avoid), i would welcome replies (please respond to me directly since I am not a registered member of this listserver). I will be happy to keep your comments confidential. In addition, if there are special features you think are essential, i would be pleased to hear of them also. thanks, tom Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400 573-882-4712 (office) 573-882-0123 (fax) PhillipsT@missouri.edu From Gui-rong.Guo <@t> biogenidec.com Fri Jul 8 18:17:26 2005 From: Gui-rong.Guo <@t> biogenidec.com (Gui-rong Guo) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Mast Cell Stain other than Toluidine Blue Message-ID: Hi, Tony, I am interesting your staining procedure of mast cell. Would you mind to send me the protocol of staining of mast cell by email. Thanks so much! Have a nice weekend! Gui-Rong Guo, Sr. Associate Scientist Antibody Discovery Dept. Biogen Idec Inc. 5200 Research Place San Diego, CA 92122 Tel: (858) 401-8273 Fax: (858)-401-5046 From jonstarr <@t> pacbell.net Tue Jul 12 12:22:31 2005 From: jonstarr <@t> pacbell.net (jonstarr) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Mohs Histotech Position in Nothern California! Message-ID: <002601c58706$496d4aa0$6901a8c0@drstarr.local> HISTOTECH We're looking for a part time to full time experienced Histotech for our dynamic and busy Mohs surgery practice in Palo Alto, California. Responsibilities include laboratory management and equipment maintenance for a 3 room surgery suite. Pay is commensurate with experience. Benefits include health insurance, paid vacation, and 401K plan with profit sharing. Jon C. Starr, M.D. Dermatologic and Mohs Micrographic Surgery 703 Welch Rd., Suite D3 Palo Alto, CA 94304 Ph 650-326-7222, FAX 650-326-7332 From gsennello <@t> osip.com Wed Jul 13 14:12:24 2005 From: gsennello <@t> osip.com (Sennello, Gina) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] automated microtomes Message-ID: I am in the market for fully automated microtome and have looked at Leica's RM2255, Thermo-Electron's Finesse ME and Microm's 355S. Does anyone have any strong feels for against any of these instruments? Thanks in advance for you opinions and help. Gina Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 From Debbie.Pepperall <@t> hnehealth.nsw.gov.au Wed Jul 13 15:59:01 2005 From: Debbie.Pepperall <@t> hnehealth.nsw.gov.au (Debbie Pepperall) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] put onto list Message-ID: Hi All You Wonderful People on The Histonet, Could someone out there help me I am trying to do immunos on paraffin sections using the Daf[CD55] ANTIBODY.Has anyone had positive staining if possible I would like to know what company you bought the antibody from and any other tips would be great. Thanks heaps from down under Deb From jbaez <@t> interscopepath.com Fri Jul 15 14:19:10 2005 From: jbaez <@t> interscopepath.com (Baez, Janet) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] AB H&E stain for frozen sections Message-ID: <9E956D8FEB06C2408B08AC16498325E90139E3@scopemx1.interscope.com> Has anyone tried the AB H&E stain for frozen sections, by Arkadiy I. Brusilovskiy, M.D., Ph.D., HT (ASCP)? If so, would you let me know what you think. Thanks, Janet From TJJ <@t> Stowers-Institute.org Fri Jul 15 14:36:08 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Vibratome sections of unfixed tissues Message-ID: How does one mount unfixed tissues for vibratome sectioning? Are they encased in any medium such as agarose or gelatin, or simply glued onto the stage and sectioned? I can't seem to find information on this anywhere. Thanks, and have a great weekend all! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From PBugelsk <@t> CNTUS.JNJ.COM Fri Jul 15 14:51:28 2005 From: PBugelsk <@t> CNTUS.JNJ.COM (Bugelski, Peter [CNTUS]) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Vibratome sections of unfixed tissues Message-ID: <7BF70FA941B9AE4783EAAF733762F1B505197D19@CNTUSMAEXS3.na.jnj.com> I've used cyanoacrylate "super glue" with great success. It works on wet tissue with no problems. Be careful of your fingers. -----Original Message----- From: Johnson, Teri [mailto:TJJ@Stowers-Institute.org] Sent: Friday, July 15, 2005 3:36 PM To: Histonet Subject: [Histonet] Vibratome sections of unfixed tissues How does one mount unfixed tissues for vibratome sectioning? Are they encased in any medium such as agarose or gelatin, or simply glued onto the stage and sectioned? I can't seem to find information on this anywhere. Thanks, and have a great weekend all! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From BoozerKA <@t> pa1.ah.org Fri Jul 15 14:55:52 2005 From: BoozerKA <@t> pa1.ah.org (Kathleen Boozer) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Full time Histology Position Message-ID: 5:30 - 2 PM Adventist Medical Center Send resume to (503) 251-6862 (503) 251-6266 ex 7856 Kathy Boozer, HT (ASCP) IHCQ From liz <@t> premierlab.com Fri Jul 15 14:59:29 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Vibratome sections of unfixed tissues Message-ID: <000c01c58977$b5d3b030$a7d48a80@AMY> Teri We have not done unfixed tissue via vibratome, but we have done paraformaldehyde fixed and we use gelatin to create a block with the tissue in it. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Friday, July 15, 2005 12:36 PM To: Histonet Subject: [Histonet] Vibratome sections of unfixed tissues How does one mount unfixed tissues for vibratome sectioning? Are they encased in any medium such as agarose or gelatin, or simply glued onto the stage and sectioned? I can't seem to find information on this anywhere. Thanks, and have a great weekend all! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From cwscouten <@t> myneurolab.com Fri Jul 15 15:09:12 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Vibratome sections of unfixed tissues Message-ID: <5784D843593D874C93E9BADCB87342AB916413@tpiserver03.Coretech-holdings.com> Embedding would take time, at a premium with unfixed tissue. When used to collect brain slices for brain slice chamber electrophsyiology, researchers either just put the brain block down on wet filter paper, and get enough adhesion from that to cut, or use an instant setting cyanoacrylate provided with the Vibratome. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Friday, July 15, 2005 2:59 PM To: 'Johnson, Teri'; 'Histonet' Subject: RE: [Histonet] Vibratome sections of unfixed tissues Teri We have not done unfixed tissue via vibratome, but we have done paraformaldehyde fixed and we use gelatin to create a block with the tissue in it. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Friday, July 15, 2005 12:36 PM To: Histonet Subject: [Histonet] Vibratome sections of unfixed tissues How does one mount unfixed tissues for vibratome sectioning? Are they encased in any medium such as agarose or gelatin, or simply glued onto the stage and sectioned? I can't seem to find information on this anywhere. Thanks, and have a great weekend all! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jfish <@t> gladstone.ucsf.edu Sat Jul 16 15:10:19 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Vibratome sections of unfixed tissues In-Reply-To: <20050715194056.AD48620000A0@gmail.gladstone.ucsf.edu> References: <20050715194056.AD48620000A0@gmail.gladstone.ucsf.edu> Message-ID: Teri, I use a type of Super Glue called Loctite 404. Works great for gluing wet tissue to metal. Hope that helps, Jo Dee At 2:36 PM -0500 7/15/05, Johnson, Teri wrote: >How does one mount unfixed tissues for vibratome sectioning? Are >they encased in any medium such as agarose or gelatin, or simply >glued onto the stage and sectioned? I can't seem to find >information on this anywhere. > >Thanks, and have a great weekend all! > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, Missouri 64110 >tjj@stowers-institute.org > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** ********** Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 734-5766 Fax: (415) 355-0824 E-mail: jfish@gladstone.ucsf.edu Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158 From BoozerKA <@t> pa1.ah.org Fri Jul 15 15:17:14 2005 From: BoozerKA <@t> pa1.ah.org (Kathleen Boozer) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Full time position Message-ID: Adventist Medical Center in Portland, Oregon has a full time Histotech opening, 5:30 AM - 2 PM. We are a small, newly automated Histology department looking for someone who enjoys a quiet, peaceful atmosphere and an opportunity to improve themselves. Contact: Kathy Boozer (503) 251-6266 ex 7856 Fax Resume to (503) 251-6862 From PMonfils <@t> Lifespan.org Fri Jul 15 15:25:16 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Vibratome sections of unfixed tissues Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717574@lsexch.lsmaster.lifespan.org> For fairly firm tissues like kidney and spleen I just cut a flat surface on the specimen, blot it on paper towels, and attach it directly to the specimen holder with "super glue" (cyanoacrylate). But for very soft tissues like embryos and most brain specimens, and for specimens with little internal support like cysts, eyes, membranes that need to be cut on edge, etc., I embed in 5 to 6% low melting point agarose (Sigma Chemical Co., cat# A-9414). I then cut a flat surface on the agarose block, and attach that to the blockholder with super glue. I also did a project vibratoming cell pellets of cells grown in liquid suspension. I spun down the cells, removed the culture medium, resuspended the cells in liquid agarose, spun them down again before the agarose solidified, then allowed the agarose block to form around the pellet. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Johnson, Teri > Sent: Friday, July 15, 2005 12:36 PM > To: Histonet > Subject: [Histonet] Vibratome sections of unfixed tissues > > <> > How does one mount unfixed tissues for vibratome sectioning? Are they > encased in any medium such as agarose or gelatin, or simply glued onto the > stage and sectioned? I can't seem to find information on this anywhere. > > Thanks, and have a great weekend all! > > Teri Johnson > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, Missouri 64110 > tjj@stowers-institute.org > > > > > From Eric <@t> ategra.com Fri Jul 15 16:35:38 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] HistoTech Job Openings Message-ID: Histonetters - My name is Eric Dye and I am the Histo Tech recruiter here at Ategra. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job opening is in Central Ohio seeking a HistoTech Manager. I also have bench histo tech jobs available in: California,Michigan, Pennsylvania, Illinois, New Hampshire, ,West Virginia,South Carolina,Georgia, Colorado and Florida. I have a opening for a pathologist's Assistant in Illinois. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------- From kccatunda <@t> terra.com.br Sat Jul 16 20:20:15 2005 From: kccatunda <@t> terra.com.br (Katia Cristina Catunda) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Dissection boards Message-ID: <019801c58a6d$b0bbc050$a279fea9@privatexx> In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia From jqb7 <@t> cdc.gov Sun Jul 17 12:12:21 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Dissection boards Message-ID: Richard Allan has some nice high-density polyethylene boards.Here is the link: http://www.rallansci.com/histology/histology.aspx?id=6 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Katia Cristina Catunda Sent: Sat 7/16/2005 9:20 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Dissection boards In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Sun Jul 17 17:52:29 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Mast Cell Stain other than Toluidine Blue Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E319@simba.kids> Dear Gui-Rong, I have attached a pdf copy of the paper that uses the alcian blue tetrakis Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Gui-rong Guo [mailto:Gui-rong.Guo@biogenidec.com] Sent: Saturday, 9 July 2005 9:17 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mast Cell Stain other than Toluidine Blue Hi, Tony, I am interesting your staining procedure of mast cell. Would you mind to send me the protocol of staining of mast cell by email. Thanks so much! Have a nice weekend! Gui-Rong Guo, Sr. Associate Scientist Antibody Discovery Dept. Biogen Idec Inc. 5200 Research Place San Diego, CA 92122 Tel: (858) 401-8273 Fax: (858)-401-5046 _______________________________________________ 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 Sun Jul 17 18:33:47 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:19 2005 Subject: [Histonet] Dissection boards Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E31A@simba.kids> Katia, Before you make your decision you may like to consider the following (from our local jounal "Histograph") Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 The Wood Chopping Board Many Histopathology Laboratories continue to use wooden cut-up boards. One advantage over plastic, glass or steel boards seems to be that there is less damage to knives and scalpels. This results in prolonged blade life. One major concern is the perceived difficulty in adequately cleaning and disinfecting wooden boards. Several years ago Dr. Dean Cliver and his associates at the University of Wisconsin-Madison, USA, presented their work on the safety of plastic and wooden cutting boards. Their research was first intended to develop means of disinfecting wooden cutting surfaces, so that they would be almost as safe as plastics. Their safety concern was that bacteria such as Escherichia coli O157:H7 (commonly known as E-coli) and Salmonella, which might contaminate a work surface when raw meat was being prepared, ought not remain on the surface to contaminate other foods that might be eaten without further cooking. They soon found that disease bacteria such as these were not recoverable from wooden surfaces in a short time after they were applied, unless very large numbers were used. New plastic surfaces allowed the bacteria to persist, but were easily cleaned and disinfected. However, wooden boards that had been used and had many knife cuts acted almost the same as new wood, whereas plastic surfaces that were knife-scarred were impossible to clean and disinfect manually, especially when food residues such as chicken fat were present. Although the bacteria that had disappeared from the wood surfaces were found alive inside the wood for some time after application, they evidently did not multiply, and they gradually died. They could be detected only by splitting or gouging the wood or by forcing water completely through from one surface to the other. If a sharp knife is used to cut into the work surfaces after used plastic or wood had been contaminated with bacteria and cleaned manually, more bacteria were recovered from a used plastic surface than from a used wood surface. "Manual cleaning", in their experiments, had been done with a sponge, hot tap water, and liquid dishwashing detergent. Mechanical cleaning with a dishwashing machine could be done successfully with plastic surfaces (even if knife-scarred) and wooden boards especially made for this. Wooden boards, but not plastics, that were small enough to fit into a microwave oven could be disinfected rapidly, but care had to be used to prevent overheating. Work surfaces that had been cleaned could be disinfected with bleach (sodium hypochlorite) solutions. This disinfecting was reliable only if cleaning had been done successfully. A case-control study of sporadic salmonellosis had been done in California and included cutting boards among many risk factors assessed (Kass et al 1992.). It revealed that those using wooden cutting boards in their home kitchens were less than half as likely on average to contract salmonellosis (odds ratio 0.42, 95% confidence interval 0.22-0.81). Those using synthetic (plastic or glass) cutting boards were about twice as likely on average to contract salmonellosis (O.R. 1.99, C.I. 1.03-3.85) and the effect of cleaning the board regularly after preparing meat on it was not statistically significant (O.R. 1.20, C.I. 0.54-2.68). Dr. Dean Cliver and his co-workers have concluded that wooden cutting boards are not a hazard to human health, but plastic cutting boards may be. As yet the natural antibiotic has not been discovered. What are your thoughts? References: http://www.peter.hemsley.btinternet.co.uk/CDB/Technical/Bacteria/bacteria.ht ml Kass, P.H., et al., (1992) "Disease determinants of sporadic salmonellosis in four northern California counties: a case control study of older children and adults" Ann. Epidemiol. 2:683-696. Ak, N. O., Cliver, D. O. Kaspar C. W., (1994) Cutting boards of plastic and wood contaminated experimentally with bacteria. J. Food Protect. 57:16- 22. Ak, N. O., Cliver D. O, Kaspar C. W., (1994) Decontamination of plastic and wooden cutting boards for kitchen use. J. Food Protect. 57:23-30,36. Galluzzo, L., Cliver D. O., (1996) Cutting boards and bacteria--oak vs. Salmonella. Dairy, Food Environ. Sanit. 16:290-293. Park, P. K., Cliver D. O., (1996) Disinfection of household cutting boards with a microwave oven. J. Food. Protect. 59:1049-1054. -----Original Message----- From: Katia Cristina Catunda [mailto:kccatunda@terra.com.br] Sent: Sunday, 17 July 2005 11:20 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dissection boards In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia _______________________________________________ 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 Jacqueline.Malam <@t> rli.mbht.nhs.uk Mon Jul 18 06:22:47 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] RE: Histonet Digest, Vol 20, Issue 17-Bcl-10 Message-ID: Have you tried any other clones? Can you get hold of some (free / cheap / small) samples? I have found that our Ventana Benchmark doesn't like the occasional clone, ie 1D5 (Ors), and certain clones for CD138 and calretinin. Jacqui -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 15 July 2005 18:05 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 17 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. microwave tissue processors (Pereira, Laurie ) 2. Re: low temp thermometer (RCHIOVETTI@aol.com) 3. Re: low temp thermometer (RCHIOVETTI@aol.com) 4. Re: low temp thermometer (Karen Weidenheim) 5. Re: Paraffin in cassette holding chamber? (Jackie M O'Connor) 6. Re: Paraffin in cassette holding chamber? (Linda Blazek) 7. Re: low temp thermometer (Philip Oshel) 8. Thanks! (wen eng) 9. RE: overdecalcified bone (Goodwin, Diana) 10. RE: Paraffin in cassette holding chamber? (Horn, Hazel V) 11. RE: Paraffin in cassette holding chamber? (Bonnie Whitaker) 12. Bcl10 (Megan Clarke) 13. Fwd: CAP (Megan Clarke) 14. Ventana Benchmark XT and ALK 1 (Malam Jacqueline) 15. RE: Paraffin in cassette holding chamber? (Molinari, Betsy) 16. Archiving of Histology Samples in Research (Dimaano, Nena) 17. RE: Ventana Benchmark XT and ALK 1 (Drew Sally A.) 18. Re: ADG & Dysferlin troubles (Jennifer Hofecker) 19. Xyless? (cfockler@mail1.vcu.edu) ---------------------------------------------------------------------- Message: 1 Date: Thu, 14 Jul 2005 10:18:42 -0700 From: "Pereira, Laurie " Subject: [Histonet] microwave tissue processors To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello Everyone, We are a small Veterinary Pathology laboratory and our Pathologist would like to look into a Microwave Tissue Processor. He would like to know the pros and cons of such a processor. We would only need a 30 specimen processor. How long do the tissues need to fix in formalin before using the microwave? Does the specimen need to be fresh or can it be 3 days up to 2 weeks old possibly autolyzed? Does the processor use more alcohols? We would appreciate any feedback from techs that have had experience with using the microwave processors. Thank you in advance. Laurie L. Pereira, RVT SDCADDL (County Veterinarian) 5555 Overland Avenue Suite 4103 San Diego, CA. 92123 Phone: 858-694-3384 Fax: 858-571-4268 ------------------------------ Message: 2 Date: Thu, 14 Jul 2005 13:34:56 EDT From: RCHIOVETTI@aol.com Subject: Re: [Histonet] low temp thermometer To: sjchtascp@yahoo.com, Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" In a message dated 7/14/2005 9:09:29 AM US Mountain Standard Time, sjchtascp@yahoo.com writes: > We're doing allot of liquid nitrogen/isopentane snap freezing and I'd > like to purchase a good low temp thermometer. Any suggestions or > advise out there? > Steve, I've worked in several low-temp "bio"- type labs, and they all used Fluke digital thermometers. You can match the thermocouple to the temperature range you need to measure. The website can steer you to a dealer or someone for tech questions, but I know the J- and K-type thermocouples can read temps down near -200 degrees C. That's around the temp of liquid nitrogen (-196) so it should do you well. Go to Fluke's homepage here. Then follow the links: Products -> thermometers -> Fluke 50 Series II Thermometers -> Specifications Cheers, Bob Robert (Bob) Chiovetti, Ph.D. The Microscope Works 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA ------------------------------ Message: 3 Date: Thu, 14 Jul 2005 13:39:08 EDT From: RCHIOVETTI@aol.com Subject: Re: [Histonet] low temp thermometer To: RCHIOVETTI@aol.com, sjchtascp@yahoo.com, Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" In a message dated 7/14/2005 10:36:00 AM US Mountain Standard Time, RCHIOVETTI@aol.com writes: > Go to Fluke's homepage here. Then follow the links: > Products -> thermometers -> Fluke 50 Series II Thermometers -> > Specifications > > Oops, sorry, can't post an active link...so...Fluke's homepage for the USA is: Bob Robert (Bob) Chiovetti, Ph.D. The Microscope Works 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA ------------------------------ Message: 4 Date: Thu, 14 Jul 2005 13:39:26 -0400 From: "Karen Weidenheim" Subject: Re: [Histonet] low temp thermometer To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Dear Steve, I presume you are freezing muscle biopsies, or possibly research material. This isopentane/liquid nitrogen method is more an art than a science. Altitude of your lab and humidity of your ambient air, are the 2 major limiting factors. The most important thing in the freezing is not measuring the temperature but rather visual assessment of the state of the isopentane. We put the isopentane in a 250 ml stainless steel beaker and lower it slowly into the liquid nitrogen. We wait to freeze, until the isopentane has formed ice all around the walls of the beaker and all the way up the wall of the beaker, and completely across the bottom of the beaker. It will be syrupy. We use 25 seconds of freezing (I know the books say 5-10 seconds). Our lab is at sea level. It is also often humid here. We discard the isopentane every 3 months when there is no major humidity and more often (every 6 weeks) when we have a run of humid weather, or when we have a problem with freezing artifact, or when I think my assistant has been sloppy with its storage. We keep the beaker very clean i.e. we scour with steel wool. We make sure there is no residual water in the beaker (I keep 2 beakers so one is freshly clean and dry at all times). I have not used the thermometer since I was a young attending a long time ago!!! Best Karen Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> Steven Coakley 7/14/2005 12:07:15 PM >>> We're doing allot of liquid nitrogen/isopentane snap freezing and I'd like to purchase a good low temp thermometer. Any suggestions or advise out there? Steve --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Thu, 14 Jul 2005 12:58:14 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] Paraffin in cassette holding chamber? To: Michele_Marggi@ssmhc.com Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" I've done this both ways, with and without. I once thought that the cassettes had to be held in molten paraffin to ensure a homogenous block - I've since changed my mind. It doesn't matter. I prefer a hot dry holding chamber - less messy. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist Discovery Cancer R4N2 AP3 Jackie.O'Connor@abbott.com Michele_Marggi@ssmhc.com Sent by: histonet-bounces@lists.utsouthwestern.edu 07/14/2005 11:47 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Thu, 14 Jul 2005 14:05:01 -0400 From: "Linda Blazek" Subject: Re: [Histonet] Paraffin in cassette holding chamber? To: histonet@lists.utsouthwestern.edu, Michele_Marggi@ssmhc.com Message-ID: Content-Type: text/plain; charset=US-ASCII I've done this both with and without melted paraffin. I prefer having a dry holding tank. Less mess. Less waste of paraffin also. As long as the holding tank keeps the cassettes from solidifying I think they can sit for quite awhile. Or at least as long as it take to embed a tank full. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> 07/14/2005 12:47 PM >>> I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Thu, 14 Jul 2005 13:29:52 -0500 From: Philip Oshel Subject: Re: [Histonet] low temp thermometer To: Histonet@Pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed; charset=us-ascii You discard your isopentane every 3 months? Is this stuff you've been using for freezing, and you keep using the same isopentane? And it stays cold the whole time? I hope not. The cold isopentane is getting enriched in oxygen as it's being used, and this heightens the fire hazard. Why not freeze into (all right, everyone who's read my rants before can ignore me now) slush nitrogen? Colder, faster freezing, better freezing, and no flammable/explosive liquids or gases involved. Phil >Dear Steve, I presume you are freezing muscle biopsies, or possibly >research material. >This isopentane/liquid nitrogen method is more an art than a science. >Altitude of your lab and humidity of your ambient air, are the 2 major >limiting factors. The most important thing in the freezing is not >measuring the temperature but rather visual assessment of the state of >the isopentane. We put the isopentane in a 250 ml stainless steel >beaker and lower it slowly into the liquid nitrogen. We wait to freeze, >until the isopentane has formed ice all around the walls of the beaker >and all the way up the wall of the beaker, and completely across the >bottom of the beaker. It will be syrupy. We use 25 seconds of freezing >(I know the books say 5-10 seconds). >Our lab is at sea level. It is also often humid here. We discard the >isopentane every 3 months when there is no major humidity and more often >(every 6 weeks) when we have a run of humid weather, or when we have a >problem with freezing artifact, or when I think my assistant has been >sloppy with its storage. >We keep the beaker very clean i.e. we scour with steel wool. We make >sure there is no residual water in the beaker (I keep 2 beakers so one >is freshly clean and dry at all times). >I have not used the thermometer since I was a young attending a long >time ago!!! >Best >Karen > >Karen M. Weidenheim, M.D. >Professor of Pathology, Clinical Neurology and Clinical Neurosurgery >Albert Einstein College of Medicine >Chief, Division of Neuropathology >Montefiore Medical Center >111 East 210th Street >Bronx, NY 10467 >(718) 920-4446 >FAX (718) 653-3409 >Beeper (917) 556 3696 >CONFIDENTIALITY NOTICE: This email, including any attachments, is for >the sole use of the intended recipient(s). The information contained in >this message may be private and confidential, and may also be subject to >the work product doctrine. Any unauthorized review, use, disclosure or >distribution is prohibited. If you are not the intended recipient, >please contact the sender by reply e-mail and destroy all copies of the >original message. Thank you. > >>>> Steven Coakley 7/14/2005 12:07:15 PM >>> >We're doing allot of liquid nitrogen/isopentane snap freezing and I'd >like to purchase a good low temp thermometer. Any suggestions or advise >out there? > >Steve > > >--------------------------------- > Start your day with Yahoo! - make it your home page >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm ------------------------------ Message: 8 Date: Thu, 14 Jul 2005 11:30:37 -0700 (PDT) From: wen eng Subject: [Histonet] Thanks! To: histonet Message-ID: <20050714183037.83786.qmail@web53403.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Thanks to all who answered my question about H&E after IHC. I really appreciate the help I got from you guys. Have a nice day! Wen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 9 Date: Thu, 14 Jul 2005 14:50:36 -0400 From: "Goodwin, Diana" Subject: RE: [Histonet] overdecalcified bone To: 'Hermina Borgerink' Cc: "Histonet \(E-mail\)" Message-ID: <992899E9EC268548AB8DDE246AF88473055F5251@PAHEX01.uphs.upenn.edu> Content-Type: text/plain; charset="iso-8859-1" Hermina: We used the 5% periodic acid overnight method with remarkable results. I am writing a procedure...do you have any idea why these methods work, ie, the chemistry behind the method? I found the original method in the AFIP manual, but no theory was given. Thanks, Diana G. Goodwin Department of Pathology Pennsylvania Hospital 800 Spruce St., Preston 655-C Philadelphia, PA 19107 ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com ----Original Message----- From: Hermina Borgerink [mailto:hborgeri@wfubmc.edu] Sent: Wednesday, July 13, 2005 1:40 PM To: Goodwin, Diana; kalschev@svm.vetmed.wisc.edu Cc: Histonet (E-mail) Subject: RE: [Histonet] overdecalcified bone >From Theory and practice of Histotechnology, Sheehan and Hrapchak, Appendix B, p. 445. To restore basophilic properties to tissues as the result of overexposure to decalcifying solution: Method 1 1. Deparaffinize and hydrate slides 2. Place slides in 5% aqueous sodium bicarbonate solution from 4 hours to overnight 3. Wash in tap water for 5 minutes 4. Stain as desired Method 2 In step 2, use 5% aqueous solution of ammonium sulfide overnight Method 3 In step 2, use 5% aqueous solution of periodic acid overnight Hopefully one of these methods will work for you. Hermina Hermina M. Borgerink, BA, HTL(ASCP)QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax. (336) 716-1515 e-mail hborgeri@wfubmc.edu "Treat a man as he appears to be, and you make him worse. But treat a man as if he already were what he potentially could be, and you make him what he should be." (Goethe) This electronic message, including any attachments, is confidential and proprietary and is solely for the intended recipient. If you are not the intended recipient, this message was sent to you in error and you are hereby advised that any review, disclosure, copying, distribution or use of this message, or any of the information included therein, is unauthorized and strictly prohibited. If you have received this electronic transmission in error, please immediately notify the sender by reply and permanently delete all copies of this message and its attachments. Thank you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Goodwin, Diana Sent: Wednesday, July 13, 2005 1:12 PM To: 'kalschev@svm.vetmed.wisc.edu' Cc: Histonet (E-mail) Subject: [Histonet] overdecalcified bone Hi, Vicki. Is there a procedure to restore the nuclear staining uptake in over-decalcified human bone? Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Thu, 14 Jul 2005 14:32:44 -0500 From: "Horn, Hazel V" Subject: RE: [Histonet] Paraffin in cassette holding chamber? To: Michele_Marggi@ssmhc.com, histonet@lists.utsouthwestern.edu Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDDA0@EMAIL.archildrens.org> Content-Type: text/plain; charset=us-ascii We do not add paraffin to our holding chamber. But they do not sit there very long, as we start embedding right away. Is there a reason you want them to sit for a time before embedding? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele_Marggi@ssmhc.com Sent: Thursday, July 14, 2005 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================ == ------------------------------ Message: 11 Date: Thu, 14 Jul 2005 14:42:53 -0500 From: "Bonnie Whitaker" Subject: RE: [Histonet] Paraffin in cassette holding chamber? To: "'Horn, Hazel V'" , , Message-ID: <000001c588ac$3b8fcf00$3601a8c0@brownpathology.net> Content-Type: text/plain; charset="us-ascii" If it's going to be a really long time (or even to hold extra tissue, unembedded for a great length of time) just pull the cassettes out and then put them back in to heat up right before embedding. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Thursday, July 14, 2005 2:33 PM To: Michele_Marggi@ssmhc.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin in cassette holding chamber? We do not add paraffin to our holding chamber. But they do not sit there very long, as we start embedding right away. Is there a reason you want them to sit for a time before embedding? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele_Marggi@ssmhc.com Sent: Thursday, July 14, 2005 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ 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. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Fri, 15 Jul 2005 12:01:38 +1000 From: "Megan Clarke" Subject: [Histonet] Bcl10 To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hello Histonetters, We would like to see if anyone can help us with an antibody optimisation for Bcl10? We have the Zymed Bcl 10 ,clone 151. We have tried HIER using citrate ph 6.0,citrate EDTA ph 8.0,and Dako TUR both on the Ventana and the Bond Max.Our dilution ranges have been from a 1 in 50 up to 1 in 2000. Our slides look "muddy" with distorted morphology.With no specific staining.We are using a reactive tonsil as our control. If anyone is familiar using this and can offer us some help we would really appreciate it. Thank you very much Megan Clarke and Zenobia Haffajee Immunohistochemisty Lab HAPS>JHH Newcastle AUSTRALIA ------------------------------ Message: 13 Date: Fri, 15 Jul 2005 12:11:54 +1000 From: "Megan Clarke" Subject: [Histonet] Fwd: CAP To: Message-ID: Content-Type: text/plain; charset=US-ASCII >>> Megan Clarke 07/15/05 12:10 PM >>> Hello Hitonetters, Could anyone please send me some information regarding you Quality Control Program on Immunohistochemisrty? I think it is called CAP?? Any information would be greatly appreciated. Thanks Megan Clarke Immunohistochemistry Lab HAPS.JHH Newcastle AUSTRALIA ------------------------------ Message: 14 Date: Fri, 15 Jul 2005 09:20:43 +0100 From: Malam Jacqueline Subject: [Histonet] Ventana Benchmark XT and ALK 1 To: "Histonet submissions (histonet@lists.utsouthwestern.edu)" Message-ID: Content-Type: text/plain Does anyone out there do ALK 1 on the Ventana Benchmark? If so, I would be grateful if you could let me know what clone you use, what the protocol and dilution are, and if you could spare me a positive control block. I have tried 2 clones - Dako's ALK 1 and Lab Vision's SP8 but to no avail; I have had experience of certain clones of some antisera (Vector's CD 138 and Dako's calretinin for example) not working on the Benchmark and it's possible that these clones are not suitable. Having said that, we only had 2 cases and possibly they may have been negative for ALK 1 anyway. I used the lowest recommended dilution, both the mild and standard heat protocols, and the 3 primary incubation temperatures (room, 37 and 42) and even tried protease and no retrieval but got nowhere. I have also tried to get a known positive control block but no one I've tried has one! All assistance appreciated! Jacqui Malam Royal Infirmary Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 15 Date: Fri, 15 Jul 2005 05:48:48 -0500 From: "Molinari, Betsy" Subject: RE: [Histonet] Paraffin in cassette holding chamber? To: Message-ID: Content-Type: text/plain; charset="us-ascii" I do not put any extra paraffin into the holding chamber. If I am not going to embed right away I take the cassettes out of the processor and let them cool off with the paraffin hardening around it. I have not had any complaints with tissue handled in this way. Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Avenue Houston, TX 77030 832-355-6524 832-355-6812 ( fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele_Marggi@ssmhc.com Sent: Thursday, July 14, 2005 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Fri, 15 Jul 2005 08:34:16 -0400 From: "Dimaano, Nena" Subject: [Histonet] Archiving of Histology Samples in Research To: Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD12A4F837@HOS2KEXCHCL.howost.strykercorp.com > Content-Type: text/plain; charset="iso-8859-1" Hi All, I was wondering if you guys can help me answer the following questions about archiving histology samples. Our Histology Lab archive the samples(including wet samples stored in 10% formalin, blocks both paraffin and plastics) indefinitely and since we are running out of space it is becoming a great deal of problem. The following are my questions: * How long do you keep the samples stored in 10% formalin? * How long do you store the blocks (paraffin and plastics) in research? * Is there any agencies like the FDA, CDC or CLIA outlining this task? thanks, Nena Nena Dimaano Orthobiologics, R&D Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com ------------------------------ Message: 17 Date: Fri, 15 Jul 2005 08:14:34 -0500 From: "Drew Sally A." Subject: RE: [Histonet] Ventana Benchmark XT and ALK 1 To: "Histonet \(Histonet\)" Message-ID: Content-Type: text/plain; charset="us-ascii" My co-worker just successfully ran a predilute polyclonal ALK from BioCare Medical on our Benchmark with an MCC1, 32"ttn and no block. We had previously only run it on the Nexes and haven't run it on our XT yet. Unfortunately, we are not flush w/ control tissue (we only have 1 positive case from 1999) so I can't send you a block. However, would a few cut slides help you get started? I know that when I first worked up this antibody I didn't have much luck w/ the rabbit monoclonals I tried. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malam Jacqueline Sent: Friday, July 15, 2005 3:21 AM To: Histonet submissions (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Ventana Benchmark XT and ALK 1 Does anyone out there do ALK 1 on the Ventana Benchmark? If so, I would be grateful if you could let me know what clone you use, what the protocol and dilution are, and if you could spare me a positive control block. I have tried 2 clones - Dako's ALK 1 and Lab Vision's SP8 but to no avail; I have had experience of certain clones of some antisera (Vector's CD 138 and Dako's calretinin for example) not working on the Benchmark and it's possible that these clones are not suitable. Having said that, we only had 2 cases and possibly they may have been negative for ALK 1 anyway. I used the lowest recommended dilution, both the mild and standard heat protocols, and the 3 primary incubation temperatures (room, 37 and 42) and even tried protease and no retrieval but got nowhere. I have also tried to get a known positive control block but no one I've tried has one! All assistance appreciated! Jacqui Malam Royal Infirmary Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Fri, 15 Jul 2005 06:32:51 -0700 (PDT) From: Jennifer Hofecker Subject: [Histonet] Re: ADG & Dysferlin troubles To: histonet@lists.utsouthwestern.edu, LewisS@ccri.net Message-ID: <20050715133251.93873.qmail@web33506.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Everyone, sorry for the delay in responding, my histonet digest was "lost in space" for a few days. We don't do ADG here, but I also use the Dysferlin antibody from Novacastra (purchased from vector) with regular IHC (no FITC) for frozen sections. I use Clone Ham1/7B6(new vector number VP-D503). Air dry frozen sections (usually 20 min or more). Dilution: 1:20 at rm temp 1 hr. I use with Vector R.T.U. vectastain kit (use secondary 30 min@37degrees C. ABC reagent 1hr. rm temp. I use DAB from Vector for 15 min. I usually do a very pale counterstain or none at all depending on pathologist. I hope this is of some help to you. Have a good weekend, Jennifer Hofecker Nashville,TN Date: Wed, 13 Jul 2005 07:43:36 -0400 From: Dana Settembre Subject: Re: [Histonet] ADG & Dysferlin troubles To: LewisS@ccri.net, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Sarah, I use Dysferlin from Novocastra on frozen sections, but not FITC, just regular IHC. If you're desperate we can talk. I use it with a detection kit for mouse, any vendor will usually do and DAB as the chromogen. No special scope is needed. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> "Lewis, Sarah" 7/12/2005 1:48:17 PM >>> Hello histoneters, I could really use any procedural information anyone has on Alpha-dystroglycan (antibody only available thru Upstate) and Dysferlin (antibody from VectorLabs). I have been attempting the stains for 2months now with no satisfying results. I have tried several different procedures/fixatives/incubation times/dilutions/. I perform a battery of 14 neuromuscular Immunoflourescence, they are all BEAUTIFUL fitc green with no background staining (picture perfect) .....except these two boogers. PLEASE HELP me make my pathologist proud!!! ALL THE CREDIT TO HISTONET!!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 19 Date: Fri, 15 Jul 2005 09:38:00 -0400 From: Subject: [Histonet] Xyless? To: Amos Brooks Cc: histonet@lists.utsouthwestern.edu, Rcartun@harthosp.org, kappeler@patho.unibe.ch Message-ID: <200507151338.JAA30123@despina.vcu.edu> Content-Type: text/plain; charset=iso-8859-1 ------------------- > Rich & Andi, > Rich, I'd like to second Andi's recommendation for Sigma as a source > for Fast Myosin although we don't bother with HIER. Give it a whirl both > ways, you never know her's might be better :-) > Andi, while you asked Rich I figured I'd chime in for Dako as a > source for Hepatitis (core & surface). Unfortunately I'm out of the lab > right now so I don't have the cat #s. If you need them just drop me a > line and I'll rummage them up. > Good luck guys, > Amos > > ////////////////////////////////////////////////////////////////////// //////////////// > > Message: 22 > Date: Tue, 12 Jul 2005 16:37:08 +0200 > From: "Andi Kappeler" > Subject: Re: [Histonet] Fast myosin IHC > To: "Richard Cartun" , "Histonet" > > Message-ID: <008801c586ef$40bf45f0$27955c82@patho.unibe.ch> > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=original > > Hi Richard > > We successfully use: > - mo-a-Myosin II (fast), clone MY-32, Sigma M 4276, working conc. approx 10 > ug Ig/ml (corresponds to 1:400 with the lot we have), HIER in citrate pH 6.0 > (pressure cooker) > - mo-a-Myosin I (slow), clone NOQ7.5.4D, Sigma M 8421, working conc. approx > 7.5 ug Ig/ml (corresponds to 1:1000 with the lot we have), HIER in citrate > pH 6.0 (pressure cooker) > Both mAbs work very well (see > http://www.pathology.unibe.ch/Diagn/speztech/spez_ihc.htm for a pic (german > website, sorry). > > Can you give me a hint in return, where you get your antibodies against > Hepatitis B surface Ag (HBs) and core Ag (HBc) from? Our source is no longer > able to supply them, due to some bureaucratic / eurocratic rules, now I'm > looking for a reliable replacement. > Many thanks in advance! > > Best regards > Andi Kappeler > Institute of Pathology, University of Bern, Switzerland > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Hello All . .. I just recently moved to a lab that uses a few changes of Xyless, (xylene substitute) in their stainer and processor. .can anyone give me any information about this product? I can't find anything useful on the web. Thanks in advance! Candyce LeTellier Histotechnologist ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 17 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Mon Jul 18 06:27:23 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] RE: Histonet Digest, Vol 20, Issue 17- Re: ALK 1 Message-ID: Would greatly appreciate a few slides. At least I can tell if my current sera will work on the Benchmark XT. If not, I will have to waste it and buy Ventana's. Many thanks Jacqui -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 15 July 2005 18:05 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 17 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. microwave tissue processors (Pereira, Laurie ) 2. Re: low temp thermometer (RCHIOVETTI@aol.com) 3. Re: low temp thermometer (RCHIOVETTI@aol.com) 4. Re: low temp thermometer (Karen Weidenheim) 5. Re: Paraffin in cassette holding chamber? (Jackie M O'Connor) 6. Re: Paraffin in cassette holding chamber? (Linda Blazek) 7. Re: low temp thermometer (Philip Oshel) 8. Thanks! (wen eng) 9. RE: overdecalcified bone (Goodwin, Diana) 10. RE: Paraffin in cassette holding chamber? (Horn, Hazel V) 11. RE: Paraffin in cassette holding chamber? (Bonnie Whitaker) 12. Bcl10 (Megan Clarke) 13. Fwd: CAP (Megan Clarke) 14. Ventana Benchmark XT and ALK 1 (Malam Jacqueline) 15. RE: Paraffin in cassette holding chamber? (Molinari, Betsy) 16. Archiving of Histology Samples in Research (Dimaano, Nena) 17. RE: Ventana Benchmark XT and ALK 1 (Drew Sally A.) 18. Re: ADG & Dysferlin troubles (Jennifer Hofecker) 19. Xyless? (cfockler@mail1.vcu.edu) ---------------------------------------------------------------------- Message: 1 Date: Thu, 14 Jul 2005 10:18:42 -0700 From: "Pereira, Laurie " Subject: [Histonet] microwave tissue processors To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello Everyone, We are a small Veterinary Pathology laboratory and our Pathologist would like to look into a Microwave Tissue Processor. He would like to know the pros and cons of such a processor. We would only need a 30 specimen processor. How long do the tissues need to fix in formalin before using the microwave? Does the specimen need to be fresh or can it be 3 days up to 2 weeks old possibly autolyzed? Does the processor use more alcohols? We would appreciate any feedback from techs that have had experience with using the microwave processors. Thank you in advance. Laurie L. Pereira, RVT SDCADDL (County Veterinarian) 5555 Overland Avenue Suite 4103 San Diego, CA. 92123 Phone: 858-694-3384 Fax: 858-571-4268 ------------------------------ Message: 2 Date: Thu, 14 Jul 2005 13:34:56 EDT From: RCHIOVETTI@aol.com Subject: Re: [Histonet] low temp thermometer To: sjchtascp@yahoo.com, Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" In a message dated 7/14/2005 9:09:29 AM US Mountain Standard Time, sjchtascp@yahoo.com writes: > We're doing allot of liquid nitrogen/isopentane snap freezing and I'd > like to purchase a good low temp thermometer. Any suggestions or > advise out there? > Steve, I've worked in several low-temp "bio"- type labs, and they all used Fluke digital thermometers. You can match the thermocouple to the temperature range you need to measure. The website can steer you to a dealer or someone for tech questions, but I know the J- and K-type thermocouples can read temps down near -200 degrees C. That's around the temp of liquid nitrogen (-196) so it should do you well. Go to Fluke's homepage here. Then follow the links: Products -> thermometers -> Fluke 50 Series II Thermometers -> Specifications Cheers, Bob Robert (Bob) Chiovetti, Ph.D. The Microscope Works 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA ------------------------------ Message: 3 Date: Thu, 14 Jul 2005 13:39:08 EDT From: RCHIOVETTI@aol.com Subject: Re: [Histonet] low temp thermometer To: RCHIOVETTI@aol.com, sjchtascp@yahoo.com, Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" In a message dated 7/14/2005 10:36:00 AM US Mountain Standard Time, RCHIOVETTI@aol.com writes: > Go to Fluke's homepage here. Then follow the links: > Products -> thermometers -> Fluke 50 Series II Thermometers -> > Specifications > > Oops, sorry, can't post an active link...so...Fluke's homepage for the USA is: Bob Robert (Bob) Chiovetti, Ph.D. The Microscope Works 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA ------------------------------ Message: 4 Date: Thu, 14 Jul 2005 13:39:26 -0400 From: "Karen Weidenheim" Subject: Re: [Histonet] low temp thermometer To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Dear Steve, I presume you are freezing muscle biopsies, or possibly research material. This isopentane/liquid nitrogen method is more an art than a science. Altitude of your lab and humidity of your ambient air, are the 2 major limiting factors. The most important thing in the freezing is not measuring the temperature but rather visual assessment of the state of the isopentane. We put the isopentane in a 250 ml stainless steel beaker and lower it slowly into the liquid nitrogen. We wait to freeze, until the isopentane has formed ice all around the walls of the beaker and all the way up the wall of the beaker, and completely across the bottom of the beaker. It will be syrupy. We use 25 seconds of freezing (I know the books say 5-10 seconds). Our lab is at sea level. It is also often humid here. We discard the isopentane every 3 months when there is no major humidity and more often (every 6 weeks) when we have a run of humid weather, or when we have a problem with freezing artifact, or when I think my assistant has been sloppy with its storage. We keep the beaker very clean i.e. we scour with steel wool. We make sure there is no residual water in the beaker (I keep 2 beakers so one is freshly clean and dry at all times). I have not used the thermometer since I was a young attending a long time ago!!! Best Karen Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> Steven Coakley 7/14/2005 12:07:15 PM >>> We're doing allot of liquid nitrogen/isopentane snap freezing and I'd like to purchase a good low temp thermometer. Any suggestions or advise out there? Steve --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Thu, 14 Jul 2005 12:58:14 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] Paraffin in cassette holding chamber? To: Michele_Marggi@ssmhc.com Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" I've done this both ways, with and without. I once thought that the cassettes had to be held in molten paraffin to ensure a homogenous block - I've since changed my mind. It doesn't matter. I prefer a hot dry holding chamber - less messy. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist Discovery Cancer R4N2 AP3 Jackie.O'Connor@abbott.com Michele_Marggi@ssmhc.com Sent by: histonet-bounces@lists.utsouthwestern.edu 07/14/2005 11:47 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Thu, 14 Jul 2005 14:05:01 -0400 From: "Linda Blazek" Subject: Re: [Histonet] Paraffin in cassette holding chamber? To: histonet@lists.utsouthwestern.edu, Michele_Marggi@ssmhc.com Message-ID: Content-Type: text/plain; charset=US-ASCII I've done this both with and without melted paraffin. I prefer having a dry holding tank. Less mess. Less waste of paraffin also. As long as the holding tank keeps the cassettes from solidifying I think they can sit for quite awhile. Or at least as long as it take to embed a tank full. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> 07/14/2005 12:47 PM >>> I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Thu, 14 Jul 2005 13:29:52 -0500 From: Philip Oshel Subject: Re: [Histonet] low temp thermometer To: Histonet@Pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed; charset=us-ascii You discard your isopentane every 3 months? Is this stuff you've been using for freezing, and you keep using the same isopentane? And it stays cold the whole time? I hope not. The cold isopentane is getting enriched in oxygen as it's being used, and this heightens the fire hazard. Why not freeze into (all right, everyone who's read my rants before can ignore me now) slush nitrogen? Colder, faster freezing, better freezing, and no flammable/explosive liquids or gases involved. Phil >Dear Steve, I presume you are freezing muscle biopsies, or possibly >research material. >This isopentane/liquid nitrogen method is more an art than a science. >Altitude of your lab and humidity of your ambient air, are the 2 major >limiting factors. The most important thing in the freezing is not >measuring the temperature but rather visual assessment of the state of >the isopentane. We put the isopentane in a 250 ml stainless steel >beaker and lower it slowly into the liquid nitrogen. We wait to freeze, >until the isopentane has formed ice all around the walls of the beaker >and all the way up the wall of the beaker, and completely across the >bottom of the beaker. It will be syrupy. We use 25 seconds of freezing >(I know the books say 5-10 seconds). >Our lab is at sea level. It is also often humid here. We discard the >isopentane every 3 months when there is no major humidity and more often >(every 6 weeks) when we have a run of humid weather, or when we have a >problem with freezing artifact, or when I think my assistant has been >sloppy with its storage. >We keep the beaker very clean i.e. we scour with steel wool. We make >sure there is no residual water in the beaker (I keep 2 beakers so one >is freshly clean and dry at all times). >I have not used the thermometer since I was a young attending a long >time ago!!! >Best >Karen > >Karen M. Weidenheim, M.D. >Professor of Pathology, Clinical Neurology and Clinical Neurosurgery >Albert Einstein College of Medicine >Chief, Division of Neuropathology >Montefiore Medical Center >111 East 210th Street >Bronx, NY 10467 >(718) 920-4446 >FAX (718) 653-3409 >Beeper (917) 556 3696 >CONFIDENTIALITY NOTICE: This email, including any attachments, is for >the sole use of the intended recipient(s). The information contained in >this message may be private and confidential, and may also be subject to >the work product doctrine. Any unauthorized review, use, disclosure or >distribution is prohibited. If you are not the intended recipient, >please contact the sender by reply e-mail and destroy all copies of the >original message. Thank you. > >>>> Steven Coakley 7/14/2005 12:07:15 PM >>> >We're doing allot of liquid nitrogen/isopentane snap freezing and I'd >like to purchase a good low temp thermometer. Any suggestions or advise >out there? > >Steve > > >--------------------------------- > Start your day with Yahoo! - make it your home page >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm ------------------------------ Message: 8 Date: Thu, 14 Jul 2005 11:30:37 -0700 (PDT) From: wen eng Subject: [Histonet] Thanks! To: histonet Message-ID: <20050714183037.83786.qmail@web53403.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Thanks to all who answered my question about H&E after IHC. I really appreciate the help I got from you guys. Have a nice day! Wen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 9 Date: Thu, 14 Jul 2005 14:50:36 -0400 From: "Goodwin, Diana" Subject: RE: [Histonet] overdecalcified bone To: 'Hermina Borgerink' Cc: "Histonet \(E-mail\)" Message-ID: <992899E9EC268548AB8DDE246AF88473055F5251@PAHEX01.uphs.upenn.edu> Content-Type: text/plain; charset="iso-8859-1" Hermina: We used the 5% periodic acid overnight method with remarkable results. I am writing a procedure...do you have any idea why these methods work, ie, the chemistry behind the method? I found the original method in the AFIP manual, but no theory was given. Thanks, Diana G. Goodwin Department of Pathology Pennsylvania Hospital 800 Spruce St., Preston 655-C Philadelphia, PA 19107 ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com ----Original Message----- From: Hermina Borgerink [mailto:hborgeri@wfubmc.edu] Sent: Wednesday, July 13, 2005 1:40 PM To: Goodwin, Diana; kalschev@svm.vetmed.wisc.edu Cc: Histonet (E-mail) Subject: RE: [Histonet] overdecalcified bone >From Theory and practice of Histotechnology, Sheehan and Hrapchak, Appendix B, p. 445. To restore basophilic properties to tissues as the result of overexposure to decalcifying solution: Method 1 1. Deparaffinize and hydrate slides 2. Place slides in 5% aqueous sodium bicarbonate solution from 4 hours to overnight 3. Wash in tap water for 5 minutes 4. Stain as desired Method 2 In step 2, use 5% aqueous solution of ammonium sulfide overnight Method 3 In step 2, use 5% aqueous solution of periodic acid overnight Hopefully one of these methods will work for you. Hermina Hermina M. Borgerink, BA, HTL(ASCP)QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax. (336) 716-1515 e-mail hborgeri@wfubmc.edu "Treat a man as he appears to be, and you make him worse. But treat a man as if he already were what he potentially could be, and you make him what he should be." (Goethe) This electronic message, including any attachments, is confidential and proprietary and is solely for the intended recipient. If you are not the intended recipient, this message was sent to you in error and you are hereby advised that any review, disclosure, copying, distribution or use of this message, or any of the information included therein, is unauthorized and strictly prohibited. If you have received this electronic transmission in error, please immediately notify the sender by reply and permanently delete all copies of this message and its attachments. Thank you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Goodwin, Diana Sent: Wednesday, July 13, 2005 1:12 PM To: 'kalschev@svm.vetmed.wisc.edu' Cc: Histonet (E-mail) Subject: [Histonet] overdecalcified bone Hi, Vicki. Is there a procedure to restore the nuclear staining uptake in over-decalcified human bone? Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Thu, 14 Jul 2005 14:32:44 -0500 From: "Horn, Hazel V" Subject: RE: [Histonet] Paraffin in cassette holding chamber? To: Michele_Marggi@ssmhc.com, histonet@lists.utsouthwestern.edu Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDDA0@EMAIL.archildrens.org> Content-Type: text/plain; charset=us-ascii We do not add paraffin to our holding chamber. But they do not sit there very long, as we start embedding right away. Is there a reason you want them to sit for a time before embedding? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele_Marggi@ssmhc.com Sent: Thursday, July 14, 2005 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================ == ------------------------------ Message: 11 Date: Thu, 14 Jul 2005 14:42:53 -0500 From: "Bonnie Whitaker" Subject: RE: [Histonet] Paraffin in cassette holding chamber? To: "'Horn, Hazel V'" , , Message-ID: <000001c588ac$3b8fcf00$3601a8c0@brownpathology.net> Content-Type: text/plain; charset="us-ascii" If it's going to be a really long time (or even to hold extra tissue, unembedded for a great length of time) just pull the cassettes out and then put them back in to heat up right before embedding. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Thursday, July 14, 2005 2:33 PM To: Michele_Marggi@ssmhc.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin in cassette holding chamber? We do not add paraffin to our holding chamber. But they do not sit there very long, as we start embedding right away. Is there a reason you want them to sit for a time before embedding? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele_Marggi@ssmhc.com Sent: Thursday, July 14, 2005 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ 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. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Fri, 15 Jul 2005 12:01:38 +1000 From: "Megan Clarke" Subject: [Histonet] Bcl10 To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hello Histonetters, We would like to see if anyone can help us with an antibody optimisation for Bcl10? We have the Zymed Bcl 10 ,clone 151. We have tried HIER using citrate ph 6.0,citrate EDTA ph 8.0,and Dako TUR both on the Ventana and the Bond Max.Our dilution ranges have been from a 1 in 50 up to 1 in 2000. Our slides look "muddy" with distorted morphology.With no specific staining.We are using a reactive tonsil as our control. If anyone is familiar using this and can offer us some help we would really appreciate it. Thank you very much Megan Clarke and Zenobia Haffajee Immunohistochemisty Lab HAPS>JHH Newcastle AUSTRALIA ------------------------------ Message: 13 Date: Fri, 15 Jul 2005 12:11:54 +1000 From: "Megan Clarke" Subject: [Histonet] Fwd: CAP To: Message-ID: Content-Type: text/plain; charset=US-ASCII >>> Megan Clarke 07/15/05 12:10 PM >>> Hello Hitonetters, Could anyone please send me some information regarding you Quality Control Program on Immunohistochemisrty? I think it is called CAP?? Any information would be greatly appreciated. Thanks Megan Clarke Immunohistochemistry Lab HAPS.JHH Newcastle AUSTRALIA ------------------------------ Message: 14 Date: Fri, 15 Jul 2005 09:20:43 +0100 From: Malam Jacqueline Subject: [Histonet] Ventana Benchmark XT and ALK 1 To: "Histonet submissions (histonet@lists.utsouthwestern.edu)" Message-ID: Content-Type: text/plain Does anyone out there do ALK 1 on the Ventana Benchmark? If so, I would be grateful if you could let me know what clone you use, what the protocol and dilution are, and if you could spare me a positive control block. I have tried 2 clones - Dako's ALK 1 and Lab Vision's SP8 but to no avail; I have had experience of certain clones of some antisera (Vector's CD 138 and Dako's calretinin for example) not working on the Benchmark and it's possible that these clones are not suitable. Having said that, we only had 2 cases and possibly they may have been negative for ALK 1 anyway. I used the lowest recommended dilution, both the mild and standard heat protocols, and the 3 primary incubation temperatures (room, 37 and 42) and even tried protease and no retrieval but got nowhere. I have also tried to get a known positive control block but no one I've tried has one! All assistance appreciated! Jacqui Malam Royal Infirmary Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 15 Date: Fri, 15 Jul 2005 05:48:48 -0500 From: "Molinari, Betsy" Subject: RE: [Histonet] Paraffin in cassette holding chamber? To: Message-ID: Content-Type: text/plain; charset="us-ascii" I do not put any extra paraffin into the holding chamber. If I am not going to embed right away I take the cassettes out of the processor and let them cool off with the paraffin hardening around it. I have not had any complaints with tissue handled in this way. Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Avenue Houston, TX 77030 832-355-6524 832-355-6812 ( fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele_Marggi@ssmhc.com Sent: Thursday, July 14, 2005 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin in cassette holding chamber? I want to get an idea of who is (and who is not, and why) adding paraffin to the cassette holding chamber of the embedding center. Once processed-How long are tissues ok to sit without paraffin on them? All thoughts, opinions, experience is appreciated...Thanks. Michele Marggi, HT Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Fri, 15 Jul 2005 08:34:16 -0400 From: "Dimaano, Nena" Subject: [Histonet] Archiving of Histology Samples in Research To: Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD12A4F837@HOS2KEXCHCL.howost.strykercorp.com > Content-Type: text/plain; charset="iso-8859-1" Hi All, I was wondering if you guys can help me answer the following questions about archiving histology samples. Our Histology Lab archive the samples(including wet samples stored in 10% formalin, blocks both paraffin and plastics) indefinitely and since we are running out of space it is becoming a great deal of problem. The following are my questions: * How long do you keep the samples stored in 10% formalin? * How long do you store the blocks (paraffin and plastics) in research? * Is there any agencies like the FDA, CDC or CLIA outlining this task? thanks, Nena Nena Dimaano Orthobiologics, R&D Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com ------------------------------ Message: 17 Date: Fri, 15 Jul 2005 08:14:34 -0500 From: "Drew Sally A." Subject: RE: [Histonet] Ventana Benchmark XT and ALK 1 To: "Histonet \(Histonet\)" Message-ID: Content-Type: text/plain; charset="us-ascii" My co-worker just successfully ran a predilute polyclonal ALK from BioCare Medical on our Benchmark with an MCC1, 32"ttn and no block. We had previously only run it on the Nexes and haven't run it on our XT yet. Unfortunately, we are not flush w/ control tissue (we only have 1 positive case from 1999) so I can't send you a block. However, would a few cut slides help you get started? I know that when I first worked up this antibody I didn't have much luck w/ the rabbit monoclonals I tried. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malam Jacqueline Sent: Friday, July 15, 2005 3:21 AM To: Histonet submissions (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Ventana Benchmark XT and ALK 1 Does anyone out there do ALK 1 on the Ventana Benchmark? If so, I would be grateful if you could let me know what clone you use, what the protocol and dilution are, and if you could spare me a positive control block. I have tried 2 clones - Dako's ALK 1 and Lab Vision's SP8 but to no avail; I have had experience of certain clones of some antisera (Vector's CD 138 and Dako's calretinin for example) not working on the Benchmark and it's possible that these clones are not suitable. Having said that, we only had 2 cases and possibly they may have been negative for ALK 1 anyway. I used the lowest recommended dilution, both the mild and standard heat protocols, and the 3 primary incubation temperatures (room, 37 and 42) and even tried protease and no retrieval but got nowhere. I have also tried to get a known positive control block but no one I've tried has one! All assistance appreciated! Jacqui Malam Royal Infirmary Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Fri, 15 Jul 2005 06:32:51 -0700 (PDT) From: Jennifer Hofecker Subject: [Histonet] Re: ADG & Dysferlin troubles To: histonet@lists.utsouthwestern.edu, LewisS@ccri.net Message-ID: <20050715133251.93873.qmail@web33506.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Everyone, sorry for the delay in responding, my histonet digest was "lost in space" for a few days. We don't do ADG here, but I also use the Dysferlin antibody from Novacastra (purchased from vector) with regular IHC (no FITC) for frozen sections. I use Clone Ham1/7B6(new vector number VP-D503). Air dry frozen sections (usually 20 min or more). Dilution: 1:20 at rm temp 1 hr. I use with Vector R.T.U. vectastain kit (use secondary 30 min@37degrees C. ABC reagent 1hr. rm temp. I use DAB from Vector for 15 min. I usually do a very pale counterstain or none at all depending on pathologist. I hope this is of some help to you. Have a good weekend, Jennifer Hofecker Nashville,TN Date: Wed, 13 Jul 2005 07:43:36 -0400 From: Dana Settembre Subject: Re: [Histonet] ADG & Dysferlin troubles To: LewisS@ccri.net, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Sarah, I use Dysferlin from Novocastra on frozen sections, but not FITC, just regular IHC. If you're desperate we can talk. I use it with a detection kit for mouse, any vendor will usually do and DAB as the chromogen. No special scope is needed. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> "Lewis, Sarah" 7/12/2005 1:48:17 PM >>> Hello histoneters, I could really use any procedural information anyone has on Alpha-dystroglycan (antibody only available thru Upstate) and Dysferlin (antibody from VectorLabs). I have been attempting the stains for 2months now with no satisfying results. I have tried several different procedures/fixatives/incubation times/dilutions/. I perform a battery of 14 neuromuscular Immunoflourescence, they are all BEAUTIFUL fitc green with no background staining (picture perfect) .....except these two boogers. PLEASE HELP me make my pathologist proud!!! ALL THE CREDIT TO HISTONET!!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 19 Date: Fri, 15 Jul 2005 09:38:00 -0400 From: Subject: [Histonet] Xyless? To: Amos Brooks Cc: histonet@lists.utsouthwestern.edu, Rcartun@harthosp.org, kappeler@patho.unibe.ch Message-ID: <200507151338.JAA30123@despina.vcu.edu> Content-Type: text/plain; charset=iso-8859-1 ------------------- > Rich & Andi, > Rich, I'd like to second Andi's recommendation for Sigma as a source > for Fast Myosin although we don't bother with HIER. Give it a whirl both > ways, you never know her's might be better :-) > Andi, while you asked Rich I figured I'd chime in for Dako as a > source for Hepatitis (core & surface). Unfortunately I'm out of the lab > right now so I don't have the cat #s. If you need them just drop me a > line and I'll rummage them up. > Good luck guys, > Amos > > ////////////////////////////////////////////////////////////////////// //////////////// > > Message: 22 > Date: Tue, 12 Jul 2005 16:37:08 +0200 > From: "Andi Kappeler" > Subject: Re: [Histonet] Fast myosin IHC > To: "Richard Cartun" , "Histonet" > > Message-ID: <008801c586ef$40bf45f0$27955c82@patho.unibe.ch> > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=original > > Hi Richard > > We successfully use: > - mo-a-Myosin II (fast), clone MY-32, Sigma M 4276, working conc. approx 10 > ug Ig/ml (corresponds to 1:400 with the lot we have), HIER in citrate pH 6.0 > (pressure cooker) > - mo-a-Myosin I (slow), clone NOQ7.5.4D, Sigma M 8421, working conc. approx > 7.5 ug Ig/ml (corresponds to 1:1000 with the lot we have), HIER in citrate > pH 6.0 (pressure cooker) > Both mAbs work very well (see > http://www.pathology.unibe.ch/Diagn/speztech/spez_ihc.htm for a pic (german > website, sorry). > > Can you give me a hint in return, where you get your antibodies against > Hepatitis B surface Ag (HBs) and core Ag (HBc) from? Our source is no longer > able to supply them, due to some bureaucratic / eurocratic rules, now I'm > looking for a reliable replacement. > Many thanks in advance! > > Best regards > Andi Kappeler > Institute of Pathology, University of Bern, Switzerland > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Hello All . .. I just recently moved to a lab that uses a few changes of Xyless, (xylene substitute) in their stainer and processor. .can anyone give me any information about this product? I can't find anything useful on the web. Thanks in advance! Candyce LeTellier Histotechnologist ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 17 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Mon Jul 18 06:39:23 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] RE: dissecting board Message-ID: Thermo Shandon do 2 colours and 2 sizes of polyethylene board. They're easy to clean and last. We actually got ours from a butchers' suppliers! -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 17 July 2005 18:07 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 19 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Dissection boards (Katia Cristina Catunda) ---------------------------------------------------------------------- Message: 1 Date: Sat, 16 Jul 2005 22:20:15 -0300 From: "Katia Cristina Catunda" Subject: [Histonet] Dissection boards To: Message-ID: <019801c58a6d$b0bbc050$a279fea9@privatexx> Content-Type: text/plain; charset="iso-8859-1" In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 19 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From n.cragg <@t> epistem.co.uk Mon Jul 18 07:39:29 2005 From: n.cragg <@t> epistem.co.uk (Nicola Cragg) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Need help with PGP9.5 on FFPE skin Message-ID: Hello, I'm posting this message with some embarrassment and apologise for raising anybody's hope of a quck solution........ I have been trying to optimise PGP9.5 on FFPE skin samples (3 micron), using a mouse monoclonal (clone 10A1) from Neuromics. Despite my initial excitement expressed in an earlier posting, I have since found some very good photos of PGP9.5 staining in published work which has put mine to shame and has made me realise that my IHC is not working well enough at all. I've found an improvement with the waterbath antigen retrieval, as the dermis remains intact and there is some specific staining (I think) in the reticular dermis, which wasn't apparent with microwave antigen retrieval. However, it appears that there seems to be more of a trend of using free-floating 50 micron sections. Is that because routine FFPE sections are difficult to stain for this antigen? Has anyone got it to work on FFPE samples? I was quite hopeful that it would work from reading the datasheet and I have also found that Dako make a Rabbit polyclonal for FFPE sections and they're usually excellent antibodies so I'm hoping to try. Any advice or suggestions will be gratefully received. Regards, Nicola Cragg From Janet.Bonner <@t> FLHOSP.ORG Mon Jul 18 07:43:37 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] automated microtomes Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4325@fh2k093.fhmis.net> We have used the Leica's with great success and few if any call backs after more than eight years!! Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet Histonet (E-mail) Sent: 7/13/2005 3:12 PM Subject: [Histonet] automated microtomes I am in the market for fully automated microtome and have looked at Leica's RM2255, Thermo-Electron's Finesse ME and Microm's 355S. Does anyone have any strong feels for against any of these instruments? Thanks in advance for you opinions and help. Gina Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Mon Jul 18 11:29:54 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Dissection boards Message-ID: I've found that kitchen cutting boards from your local department store works well, and it's cheaper than from a medical supplier. Fred >>> "Katia Cristina Catunda" 07/16/05 09:20PM >>> In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histo20 <@t> hotmail.com Mon Jul 18 11:37:34 2005 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Bodies in morgue - who does the checking? Message-ID: Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 From Charles.Embrey <@t> carle.com Mon Jul 18 12:01:38 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Bodies in morgue - who does the checking? Message-ID: It depends what you mean. Check bodies into the morgue or check bodies that are in the morgue. Here we have a house officer (nurse) that coordinates the release of bodies from the hospital through security. Sometimes if the funeral home can come quickly enough then they take them from the hospital room. If they can't come quickly the ward staff put the body into the morgue with the help of security. I keep an eye on who is in my morgue so I can get a jump on any autopsy that might be required. Since I do all the weekday autopsies I don't like surprises. My dieners are all part-time (as needed) and my pathology assistant is only concerned with setting up gross and logging specimens into the computer (I can't get her to go near the morgue). I did work at one place in Ohio where the diener was full time and actually had his office in the morgue. Hope this helps.... Charles Embrey, PA(ASCP) Pathologists' Assistant Histology Manager -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Wilder Sent: Monday, July 18, 2005 11:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bodies in morgue - who does the checking? Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BZIMMERM <@t> mail.mcg.edu Mon Jul 18 12:09:53 2005 From: BZIMMERM <@t> mail.mcg.edu (Billie Zimmerman) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the Message-ID: How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the technical quality of histologic preparations by the pathologist? This req. came out in April so I was wondering the best way to handle this. Thanks, Billie Zimmerman Medical College of GA From HornHV <@t> archildrens.org Mon Jul 18 12:13:02 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Bodies in morgue - who does the checking? Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDDA4@EMAIL.archildrens.org> At our hospital is is histology's responsibility to check the morgue on day shift, Monday-Friday. On evenings, nights and weekends our Nursing service does it. BUT we can check the morgue, via computer. We have the option to pull up and expired patient list. If we know someone has expired we can go down to the morgue and check to see if there is autopsy. We also release bodies on dayshift to the funeral homes. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, July 18, 2005 12:02 PM To: Paula Wilder; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Bodies in morgue - who does the checking? It depends what you mean. Check bodies into the morgue or check bodies that are in the morgue. Here we have a house officer (nurse) that coordinates the release of bodies from the hospital through security. Sometimes if the funeral home can come quickly enough then they take them from the hospital room. If they can't come quickly the ward staff put the body into the morgue with the help of security. I keep an eye on who is in my morgue so I can get a jump on any autopsy that might be required. Since I do all the weekday autopsies I don't like surprises. My dieners are all part-time (as needed) and my pathology assistant is only concerned with setting up gross and logging specimens into the computer (I can't get her to go near the morgue). I did work at one place in Ohio where the diener was full time and actually had his office in the morgue. Hope this helps.... Charles Embrey, PA(ASCP) Pathologists' Assistant Histology Manager -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Wilder Sent: Monday, July 18, 2005 11:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bodies in morgue - who does the checking? Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From galalmkh <@t> yahoo.com Sat Jul 16 07:06:46 2005 From: galalmkh <@t> yahoo.com (manal galal) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] liquid nitrogen Message-ID: <20050716120646.93890.qmail@web50204.mail.yahoo.com> Hi all, I wanted to ask about how to use liquid nitogen in freezing muscle biopsies. I mean what does it come in? How much do I use? Do I discard it after I use it? What kind of container do I use? How long do I put the biopsy in it? How do I store it? By the way I freeze muscle in the quick freezing chamber of the cryostat, and am getting very little to no artifacts. But anyone that learns of how I freeeze biopsies says that it will give me disasterous resuls. Do you think liquid nitrogen will be superior to my old method. Thanks in advance manal __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From la.sebree <@t> hosp.wisc.edu Mon Jul 18 12:36:12 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the Message-ID: As technologists, we sign off on the run sheets of each instrument batch of slides. This indicates that the positive and negative controls have stained as expected and that the patient unknowns are of high quality. If something is not as it should be at this point, we may decide on our own to rerun a case or slides and inform the pathologist or we discuss it with the requesting pathologist and come up with a plan of action, which is documented on the run sheet. When the slides are delivered to the pathologists, a QA form accompanies them and the pathologist is required to indicate "satisfactory" or "unsatisfactory" and sign his name. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Billie Zimmerman Sent: Monday, July 18, 2005 12:10 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the technical quality of histologic preparations by the pathologist? This req. came out in April so I was wondering the best way to handle this. Thanks, Billie Zimmerman Medical College of GA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sluhisto <@t> yahoo.com Mon Jul 18 12:38:19 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] CAP inspections Message-ID: <20050718173819.2450.qmail@web51011.mail.yahoo.com> Hello All: This is directed at those of you that may have had your CAP inspection in the last year. I have been hearing that the inspectors have been, shall we say, interpreting some of the checklist questions in an unusual manner. My question to you all is, were there any questions where you got "tagged" for something that you had always done and had passed previously? Also, can you share ANY experiences that you had through your CAP inspection good or bad? As always, thank you for your insights and opinions. This is a super group and I always appreciate your comments. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From GoodwinD <@t> pahosp.com Mon Jul 18 10:56:14 2005 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] seeking part time position Message-ID: <992899E9EC268548AB8DDE246AF88473055F5266@PAHEX01.uphs.upenn.edu> Experienced histo tech seeking part time position in Philadelphia area. If interested, contact Coleen Kiehl at 215-829-6940. From DMBCMP <@t> aol.com Mon Jul 18 13:30:48 2005 From: DMBCMP <@t> aol.com (DMBCMP@aol.com) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] How is everyone carrying.... Message-ID: <129.6119d036.300d4f58@aol.com> Hi: Just wanted to ad that we, also, send a QC sheet to each of our pathologists every morning with their first run of slides. They respond on the sheet, even though they speak to us about any problem, of course. This is a way for us to keep a record for CAP. Dannie Blake, HT Community Medical Centers Fresno, Ca From mweirauch <@t> crittenton.com Mon Jul 18 13:44:41 2005 From: mweirauch <@t> crittenton.com (Maray Weirauch) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Workloads vs Hiso staffing Message-ID: ** PRIVATE ** Our hospital is conducting an analysis of laboratory workload, workflow and staffing. This will include Histology as well, and we are being asked for input with any national benchmarks available for surgical histology tasks (i.e. average number cases accessioned, blocks embedded, slides cut../some type of time unit) Does anyone know of some realistic published values or ranges? Thanks! From mcauliff <@t> umdnj.edu Mon Jul 18 13:42:07 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] mouse brain In-Reply-To: <6.0.1.1.0.20050713102532.01b7d838@mailhost.vetmed.auburn.edu> References: <6.0.1.1.0.20050713102532.01b7d838@mailhost.vetmed.auburn.edu> Message-ID: <42DBF7FF.8080406@umdnj.edu> Hi Atoska: Hydrocephalus is not uncommon in mice, perhaps that is the source of your concern? I suppose it could be manifested more on one side than the other? Maybe you could post a photo? Geoff Atoska S. Gentry wrote: > Hello, please have any of you who process & section paraformaldehyde > fixed, paraffin embedded mouse brain experienced asymmetry of the > right & left hemispheres? Recently our coronal mouse brain sections > which appear symmetrical upon gross trim and after initial facing of > paraffin are asymmetrical after staining. I've rotated the block > holder to the limit and sectioned the most rostral hemisphere at > higher micron thickness to accommodate. However, to my dismay and > disappointment none of this has helped very much. Any pointers in > remedying this situation ASAP will be greatly appreciated. Thanks, Atoska > > _______________________________________________ > 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 GoodwinD <@t> pahosp.com Mon Jul 18 14:27:58 2005 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] RE: Histonet Digest, Vol 20, Issue 21 Message-ID: <992899E9EC268548AB8DDE246AF88473055F526B@PAHEX01.uphs.upenn.edu> At our institution, the Anatomic Path dept. is responsible for "morgue patient inventory" and releasing bodies. This has been a point of contention for years, ever since they eliminated the position of "morgue attendant", but one that our administration insists is the pathology dept's responsibility. At the institution where I previously worked, body release was handled by the security dept, with the central switchboard in control of the status of decedents in the hospital via the HIS. Good luck! Diana G. Goodwin Department of Pathology Pennsylvania Hospital 800 Spruce St., Preston 655-C Philadelphia, PA? 19107 ? ph:? 215-829-6532 fax:? 215-829-7564 e-mail:? goodwind@pahosp.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Sent: Monday, July 18, 2005 1:01 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 21 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: dissecting board (Malam Jacqueline) 2. Need help with PGP9.5 on FFPE skin (Nicola Cragg) 3. RE: automated microtomes (Bonner, Janet) 4. Re: Dissection boards (Fred Underwood) 5. Bodies in morgue - who does the checking? (Paula Wilder) ---------------------------------------------------------------------- Message: 1 Date: Mon, 18 Jul 2005 12:39:23 +0100 From: Malam Jacqueline Subject: [Histonet] RE: dissecting board To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Thermo Shandon do 2 colours and 2 sizes of polyethylene board. They're easy to clean and last. We actually got ours from a butchers' suppliers! -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 17 July 2005 18:07 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 19 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Dissection boards (Katia Cristina Catunda) ---------------------------------------------------------------------- Message: 1 Date: Sat, 16 Jul 2005 22:20:15 -0300 From: "Katia Cristina Catunda" Subject: [Histonet] Dissection boards To: Message-ID: <019801c58a6d$b0bbc050$a279fea9@privatexx> Content-Type: text/plain; charset="iso-8859-1" In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 19 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 2 Date: Mon, 18 Jul 2005 13:39:29 +0100 From: "Nicola Cragg" Subject: [Histonet] Need help with PGP9.5 on FFPE skin To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello, I'm posting this message with some embarrassment and apologise for raising anybody's hope of a quck solution........ I have been trying to optimise PGP9.5 on FFPE skin samples (3 micron), using a mouse monoclonal (clone 10A1) from Neuromics. Despite my initial excitement expressed in an earlier posting, I have since found some very good photos of PGP9.5 staining in published work which has put mine to shame and has made me realise that my IHC is not working well enough at all. I've found an improvement with the waterbath antigen retrieval, as the dermis remains intact and there is some specific staining (I think) in the reticular dermis, which wasn't apparent with microwave antigen retrieval. However, it appears that there seems to be more of a trend of using free-floating 50 micron sections. Is that because routine FFPE sections are difficult to stain for this antigen? Has anyone got it to work on FFPE samples? I was quite hopeful that it would work from reading the datasheet and I have also found that Dako make a Rabbit polyclonal for FFPE sections and they're usually excellent antibodies so I'm hoping to try. Any advice or suggestions will be gratefully received. Regards, Nicola Cragg ------------------------------ Message: 3 Date: Mon, 18 Jul 2005 08:43:37 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] automated microtomes To: "'Sennello, Gina '" , "'histonet-bounces@lists.utsouthwestern.edu '" , "'Histonet Histonet (E-mail) '" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4325@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 We have used the Leica's with great success and few if any call backs after more than eight years!! Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet Histonet (E-mail) Sent: 7/13/2005 3:12 PM Subject: [Histonet] automated microtomes I am in the market for fully automated microtome and have looked at Leica's RM2255, Thermo-Electron's Finesse ME and Microm's 355S. Does anyone have any strong feels for against any of these instruments? Thanks in advance for you opinions and help. Gina Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 18 Jul 2005 12:29:54 -0400 From: "Fred Underwood" Subject: Re: [Histonet] Dissection boards To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I've found that kitchen cutting boards from your local department store works well, and it's cheaper than from a medical supplier. Fred >>> "Katia Cristina Catunda" 07/16/05 09:20PM >>> In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 18 Jul 2005 16:37:34 +0000 From: "Paula Wilder" Subject: [Histonet] Bodies in morgue - who does the checking? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 21 **************************************** From CDodson <@t> clarian.org Mon Jul 18 15:26:05 2005 From: CDodson <@t> clarian.org (Dodson, Cecelia) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] PGP9.5 Message-ID: <6A65BA49B1BEEA419A3B4D16DFA8EC5A08283D@CHEX1.chp.clarian.org> We are staining PGP9.5 on a regular bases using a polyclonal antibody we purchase from Biogenesis. It is antigen retrieved in a steamer for 15 minutes then stained with LSAB2 from Dako. Cecelia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, July 18, 2005 12:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 21 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: dissecting board (Malam Jacqueline) 2. Need help with PGP9.5 on FFPE skin (Nicola Cragg) 3. RE: automated microtomes (Bonner, Janet) 4. Re: Dissection boards (Fred Underwood) 5. Bodies in morgue - who does the checking? (Paula Wilder) ---------------------------------------------------------------------- Message: 1 Date: Mon, 18 Jul 2005 12:39:23 +0100 From: Malam Jacqueline Subject: [Histonet] RE: dissecting board To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Thermo Shandon do 2 colours and 2 sizes of polyethylene board. They're easy to clean and last. We actually got ours from a butchers' suppliers! -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 17 July 2005 18:07 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 19 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Dissection boards (Katia Cristina Catunda) ---------------------------------------------------------------------- Message: 1 Date: Sat, 16 Jul 2005 22:20:15 -0300 From: "Katia Cristina Catunda" Subject: [Histonet] Dissection boards To: Message-ID: <019801c58a6d$b0bbc050$a279fea9@privatexx> Content-Type: text/plain; charset="iso-8859-1" In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 19 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 2 Date: Mon, 18 Jul 2005 13:39:29 +0100 From: "Nicola Cragg" Subject: [Histonet] Need help with PGP9.5 on FFPE skin To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello, I'm posting this message with some embarrassment and apologise for raising anybody's hope of a quck solution........ I have been trying to optimise PGP9.5 on FFPE skin samples (3 micron), using a mouse monoclonal (clone 10A1) from Neuromics. Despite my initial excitement expressed in an earlier posting, I have since found some very good photos of PGP9.5 staining in published work which has put mine to shame and has made me realise that my IHC is not working well enough at all. I've found an improvement with the waterbath antigen retrieval, as the dermis remains intact and there is some specific staining (I think) in the reticular dermis, which wasn't apparent with microwave antigen retrieval. However, it appears that there seems to be more of a trend of using free-floating 50 micron sections. Is that because routine FFPE sections are difficult to stain for this antigen? Has anyone got it to work on FFPE samples? I was quite hopeful that it would work from reading the datasheet and I have also found that Dako make a Rabbit polyclonal for FFPE sections and they're usually excellent antibodies so I'm hoping to try. Any advice or suggestions will be gratefully received. Regards, Nicola Cragg ------------------------------ Message: 3 Date: Mon, 18 Jul 2005 08:43:37 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] automated microtomes To: "'Sennello, Gina '" , "'histonet-bounces@lists.utsouthwestern.edu '" , "'Histonet Histonet (E-mail) '" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4325@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 We have used the Leica's with great success and few if any call backs after more than eight years!! Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet Histonet (E-mail) Sent: 7/13/2005 3:12 PM Subject: [Histonet] automated microtomes I am in the market for fully automated microtome and have looked at Leica's RM2255, Thermo-Electron's Finesse ME and Microm's 355S. Does anyone have any strong feels for against any of these instruments? Thanks in advance for you opinions and help. Gina Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 18 Jul 2005 12:29:54 -0400 From: "Fred Underwood" Subject: Re: [Histonet] Dissection boards To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I've found that kitchen cutting boards from your local department store works well, and it's cheaper than from a medical supplier. Fred >>> "Katia Cristina Catunda" 07/16/05 09:20PM >>> In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 18 Jul 2005 16:37:34 +0000 From: "Paula Wilder" Subject: [Histonet] Bodies in morgue - who does the checking? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 21 **************************************** From kweidenh <@t> montefiore.org Mon Jul 18 15:51:26 2005 From: kweidenh <@t> montefiore.org (Karen Weidenheim) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] liquid nitrogen Message-ID: Hi Manal, The group has been discussing this topic lately. Our lab does classical muscle biopsy freezing with liquid nitrogen and isopentane. (I am happy to hear that you are successful with the cryostat and will have to check this out, but anyway, here is a little synopsis): 1. Liquid nitrogen comes in a large pressurized tank that one orders from a company that provides compressed gases. 2. For freezing, We fill a 2 liter Dewar flask , Cat # 2119, Lab Line Instruments Inc, Melrose Park, IL about 75% full of liquid nitrogen and we suspend our 250 ml stainless steel, very clean beaker that is around 65% full of isopentane in it using a home-made wire contraption made from coat hangers (!) that encircles the beaker and allows us to lower it slowly in to the isopentane (this whole procedure requires 2 people, 1 to handle the isopentane and 1 to do the specimen.) If you are alone, say, on call at night, you can use a ring stand to hold the beaker suspended by the wires. Or you can bend the wire contraption so it hooks on the side of the beaker. When ordering your equipment, you have to be sure, by measuring the internal diameter of the Dewar flask and the external diameter of your stainless beaker, that you can suspend the beaker in the flask comfortably. You have to lower the beaker of isopentane slowly into the liquid nitrogen to avoid the liquid nitrogen boiling into the isopentane and making a mess, at which point you have to start over. 3. We freeze a 0.8 x 0.5 cm portion of oriented skeletal muscle 25 seconds in syrupy isopentane that has ice all around the sides and the bottom. THe textbooks say 5-10 seconds, but we find we need 25 seconds. I think lots of this method is empiric and depends on your location, altitude and humidity. We never reuse the isopentane which, by the way, has to be kept in a flammable cabinet according to regulations. I might do 4 blocks in one aliquot of isopentane but I change it frequently and I think this helps me avoid artifacts. As we cannot put the liquid nitrogen back in the tank, it is not reused either, though if we have several biopsies in a day we use the same liquid nitrogen. Frozen biopsies are stored in a little white box (EM sciences, I think)labeled with patient name and number, in the ultrafreezer. Best of luck with your lab. If your pathologist is satisfied with the method you use now you should stick to it, as I said, I will investigate it with our cryostats here. Karen Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> manal galal 7/16/2005 8:06:46 AM >>> Hi all, I wanted to ask about how to use liquid nitogen in freezing muscle biopsies. I mean what does it come in? How much do I use? Do I discard it after I use it? What kind of container do I use? How long do I put the biopsy in it? How do I store it? By the way I freeze muscle in the quick freezing chamber of the cryostat, and am getting very little to no artifacts. But anyone that learns of how I freeeze biopsies says that it will give me disasterous resuls. Do you think liquid nitrogen will be superior to my old method. Thanks in advance manal __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Mon Jul 18 17:33:16 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] liquid nitrogen Message-ID: <5D2189E74151CC42BEC02906BA8996322B90CB@exchsrv01.barrynet.barry.edu> Before trying to handle liquid nitrogen, get someone who uses it to show you how to handle it! Liquid nitrogen is very dangerous when improperly handled. I know of 2 deaths from the abuse of liquid nitrogen. Liquid nitrogen comes in a high pressure cylinder holding anything from 200 ml to 50 liters of liquid nitrogen (which will expand to 0.025 to 6 cubic meters of nitrogen gas). The high pressure cylinder itself is dangerous. If it falls and the valve breaks, escaping gas will turn the cylinder into a missile. The escaping gas can cause severe frostbite if you are near it. Even at a distance, large amounts of escaping nitrogen can displace enough air to suffocate you. Small amounts of liquid nitrogen spilled on yourself will cause severe frostbite. Large amounts can freeze a limb solid and make it as prone to shattering as an ice cube. If you must learn by yourself, buy one of the small bottles sold for dermatological or lecture demonstration use. They usually contain about 50 liters of nitrogen gas compressed down to about 400 ml at 120 Atm. Wear nitrile gloves over leather gloves. Make sure your work area is well ventilated. Clamp a Dewar flask in place and release the nitrogen quickly into the Dewar flask. Half of the nitrogen will escape as a very cold gas. The other half will run into the Dewar flask as a liquid. When the Dewar flask is half full, shut off the nitrogen. Drop the tissue into the liquid nitrogen in the bottom of the Dewar flask. After a few minutes retrieve the tissue with long forceps and put it in your cryostat. Let the liquid nitrogen evaporate. Personally, I don't like liquid nitrogen. The layer of nitrogen gas that forms between the tissue and the liquid nitrogen prevents efficient freezing. If the piece of tissue is large, the outside freezes first; then the freezing of the inside of the tissue cracks the frozen outside. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of manal galal Sent: Saturday, July 16, 2005 8:07 AM To: Histonet@Pathology.swmed.edu Subject: [Histonet] liquid nitrogen Hi all, I wanted to ask about how to use liquid nitogen in freezing muscle biopsies. I mean what does it come in? How much do I use? Do I discard it after I use it? What kind of container do I use? How long do I put the biopsy in it? How do I store it? By the way I freeze muscle in the quick freezing chamber of the cryostat, and am getting very little to no artifacts. But anyone that learns of how I freeeze biopsies says that it will give me disasterous resuls. Do you think liquid nitrogen will be superior to my old method. Thanks in advance manal __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information 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 jnocito <@t> satx.rr.com Mon Jul 18 19:09:37 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] CAP inspections References: <20050718173819.2450.qmail@web51011.mail.yahoo.com> Message-ID: <000501c58bf6$26c29830$b4bd0b43@yourxhtr8hvc4p> Susan, I had my inspection in January. Can't speak for others, but, we received Accreditation with Accommodation on the last two inspections. This year we got slammed pretty hard. I think it's because of the incident last year concerning a lab in Maryland. CAP gave the lab an outstanding inspection, but there were many QA/QC issues that the some lab employees complained to CLIA. From what I was told, CAP had to explain itself to Congress. My QA program was torn apart (received compliments on the same programs during the last two inspections). Was written up because some of the temperature charts had missing dates (would you rather me pencil whip these?).Really took it the you know what because a couple of water cultures were not documented. As a matter of fact, we're giving a lecture on "Preparing for a CAP Inspection" at he NSH. It's number 99, the last lecture. Good luck. If I can help, just drop me a line. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Histology SLU" To: Sent: Monday, July 18, 2005 12:38 PM Subject: [Histonet] CAP inspections > Hello All: > > This is directed at those of you that may have had your CAP inspection in > the last year. I have been hearing that the inspectors have been, shall > we say, interpreting some of the checklist questions in an unusual manner. > My question to you all is, were there any questions where you got "tagged" > for something that you had always done and had passed previously? Also, > can you share ANY experiences that you had through your CAP inspection > good or bad? As always, thank you for your insights and opinions. This > is a super group and I always appreciate your comments. > > Susan > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.9.1/51 - Release Date: 7/18/2005 > > From jnocito <@t> satx.rr.com Mon Jul 18 19:14:31 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the References: Message-ID: <008801c58bf7$1d4807d0$b4bd0b43@yourxhtr8hvc4p> Billie, we send a QC sheet and an H&E control with the first set of slides. The sheet has a large "comments" section. The pathologists write anything in that space from "too many wrinkles" to "not enough hematoxylin" This was accepted by the nice inspector who inspected my lab this January. One of the few items he did like. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Billie Zimmerman" To: Sent: Monday, July 18, 2005 12:09 PM Subject: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the > How is everyone carrying out the new CAP req.? Is there documented > evidence of daily review of the technical quality of histologic > preparations by the pathologist? > > This req. came out in April so I was wondering the best way to handle > this. > > Thanks, > Billie Zimmerman > Medical College of GA > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.9.1/51 - Release Date: 7/18/2005 > > From Kemlo.Rogerson <@t> elht.nhs.uk Tue Jul 19 01:49:00 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Bodies in morgue - who does the checking? Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD49F@elht-exch1.xelht.nhs.uk> In the UK I believe that it is the Police's responsibility to check in the body, strip and take valuables (with a Porter in attendance); these are obviously BID's (Brought in Dead). If 'more expert help' is needed, for example a badly decomposed body or a mutilated one, then an APT (Anatomical Pathology Technician) would be called. Hospital cases are dealt with by the Nursing Staff and placed in the Mortuary by the Porters. Is this a Universal UK procedure? -----Original Message----- From: Paula Wilder [mailto:histo20@hotmail.com] Sent: 18 July 2005 17:38 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bodies in morgue - who does the checking? Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From padunnje <@t> iupui.edu Tue Jul 19 09:29:42 2005 From: padunnje <@t> iupui.edu (Dunn-Jena, Patsy A) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] unsubscribe Message-ID: From jean.gillson <@t> elht.nhs.uk Tue Jul 19 09:40:51 2005 From: jean.gillson <@t> elht.nhs.uk (Jean Gillson) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Cytology Immuno's Message-ID: Can I ask you all, what do you use as controls when doing ICC on cytology cytospin preps or smears? Since there isn't much sample, do you use paraffin sections as positive controls and is that adequate control measures? We currently only use paraffin cytoblocks but considering ICC on thin layer cytology preps and wondering what control material to use. Thanks for your help. From Michael.Habitzruther <@t> RoswellPark.org Tue Jul 19 09:56:39 2005 From: Michael.Habitzruther <@t> RoswellPark.org (Habitzruther, Michael) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Problem with Eosin Y staining Message-ID: <97101976F8A044468CA74FE11883B90E25EAAF@VISTA.roswellpark.org> Our laboratory is having a problem with the staining of mouse tumor samples using hemotoxylin and Eosin Y (H&E Stain). Everything was going fine until recently. The specific problem we are having is after the cover-slip is put on and the acrymount dries, the Eosin Y stain seems to be bleeding out, leaving only the hemotoxylin staining. A histologist from the institute suggested it may have something to do with the relative humidity in our lab, and therefore allowing the slides to set under a flow-hood may help. Any other suggestions or protocol modifications would be greatly appreciated. Thank you very much. This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. From Kemlo.Rogerson <@t> elht.nhs.uk Tue Jul 19 10:05:25 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Problem with Eosin Y staining Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698DCB@elht-exch1.xelht.nhs.uk> Eosin is soluble in both water and alcohol isn't it? Well if it bleeds out one of these must be present mustn't it? Solution (forgive the pun) is to eradicate both; I suspect it's alcohol. -----Original Message----- From: Habitzruther, Michael [mailto:Michael.Habitzruther@RoswellPark.org] Sent: 19 July 2005 15:57 To: histonet@lists.utsouthwestern.edu Cc: Demant, Peter Subject: [Histonet] Problem with Eosin Y staining Our laboratory is having a problem with the staining of mouse tumor samples using hemotoxylin and Eosin Y (H&E Stain). Everything was going fine until recently. The specific problem we are having is after the cover-slip is put on and the acrymount dries, the Eosin Y stain seems to be bleeding out, leaving only the hemotoxylin staining. A histologist from the institute suggested it may have something to do with the relative humidity in our lab, and therefore allowing the slides to set under a flow-hood may help. Any other suggestions or protocol modifications would be greatly appreciated. Thank you very much. This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Tue Jul 19 10:20:03 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Cytology Immuno's Message-ID: I use paraffin sections for positive controls and, yes, I consider it adequate for confirming that the test worked. Obviously, an internal positive control is best, but is not always present. Each laboratory needs to validate the individual antibodies on cytologic specimens using known cases. In our experience, some antibodies don't work very well on alcohol-fixed cytology specimens and, therefore, should be used with caution. I have not been able to confirm this, but I don't think "IVD" labels for antibodies used on formalin-fixed, paraffin-embedded tissue sections apply when used on cytology specimens. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> Jean Gillson 07/19/05 10:40AM >>> Can I ask you all, what do you use as controls when doing ICC on cytology cytospin preps or smears? Since there isn't much sample, do you use paraffin sections as positive controls and is that adequate control measures? We currently only use paraffin cytoblocks but considering ICC on thin layer cytology preps and wondering what control material to use. Thanks for your help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From chiggerson <@t> memhosp.com Tue Jul 19 11:35:58 2005 From: chiggerson <@t> memhosp.com (chiggerson@memhosp.com) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Re: Bodies in Morgue Message-ID: Bodies are checked in and released by Security. Cindy ----- Forwarded by Cindy Higgerson/6550/Pmmc on 07/19/2005 11:33 AM ----- Date: Mon, 18 Jul 2005 16:37:34 +0000 From: "Paula Wilder" Subject: [Histonet] Bodies in morgue - who does the checking? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 21 **************************************** From jcline <@t> wchsys.org Tue Jul 19 12:08:32 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Problem with Eosin Y staining In-Reply-To: <53A22BBB01D6184EBF7A4DC18A021E9E698DCB@elht-exch1.xelht.nhs.uk> Message-ID: <000601c58c84$7e1b3090$1d2a14ac@wchsys.org> We have a high humidity problem and I use H-2 Blue beads to remove water from our clearing agent. We check and change our alcohols after our eosin everyday. Our clearing agent will be opaque if not checked frequently. (water in the clearing agent will cause Eosin to bleed) ********************************************************************* Our laboratory is having a problem with the staining of mouse tumor samples using hemotoxylin and Eosin Y (H&E Stain). Everything was going fine until recently. The specific problem we are having is after the cover-slip is put on and the acrymount dries, the Eosin Y stain seems to be bleeding out, leaving only the hemotoxylin staining. A histologist from the institute suggested it may have something to do with the relative humidity in our lab, and therefore allowing the slides to set under a flow-hood may help. Any other suggestions or protocol modifications would be greatly appreciated. Thank you very much. This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message 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 ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From ljb <@t> medicine.wisc.edu Tue Jul 19 12:16:56 2005 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] culture on coverslips troubles Message-ID: Greetings Histonetters: I'm being asked to give advice about how to keep cultured cells on their cover slips during IHC for Confocal Microscopy. Is there a specific technique for washing the cover slips without losing cells? Should the cover slips be "air-dried" at some point? What fixative is best for confocal microscopy? Are there positively charged cover slips out there? Last question: Does anyone know of a list server for culture people? Marketing people please feel free to respond to me regarding these questions if you think you can be of service. I thank you all, and I send thanks for my questioning friend. 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 katri <@t> cogeco.ca Tue Jul 19 12:32:15 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Problem with Eosin Y staining References: <97101976F8A044468CA74FE11883B90E25EAAF@VISTA.roswellpark.org> Message-ID: <002101c58c87$ce4eee00$6a9a9618@Katri> What is this acrymount you coverslip with? It sounds like a waterbased mounting medium meant for applications, where you can't dehydrate and clear with xylene. Do you dehydrate and clear your H&E before mounting? Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Habitzruther, Michael" To: Cc: "Demant, Peter" Sent: Tuesday, July 19, 2005 10:56 AM Subject: [Histonet] Problem with Eosin Y staining Our laboratory is having a problem with the staining of mouse tumor samples using hemotoxylin and Eosin Y (H&E Stain). Everything was going fine until recently. The specific problem we are having is after the cover-slip is put on and the acrymount dries, the Eosin Y stain seems to be bleeding out, leaving only the hemotoxylin staining. A histologist from the institute suggested it may have something to do with the relative humidity in our lab, and therefore allowing the slides to set under a flow-hood may help. Any other suggestions or protocol modifications would be greatly appreciated. Thank you very much. This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From specialstainsqueen <@t> hotmail.com Tue Jul 19 13:03:56 2005 From: specialstainsqueen <@t> hotmail.com (Sherri Anderson) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Problem with Eosin Y staining In-Reply-To: <97101976F8A044468CA74FE11883B90E25EAAF@VISTA.roswellpark.org> Message-ID: Michael- Eosin "bleeding" of any sort is generally caused by the presence of an aqueous contaminant (i.e. atmospheric moisture, inadequate dehydration after the eosin, etc). So, it very well could be due to a humid environment. However, it would also behoove you to check the concentrations of the alcohols after your eosin to be sure that you have at least a couple of changes of 100% before the slides proceed to the clearant. Personally, I always had (3) 100% alcohol positions....but since you're in a humid environment, you may want to increase that to four. If you are not doing so already, I would also suggest covering the reagent dishes when not actively staining...so that the absolute alcohol does not absorb any more moisture from the air than it absolutely has to absorb (I would use individual dish covers rather than just the overall lid of the stainer...if such covers are available to you). In addition, increasing the frequency with which you change your last series of alcohols might help (or if you "rotate", perhaps a total "changeout" would be in order). Finally, I would make sure that your mountant is tightly capped when not in use (for the same reason as covering the reagent dishes when not in use -- to prevent the absorption of atmospheric moisture). Most people, in their staining protocols, follow their eosin with a 95% alcohol. It's the 5% water that differentiates the eosin -- increase the water, and you'll increase the amount of eosin that is removed from the tissue....decrease the water, and you'll decrease the amount of eosin that's removed. The presence of water is almost certainly the culprit to the issue you are describing -- the challenge is finding and eliminating the source. I hope that you find this to be somewhat helpful -- good luck! Sincerely, Sherri L. Anderson, BS, HTL(ASCP) Product Specialist Anatomical Pathology Thermo Electron Corp. >From: "Habitzruther, Michael" >To: >CC: "Demant, Peter" >Subject: [Histonet] Problem with Eosin Y staining >Date: Tue, 19 Jul 2005 10:56:39 -0400 > >Our laboratory is having a problem with the staining of mouse tumor samples >using hemotoxylin and Eosin Y (H&E Stain). Everything was going fine until >recently. The specific problem we are having is after the cover-slip is >put on and the acrymount dries, the Eosin Y stain seems to be bleeding out, >leaving only the hemotoxylin staining. A histologist from the institute >suggested it may have something to do with the relative humidity in our >lab, and therefore allowing the slides to set under a flow-hood may help. >Any other suggestions or protocol modifications would be greatly >appreciated. Thank you very much. > > >This email message may contain legally privileged and/or confidential >information. If you are not the intended recipient(s), or the employee or >agent responsible for the delivery of this message to the intended >recipient(s), you are hereby notified that any disclosure, copying, >distribution, or use of this email message is prohibited. If you have >received this message in error, please notify the sender immediately by >e-mail and delete this email message from your computer. Thank you. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From PatPatterson <@t> mhd.com Tue Jul 19 13:13:40 2005 From: PatPatterson <@t> mhd.com (Patterson, Pat) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] RE: Bodies in Morgue Message-ID: <293C7C19EFF7D611AE1A0002A53F81140FA63645@omega.mhd.com> Paula - Here bodies are released by Pastoral Care - they handle the entire process - from speaking with the families to release. Pat > Date: Mon, 18 Jul 2005 16:37:34 +0000 > From: "Paula Wilder" > Subject: [Histonet] Bodies in morgue - who does the checking? > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; format=flowed > > Hi everyone! > > Any feedback on whose responsibility it is to check bodies in the morgue > at > your respective instituition would be greatly appreciated. So far, by > phoning neighboring hospitals, I have found that Security, a diener > service, > or the Pathology Assistants are the ones responsible. Any help in this > would truly be greatly appreciated! Thanks so much! > > Paula Wilder > St.Joseph Medical Center > Towson, MD 21204 > 410-337-1741 > > > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 20, Issue 21 > **************************************** > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 20, Issue 23 > **************************************** *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. From herme013 <@t> umn.edu Tue Jul 19 13:14:08 2005 From: herme013 <@t> umn.edu (Yves Heremans) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] mixing up primary and secondary antibodies for double/triple staining Message-ID: Dear Histonetters, I have a general question on double or triple fluorescence stainings. Jackson Immunoresearch recommends doing sequential stainings, i.e. complete staining for antigen A first, then antigen B and then antigen C. Is there any danger in mixing up all the primary antibodies in one mix instead of using the sequential approach ? And, if using secondary antibodies derived from the same species, is there any danger in doing the same for the secondaries ? Yves From emry <@t> u.washington.edu Tue Jul 19 13:15:57 2005 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Fumeguard filters Message-ID: Hi, We have an older Fumeguard bench top hood. It has a charcole filter that sits in front of the fan. I am trying to find a vendor that can supply the replacement filters. Thanks, Trisha U of WA Seattle From algranth <@t> u.arizona.edu Tue Jul 19 13:23:32 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] culture on coverslips troubles In-Reply-To: Message-ID: <4.3.2.7.2.20050719111049.02506ea8@algranth.inbox.email.arizona.edu> Linda, When an investigator requests this we dip our coverslips in a chrome-alum-gelatin mixture and let them dry then pick up frozen sections on the coverslips. We use it for slides as well sometimes. 10 gm Gelatin 1000 ml D. H2O 5 gm Chromium Potassium Sulfate optional - few crystals of Thymol Dissolve the gelatin in hot distilled water - not boiling. Cool and dissolve the Chromium Potassium Sulfate in the gelatin mixture. A magnetic stirrer is helpful. Add a few crystals of thymol if you want. Store at RT in clean bottles. If you put it in the refrigerator the gelatin will harden (duh!). Make sure your coverslips are clean, clean, clean. Drain on paper toweling and allow to air dry. Store them in a dust free box when dry. Wear gloves - this is a possible carcinogen. You might be able to use a Poly-L-lysine Solution too. Got mine from Sigma and dilute it 1ml to 10 in D. H2O. Andi At 12:16 PM 7/19/2005 -0500, LaCinda Burchell wrote: >Greetings Histonetters: > I'm being asked to give advice about how to keep cultured cells on > their cover slips during IHC for Confocal Microscopy. Is there a > specific technique for washing the cover slips without losing > cells? Should the cover slips be "air-dried" at some point? What > fixative is best for confocal microscopy? Are there positively charged > cover slips out there? Last question: Does anyone know of a list server > for culture people? Marketing people please feel free to respond to me > regarding these questions if you think you can be of service. I thank > you all, and I send thanks for my questioning friend. > >LaCinda Burchell, BA, AS, HT(ASCP) >University of Wisconsin-Madison, Medical School >Asthma and Allergy Research IHC Lab >600 Highland Ave. CSC K4/913 >Madison, Wisconsin 53792 > >Phone: 608-262-3518 >FAX: 608-263-3746 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : 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 liz <@t> premierlab.com Tue Jul 19 13:35:21 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] mixing up primary and secondary antibodies fordouble/triple staining In-Reply-To: Message-ID: <000701c58c90$9efbf9a0$a7d48a80@AMY> Yves You can combine the primaries if all of them are from different species. If they are from the same species you will have to perform the staining sequentially. We have some images posted on our web page of immunofluorescent staining on 50 micron vibratome sections that have been done by combining the primary and secondary reagents. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Yves Heremans Sent: Tuesday, July 19, 2005 11:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] mixing up primary and secondary antibodies fordouble/triple staining Dear Histonetters, I have a general question on double or triple fluorescence stainings. Jackson Immunoresearch recommends doing sequential stainings, i.e. complete staining for antigen A first, then antigen B and then antigen C. Is there any danger in mixing up all the primary antibodies in one mix instead of using the sequential approach ? And, if using secondary antibodies derived from the same species, is there any danger in doing the same for the secondaries ? Yves _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Tue Jul 19 14:38:26 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Cytology Immuno's In-Reply-To: Message-ID: Jean, we use paraffin controls for any cytology specimen. I wish we could keep some cyto prep controls available, but we just don't have the room, nor the specimens. During my CAP inspection in January, it wasn't an issue, 50 other items were an issue, but not using FFPE controls with cytology Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jean Gillson Sent: Tuesday, July 19, 2005 9:41 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Cytology Immuno's Can I ask you all, what do you use as controls when doing ICC on cytology cytospin preps or smears? Since there isn't much sample, do you use paraffin sections as positive controls and is that adequate control measures? We currently only use paraffin cytoblocks but considering ICC on thin layer cytology preps and wondering what control material to use. Thanks for your help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From renafail <@t> bellsouth.net Tue Jul 19 14:56:51 2005 From: renafail <@t> bellsouth.net (renafail@bellsouth.net) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Fumeguard filters In-Reply-To: Message-ID: <000001c58c9c$01a514f0$0301a8c0@RENAD4YK9B8ABE> Trisha, You can get them from Shandon Lipshaw Rena -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Trisha Emry Sent: Tuesday, July 19, 2005 2:16 PM To: histo Subject: [Histonet] Fumeguard filters Hi, We have an older Fumeguard bench top hood. It has a charcole filter that sits in front of the fan. I am trying to find a vendor that can supply the replacement filters. Thanks, Trisha U of WA Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From renafail <@t> bellsouth.net Tue Jul 19 15:00:23 2005 From: renafail <@t> bellsouth.net (renafail@bellsouth.net) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Fumeguard filters In-Reply-To: Message-ID: <000101c58c9c$80b3c8e0$0301a8c0@RENAD4YK9B8ABE> Trisha, You can get them from Shandon Lipshaw Rena -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Trisha Emry Sent: Tuesday, July 19, 2005 2:16 PM To: histo Subject: [Histonet] Fumeguard filters Hi, We have an older Fumeguard bench top hood. It has a charcole filter that sits in front of the fan. I am trying to find a vendor that can supply the replacement filters. Thanks, Trisha U of WA Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Tue Jul 19 15:01:13 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] CAP & Antibody Expiration Date Message-ID: Hi all, since CAP never answered my email, I decided to stir up the pot again. According to Ed Groober in the Accreditation office, Federal Register dated 1/23/2003 section 493.1252d has the requirements about using expired reagents, or in this case, not using them. Now I need some help. Does anyone know where I go from here? I tried contacting my state and national representatives on another matter and received a nice little letter on fancy letterhead that basically said too bad or as we say here in south Texas "pass the cow patties". As John Paul Jones stated during the American Revolution (sorry my Brit friends) "I have yet not begun to fight". Tim, I know your hands are tied because of Lab Vision, but what are your thought on this? Joe From PMonfils <@t> Lifespan.org Tue Jul 19 15:59:04 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] culture on coverslips troubles Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717578@lsexch.lsmaster.lifespan.org> It depends a lot on what kind of cells are being cultured. When cells are grown in a culture flask, some kinds will come loose into suspension if you just hit the flask with your hand, while other cells require trypsin or other enzymatic treatment to get them loose. If you are dealing with a culture that has poor adherence, the cells can be firmly attached to the glass by using a coagulative fixative like Bouin's solution or alcoholic formalin. This works well if you are going to do a standard chemical stain on the cells. But for IHC it's a bit more complicated. Coagulative fixatives often denature antigens to the point where their antibodies no longer recognize them. And fixatives commonly used for IHC, like acetone or aqueous formalin, don't coagulate proteins enough to make the cells adhere. Air drying the coverslips without fixation will cause the cells to stick fast, but not without severe loss of morphology. If your antibody will tolerate formalin fixation, I would try brief (1 minute) immersion in 5% alcoholic formalin (1 part formaldehyde to 19 parts 70% ethanol). If this interferes with your antibody binding, try brief fixation in plain 70% ethanol. In fixing cultures on coverslips, drain off the excess fluid from the coverslips, and put them into the fixative when they are just "damp". > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > LaCinda Burchell > Sent: Tuesday, July 19, 2005 10:16 AM > To: histonet@pathology.swmed.edu > Subject: [Histonet] culture on coverslips troubles > > Greetings Histonetters: > I'm being asked to give advice about how to keep cultured cells on their > cover slips during IHC for Confocal Microscopy. Is there a specific > technique for washing the cover slips without losing cells? Should the > cover slips be "air-dried" at some point? What fixative is best for > confocal microscopy? Are there positively charged cover slips out there? > Last question: Does anyone know of a list server for culture people? > Marketing people please feel free to respond to me regarding these > questions if you think you can be of service. I thank you all, and I > send thanks for my questioning friend. > > LaCinda Burchell, BA, AS, HT(ASCP) > University of Wisconsin-Madison, Medical School > Asthma and Allergy Research IHC Lab > 600 Highland Ave. CSC K4/913 > Madison, Wisconsin 53792 > > Phone: 608-262-3518 > FAX: 608-263-3746 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From dusko.trajkovic <@t> pfizer.com Tue Jul 19 16:11:13 2005 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Confusing IHC results? Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B2D89551@lajamrexm01.amer.pfizer.com> Hi everyone, I just completed an IHC run for Calpain 1 (Santa Cruz), using dilutions 1:800, 1:1000 and 1:2000. I also ran previous runs where the dilutions were 1:100, 1:200, 1:400 and 1:800. I am using a Universal secondary from vector at 1:200 on all of the above slides. My staining results do not vary at all. As a matter of fact, my 1:2000 looks more intense and has more non specific staining than my slide at 1:100. When I run my negative at any of the concentrations to match my primary dilution, it is completely clean. What is going on here? Has anyone ever experienced something like this. Primary is a goat polyclonal. I have tried other secondaries such as, bovine anti goat, horse anti goat, rabbit anti goat, donkey anti goat, with either negative results or sections staining completely brown. Any suggestions, advice, email lashings would be greatly appreciated. Person to come up with a solution will receive a hug and a kiss, OR a couple of drinks at the bar, during the NSH symposium. It's your choice. Hell, you can take a chance and go for both. See, I'm going crazy trying to figure this out!!! Thank you, Dusko Trajkovic 1-858-638-6202 LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From JMacDonald <@t> mtsac.edu Tue Jul 19 16:22:31 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Problem with Eosin Y staining In-Reply-To: <97101976F8A044468CA74FE11883B90E25EAAF@VISTA.roswellpark.org> Message-ID: Is this a new mounting medium that you are using, or a new clearing agent? We had problems with eosin bleeding when the mounting medium and the clearing agent were not compatible. Jennifer MacDonald "Habitzruther, Michael" Sent by: histonet-bounces@lists.utsouthwestern.edu 07/19/2005 07:56 AM To cc "Demant, Peter" Subject [Histonet] Problem with Eosin Y staining Our laboratory is having a problem with the staining of mouse tumor samples using hemotoxylin and Eosin Y (H&E Stain). Everything was going fine until recently. The specific problem we are having is after the cover-slip is put on and the acrymount dries, the Eosin Y stain seems to be bleeding out, leaving only the hemotoxylin staining. A histologist from the institute suggested it may have something to do with the relative humidity in our lab, and therefore allowing the slides to set under a flow-hood may help. Any other suggestions or protocol modifications would be greatly appreciated. Thank you very much. This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gsennello <@t> osip.com Tue Jul 19 16:49:08 2005 From: gsennello <@t> osip.com (Sennello, Gina) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] job opening Message-ID: OSI Pharmaceuticals in Boulder, Colorado has a Research Associate/ Histotechnologist position open. For details go to: www.osip.com/careers Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 From vbaker60 <@t> yahoo.com Tue Jul 19 17:02:44 2005 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] COLA5-A2 Message-ID: <20050719220244.41627.qmail@web52515.mail.yahoo.com> Hi, I'm looking for the antibody that would correspond to this gene. I've found Collagen V antibodies, but not the right subunit. Has anyone done any work with this gene? Thanks in advance. Vikki Baker Mt. Sinai School of Medicine NY, NY __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From roy.ellis <@t> imvs.sa.gov.au Tue Jul 19 17:20:52 2005 From: roy.ellis <@t> imvs.sa.gov.au (Roy Ellis) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Problem with Eosin Y staining In-Reply-To: <97101976F8A044468CA74FE11883B90E25EAAF@VISTA.roswellpark.org> Message-ID: <000401c58cb0$1fa02990$c28a140a@iqe36417> Hi Michael Alkaline mountants will bleed eosin from the section into the mounting medium. I suggest you change your mounting medium for one that is known to be neutral. Regards Roy Ellis mailto:roy.ellis@imvs.sa.gov.au > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of > Habitzruther, Michael > Sent: Wednesday, 20 July 2005 12:27 AM > To: histonet@lists.utsouthwestern.edu > Cc: Demant, Peter > Subject: [Histonet] Problem with Eosin Y staining > > > Our laboratory is having a problem with the staining of mouse > tumor samples using hemotoxylin and Eosin Y (H&E Stain). > Everything was going fine until recently. The specific problem > we are having is after the cover-slip is put on and the acrymount > dries, the Eosin Y stain seems to be bleeding out, leaving only > the hemotoxylin staining. A histologist from the institute > suggested it may have something to do with the relative humidity > in our lab, and therefore allowing the slides to set under a > flow-hood may help. Any other suggestions or protocol > modifications would be greatly appreciated. Thank you very much. > > > This email message may contain legally privileged and/or > confidential information. If you are not the intended > recipient(s), or the employee or agent responsible for the > delivery of this message to the intended recipient(s), you are > hereby notified that any disclosure, copying, distribution, or > use of this email message is prohibited. If you have received > this message in error, please notify the sender immediately by > e-mail and delete this email message from your computer. Thank you. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ynwang <@t> u.washington.edu Tue Jul 19 18:19:41 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] postmortem tissue collection Message-ID: Dear histonetters, I have a question regarding collection of human tissue. A colleague would like to collect human tissue (skin and underlying adipose tissue) for mechanical testing, histological analysis (cellular and structural evaluation), IHC and protein analysis (extracellular matrix structure and concentrations). They asked what would be a fair cut off time for tissue collection so that the effects of decay would not be a factor. Currently they have given the tissue bank a time of 12 hours postmortem (I'm not very sure how the body is stored during this time or how this is calculated). I've read that human decomposition starts approx. 4 min after death and autolysis is quicker in tissues with high enzyme and water content. However, in terms of the skin and underlying adipose tissue I didn't know if there was an accepted 'cut off time' postmortem after which tissue is considered far from representative of 'live' tissue and not worth analysis. Can anyone give some insight? Any information or references would be greatly appreciated. Thank you Yak-Nam Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 From lpwenk <@t> sbcglobal.net Tue Jul 19 18:19:31 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] CAP & Antibody Expiration Date In-Reply-To: Message-ID: <1090307313-7411177@pathology.swmed.edu> The exact source is in the Code of Federal Register 42CFR493.1252.d http://www.gpoaccess.gov/cft/retrieve.html Go to this site, then type in the title (42), part (493) and section number (1252). I find it works best if sub-numbers and sub-letters that might be found after the section number (for example, your "d") are not used. Then hit "Go". (GPO in the web address is the Government Printing Office. CFR are all the laws of the US that must be followed - OSHA, EPA, etc.) This particular line reads: "(d) Reagents, solutions, culture media, control materials, calibration materials, and other supplies must not be used when they have exceeded their expiration date, have deteriorated, or are of substandard quality." Hope that helps. Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, July 19, 2005 4:01 PM To: Histonet Subject: [Histonet] CAP & Antibody Expiration Date Hi all, since CAP never answered my email, I decided to stir up the pot again. According to Ed Groober in the Accreditation office, Federal Register dated 1/23/2003 section 493.1252d has the requirements about using expired reagents, or in this case, not using them. Now I need some help. Does anyone know where I go from here? I tried contacting my state and national representatives on another matter and received a nice little letter on fancy letterhead that basically said too bad or as we say here in south Texas "pass the cow patties". As John Paul Jones stated during the American Revolution (sorry my Brit friends) "I have yet not begun to fight". Tim, I know your hands are tied because of Lab Vision, but what are your thought on this? Joe _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Tue Jul 19 18:48:26 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] CAP & Antibody Expiration Date References: <1090307313-7411177@pathology.swmed.edu> Message-ID: <001c01c58cbc$5cd3eca0$b4bd0b43@yourxhtr8hvc4p> See, people at CAP make mistakes to, and they think they are infallible. Yeah, right Joe ----- Original Message ----- From: "Lee & Peggy Wenk" To: "'Joe Nocito'" ; "'Histonet'" Sent: Tuesday, July 19, 2005 6:19 PM Subject: RE: [Histonet] CAP & Antibody Expiration Date > The exact source is in the Code of Federal Register 42CFR493.1252.d > > http://www.gpoaccess.gov/cft/retrieve.html > > Go to this site, then type in the title (42), part (493) and section > number > (1252). I find it works best if sub-numbers and sub-letters that might be > found after the section number (for example, your "d") are not used. > > Then hit "Go". > > (GPO in the web address is the Government Printing Office. CFR are all the > laws of the US that must be followed - OSHA, EPA, etc.) > > This particular line reads: "(d) Reagents, solutions, culture media, > control > materials, calibration materials, and other supplies must not be used when > they have exceeded their expiration date, have deteriorated, or are of > substandard quality." > > Hope that helps. > > Peggy Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito > Sent: Tuesday, July 19, 2005 4:01 PM > To: Histonet > Subject: [Histonet] CAP & Antibody Expiration Date > > Hi all, > since CAP never answered my email, I decided to stir up the pot again. > According to Ed Groober in the Accreditation office, Federal > Register dated > 1/23/2003 section 493.1252d has the requirements about using expired > reagents, or in this case, not using them. Now I need some help. Does > anyone > know where I go from here? > I tried contacting my state and national representatives on another > matter and received a nice little letter on fancy letterhead that > basically > said too bad or as we say here in south Texas "pass the cow patties". > As John Paul Jones stated during the American Revolution (sorry my > Brit > friends) "I have yet not begun to fight". > Tim, I know your hands are tied because of Lab Vision, but what are > your thought on this? > > Joe > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.9.1/51 - Release Date: 7/18/2005 > > From kccatunda <@t> terra.com.br Tue Jul 19 20:21:39 2005 From: kccatunda <@t> terra.com.br (Katia Cristina Catunda) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Dissection boards References: Message-ID: <003001c58cc9$61da9fc0$a279fea9@privatexx> Thank you all for the answers about the Dissection boards... your ideas and great informations will be very useful Soon I'll give you some news... Katia From raestask <@t> galesburg.net Tue Jul 19 23:11:05 2005 From: raestask <@t> galesburg.net (Rae Staskiewiez) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Request Message-ID: <001c01c58ce1$0d7c5730$479a46c6@rae1ktsbyhej72> I am posting this for my supervisor. I have a reference: Brinn N: Microwave Techniques. Histotechnologic teleconference, Univ Texas Does anyone have more information? Date? Published? Pages? Etc.? Thanks, DW If anyone has this info, he really would appreciate it. Thanks, Rae Ann Staskiewicz HT(ASCP) Section Head--Histology Galesburg Animal Disease Lab Galesburg, IL From int09018 <@t> alphahunt.com Wed Jul 20 01:16:35 2005 From: int09018 <@t> alphahunt.com (HCS) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Can anyone help answer the question? Message-ID: <000601c58cf2$97270be0$6601a8c0@hp> From: holling@med-in.uni-saarland.de (Nicole Hollinger) Sent: Wed, 20 Jul 2005 7:10:02 I?m a medical technologist in research. I need help. I?m searching a dyeing instruction for the two special stains: Carstair?s and Fraser Lendrum. Maybe you can help me.Sincerely yours,Nicole Hollinger Please reply to histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Jul 20 02:29:01 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] postmortem tissue collection Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4A5@elht-exch1.xelht.nhs.uk> Interestingly I was reading about tissue death after death (so as to speak) on a Web Site dealing with Death (I'm strange like that); I was interested in rigor. I hadn't realised that some tissue deep within the cadaver can survive for many hours after death. I suppose it's those tissues that require little or no oxygen to survive; brain dies very rapidly. If I could remember the Site then I'd tell you but a search in Google under rigor may help. Wonder which bit of you dies last? Same in men and women? -----Original Message----- From: Y. Wang [mailto:ynwang@u.washington.edu] Sent: 20 July 2005 00:20 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] postmortem tissue collection Dear histonetters, I have a question regarding collection of human tissue. A colleague would like to collect human tissue (skin and underlying adipose tissue) for mechanical testing, histological analysis (cellular and structural evaluation), IHC and protein analysis (extracellular matrix structure and concentrations). They asked what would be a fair cut off time for tissue collection so that the effects of decay would not be a factor. Currently they have given the tissue bank a time of 12 hours postmortem (I'm not very sure how the body is stored during this time or how this is calculated). I've read that human decomposition starts approx. 4 min after death and autolysis is quicker in tissues with high enzyme and water content. However, in terms of the skin and underlying adipose tissue I didn't know if there was an accepted 'cut off time' postmortem after which tissue is considered far from representative of 'live' tissue and not worth analysis. Can anyone give some insight? Any information or references would be greatly appreciated. Thank you Yak-Nam Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Jul 20 04:10:29 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] RE: ICC on cytology preps Message-ID: We occasionally get asked for ICC on cyt preps and, as these don't need ag. retrieval, I felt the positive controls should also be a cyt prep to be valid. We have found that if you receive cellular fluids (eg malig pleural or ascitic) in cyt fix, after they have been diagnosed you can test their positivity for the antigen in question and, instead of disposing of them, they can be kept to use as positive control material. The dilution of the primary antisera may need adjusting so it's worth doing a titration. Also, we suspend the fluid in saline, spin down, discard the supernatant and resuspend x3 to get rid of the protein in the fluid which can cause background staining and interfere with the picture. The downside is if you can't get hold of a fluid which is positive for the antigen you need, or if your test arrives ready prepared on the slide, or is a fluid which only has scanty cells - you may lose some in washing with saline. You could always make a cell block with agar (if you have enough cells) and fix in formalin etc then you can do your routine protocols. Good luck! Jacqui -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 19 July 2005 18:02 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 23 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re: Cytology Immuno's (Richard Cartun) 2. Re: Bodies in Morgue (chiggerson@memhosp.com) ---------------------------------------------------------------------- Message: 1 Date: Tue, 19 Jul 2005 11:20:03 -0400 From: "Richard Cartun" Subject: Re: [Histonet] Cytology Immuno's To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I use paraffin sections for positive controls and, yes, I consider it adequate for confirming that the test worked. Obviously, an internal positive control is best, but is not always present. Each laboratory needs to validate the individual antibodies on cytologic specimens using known cases. In our experience, some antibodies don't work very well on alcohol-fixed cytology specimens and, therefore, should be used with caution. I have not been able to confirm this, but I don't think "IVD" labels for antibodies used on formalin-fixed, paraffin-embedded tissue sections apply when used on cytology specimens. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> Jean Gillson 07/19/05 10:40AM >>> Can I ask you all, what do you use as controls when doing ICC on cytology cytospin preps or smears? Since there isn't much sample, do you use paraffin sections as positive controls and is that adequate control measures? We currently only use paraffin cytoblocks but considering ICC on thin layer cytology preps and wondering what control material to use. Thanks for your help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 2 Date: Tue, 19 Jul 2005 11:35:58 -0500 From: chiggerson@memhosp.com Subject: [Histonet] Re: Bodies in Morgue To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Bodies are checked in and released by Security. Cindy ----- Forwarded by Cindy Higgerson/6550/Pmmc on 07/19/2005 11:33 AM ----- Date: Mon, 18 Jul 2005 16:37:34 +0000 From: "Paula Wilder" Subject: [Histonet] Bodies in morgue - who does the checking? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 21 **************************************** ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 23 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From renafail <@t> bellsouth.net Wed Jul 20 04:36:12 2005 From: renafail <@t> bellsouth.net (renafail@bellsouth.net) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] loss of cells on cytology preps Message-ID: <000201c58d0e$783e1400$0301a8c0@RENAD4YK9B8ABE> We have the same problems with controls and have to use our paraffin sections for controls. We also have an issue with loss of cells on cytology preps. Some are brought to us already stained and coverslipped others in 95%. We air dry the slides and put them in the oven an hour before staining Since doing this I have had only one case where all the cells washed off and one where some large patches washed off. Antigen retrieval is not performed on cytology preps. One of the pathologist is vehemently opposed to placing the specimens on the autostainer, she fells it blows the cells off. But slides hand stained are subjected to more vigorous washing from the wash bottles. Besides the problem of cells washing off, she is on one side insisting on hand runs, the manager is on the other saying absolutely no hand runs. With the volume we have I would prefer they go on the autostainer. Any other ideas for keeping the cells on the slides? Rena Fail From c.m.vanderloos <@t> amc.uva.nl Wed Jul 20 04:42:59 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] RE: Cytology Immuno's Message-ID: <11dbeab11d9b57.11d9b5711dbeab@amc.uva.nl> Dear Jean, Despite previous posts on this subject, to my opinion using paraffin sections as positive control along with cytology cytospin preps is not such a good idea! That also accounts for paraffin cytoblocks as positive control. I do understand that finding appropriate positive controls here isn't easy from practical point of view. My point is that a cyotology cytospin prep consists of cells that are (more or less) intact with an outer membrane, whereas paraffin sections from tissue or cytoblocks contain cut-open cells and no outer membrane. We have observed here quite dramatic differences between intact cells and cut-open cells when performing IHC techniques. This has everything to do with the cellular outer membrane as barrier for your antibodies. I think that immunostaining of intact cells, needs intact cells as positive control. And, immunostaining of sections, needs sections as positive control. Having said that, I also think we have to go back to the original terminology: "immunocytochemistry" for cells and "immunohistochemistry" for tissue sections. Mixing up those terms long time ago was a pretty bad idea. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Tue, 19 Jul 2005 15:40:51 +0100 From: Jean Gillson Subject: [Histonet] Cytology Immuno's To: "'histonet@lists.utsouthwestern.edu'" Can I ask you all, what do you use as controls when doing ICC on cytology cytospin preps or smears? Since there isn't much sample, do you use paraffin sections as positive controls and is that adequate control measures? We currently only use paraffin cytoblocks but considering ICC on thin layer cytology preps and wondering what control material to use. Thanks for your help. From IKirbis <@t> onko-i.si Wed Jul 20 04:49:44 2005 From: IKirbis <@t> onko-i.si (=?iso-8859-2?Q?Kirbi=B9_Srebotnik_Irena?=) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] loss of cells on cytology preps Message-ID: 3000 ICC/per year are performed in our lab on cytospins (80%)or smears (20%),since 2001 all stainings are performed by Ventana Nexes immunostainer,we strictly use cytology samples/cytospins for positive and negative controls, prepared and fixed in the same manner as samples. I strongly believe that FFPE sections are NOT APPROPRIATE controls for cytology samples we prepare cytospins for controls from: - leftovers of high cellular diagnostics samples - cell lines (MCF-7, HeLa, SK-MEL) - FNAB of resected tumours we don't have ANY problems with loss of cells! not at all, nor with any background Irena Srebotnik Kirbi?, MSc Institute of Oncology Dept. of Cytopathology Zalo?ka 2 1000 Ljubljana Slovenia phone +386 1 522 3826 fax +38615879400 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Sent: Wednesday, July 20, 2005 11:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] loss of cells on cytology preps We have the same problems with controls and have to use our paraffin sections for controls. We also have an issue with loss of cells on cytology preps. Some are brought to us already stained and coverslipped others in 95%. We air dry the slides and put them in the oven an hour before staining Since doing this I have had only one case where all the cells washed off and one where some large patches washed off. Antigen retrieval is not performed on cytology preps. One of the pathologist is vehemently opposed to placing the specimens on the autostainer, she fells it blows the cells off. But slides hand stained are subjected to more vigorous washing from the wash bottles. Besides the problem of cells washing off, she is on one side insisting on hand runs, the manager is on the other saying absolutely no hand runs. With the volume we have I would prefer they go on the autostainer. Any other ideas for keeping the cells on the slides? Rena Fail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Wed Jul 20 04:58:03 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] RE: Confusing IHC results? Message-ID: <10e71f310e56e1.10e56e110e71f3@amc.uva.nl> Hello Dusco, When antibodies are applied at a far too high concentration staining will get weaker. This is called the "prozone effect". I would suggest to go up with your dilutions like: 1:2000 - 5000 - 10,000 - 20,000 - 50,000. At least until the specific staining disappears again. Lately, I came across the same situation and ended up unexpectedly at 1:200,000-500,000! Stupid remark perhaps, but did you take care that your blocking serum isn't normal goat since your secondary is anti-goat? As far as the reward: I keep it to the drink! Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Tue, 19 Jul 2005 17:11:13 -0400 From: "Trajkovic, Dusko" Subject: [Histonet] Confusing IHC results? To: "'histonet@lists.utsouthwestern.edu'" Hi everyone, I just completed an IHC run for Calpain 1 (Santa Cruz), using dilutions 1:800, 1:1000 and 1:2000. I also ran previous runs where the dilutions were 1:100, 1:200, 1:400 and 1:800. I am using a Universal secondary from vector at 1:200 on all of the above slides. My staining results do not vary at all. As a matter of fact, my 1:2000 looks more intense and has more non specific staining than my slide at 1:100. When I run my negative at any of the concentrations to match my primary dilution, it is completely clean. What is going on here? Has anyone ever experienced something like this. Primary is a goat polyclonal. I have tried other secondaries such as, bovine anti goat, horse anti goat, rab! bit anti From zyu <@t> mail.med.upenn.edu Wed Jul 20 07:50:20 2005 From: zyu <@t> mail.med.upenn.edu (Zhenming Yu) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] chatter in fly head paraffin section Message-ID: <1121863820.42de488c68b81@webmail.pobox.upenn.edu> Dear Histonetters, I'm looking for some guidance in obtaining good paraffin sections from fruitfly brain/eye. We repeated saw chatter in the brain region and are desperately trying to troubleshoot the problem. We have tried 10% NBF, 4% paraformaldehyde, Bourin's for fixation. And for tissue processing, we used the following schedule: 70% Alcohol 3 hours 95% Alcohol 2 hours 100% Alcohol 2 hours 100% Alcohol 2 hours ? 100% Alcohol 1 hours Xylene 1 hour Xylene 1 hour Xylene 1 hour Paraffin(60C): 2 hours Paraffin(60C): 2 hours---we didn't use vacuum at this step considering fly heads are quite small. Could this cause inadequate praffin infiltration and chatter in sections? The technician did leave paraffin in the processor melting at 60C for long period of time--she never shut off the power after the cycle is done. She also always left the embedding station on with paraffin melting at 62C. Could this cause deterioration of paraffin? For sectioning, we've tried using brand new blade, adjusting clearance angle, slowing down cutting speed etc. None seems to help. ?? Any suggestion would be greatly appreciated. Thank you all. **************************** Zhenming Yu Univ. of Penn Philadelphia, PA **************************** From JNocito <@t> Pathreflab.com Wed Jul 20 08:09:23 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] Confusing IHC results? In-Reply-To: <3AD0BD3142459B4E9B12CBEAFF2B89B2D89551@lajamrexm01.amer.pfizer.com> Message-ID: Dusko, some suggestions would be to titer out your secondary antibody further, say 1:400 and 1:800, and/or titer Calpain 1 out to 1:4000 and 1:8000 you might have to perform a checkerboard titration. i.e. Calpain 1 1:4000 Secondary 1:400 & 1:800 Calpain 1 1:8000 Secondary 1:400 & 1:800 I know it's a pain to titer out your secondary when it works on other antibodies, but I had to do this when I was at the AFIP some time ago. Just some thoughts. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Trajkovic, Dusko Sent: Tuesday, July 19, 2005 4:11 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Confusing IHC results? Hi everyone, I just completed an IHC run for Calpain 1 (Santa Cruz), using dilutions 1:800, 1:1000 and 1:2000. I also ran previous runs where the dilutions were 1:100, 1:200, 1:400 and 1:800. I am using a Universal secondary from vector at 1:200 on all of the above slides. My staining results do not vary at all. As a matter of fact, my 1:2000 looks more intense and has more non specific staining than my slide at 1:100. When I run my negative at any of the concentrations to match my primary dilution, it is completely clean. What is going on here? Has anyone ever experienced something like this. Primary is a goat polyclonal. I have tried other secondaries such as, bovine anti goat, horse anti goat, rabbit anti goat, donkey anti goat, with either negative results or sections staining completely brown. Any suggestions, advice, email lashings would be greatly appreciated. Person to come up with a solution will receive a hug and a kiss, OR a couple of drinks at the bar, during the NSH symposium. It's your choice. Hell, you can take a chance and go for both. See, I'm going crazy trying to figure this out!!! Thank you, Dusko Trajkovic 1-858-638-6202 LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MadaryJ <@t> MedImmune.com Wed Jul 20 08:30:53 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] RE: Histonet Digest, Vol 20, Issue 23 Message-ID: <746FDB897740814EA52BDDCB5ED1DDBCBB16FC@medimmune4.medimmune.com> Checked in by medical personnel with security right there, and checked out by security. After many arguments with security I was finally able to get them to assist with lifting the bodies on and off the table. It was in their job description but I guess they thought no was an acceptable answer. When could we start saying that? I missed the boat again! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, July 19, 2005 1:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 23 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re: Cytology Immuno's (Richard Cartun) 2. Re: Bodies in Morgue (chiggerson@memhosp.com) ---------------------------------------------------------------------- Message: 1 Date: Tue, 19 Jul 2005 11:20:03 -0400 From: "Richard Cartun" Subject: Re: [Histonet] Cytology Immuno's To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I use paraffin sections for positive controls and, yes, I consider it adequate for confirming that the test worked. Obviously, an internal positive control is best, but is not always present. Each laboratory needs to validate the individual antibodies on cytologic specimens using known cases. In our experience, some antibodies don't work very well on alcohol-fixed cytology specimens and, therefore, should be used with caution. I have not been able to confirm this, but I don't think "IVD" labels for antibodies used on formalin-fixed, paraffin-embedded tissue sections apply when used on cytology specimens. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> Jean Gillson 07/19/05 10:40AM >>> Can I ask you all, what do you use as controls when doing ICC on cytology cytospin preps or smears? Since there isn't much sample, do you use paraffin sections as positive controls and is that adequate control measures? We currently only use paraffin cytoblocks but considering ICC on thin layer cytology preps and wondering what control material to use. Thanks for your help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 2 Date: Tue, 19 Jul 2005 11:35:58 -0500 From: chiggerson@memhosp.com Subject: [Histonet] Re: Bodies in Morgue To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Bodies are checked in and released by Security. Cindy ----- Forwarded by Cindy Higgerson/6550/Pmmc on 07/19/2005 11:33 AM ----- Date: Mon, 18 Jul 2005 16:37:34 +0000 From: "Paula Wilder" Subject: [Histonet] Bodies in morgue - who does the checking? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 21 **************************************** ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 23 **************************************** From peoshel <@t> wisc.edu Wed Jul 20 08:33:54 2005 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] postmortem tissue collection In-Reply-To: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4A5@elht-exch1.xelht.nhs.uk> References: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4A5@elht-exch1.xelht.nhs.uk> Message-ID: Interesting question. My bet is tendon. Chrondrocytes in dense cartilage would seem to be likely, but experience here has shown that they have to be freeze-fixed by high-pressure freezing to avoid cell death artifacts. Any other way is too slow. Or ... dormant stem cells? Phil >Interestingly I was reading about tissue death after death (so as to >speak) on a Web Site dealing with Death (I'm strange like that); I was >interested in rigor. I hadn't realised that some tissue deep within the >cadaver can survive for many hours after death. I suppose it's those >tissues that require little or no oxygen to survive; brain dies very >rapidly. If I could remember the Site then I'd tell you but a search in >Google under rigor may help. Wonder which bit of you dies last? Same in >men and women? > >-----Original Message----- >From: Y. Wang [mailto:ynwang@u.washington.edu] >Sent: 20 July 2005 00:20 >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] postmortem tissue collection > >Dear histonetters, > >I have a question regarding collection of human tissue. A colleague >would >like to collect human tissue (skin and underlying adipose tissue) for >mechanical testing, histological analysis (cellular and structural >evaluation), IHC and protein analysis (extracellular matrix structure >and >concentrations). They asked what would be a fair cut off time for tissue > >collection so that the effects of decay would not be a factor. Currently > >they have given the tissue bank a time of 12 hours postmortem (I'm not >very sure how the body is stored during this time or how this is >calculated). > >I've read that human decomposition starts approx. 4 min after death and >autolysis is quicker in tissues with high enzyme and water content. >However, in terms of the skin and underlying adipose tissue I didn't >know >if there was an accepted 'cut off time' postmortem after which tissue >is considered far from representative of 'live' tissue and not worth >analysis. > >Can anyone give some insight? Any information or references would be >greatly appreciated. > >Thank you >Yak-Nam > >Senior Fellow >Department of Bioengineering >University of Washington >Box 357962 >Seattle, WA 98195 > >Tel.: (206)-221-5873 >Fax.: (206)-221-5874 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm From pmarcum <@t> vet.upenn.edu Wed Jul 20 08:34:09 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:20 2005 Subject: [Histonet] RE: Histonet Digest, Vol 20, Issue 23 In-Reply-To: <746FDB897740814EA52BDDCB5ED1DDBCBB16FC@medimmune4.medimmun e.com> References: <746FDB897740814EA52BDDCB5ED1DDBCBB16FC@medimmune4.medimmune.com> Message-ID: <6.1.1.1.2.20050720093332.019ace10@mail.vet.upenn.edu> We could always say NO it just isn't paid any attention too by most other people. Pam Marcum At 09:30 AM 7/20/2005, Madary, Joseph wrote: >Checked in by medical personnel with security right there, and checked out >by security. After many arguments with security I was finally able to get >them to assist with lifting the bodies on and off the table. It was in >their job description but I guess they thought no was an acceptable >answer. When could we start saying that? I missed the boat again! > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >histonet-request@lists.utsouthwestern.edu >Sent: Tuesday, July 19, 2005 1:05 PM >To: histonet@lists.utsouthwestern.edu >Subject: Histonet Digest, Vol 20, Issue 23 > > >Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > >To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet >or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > >You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > >When replying, please edit your Subject line so it is more specific >than "Re: Contents of Histonet digest..." > > >Today's Topics: > > 1. Re: Cytology Immuno's (Richard Cartun) > 2. Re: Bodies in Morgue (chiggerson@memhosp.com) > > >---------------------------------------------------------------------- > >Message: 1 >Date: Tue, 19 Jul 2005 11:20:03 -0400 >From: "Richard Cartun" >Subject: Re: [Histonet] Cytology Immuno's >To: , >Message-ID: >Content-Type: text/plain; charset=US-ASCII > >I use paraffin sections for positive controls and, yes, I consider it >adequate for confirming that the test worked. Obviously, an internal >positive control is best, but is not always present. Each laboratory >needs to validate the individual antibodies on cytologic specimens using >known cases. In our experience, some antibodies don't work very well on >alcohol-fixed cytology specimens and, therefore, should be used with >caution. I have not been able to confirm this, but I don't think "IVD" >labels for antibodies used on formalin-fixed, paraffin-embedded tissue >sections apply when used on cytology specimens. > >Richard > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 >(860) 545-1596 >(860) 545-0174 Fax > > >>> Jean Gillson 07/19/05 10:40AM >>> >Can I ask you all, what do you use as controls when doing ICC on cytology >cytospin preps or smears? Since there isn't much sample, do you use >paraffin sections as positive controls and is that adequate control >measures? We currently only use paraffin cytoblocks but considering ICC on >thin layer cytology preps and wondering what control material to use. >Thanks for your help. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >----------------------------------------------- >Confidentiality Notice > >This e-mail message, including any attachments, is for the sole use of the >intended recipient(s) and may contain confidential or proprietary >information which is legally privileged. Any unauthorized review, use, >disclosure, or distribution is prohibited. If you are not the intended >recipient, please promptly contact the sender by reply e-mail and destroy >all copies of the original message. > > > >------------------------------ > >Message: 2 >Date: Tue, 19 Jul 2005 11:35:58 -0500 >From: chiggerson@memhosp.com >Subject: [Histonet] Re: Bodies in Morgue >To: histonet@lists.utsouthwestern.edu >Message-ID: > >Content-Type: text/plain; charset="US-ASCII" > >Bodies are checked in and released by Security. > >Cindy > >----- Forwarded by Cindy Higgerson/6550/Pmmc on 07/19/2005 11:33 AM ----- > > > > > > > > > > > > > > > > >Date: Mon, 18 Jul 2005 16:37:34 +0000 >From: "Paula Wilder" >Subject: [Histonet] Bodies in morgue - who does the checking? >To: histonet@lists.utsouthwestern.edu >Message-ID: >Content-Type: text/plain; format=flowed > >Hi everyone! > >Any feedback on whose responsibility it is to check bodies in the morgue >at >your respective instituition would be greatly appreciated. So far, by >phoning neighboring hospitals, I have found that Security, a diener >service, >or the Pathology Assistants are the ones responsible. Any help in this >would truly be greatly appreciated! Thanks so much! > >Paula Wilder >St.Joseph Medical Center >Towson, MD 21204 >410-337-1741 > > > > > >------------------------------ > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >End of Histonet Digest, Vol 20, Issue 21 >**************************************** > > >------------------------------ > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >End of Histonet Digest, Vol 20, Issue 23 >**************************************** > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From eva.alstromer <@t> histolab.se Wed Jul 20 08:40:25 2005 From: eva.alstromer <@t> histolab.se (=?iso-8859-1?Q?Eva_Alstr=F6mer?=) Date: Fri Sep 16 15:25:20 2005 Subject: SV: [Histonet] chatter in fly head paraffin section In-Reply-To: <1121863820.42de488c68b81@webmail.pobox.upenn.edu> Message-ID: <002301c58d30$95914780$70b6a8c0@HISSWE.lokal> If you use 4% buffered NBF be sure that you have washed out the salt crystals from the 4% buffered NBF as this cristals can cause scratehes in cutting. Best regards, Eva Alstromer -----Ursprungligt meddelande----- Fr?n: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] F?r Zhenming Yu Skickat: den 20 juli 2005 14:50 Till: histonet@lists.utsouthwestern.edu ?mne: [Histonet] chatter in fly head paraffin section Dear Histonetters, I'm looking for some guidance in obtaining good paraffin sections from fruitfly brain/eye. We repeated saw chatter in the brain region and are desperately trying to troubleshoot the problem. We have tried 10% NBF, 4% paraformaldehyde, Bourin's for fixation. And for tissue processing, we used the following schedule: 70% Alcohol 3 hours 95% Alcohol 2 hours 100% Alcohol 2 hours 100% Alcohol 2 hours ? 100% Alcohol 1 hours Xylene 1 hour Xylene 1 hour Xylene 1 hour Paraffin(60C): 2 hours Paraffin(60C): 2 hours---we didn't use vacuum at this step considering fly heads are quite small. Could this cause inadequate praffin infiltration and chatter in sections? The technician did leave paraffin in the processor melting at 60C for long period of time--she never shut off the power after the cycle is done. She also always left the embedding station on with paraffin melting at 62C. Could this cause deterioration of paraffin? For sectioning, we've tried using brand new blade, adjusting clearance angle, slowing down cutting speed etc. None seems to help. ?? Any suggestion would be greatly appreciated. Thank you all. **************************** Zhenming Yu Univ. of Penn Philadelphia, PA **************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MadaryJ <@t> MedImmune.com Wed Jul 20 08:41:05 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Hema tek wright giemsa stainer calling vendors! Message-ID: <746FDB897740814EA52BDDCB5ED1DDBCBB16FE@medimmune4.medimmune.com> Any vendor or anyone out there that can get me a quote on a wright giemsa stainer, we are in the market. Specifically the Hema Tek 2000 unless someone knows of a better one. I am in the DC MD area. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, July 19, 2005 11:10 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 22 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: Bodies in morgue - who does the checking? (Charles.Embrey) 2. How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the (Billie Zimmerman) 3. RE: Bodies in morgue - who does the checking? (Horn, Hazel V) 4. liquid nitrogen (manal galal) 5. RE: How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the (Sebree Linda A.) 6. CAP inspections (Histology SLU) 7. seeking part time position (Goodwin, Diana) 8. How is everyone carrying.... (DMBCMP@aol.com) 9. Workloads vs Hiso staffing (Maray Weirauch) 10. Re: mouse brain (Geoff McAuliffe) 11. RE: Histonet Digest, Vol 20, Issue 21 (Goodwin, Diana) 12. PGP9.5 (Dodson, Cecelia) 13. Re: liquid nitrogen (Karen Weidenheim) 14. RE: liquid nitrogen (Smith, Allen) 15. Re: CAP inspections (Joe Nocito) 16. Re: How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the (Joe Nocito) 17. RE: Bodies in morgue - who does the checking? (Rogerson Kemlo (ELHT) Pathology) 18. unsubscribe (Dunn-Jena, Patsy A) 19. Cytology Immuno's (Jean Gillson) 20. Problem with Eosin Y staining (Habitzruther, Michael) 21. RE: Problem with Eosin Y staining (Rogerson Kemlo (ELHT) Pathology) ---------------------------------------------------------------------- Message: 1 Date: Mon, 18 Jul 2005 12:01:38 -0500 From: "Charles.Embrey" Subject: RE: [Histonet] Bodies in morgue - who does the checking? To: "Paula Wilder" , Message-ID: Content-Type: text/plain; charset="us-ascii" It depends what you mean. Check bodies into the morgue or check bodies that are in the morgue. Here we have a house officer (nurse) that coordinates the release of bodies from the hospital through security. Sometimes if the funeral home can come quickly enough then they take them from the hospital room. If they can't come quickly the ward staff put the body into the morgue with the help of security. I keep an eye on who is in my morgue so I can get a jump on any autopsy that might be required. Since I do all the weekday autopsies I don't like surprises. My dieners are all part-time (as needed) and my pathology assistant is only concerned with setting up gross and logging specimens into the computer (I can't get her to go near the morgue). I did work at one place in Ohio where the diener was full time and actually had his office in the morgue. Hope this helps.... Charles Embrey, PA(ASCP) Pathologists' Assistant Histology Manager -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Wilder Sent: Monday, July 18, 2005 11:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bodies in morgue - who does the checking? Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Mon, 18 Jul 2005 13:09:53 -0400 From: "Billie Zimmerman" Subject: [Histonet] How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the To: Message-ID: Content-Type: text/plain; charset=US-ASCII How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the technical quality of histologic preparations by the pathologist? This req. came out in April so I was wondering the best way to handle this. Thanks, Billie Zimmerman Medical College of GA ------------------------------ Message: 3 Date: Mon, 18 Jul 2005 12:13:02 -0500 From: "Horn, Hazel V" Subject: RE: [Histonet] Bodies in morgue - who does the checking? To: "Charles.Embrey" , "Paula Wilder" , histonet@lists.utsouthwestern.edu Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDDA4@EMAIL.archildrens.org> Content-Type: text/plain; charset=us-ascii At our hospital is is histology's responsibility to check the morgue on day shift, Monday-Friday. On evenings, nights and weekends our Nursing service does it. BUT we can check the morgue, via computer. We have the option to pull up and expired patient list. If we know someone has expired we can go down to the morgue and check to see if there is autopsy. We also release bodies on dayshift to the funeral homes. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, July 18, 2005 12:02 PM To: Paula Wilder; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Bodies in morgue - who does the checking? It depends what you mean. Check bodies into the morgue or check bodies that are in the morgue. Here we have a house officer (nurse) that coordinates the release of bodies from the hospital through security. Sometimes if the funeral home can come quickly enough then they take them from the hospital room. If they can't come quickly the ward staff put the body into the morgue with the help of security. I keep an eye on who is in my morgue so I can get a jump on any autopsy that might be required. Since I do all the weekday autopsies I don't like surprises. My dieners are all part-time (as needed) and my pathology assistant is only concerned with setting up gross and logging specimens into the computer (I can't get her to go near the morgue). I did work at one place in Ohio where the diener was full time and actually had his office in the morgue. Hope this helps.... Charles Embrey, PA(ASCP) Pathologists' Assistant Histology Manager -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Wilder Sent: Monday, July 18, 2005 11:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bodies in morgue - who does the checking? Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== ------------------------------ Message: 4 Date: Sat, 16 Jul 2005 05:06:46 -0700 (PDT) From: manal galal Subject: [Histonet] liquid nitrogen To: Histonet@Pathology.swmed.edu Message-ID: <20050716120646.93890.qmail@web50204.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi all, I wanted to ask about how to use liquid nitogen in freezing muscle biopsies. I mean what does it come in? How much do I use? Do I discard it after I use it? What kind of container do I use? How long do I put the biopsy in it? How do I store it? By the way I freeze muscle in the quick freezing chamber of the cryostat, and am getting very little to no artifacts. But anyone that learns of how I freeeze biopsies says that it will give me disasterous resuls. Do you think liquid nitrogen will be superior to my old method. Thanks in advance manal __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 5 Date: Mon, 18 Jul 2005 12:36:12 -0500 From: "Sebree Linda A." Subject: RE: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the To: "Billie Zimmerman" , Message-ID: Content-Type: text/plain; charset="us-ascii" As technologists, we sign off on the run sheets of each instrument batch of slides. This indicates that the positive and negative controls have stained as expected and that the patient unknowns are of high quality. If something is not as it should be at this point, we may decide on our own to rerun a case or slides and inform the pathologist or we discuss it with the requesting pathologist and come up with a plan of action, which is documented on the run sheet. When the slides are delivered to the pathologists, a QA form accompanies them and the pathologist is required to indicate "satisfactory" or "unsatisfactory" and sign his name. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Billie Zimmerman Sent: Monday, July 18, 2005 12:10 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the technical quality of histologic preparations by the pathologist? This req. came out in April so I was wondering the best way to handle this. Thanks, Billie Zimmerman Medical College of GA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 18 Jul 2005 10:38:19 -0700 (PDT) From: Histology SLU Subject: [Histonet] CAP inspections To: histonet@lists.utsouthwestern.edu Message-ID: <20050718173819.2450.qmail@web51011.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello All: This is directed at those of you that may have had your CAP inspection in the last year. I have been hearing that the inspectors have been, shall we say, interpreting some of the checklist questions in an unusual manner. My question to you all is, were there any questions where you got "tagged" for something that you had always done and had passed previously? Also, can you share ANY experiences that you had through your CAP inspection good or bad? As always, thank you for your insights and opinions. This is a super group and I always appreciate your comments. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 7 Date: Mon, 18 Jul 2005 11:56:14 -0400 From: "Goodwin, Diana" Subject: [Histonet] seeking part time position To: "Histonet (E-mail)" Message-ID: <992899E9EC268548AB8DDE246AF88473055F5266@PAHEX01.uphs.upenn.edu> Content-Type: text/plain; charset="iso-8859-1" Experienced histo tech seeking part time position in Philadelphia area. If interested, contact Coleen Kiehl at 215-829-6940. ------------------------------ Message: 8 Date: Mon, 18 Jul 2005 14:30:48 EDT From: DMBCMP@aol.com Subject: [Histonet] How is everyone carrying.... To: Histonet@lists.utsouthwestern.edu Message-ID: <129.6119d036.300d4f58@aol.com> Content-Type: text/plain; charset="US-ASCII" Hi: Just wanted to ad that we, also, send a QC sheet to each of our pathologists every morning with their first run of slides. They respond on the sheet, even though they speak to us about any problem, of course. This is a way for us to keep a record for CAP. Dannie Blake, HT Community Medical Centers Fresno, Ca ------------------------------ Message: 9 Date: Mon, 18 Jul 2005 14:44:41 -0400 From: "Maray Weirauch" Subject: [Histonet] Workloads vs Hiso staffing To: Message-ID: Content-Type: text/plain; charset=US-ASCII ** PRIVATE ** Our hospital is conducting an analysis of laboratory workload, workflow and staffing. This will include Histology as well, and we are being asked for input with any national benchmarks available for surgical histology tasks (i.e. average number cases accessioned, blocks embedded, slides cut../some type of time unit) Does anyone know of some realistic published values or ranges? Thanks! ------------------------------ Message: 10 Date: Mon, 18 Jul 2005 14:42:07 -0400 From: Geoff McAuliffe Subject: Re: [Histonet] mouse brain To: "Atoska S. Gentry" Cc: Histonet Message-ID: <42DBF7FF.8080406@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi Atoska: Hydrocephalus is not uncommon in mice, perhaps that is the source of your concern? I suppose it could be manifested more on one side than the other? Maybe you could post a photo? Geoff Atoska S. Gentry wrote: > Hello, please have any of you who process & section paraformaldehyde > fixed, paraffin embedded mouse brain experienced asymmetry of the > right & left hemispheres? Recently our coronal mouse brain sections > which appear symmetrical upon gross trim and after initial facing of > paraffin are asymmetrical after staining. I've rotated the block > holder to the limit and sectioned the most rostral hemisphere at > higher micron thickness to accommodate. However, to my dismay and > disappointment none of this has helped very much. Any pointers in > remedying this situation ASAP will be greatly appreciated. Thanks, Atoska > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** ------------------------------ Message: 11 Date: Mon, 18 Jul 2005 15:27:58 -0400 From: "Goodwin, Diana" Subject: [Histonet] RE: Histonet Digest, Vol 20, Issue 21 To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <992899E9EC268548AB8DDE246AF88473055F526B@PAHEX01.uphs.upenn.edu> Content-Type: text/plain; charset="iso-8859-1" At our institution, the Anatomic Path dept. is responsible for "morgue patient inventory" and releasing bodies. This has been a point of contention for years, ever since they eliminated the position of "morgue attendant", but one that our administration insists is the pathology dept's responsibility. At the institution where I previously worked, body release was handled by the security dept, with the central switchboard in control of the status of decedents in the hospital via the HIS. Good luck! Diana G. Goodwin Department of Pathology Pennsylvania Hospital 800 Spruce St., Preston 655-C Philadelphia, PA? 19107 ? ph:? 215-829-6532 fax:? 215-829-7564 e-mail:? goodwind@pahosp.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Sent: Monday, July 18, 2005 1:01 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 21 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: dissecting board (Malam Jacqueline) 2. Need help with PGP9.5 on FFPE skin (Nicola Cragg) 3. RE: automated microtomes (Bonner, Janet) 4. Re: Dissection boards (Fred Underwood) 5. Bodies in morgue - who does the checking? (Paula Wilder) ---------------------------------------------------------------------- Message: 1 Date: Mon, 18 Jul 2005 12:39:23 +0100 From: Malam Jacqueline Subject: [Histonet] RE: dissecting board To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Thermo Shandon do 2 colours and 2 sizes of polyethylene board. They're easy to clean and last. We actually got ours from a butchers' suppliers! -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 17 July 2005 18:07 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 19 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Dissection boards (Katia Cristina Catunda) ---------------------------------------------------------------------- Message: 1 Date: Sat, 16 Jul 2005 22:20:15 -0300 From: "Katia Cristina Catunda" Subject: [Histonet] Dissection boards To: Message-ID: <019801c58a6d$b0bbc050$a279fea9@privatexx> Content-Type: text/plain; charset="iso-8859-1" In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 19 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 2 Date: Mon, 18 Jul 2005 13:39:29 +0100 From: "Nicola Cragg" Subject: [Histonet] Need help with PGP9.5 on FFPE skin To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello, I'm posting this message with some embarrassment and apologise for raising anybody's hope of a quck solution........ I have been trying to optimise PGP9.5 on FFPE skin samples (3 micron), using a mouse monoclonal (clone 10A1) from Neuromics. Despite my initial excitement expressed in an earlier posting, I have since found some very good photos of PGP9.5 staining in published work which has put mine to shame and has made me realise that my IHC is not working well enough at all. I've found an improvement with the waterbath antigen retrieval, as the dermis remains intact and there is some specific staining (I think) in the reticular dermis, which wasn't apparent with microwave antigen retrieval. However, it appears that there seems to be more of a trend of using free-floating 50 micron sections. Is that because routine FFPE sections are difficult to stain for this antigen? Has anyone got it to work on FFPE samples? I was quite hopeful that it would work from reading the datasheet and I have also found that Dako make a Rabbit polyclonal for FFPE sections and they're usually excellent antibodies so I'm hoping to try. Any advice or suggestions will be gratefully received. Regards, Nicola Cragg ------------------------------ Message: 3 Date: Mon, 18 Jul 2005 08:43:37 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] automated microtomes To: "'Sennello, Gina '" , "'histonet-bounces@lists.utsouthwestern.edu '" , "'Histonet Histonet (E-mail) '" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4325@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 We have used the Leica's with great success and few if any call backs after more than eight years!! Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet Histonet (E-mail) Sent: 7/13/2005 3:12 PM Subject: [Histonet] automated microtomes I am in the market for fully automated microtome and have looked at Leica's RM2255, Thermo-Electron's Finesse ME and Microm's 355S. Does anyone have any strong feels for against any of these instruments? Thanks in advance for you opinions and help. Gina Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 18 Jul 2005 12:29:54 -0400 From: "Fred Underwood" Subject: Re: [Histonet] Dissection boards To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I've found that kitchen cutting boards from your local department store works well, and it's cheaper than from a medical supplier. Fred >>> "Katia Cristina Catunda" 07/16/05 09:20PM >>> In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 18 Jul 2005 16:37:34 +0000 From: "Paula Wilder" Subject: [Histonet] Bodies in morgue - who does the checking? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 21 **************************************** ------------------------------ Message: 12 Date: Mon, 18 Jul 2005 15:26:05 -0500 From: "Dodson, Cecelia" Subject: [Histonet] PGP9.5 To: Message-ID: <6A65BA49B1BEEA419A3B4D16DFA8EC5A08283D@CHEX1.chp.clarian.org> Content-Type: text/plain; charset="iso-8859-1" We are staining PGP9.5 on a regular bases using a polyclonal antibody we purchase from Biogenesis. It is antigen retrieved in a steamer for 15 minutes then stained with LSAB2 from Dako. Cecelia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, July 18, 2005 12:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 21 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: dissecting board (Malam Jacqueline) 2. Need help with PGP9.5 on FFPE skin (Nicola Cragg) 3. RE: automated microtomes (Bonner, Janet) 4. Re: Dissection boards (Fred Underwood) 5. Bodies in morgue - who does the checking? (Paula Wilder) ---------------------------------------------------------------------- Message: 1 Date: Mon, 18 Jul 2005 12:39:23 +0100 From: Malam Jacqueline Subject: [Histonet] RE: dissecting board To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Thermo Shandon do 2 colours and 2 sizes of polyethylene board. They're easy to clean and last. We actually got ours from a butchers' suppliers! -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 17 July 2005 18:07 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 19 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Dissection boards (Katia Cristina Catunda) ---------------------------------------------------------------------- Message: 1 Date: Sat, 16 Jul 2005 22:20:15 -0300 From: "Katia Cristina Catunda" Subject: [Histonet] Dissection boards To: Message-ID: <019801c58a6d$b0bbc050$a279fea9@privatexx> Content-Type: text/plain; charset="iso-8859-1" In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 19 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 2 Date: Mon, 18 Jul 2005 13:39:29 +0100 From: "Nicola Cragg" Subject: [Histonet] Need help with PGP9.5 on FFPE skin To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello, I'm posting this message with some embarrassment and apologise for raising anybody's hope of a quck solution........ I have been trying to optimise PGP9.5 on FFPE skin samples (3 micron), using a mouse monoclonal (clone 10A1) from Neuromics. Despite my initial excitement expressed in an earlier posting, I have since found some very good photos of PGP9.5 staining in published work which has put mine to shame and has made me realise that my IHC is not working well enough at all. I've found an improvement with the waterbath antigen retrieval, as the dermis remains intact and there is some specific staining (I think) in the reticular dermis, which wasn't apparent with microwave antigen retrieval. However, it appears that there seems to be more of a trend of using free-floating 50 micron sections. Is that because routine FFPE sections are difficult to stain for this antigen? Has anyone got it to work on FFPE samples? I was quite hopeful that it would work from reading the datasheet and I have also found that Dako make a Rabbit polyclonal for FFPE sections and they're usually excellent antibodies so I'm hoping to try. Any advice or suggestions will be gratefully received. Regards, Nicola Cragg ------------------------------ Message: 3 Date: Mon, 18 Jul 2005 08:43:37 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] automated microtomes To: "'Sennello, Gina '" , "'histonet-bounces@lists.utsouthwestern.edu '" , "'Histonet Histonet (E-mail) '" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4325@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 We have used the Leica's with great success and few if any call backs after more than eight years!! Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet Histonet (E-mail) Sent: 7/13/2005 3:12 PM Subject: [Histonet] automated microtomes I am in the market for fully automated microtome and have looked at Leica's RM2255, Thermo-Electron's Finesse ME and Microm's 355S. Does anyone have any strong feels for against any of these instruments? Thanks in advance for you opinions and help. Gina Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 18 Jul 2005 12:29:54 -0400 From: "Fred Underwood" Subject: Re: [Histonet] Dissection boards To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I've found that kitchen cutting boards from your local department store works well, and it's cheaper than from a medical supplier. Fred >>> "Katia Cristina Catunda" 07/16/05 09:20PM >>> In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 18 Jul 2005 16:37:34 +0000 From: "Paula Wilder" Subject: [Histonet] Bodies in morgue - who does the checking? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 21 **************************************** ------------------------------ Message: 13 Date: Mon, 18 Jul 2005 16:51:26 -0400 From: "Karen Weidenheim" Subject: Re: [Histonet] liquid nitrogen To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Manal, The group has been discussing this topic lately. Our lab does classical muscle biopsy freezing with liquid nitrogen and isopentane. (I am happy to hear that you are successful with the cryostat and will have to check this out, but anyway, here is a little synopsis): 1. Liquid nitrogen comes in a large pressurized tank that one orders from a company that provides compressed gases. 2. For freezing, We fill a 2 liter Dewar flask , Cat # 2119, Lab Line Instruments Inc, Melrose Park, IL about 75% full of liquid nitrogen and we suspend our 250 ml stainless steel, very clean beaker that is around 65% full of isopentane in it using a home-made wire contraption made from coat hangers (!) that encircles the beaker and allows us to lower it slowly in to the isopentane (this whole procedure requires 2 people, 1 to handle the isopentane and 1 to do the specimen.) If you are alone, say, on call at night, you can use a ring stand to hold the beaker suspended by the wires. Or you can bend the wire contraption so it hooks on the side of the beaker. When ordering your equipment, you have to be sure, by measuring the internal diameter of the Dewar flask and the external diameter of your stainless beaker, that you can suspend the beaker in the flask comfortably. You have to lower the beaker of isopentane slowly into the liquid nitrogen to avoid the liquid nitrogen boiling into the isopentane and making a mess, at which point you have to start over. 3. We freeze a 0.8 x 0.5 cm portion of oriented skeletal muscle 25 seconds in syrupy isopentane that has ice all around the sides and the bottom. THe textbooks say 5-10 seconds, but we find we need 25 seconds. I think lots of this method is empiric and depends on your location, altitude and humidity. We never reuse the isopentane which, by the way, has to be kept in a flammable cabinet according to regulations. I might do 4 blocks in one aliquot of isopentane but I change it frequently and I think this helps me avoid artifacts. As we cannot put the liquid nitrogen back in the tank, it is not reused either, though if we have several biopsies in a day we use the same liquid nitrogen. Frozen biopsies are stored in a little white box (EM sciences, I think)labeled with patient name and number, in the ultrafreezer. Best of luck with your lab. If your pathologist is satisfied with the method you use now you should stick to it, as I said, I will investigate it with our cryostats here. Karen Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> manal galal 7/16/2005 8:06:46 AM >>> Hi all, I wanted to ask about how to use liquid nitogen in freezing muscle biopsies. I mean what does it come in? How much do I use? Do I discard it after I use it? What kind of container do I use? How long do I put the biopsy in it? How do I store it? By the way I freeze muscle in the quick freezing chamber of the cryostat, and am getting very little to no artifacts. But anyone that learns of how I freeeze biopsies says that it will give me disasterous resuls. Do you think liquid nitrogen will be superior to my old method. Thanks in advance manal __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Mon, 18 Jul 2005 18:33:16 -0400 From: "Smith, Allen" Subject: RE: [Histonet] liquid nitrogen To: "manal galal" Cc: histonet@lists.utsouthwestern.edu Message-ID: <5D2189E74151CC42BEC02906BA8996322B90CB@exchsrv01.barrynet.barry.edu> Content-Type: text/plain; charset="us-ascii" Before trying to handle liquid nitrogen, get someone who uses it to show you how to handle it! Liquid nitrogen is very dangerous when improperly handled. I know of 2 deaths from the abuse of liquid nitrogen. Liquid nitrogen comes in a high pressure cylinder holding anything from 200 ml to 50 liters of liquid nitrogen (which will expand to 0.025 to 6 cubic meters of nitrogen gas). The high pressure cylinder itself is dangerous. If it falls and the valve breaks, escaping gas will turn the cylinder into a missile. The escaping gas can cause severe frostbite if you are near it. Even at a distance, large amounts of escaping nitrogen can displace enough air to suffocate you. Small amounts of liquid nitrogen spilled on yourself will cause severe frostbite. Large amounts can freeze a limb solid and make it as prone to shattering as an ice cube. If you must learn by yourself, buy one of the small bottles sold for dermatological or lecture demonstration use. They usually contain about 50 liters of nitrogen gas compressed down to about 400 ml at 120 Atm. Wear nitrile gloves over leather gloves. Make sure your work area is well ventilated. Clamp a Dewar flask in place and release the nitrogen quickly into the Dewar flask. Half of the nitrogen will escape as a very cold gas. The other half will run into the Dewar flask as a liquid. When the Dewar flask is half full, shut off the nitrogen. Drop the tissue into the liquid nitrogen in the bottom of the Dewar flask. After a few minutes retrieve the tissue with long forceps and put it in your cryostat. Let the liquid nitrogen evaporate. Personally, I don't like liquid nitrogen. The layer of nitrogen gas that forms between the tissue and the liquid nitrogen prevents efficient freezing. If the piece of tissue is large, the outside freezes first; then the freezing of the inside of the tissue cracks the frozen outside. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of manal galal Sent: Saturday, July 16, 2005 8:07 AM To: Histonet@Pathology.swmed.edu Subject: [Histonet] liquid nitrogen Hi all, I wanted to ask about how to use liquid nitogen in freezing muscle biopsies. I mean what does it come in? How much do I use? Do I discard it after I use it? What kind of container do I use? How long do I put the biopsy in it? How do I store it? By the way I freeze muscle in the quick freezing chamber of the cryostat, and am getting very little to no artifacts. But anyone that learns of how I freeeze biopsies says that it will give me disasterous resuls. Do you think liquid nitrogen will be superior to my old method. Thanks in advance manal __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) ------------------------------ Message: 15 Date: Mon, 18 Jul 2005 19:09:37 -0500 From: "Joe Nocito" Subject: Re: [Histonet] CAP inspections To: "Histology SLU" , Message-ID: <000501c58bf6$26c29830$b4bd0b43@yourxhtr8hvc4p> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Susan, I had my inspection in January. Can't speak for others, but, we received Accreditation with Accommodation on the last two inspections. This year we got slammed pretty hard. I think it's because of the incident last year concerning a lab in Maryland. CAP gave the lab an outstanding inspection, but there were many QA/QC issues that the some lab employees complained to CLIA. From what I was told, CAP had to explain itself to Congress. My QA program was torn apart (received compliments on the same programs during the last two inspections). Was written up because some of the temperature charts had missing dates (would you rather me pencil whip these?).Really took it the you know what because a couple of water cultures were not documented. As a matter of fact, we're giving a lecture on "Preparing for a CAP Inspection" at he NSH. It's number 99, the last lecture. Good luck. If I can help, just drop me a line. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Histology SLU" To: Sent: Monday, July 18, 2005 12:38 PM Subject: [Histonet] CAP inspections > Hello All: > > This is directed at those of you that may have had your CAP inspection in > the last year. I have been hearing that the inspectors have been, shall > we say, interpreting some of the checklist questions in an unusual manner. > My question to you all is, were there any questions where you got "tagged" > for something that you had always done and had passed previously? Also, > can you share ANY experiences that you had through your CAP inspection > good or bad? As always, thank you for your insights and opinions. This > is a super group and I always appreciate your comments. > > Susan > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.9.1/51 - Release Date: 7/18/2005 > > ------------------------------ Message: 16 Date: Mon, 18 Jul 2005 19:14:31 -0500 From: "Joe Nocito" Subject: Re: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the To: "Billie Zimmerman" , Message-ID: <008801c58bf7$1d4807d0$b4bd0b43@yourxhtr8hvc4p> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Billie, we send a QC sheet and an H&E control with the first set of slides. The sheet has a large "comments" section. The pathologists write anything in that space from "too many wrinkles" to "not enough hematoxylin" This was accepted by the nice inspector who inspected my lab this January. One of the few items he did like. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Billie Zimmerman" To: Sent: Monday, July 18, 2005 12:09 PM Subject: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the > How is everyone carrying out the new CAP req.? Is there documented > evidence of daily review of the technical quality of histologic > preparations by the pathologist? > > This req. came out in April so I was wondering the best way to handle > this. > > Thanks, > Billie Zimmerman > Medical College of GA > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.9.1/51 - Release Date: 7/18/2005 > > ------------------------------ Message: 17 Date: Tue, 19 Jul 2005 07:49:00 +0100 From: "Rogerson Kemlo (ELHT) Pathology" Subject: RE: [Histonet] Bodies in morgue - who does the checking? To: "Paula Wilder" , Cc: "Richardson Lorraine \(REU\) BurnleyHC" Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD49F@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" In the UK I believe that it is the Police's responsibility to check in the body, strip and take valuables (with a Porter in attendance); these are obviously BID's (Brought in Dead). If 'more expert help' is needed, for example a badly decomposed body or a mutilated one, then an APT (Anatomical Pathology Technician) would be called. Hospital cases are dealt with by the Nursing Staff and placed in the Mortuary by the Porters. Is this a Universal UK procedure? -----Original Message----- From: Paula Wilder [mailto:histo20@hotmail.com] Sent: 18 July 2005 17:38 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bodies in morgue - who does the checking? Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Tue, 19 Jul 2005 09:29:42 -0500 From: "Dunn-Jena, Patsy A" Subject: [Histonet] unsubscribe To: "HistoNet Server" Message-ID: Content-Type: text/plain; charset="us-ascii" ------------------------------ Message: 19 Date: Tue, 19 Jul 2005 15:40:51 +0100 From: Jean Gillson Subject: [Histonet] Cytology Immuno's To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Can I ask you all, what do you use as controls when doing ICC on cytology cytospin preps or smears? Since there isn't much sample, do you use paraffin sections as positive controls and is that adequate control measures? We currently only use paraffin cytoblocks but considering ICC on thin layer cytology preps and wondering what control material to use. Thanks for your help. ------------------------------ Message: 20 Date: Tue, 19 Jul 2005 10:56:39 -0400 From: "Habitzruther, Michael" Subject: [Histonet] Problem with Eosin Y staining To: Cc: "Demant, Peter" Message-ID: <97101976F8A044468CA74FE11883B90E25EAAF@VISTA.roswellpark.org> Content-Type: text/plain; charset="iso-8859-1" Our laboratory is having a problem with the staining of mouse tumor samples using hemotoxylin and Eosin Y (H&E Stain). Everything was going fine until recently. The specific problem we are having is after the cover-slip is put on and the acrymount dries, the Eosin Y stain seems to be bleeding out, leaving only the hemotoxylin staining. A histologist from the institute suggested it may have something to do with the relative humidity in our lab, and therefore allowing the slides to set under a flow-hood may help. Any other suggestions or protocol modifications would be greatly appreciated. Thank you very much. This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. ------------------------------ Message: 21 Date: Tue, 19 Jul 2005 16:05:25 +0100 From: "Rogerson Kemlo (ELHT) Pathology" Subject: RE: [Histonet] Problem with Eosin Y staining To: "Habitzruther, Michael" , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698DCB@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" Eosin is soluble in both water and alcohol isn't it? Well if it bleeds out one of these must be present mustn't it? Solution (forgive the pun) is to eradicate both; I suspect it's alcohol. -----Original Message----- From: Habitzruther, Michael [mailto:Michael.Habitzruther@RoswellPark.org] Sent: 19 July 2005 15:57 To: histonet@lists.utsouthwestern.edu Cc: Demant, Peter Subject: [Histonet] Problem with Eosin Y staining Our laboratory is having a problem with the staining of mouse tumor samples using hemotoxylin and Eosin Y (H&E Stain). Everything was going fine until recently. The specific problem we are having is after the cover-slip is put on and the acrymount dries, the Eosin Y stain seems to be bleeding out, leaving only the hemotoxylin staining. A histologist from the institute suggested it may have something to do with the relative humidity in our lab, and therefore allowing the slides to set under a flow-hood may help. Any other suggestions or protocol modifications would be greatly appreciated. Thank you very much. This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message 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 End of Histonet Digest, Vol 20, Issue 22 **************************************** From bcgpath <@t> rediffmail.com Wed Jul 20 08:54:40 2005 From: bcgpath <@t> rediffmail.com (bcgirish) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] muscle histochemistry Message-ID: <20050720135440.9641.qmail@webmail46.rediffmail.com> ? Hi all I am Dr Girish,toxicological pathologist working for a pharmaceutical company in India. We are recently standardizing myosin atpse histochemistry to study the effect of ppar compounds on skeletal muscle. We are using liquid nitrogen without isopentane to collect the muscle gastrocnemius from rats.The reaction is givinig nonspecific reaction and artifacts. Is using of isopentane must? Is there a better method than the one described by Bancroft? How to preserve the enzyme activity?How long we have to incubate in incubating solution if we are using Bancroft method?we are using atp at the concentration of 5mg/5ml solution b of bancroft method.How long this solution can be kept and at what temperature? Is the pH maintained to be 9.4 or 10.4 (alkaline)? We are getting a sort of cavity in the muscle fibers, is it because of ice crystal artifacts? I am waiting for quick suggestions.Do share ur experience . .......................... REGARDS DR.GIRISH B.CHANDRASHEKAR DEPT OF PRECLINICAL SAFETY EVALUATION DR REDDYS LAB HYDERABAD-49 From PMonfils <@t> Lifespan.org Wed Jul 20 09:04:09 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Can anyone help answer the question? Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717579@lsexch.lsmaster.lifespan.org> Carstairs Stain can be found in Dezna Sheehan's book, Theory and Practice of Histotechnology. I do this stain in my lab. If you don't have the book, I can send you the procedure. I don't do the Fraser Lendrum stain, but it can be found here: http://www.histosearch.com/histonet/Dec03A/Re.HistonetFraser-Lendrum.html > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of HCS > Sent: Tuesday, July 19, 2005 11:16 PM > To: Histonet > Cc: holling@med-in.uni-saarland.de > Subject: [Histonet] Can anyone help answer the question? > > > From: holling@med-in.uni-saarland.de (Nicole Hollinger) > Sent: Wed, 20 Jul 2005 7:10:02 > > I?m a medical technologist in research. I need help. I?m searching a > dyeing instruction for the two special stains: Carstair?s and Fraser > Lendrum. Maybe you can help me.Sincerely yours,Nicole Hollinger > > Please reply to histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From linhines <@t> yahoo.com Wed Jul 20 09:18:58 2005 From: linhines <@t> yahoo.com (Linda Hines) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Rapid geimsa Message-ID: <20050720141858.36383.qmail@web33005.mail.mud.yahoo.com> HI! All I am searching for the chemicals, and maker of the rapid geimsa staining for Helicobacter pylori. Anyone using this stain can let me know it is appreciated. Thanx and Have a Great Day!!! Linda Hines HT Histology Supervisor Specialty Diagnostics Phx. AZ 602-241-5134 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From rgrow <@t> bmnet.com Wed Jul 20 09:26:31 2005 From: rgrow <@t> bmnet.com (rgrow@bmnet.com) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Cytology Immuno's In-Reply-To: <200507200654.j6K6svc0021107@dns1.bmnet.com> Message-ID: In our lab, we are making icc controls to mirror conditions for same type of cytology specimen. Normal tonsil, appendix, and skin (from a breast reduction) are good controls for many IHC's. We just call surgery and ask them to send us some fresh when we see it on the schedule. After gross dictation and sectioning, we take positive charged slides and make touch preps, smears, and swish a section in a thin prep bottle. After slides have dried, I package a few at a time in transport tubes, seal with parafilm and place in a freezer. The thin prep bottle is refrigerated. We document the case number it was collected from, then validate. When we have an ICC on something we don't have cyto controls on, we use standard FFPE controls. So far, it has worked great. Any comments? We also use a daily stain validation sheet with these catagories: acceptable, unacceptable, comments, signature, action taken, and tech. One side is for routine and special stains, flip over and the other side is for IHC's. We take to the pathologist in charge for that week with the first slides of the day. Renee Grow, BA., HT (ASCP) rgrow@bmnet.com Histology Supervisor Blount Memorial Hospital 907 E. Lamar Alexander Pkwy. Maryville, TN 37804-5016 (865) 977-4744 (865) 977-5766 Fax Message: 10 Date: Tue, 19 Jul 2005 14:38:26 -0500 From: "Joe Nocito" Subject: RE: [Histonet] Cytology Immuno's To: "Jean Gillson" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Jean, we use paraffin controls for any cytology specimen. I wish we could keep some cyto prep controls available, but we just don't have the room, nor the specimens. During my CAP inspection in January, it wasn't an issue, 50 other items were an issue, but not using FFPE controls with cytology Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jean Gillson Sent: Tuesday, July 19, 2005 9:41 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Cytology Immuno's Can I ask you all, what do you use as controls when doing ICC on cytology cytospin preps or smears? Since there isn't much sample, do you use paraffin sections as positive controls and is that adequate control measures? We currently only use paraffin cytoblocks but considering ICC on thin layer cytology preps and wondering what control material to use. Thanks for your help. From gu.lang <@t> gmx.at Wed Jul 20 09:36:11 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:21 2005 Subject: AW: [Histonet] Confusing IHC results? In-Reply-To: <3AD0BD3142459B4E9B12CBEAFF2B89B2D89551@lajamrexm01.amer.pfizer.com> Message-ID: Do you use the PAP-technique? I have read, that in some circumstances in the lower dilution the antibodies are so many, that the bridge-antibody is bound with both fab's and there is nothing left for the PAP-complex (bridge doesnt work). So staining will be weaker than in a higher dilution. Just a thought. Gudrun -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Trajkovic, Dusko Gesendet: Dienstag, 19. Juli 2005 23:11 An: 'histonet@lists.utsouthwestern.edu' Betreff: [Histonet] Confusing IHC results? Hi everyone, I just completed an IHC run for Calpain 1 (Santa Cruz), using dilutions 1:800, 1:1000 and 1:2000. I also ran previous runs where the dilutions were 1:100, 1:200, 1:400 and 1:800. I am using a Universal secondary from vector at 1:200 on all of the above slides. My staining results do not vary at all. As a matter of fact, my 1:2000 looks more intense and has more non specific staining than my slide at 1:100. When I run my negative at any of the concentrations to match my primary dilution, it is completely clean. What is going on here? Has anyone ever experienced something like this. Primary is a goat polyclonal. I have tried other secondaries such as, bovine anti goat, horse anti goat, rabbit anti goat, donkey anti goat, with either negative results or sections staining completely brown. Any suggestions, advice, email lashings would be greatly appreciated. Person to come up with a solution will receive a hug and a kiss, OR a couple of drinks at the bar, during the NSH symposium. It's your choice. Hell, you can take a chance and go for both. See, I'm going crazy trying to figure this out!!! Thank you, Dusko Trajkovic 1-858-638-6202 LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Jul 20 09:37:06 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Cytology Immuno's Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698DD8@elht-exch1.xelht.nhs.uk> I assume in the UK we would have to get permission to sample and retain these controls? Organ retention et al is something we Brits must be very aware of. -----Original Message----- From: rgrow@bmnet.com [mailto:rgrow@bmnet.com] Sent: 20 July 2005 15:27 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Cytology Immuno's In our lab, we are making icc controls to mirror conditions for same type of cytology specimen. Normal tonsil, appendix, and skin (from a breast reduction) are good controls for many IHC's. We just call surgery and ask them to send us some fresh when we see it on the schedule. After gross dictation and sectioning, we take positive charged slides and make touch preps, smears, and swish a section in a thin prep bottle. After slides have dried, I package a few at a time in transport tubes, seal with parafilm and place in a freezer. The thin prep bottle is refrigerated. We document the case number it was collected from, then validate. When we have an ICC on something we don't have cyto controls on, we use standard FFPE controls. So far, it has worked great. Any comments? We also use a daily stain validation sheet with these catagories: acceptable, unacceptable, comments, signature, action taken, and tech. One side is for routine and special stains, flip over and the other side is for IHC's. We take to the pathologist in charge for that week with the first slides of the day. Renee Grow, BA., HT (ASCP) rgrow@bmnet.com Histology Supervisor Blount Memorial Hospital 907 E. Lamar Alexander Pkwy. Maryville, TN 37804-5016 (865) 977-4744 (865) 977-5766 Fax Message: 10 Date: Tue, 19 Jul 2005 14:38:26 -0500 From: "Joe Nocito" Subject: RE: [Histonet] Cytology Immuno's To: "Jean Gillson" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Jean, we use paraffin controls for any cytology specimen. I wish we could keep some cyto prep controls available, but we just don't have the room, nor the specimens. During my CAP inspection in January, it wasn't an issue, 50 other items were an issue, but not using FFPE controls with cytology Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jean Gillson Sent: Tuesday, July 19, 2005 9:41 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Cytology Immuno's Can I ask you all, what do you use as controls when doing ICC on cytology cytospin preps or smears? Since there isn't much sample, do you use paraffin sections as positive controls and is that adequate control measures? We currently only use paraffin cytoblocks but considering ICC on thin layer cytology preps and wondering what control material to use. Thanks for your help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jhnspam <@t> aol.com Wed Jul 20 09:55:35 2005 From: jhnspam <@t> aol.com (jhnspam@aol.com) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Fumeguard filters In-Reply-To: <000001c58c9c$01a514f0$0301a8c0@RENAD4YK9B8ABE> Message-ID: <8C75B5237E2DF85-588-2A696@MBLK-M09.sysops.aol.com> The filters can be purchased at Thermo Electron (formally Shandon Lipshaw). The phone number is 1-800-245-6212. The web site is thermo.com. -----Original Message----- From: renafail@bellsouth.net To: 'Trisha Emry' ; 'histo' Sent: Tue, 19 Jul 2005 15:56:51 -0400 Subject: RE: [Histonet] Fumeguard filters Trisha, You can get them from Shandon Lipshaw Rena -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Trisha Emry Sent: Tuesday, July 19, 2005 2:16 PM To: histo Subject: [Histonet] Fumeguard filters Hi, We have an older Fumeguard bench top hood. It has a charcole filter that sits in front of the fan. I am trying to find a vendor that can supply the replacement filters. Thanks, Trisha U of WA Seattle _______________________________________________ Histonet mailing list Histonet@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 wecare <@t> qualityhistology.com Wed Jul 20 10:12:17 2005 From: wecare <@t> qualityhistology.com (wecare@qualityhistology.com) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Hema tek wright giemsa stainer calling vendors! References: <746FDB897740814EA52BDDCB5ED1DDBCBB16FE@medimmune4.medimmune.com> Message-ID: <00ee01c58d3d$6b986c80$0300a8c0@internetconnect.net> Dear Mr. Madary, Please consider Midas III Automated slide stainer. It can perform Write-Giemsa Stains. You can get some preliminary information at http://www.emdchemicals.com/analytics/literature/displaylit.asp?location=ar&litfile=dyes_stainers_midas3.htm&Search=midas and also by calling 1-800-222-0342 and asking for Technical Support personnel at EMD Chemicals. It costs about $8,000.00 - $8,500.00 each and can be purchased through Fisher Scientific or VWR catalog. Please contact me at 215-491-0081 if I can be of further help in getting you a quote or additional information. Best regards, Preyas Shah ----- Original Message ----- From: "Madary, Joseph" To: Sent: Wednesday, July 20, 2005 9:41 AM Subject: [Histonet] Hema tek wright giemsa stainer calling vendors! Any vendor or anyone out there that can get me a quote on a wright giemsa stainer, we are in the market. Specifically the Hema Tek 2000 unless someone knows of a better one. I am in the DC MD area. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, July 19, 2005 11:10 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 22 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: Bodies in morgue - who does the checking? (Charles.Embrey) 2. How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the (Billie Zimmerman) 3. RE: Bodies in morgue - who does the checking? (Horn, Hazel V) 4. liquid nitrogen (manal galal) 5. RE: How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the (Sebree Linda A.) 6. CAP inspections (Histology SLU) 7. seeking part time position (Goodwin, Diana) 8. How is everyone carrying.... (DMBCMP@aol.com) 9. Workloads vs Hiso staffing (Maray Weirauch) 10. Re: mouse brain (Geoff McAuliffe) 11. RE: Histonet Digest, Vol 20, Issue 21 (Goodwin, Diana) 12. PGP9.5 (Dodson, Cecelia) 13. Re: liquid nitrogen (Karen Weidenheim) 14. RE: liquid nitrogen (Smith, Allen) 15. Re: CAP inspections (Joe Nocito) 16. Re: How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the (Joe Nocito) 17. RE: Bodies in morgue - who does the checking? (Rogerson Kemlo (ELHT) Pathology) 18. unsubscribe (Dunn-Jena, Patsy A) 19. Cytology Immuno's (Jean Gillson) 20. Problem with Eosin Y staining (Habitzruther, Michael) 21. RE: Problem with Eosin Y staining (Rogerson Kemlo (ELHT) Pathology) ---------------------------------------------------------------------- Message: 1 Date: Mon, 18 Jul 2005 12:01:38 -0500 From: "Charles.Embrey" Subject: RE: [Histonet] Bodies in morgue - who does the checking? To: "Paula Wilder" , Message-ID: Content-Type: text/plain; charset="us-ascii" It depends what you mean. Check bodies into the morgue or check bodies that are in the morgue. Here we have a house officer (nurse) that coordinates the release of bodies from the hospital through security. Sometimes if the funeral home can come quickly enough then they take them from the hospital room. If they can't come quickly the ward staff put the body into the morgue with the help of security. I keep an eye on who is in my morgue so I can get a jump on any autopsy that might be required. Since I do all the weekday autopsies I don't like surprises. My dieners are all part-time (as needed) and my pathology assistant is only concerned with setting up gross and logging specimens into the computer (I can't get her to go near the morgue). I did work at one place in Ohio where the diener was full time and actually had his office in the morgue. Hope this helps.... Charles Embrey, PA(ASCP) Pathologists' Assistant Histology Manager -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Wilder Sent: Monday, July 18, 2005 11:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bodies in morgue - who does the checking? Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Mon, 18 Jul 2005 13:09:53 -0400 From: "Billie Zimmerman" Subject: [Histonet] How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the To: Message-ID: Content-Type: text/plain; charset=US-ASCII How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the technical quality of histologic preparations by the pathologist? This req. came out in April so I was wondering the best way to handle this. Thanks, Billie Zimmerman Medical College of GA ------------------------------ Message: 3 Date: Mon, 18 Jul 2005 12:13:02 -0500 From: "Horn, Hazel V" Subject: RE: [Histonet] Bodies in morgue - who does the checking? To: "Charles.Embrey" , "Paula Wilder" , histonet@lists.utsouthwestern.edu Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDDA4@EMAIL.archildrens.org> Content-Type: text/plain; charset=us-ascii At our hospital is is histology's responsibility to check the morgue on day shift, Monday-Friday. On evenings, nights and weekends our Nursing service does it. BUT we can check the morgue, via computer. We have the option to pull up and expired patient list. If we know someone has expired we can go down to the morgue and check to see if there is autopsy. We also release bodies on dayshift to the funeral homes. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, July 18, 2005 12:02 PM To: Paula Wilder; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Bodies in morgue - who does the checking? It depends what you mean. Check bodies into the morgue or check bodies that are in the morgue. Here we have a house officer (nurse) that coordinates the release of bodies from the hospital through security. Sometimes if the funeral home can come quickly enough then they take them from the hospital room. If they can't come quickly the ward staff put the body into the morgue with the help of security. I keep an eye on who is in my morgue so I can get a jump on any autopsy that might be required. Since I do all the weekday autopsies I don't like surprises. My dieners are all part-time (as needed) and my pathology assistant is only concerned with setting up gross and logging specimens into the computer (I can't get her to go near the morgue). I did work at one place in Ohio where the diener was full time and actually had his office in the morgue. Hope this helps.... Charles Embrey, PA(ASCP) Pathologists' Assistant Histology Manager -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Wilder Sent: Monday, July 18, 2005 11:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bodies in morgue - who does the checking? Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================ == ------------------------------ Message: 4 Date: Sat, 16 Jul 2005 05:06:46 -0700 (PDT) From: manal galal Subject: [Histonet] liquid nitrogen To: Histonet@Pathology.swmed.edu Message-ID: <20050716120646.93890.qmail@web50204.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi all, I wanted to ask about how to use liquid nitogen in freezing muscle biopsies. I mean what does it come in? How much do I use? Do I discard it after I use it? What kind of container do I use? How long do I put the biopsy in it? How do I store it? By the way I freeze muscle in the quick freezing chamber of the cryostat, and am getting very little to no artifacts. But anyone that learns of how I freeeze biopsies says that it will give me disasterous resuls. Do you think liquid nitrogen will be superior to my old method. Thanks in advance manal __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 5 Date: Mon, 18 Jul 2005 12:36:12 -0500 From: "Sebree Linda A." Subject: RE: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the To: "Billie Zimmerman" , Message-ID: Content-Type: text/plain; charset="us-ascii" As technologists, we sign off on the run sheets of each instrument batch of slides. This indicates that the positive and negative controls have stained as expected and that the patient unknowns are of high quality. If something is not as it should be at this point, we may decide on our own to rerun a case or slides and inform the pathologist or we discuss it with the requesting pathologist and come up with a plan of action, which is documented on the run sheet. When the slides are delivered to the pathologists, a QA form accompanies them and the pathologist is required to indicate "satisfactory" or "unsatisfactory" and sign his name. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Billie Zimmerman Sent: Monday, July 18, 2005 12:10 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the How is everyone carrying out the new CAP req.? Is there documented evidence of daily review of the technical quality of histologic preparations by the pathologist? This req. came out in April so I was wondering the best way to handle this. Thanks, Billie Zimmerman Medical College of GA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 18 Jul 2005 10:38:19 -0700 (PDT) From: Histology SLU Subject: [Histonet] CAP inspections To: histonet@lists.utsouthwestern.edu Message-ID: <20050718173819.2450.qmail@web51011.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello All: This is directed at those of you that may have had your CAP inspection in the last year. I have been hearing that the inspectors have been, shall we say, interpreting some of the checklist questions in an unusual manner. My question to you all is, were there any questions where you got "tagged" for something that you had always done and had passed previously? Also, can you share ANY experiences that you had through your CAP inspection good or bad? As always, thank you for your insights and opinions. This is a super group and I always appreciate your comments. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 7 Date: Mon, 18 Jul 2005 11:56:14 -0400 From: "Goodwin, Diana" Subject: [Histonet] seeking part time position To: "Histonet (E-mail)" Message-ID: <992899E9EC268548AB8DDE246AF88473055F5266@PAHEX01.uphs.upenn.edu> Content-Type: text/plain; charset="iso-8859-1" Experienced histo tech seeking part time position in Philadelphia area. If interested, contact Coleen Kiehl at 215-829-6940. ------------------------------ Message: 8 Date: Mon, 18 Jul 2005 14:30:48 EDT From: DMBCMP@aol.com Subject: [Histonet] How is everyone carrying.... To: Histonet@lists.utsouthwestern.edu Message-ID: <129.6119d036.300d4f58@aol.com> Content-Type: text/plain; charset="US-ASCII" Hi: Just wanted to ad that we, also, send a QC sheet to each of our pathologists every morning with their first run of slides. They respond on the sheet, even though they speak to us about any problem, of course. This is a way for us to keep a record for CAP. Dannie Blake, HT Community Medical Centers Fresno, Ca ------------------------------ Message: 9 Date: Mon, 18 Jul 2005 14:44:41 -0400 From: "Maray Weirauch" Subject: [Histonet] Workloads vs Hiso staffing To: Message-ID: Content-Type: text/plain; charset=US-ASCII ** PRIVATE ** Our hospital is conducting an analysis of laboratory workload, workflow and staffing. This will include Histology as well, and we are being asked for input with any national benchmarks available for surgical histology tasks (i.e. average number cases accessioned, blocks embedded, slides cut../some type of time unit) Does anyone know of some realistic published values or ranges? Thanks! ------------------------------ Message: 10 Date: Mon, 18 Jul 2005 14:42:07 -0400 From: Geoff McAuliffe Subject: Re: [Histonet] mouse brain To: "Atoska S. Gentry" Cc: Histonet Message-ID: <42DBF7FF.8080406@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi Atoska: Hydrocephalus is not uncommon in mice, perhaps that is the source of your concern? I suppose it could be manifested more on one side than the other? Maybe you could post a photo? Geoff Atoska S. Gentry wrote: > Hello, please have any of you who process & section paraformaldehyde > fixed, paraffin embedded mouse brain experienced asymmetry of the > right & left hemispheres? Recently our coronal mouse brain sections > which appear symmetrical upon gross trim and after initial facing of > paraffin are asymmetrical after staining. I've rotated the block > holder to the limit and sectioned the most rostral hemisphere at > higher micron thickness to accommodate. However, to my dismay and > disappointment none of this has helped very much. Any pointers in > remedying this situation ASAP will be greatly appreciated. Thanks, Atoska > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** ------------------------------ Message: 11 Date: Mon, 18 Jul 2005 15:27:58 -0400 From: "Goodwin, Diana" Subject: [Histonet] RE: Histonet Digest, Vol 20, Issue 21 To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <992899E9EC268548AB8DDE246AF88473055F526B@PAHEX01.uphs.upenn.edu> Content-Type: text/plain; charset="iso-8859-1" At our institution, the Anatomic Path dept. is responsible for "morgue patient inventory" and releasing bodies. This has been a point of contention for years, ever since they eliminated the position of "morgue attendant", but one that our administration insists is the pathology dept's responsibility. At the institution where I previously worked, body release was handled by the security dept, with the central switchboard in control of the status of decedents in the hospital via the HIS. Good luck! Diana G. Goodwin Department of Pathology Pennsylvania Hospital 800 Spruce St., Preston 655-C Philadelphia, PA 19107 ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Sent: Monday, July 18, 2005 1:01 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 21 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: dissecting board (Malam Jacqueline) 2. Need help with PGP9.5 on FFPE skin (Nicola Cragg) 3. RE: automated microtomes (Bonner, Janet) 4. Re: Dissection boards (Fred Underwood) 5. Bodies in morgue - who does the checking? (Paula Wilder) ---------------------------------------------------------------------- Message: 1 Date: Mon, 18 Jul 2005 12:39:23 +0100 From: Malam Jacqueline Subject: [Histonet] RE: dissecting board To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Thermo Shandon do 2 colours and 2 sizes of polyethylene board. They're easy to clean and last. We actually got ours from a butchers' suppliers! -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 17 July 2005 18:07 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 19 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Dissection boards (Katia Cristina Catunda) ---------------------------------------------------------------------- Message: 1 Date: Sat, 16 Jul 2005 22:20:15 -0300 From: "Katia Cristina Catunda" Subject: [Histonet] Dissection boards To: Message-ID: <019801c58a6d$b0bbc050$a279fea9@privatexx> Content-Type: text/plain; charset="iso-8859-1" In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 19 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 2 Date: Mon, 18 Jul 2005 13:39:29 +0100 From: "Nicola Cragg" Subject: [Histonet] Need help with PGP9.5 on FFPE skin To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello, I'm posting this message with some embarrassment and apologise for raising anybody's hope of a quck solution........ I have been trying to optimise PGP9.5 on FFPE skin samples (3 micron), using a mouse monoclonal (clone 10A1) from Neuromics. Despite my initial excitement expressed in an earlier posting, I have since found some very good photos of PGP9.5 staining in published work which has put mine to shame and has made me realise that my IHC is not working well enough at all. I've found an improvement with the waterbath antigen retrieval, as the dermis remains intact and there is some specific staining (I think) in the reticular dermis, which wasn't apparent with microwave antigen retrieval. However, it appears that there seems to be more of a trend of using free-floating 50 micron sections. Is that because routine FFPE sections are difficult to stain for this antigen? Has anyone got it to work on FFPE samples? I was quite hopeful that it would work from reading the datasheet and I have also found that Dako make a Rabbit polyclonal for FFPE sections and they're usually excellent antibodies so I'm hoping to try. Any advice or suggestions will be gratefully received. Regards, Nicola Cragg ------------------------------ Message: 3 Date: Mon, 18 Jul 2005 08:43:37 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] automated microtomes To: "'Sennello, Gina '" , "'histonet-bounces@lists.utsouthwestern.edu '" , "'Histonet Histonet (E-mail) '" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4325@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 We have used the Leica's with great success and few if any call backs after more than eight years!! Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet Histonet (E-mail) Sent: 7/13/2005 3:12 PM Subject: [Histonet] automated microtomes I am in the market for fully automated microtome and have looked at Leica's RM2255, Thermo-Electron's Finesse ME and Microm's 355S. Does anyone have any strong feels for against any of these instruments? Thanks in advance for you opinions and help. Gina Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 18 Jul 2005 12:29:54 -0400 From: "Fred Underwood" Subject: Re: [Histonet] Dissection boards To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I've found that kitchen cutting boards from your local department store works well, and it's cheaper than from a medical supplier. Fred >>> "Katia Cristina Catunda" 07/16/05 09:20PM >>> In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 18 Jul 2005 16:37:34 +0000 From: "Paula Wilder" Subject: [Histonet] Bodies in morgue - who does the checking? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 21 **************************************** ------------------------------ Message: 12 Date: Mon, 18 Jul 2005 15:26:05 -0500 From: "Dodson, Cecelia" Subject: [Histonet] PGP9.5 To: Message-ID: <6A65BA49B1BEEA419A3B4D16DFA8EC5A08283D@CHEX1.chp.clarian.org> Content-Type: text/plain; charset="iso-8859-1" We are staining PGP9.5 on a regular bases using a polyclonal antibody we purchase from Biogenesis. It is antigen retrieved in a steamer for 15 minutes then stained with LSAB2 from Dako. Cecelia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, July 18, 2005 12:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 21 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: dissecting board (Malam Jacqueline) 2. Need help with PGP9.5 on FFPE skin (Nicola Cragg) 3. RE: automated microtomes (Bonner, Janet) 4. Re: Dissection boards (Fred Underwood) 5. Bodies in morgue - who does the checking? (Paula Wilder) ---------------------------------------------------------------------- Message: 1 Date: Mon, 18 Jul 2005 12:39:23 +0100 From: Malam Jacqueline Subject: [Histonet] RE: dissecting board To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Thermo Shandon do 2 colours and 2 sizes of polyethylene board. They're easy to clean and last. We actually got ours from a butchers' suppliers! -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 17 July 2005 18:07 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 19 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Dissection boards (Katia Cristina Catunda) ---------------------------------------------------------------------- Message: 1 Date: Sat, 16 Jul 2005 22:20:15 -0300 From: "Katia Cristina Catunda" Subject: [Histonet] Dissection boards To: Message-ID: <019801c58a6d$b0bbc050$a279fea9@privatexx> Content-Type: text/plain; charset="iso-8859-1" In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 19 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 2 Date: Mon, 18 Jul 2005 13:39:29 +0100 From: "Nicola Cragg" Subject: [Histonet] Need help with PGP9.5 on FFPE skin To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello, I'm posting this message with some embarrassment and apologise for raising anybody's hope of a quck solution........ I have been trying to optimise PGP9.5 on FFPE skin samples (3 micron), using a mouse monoclonal (clone 10A1) from Neuromics. Despite my initial excitement expressed in an earlier posting, I have since found some very good photos of PGP9.5 staining in published work which has put mine to shame and has made me realise that my IHC is not working well enough at all. I've found an improvement with the waterbath antigen retrieval, as the dermis remains intact and there is some specific staining (I think) in the reticular dermis, which wasn't apparent with microwave antigen retrieval. However, it appears that there seems to be more of a trend of using free-floating 50 micron sections. Is that because routine FFPE sections are difficult to stain for this antigen? Has anyone got it to work on FFPE samples? I was quite hopeful that it would work from reading the datasheet and I have also found that Dako make a Rabbit polyclonal for FFPE sections and they're usually excellent antibodies so I'm hoping to try. Any advice or suggestions will be gratefully received. Regards, Nicola Cragg ------------------------------ Message: 3 Date: Mon, 18 Jul 2005 08:43:37 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] automated microtomes To: "'Sennello, Gina '" , "'histonet-bounces@lists.utsouthwestern.edu '" , "'Histonet Histonet (E-mail) '" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4325@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 We have used the Leica's with great success and few if any call backs after more than eight years!! Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet Histonet (E-mail) Sent: 7/13/2005 3:12 PM Subject: [Histonet] automated microtomes I am in the market for fully automated microtome and have looked at Leica's RM2255, Thermo-Electron's Finesse ME and Microm's 355S. Does anyone have any strong feels for against any of these instruments? Thanks in advance for you opinions and help. Gina Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 18 Jul 2005 12:29:54 -0400 From: "Fred Underwood" Subject: Re: [Histonet] Dissection boards To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I've found that kitchen cutting boards from your local department store works well, and it's cheaper than from a medical supplier. Fred >>> "Katia Cristina Catunda" 07/16/05 09:20PM >>> In our lab we still use wood-dissecting boards but we really want to change it!! (wood can be very very very dirty after some years even if we submit the boards to an intensive descontaminating process). Would like some tips about what kind of material we should use it and what is the best option, buy it from distributors like Mopec or to pay for someone to make them? Some simple questions that makes a lot of difference for us... Thanks Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 18 Jul 2005 16:37:34 +0000 From: "Paula Wilder" Subject: [Histonet] Bodies in morgue - who does the checking? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 21 **************************************** ------------------------------ Message: 13 Date: Mon, 18 Jul 2005 16:51:26 -0400 From: "Karen Weidenheim" Subject: Re: [Histonet] liquid nitrogen To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Manal, The group has been discussing this topic lately. Our lab does classical muscle biopsy freezing with liquid nitrogen and isopentane. (I am happy to hear that you are successful with the cryostat and will have to check this out, but anyway, here is a little synopsis): 1. Liquid nitrogen comes in a large pressurized tank that one orders from a company that provides compressed gases. 2. For freezing, We fill a 2 liter Dewar flask , Cat # 2119, Lab Line Instruments Inc, Melrose Park, IL about 75% full of liquid nitrogen and we suspend our 250 ml stainless steel, very clean beaker that is around 65% full of isopentane in it using a home-made wire contraption made from coat hangers (!) that encircles the beaker and allows us to lower it slowly in to the isopentane (this whole procedure requires 2 people, 1 to handle the isopentane and 1 to do the specimen.) If you are alone, say, on call at night, you can use a ring stand to hold the beaker suspended by the wires. Or you can bend the wire contraption so it hooks on the side of the beaker. When ordering your equipment, you have to be sure, by measuring the internal diameter of the Dewar flask and the external diameter of your stainless beaker, that you can suspend the beaker in the flask comfortably. You have to lower the beaker of isopentane slowly into the liquid nitrogen to avoid the liquid nitrogen boiling into the isopentane and making a mess, at which point you have to start over. 3. We freeze a 0.8 x 0.5 cm portion of oriented skeletal muscle 25 seconds in syrupy isopentane that has ice all around the sides and the bottom. THe textbooks say 5-10 seconds, but we find we need 25 seconds. I think lots of this method is empiric and depends on your location, altitude and humidity. We never reuse the isopentane which, by the way, has to be kept in a flammable cabinet according to regulations. I might do 4 blocks in one aliquot of isopentane but I change it frequently and I think this helps me avoid artifacts. As we cannot put the liquid nitrogen back in the tank, it is not reused either, though if we have several biopsies in a day we use the same liquid nitrogen. Frozen biopsies are stored in a little white box (EM sciences, I think)labeled with patient name and number, in the ultrafreezer. Best of luck with your lab. If your pathologist is satisfied with the method you use now you should stick to it, as I said, I will investigate it with our cryostats here. Karen Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> manal galal 7/16/2005 8:06:46 AM >>> Hi all, I wanted to ask about how to use liquid nitogen in freezing muscle biopsies. I mean what does it come in? How much do I use? Do I discard it after I use it? What kind of container do I use? How long do I put the biopsy in it? How do I store it? By the way I freeze muscle in the quick freezing chamber of the cryostat, and am getting very little to no artifacts. But anyone that learns of how I freeeze biopsies says that it will give me disasterous resuls. Do you think liquid nitrogen will be superior to my old method. Thanks in advance manal __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Mon, 18 Jul 2005 18:33:16 -0400 From: "Smith, Allen" Subject: RE: [Histonet] liquid nitrogen To: "manal galal" Cc: histonet@lists.utsouthwestern.edu Message-ID: <5D2189E74151CC42BEC02906BA8996322B90CB@exchsrv01.barrynet.barry.edu> Content-Type: text/plain; charset="us-ascii" Before trying to handle liquid nitrogen, get someone who uses it to show you how to handle it! Liquid nitrogen is very dangerous when improperly handled. I know of 2 deaths from the abuse of liquid nitrogen. Liquid nitrogen comes in a high pressure cylinder holding anything from 200 ml to 50 liters of liquid nitrogen (which will expand to 0.025 to 6 cubic meters of nitrogen gas). The high pressure cylinder itself is dangerous. If it falls and the valve breaks, escaping gas will turn the cylinder into a missile. The escaping gas can cause severe frostbite if you are near it. Even at a distance, large amounts of escaping nitrogen can displace enough air to suffocate you. Small amounts of liquid nitrogen spilled on yourself will cause severe frostbite. Large amounts can freeze a limb solid and make it as prone to shattering as an ice cube. If you must learn by yourself, buy one of the small bottles sold for dermatological or lecture demonstration use. They usually contain about 50 liters of nitrogen gas compressed down to about 400 ml at 120 Atm. Wear nitrile gloves over leather gloves. Make sure your work area is well ventilated. Clamp a Dewar flask in place and release the nitrogen quickly into the Dewar flask. Half of the nitrogen will escape as a very cold gas. The other half will run into the Dewar flask as a liquid. When the Dewar flask is half full, shut off the nitrogen. Drop the tissue into the liquid nitrogen in the bottom of the Dewar flask. After a few minutes retrieve the tissue with long forceps and put it in your cryostat. Let the liquid nitrogen evaporate. Personally, I don't like liquid nitrogen. The layer of nitrogen gas that forms between the tissue and the liquid nitrogen prevents efficient freezing. If the piece of tissue is large, the outside freezes first; then the freezing of the inside of the tissue cracks the frozen outside. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of manal galal Sent: Saturday, July 16, 2005 8:07 AM To: Histonet@Pathology.swmed.edu Subject: [Histonet] liquid nitrogen Hi all, I wanted to ask about how to use liquid nitogen in freezing muscle biopsies. I mean what does it come in? How much do I use? Do I discard it after I use it? What kind of container do I use? How long do I put the biopsy in it? How do I store it? By the way I freeze muscle in the quick freezing chamber of the cryostat, and am getting very little to no artifacts. But anyone that learns of how I freeeze biopsies says that it will give me disasterous resuls. Do you think liquid nitrogen will be superior to my old method. Thanks in advance manal __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) ------------------------------ Message: 15 Date: Mon, 18 Jul 2005 19:09:37 -0500 From: "Joe Nocito" Subject: Re: [Histonet] CAP inspections To: "Histology SLU" , Message-ID: <000501c58bf6$26c29830$b4bd0b43@yourxhtr8hvc4p> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Susan, I had my inspection in January. Can't speak for others, but, we received Accreditation with Accommodation on the last two inspections. This year we got slammed pretty hard. I think it's because of the incident last year concerning a lab in Maryland. CAP gave the lab an outstanding inspection, but there were many QA/QC issues that the some lab employees complained to CLIA. From what I was told, CAP had to explain itself to Congress. My QA program was torn apart (received compliments on the same programs during the last two inspections). Was written up because some of the temperature charts had missing dates (would you rather me pencil whip these?).Really took it the you know what because a couple of water cultures were not documented. As a matter of fact, we're giving a lecture on "Preparing for a CAP Inspection" at he NSH. It's number 99, the last lecture. Good luck. If I can help, just drop me a line. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Histology SLU" To: Sent: Monday, July 18, 2005 12:38 PM Subject: [Histonet] CAP inspections > Hello All: > > This is directed at those of you that may have had your CAP inspection in > the last year. I have been hearing that the inspectors have been, shall > we say, interpreting some of the checklist questions in an unusual manner. > My question to you all is, were there any questions where you got "tagged" > for something that you had always done and had passed previously? Also, > can you share ANY experiences that you had through your CAP inspection > good or bad? As always, thank you for your insights and opinions. This > is a super group and I always appreciate your comments. > > Susan > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.9.1/51 - Release Date: 7/18/2005 > > ------------------------------ Message: 16 Date: Mon, 18 Jul 2005 19:14:31 -0500 From: "Joe Nocito" Subject: Re: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the To: "Billie Zimmerman" , Message-ID: <008801c58bf7$1d4807d0$b4bd0b43@yourxhtr8hvc4p> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Billie, we send a QC sheet and an H&E control with the first set of slides. The sheet has a large "comments" section. The pathologists write anything in that space from "too many wrinkles" to "not enough hematoxylin" This was accepted by the nice inspector who inspected my lab this January. One of the few items he did like. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Billie Zimmerman" To: Sent: Monday, July 18, 2005 12:09 PM Subject: [Histonet] How is everyone carrying out the new CAP req.? Is theredocumented evidence of daily review of the > How is everyone carrying out the new CAP req.? Is there documented > evidence of daily review of the technical quality of histologic > preparations by the pathologist? > > This req. came out in April so I was wondering the best way to handle > this. > > Thanks, > Billie Zimmerman > Medical College of GA > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.323 / Virus Database: 267.9.1/51 - Release Date: 7/18/2005 > > ------------------------------ Message: 17 Date: Tue, 19 Jul 2005 07:49:00 +0100 From: "Rogerson Kemlo (ELHT) Pathology" Subject: RE: [Histonet] Bodies in morgue - who does the checking? To: "Paula Wilder" , Cc: "Richardson Lorraine \(REU\) BurnleyHC" Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD49F@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" In the UK I believe that it is the Police's responsibility to check in the body, strip and take valuables (with a Porter in attendance); these are obviously BID's (Brought in Dead). If 'more expert help' is needed, for example a badly decomposed body or a mutilated one, then an APT (Anatomical Pathology Technician) would be called. Hospital cases are dealt with by the Nursing Staff and placed in the Mortuary by the Porters. Is this a Universal UK procedure? -----Original Message----- From: Paula Wilder [mailto:histo20@hotmail.com] Sent: 18 July 2005 17:38 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bodies in morgue - who does the checking? Hi everyone! Any feedback on whose responsibility it is to check bodies in the morgue at your respective instituition would be greatly appreciated. So far, by phoning neighboring hospitals, I have found that Security, a diener service, or the Pathology Assistants are the ones responsible. Any help in this would truly be greatly appreciated! Thanks so much! Paula Wilder St.Joseph Medical Center Towson, MD 21204 410-337-1741 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Tue, 19 Jul 2005 09:29:42 -0500 From: "Dunn-Jena, Patsy A" Subject: [Histonet] unsubscribe To: "HistoNet Server" Message-ID: Content-Type: text/plain; charset="us-ascii" ------------------------------ Message: 19 Date: Tue, 19 Jul 2005 15:40:51 +0100 From: Jean Gillson Subject: [Histonet] Cytology Immuno's To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Can I ask you all, what do you use as controls when doing ICC on cytology cytospin preps or smears? Since there isn't much sample, do you use paraffin sections as positive controls and is that adequate control measures? We currently only use paraffin cytoblocks but considering ICC on thin layer cytology preps and wondering what control material to use. Thanks for your help. ------------------------------ Message: 20 Date: Tue, 19 Jul 2005 10:56:39 -0400 From: "Habitzruther, Michael" Subject: [Histonet] Problem with Eosin Y staining To: Cc: "Demant, Peter" Message-ID: <97101976F8A044468CA74FE11883B90E25EAAF@VISTA.roswellpark.org> Content-Type: text/plain; charset="iso-8859-1" Our laboratory is having a problem with the staining of mouse tumor samples using hemotoxylin and Eosin Y (H&E Stain). Everything was going fine until recently. The specific problem we are having is after the cover-slip is put on and the acrymount dries, the Eosin Y stain seems to be bleeding out, leaving only the hemotoxylin staining. A histologist from the institute suggested it may have something to do with the relative humidity in our lab, and therefore allowing the slides to set under a flow-hood may help. Any other suggestions or protocol modifications would be greatly appreciated. Thank you very much. This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. ------------------------------ Message: 21 Date: Tue, 19 Jul 2005 16:05:25 +0100 From: "Rogerson Kemlo (ELHT) Pathology" Subject: RE: [Histonet] Problem with Eosin Y staining To: "Habitzruther, Michael" , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698DCB@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" Eosin is soluble in both water and alcohol isn't it? Well if it bleeds out one of these must be present mustn't it? Solution (forgive the pun) is to eradicate both; I suspect it's alcohol. -----Original Message----- From: Habitzruther, Michael [mailto:Michael.Habitzruther@RoswellPark.org] Sent: 19 July 2005 15:57 To: histonet@lists.utsouthwestern.edu Cc: Demant, Peter Subject: [Histonet] Problem with Eosin Y staining Our laboratory is having a problem with the staining of mouse tumor samples using hemotoxylin and Eosin Y (H&E Stain). Everything was going fine until recently. The specific problem we are having is after the cover-slip is put on and the acrymount dries, the Eosin Y stain seems to be bleeding out, leaving only the hemotoxylin staining. A histologist from the institute suggested it may have something to do with the relative humidity in our lab, and therefore allowing the slides to set under a flow-hood may help. Any other suggestions or protocol modifications would be greatly appreciated. Thank you very much. This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message 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 End of Histonet Digest, Vol 20, Issue 22 **************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Wed Jul 20 11:07:06 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Re: chatter in fly head paraffin section Message-ID: Zhenming, I suspect your samples are being exposed way too long in the processing solutions. Fly heads and brains are extremely small. I would recommend cutting the times in each your processing solutions to 20-30 minutes maximum. To section the ones you already have processed, try facing into the tissue and then letting them sit for about 5 minutes in a crushed ice/ice water bath. Blot them and try sectioning again. You may have to repeat this every several sections because you will eventually cut past the area that the water has penetrated into the sample. You never mentioned how thick you are trying to section these as well. If you are sectioning thicker than 7 microns, you might also try using room temperature or warmer water. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From ian.montgomery <@t> bio.gla.ac.uk Wed Jul 20 11:11:09 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] One Step Trichrome. Message-ID: <6.2.1.2.2.20050720170629.037ab270@udcf.gla.ac.uk> I'm considering using a one step trichrome for a zoology undergraduate teaching class. As the material will come from invertebrates is there a one step technique which would offer the best and most consistent result. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From Janet.Bonner <@t> FLHOSP.ORG Wed Jul 20 10:52:18 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Rapid Giemsa Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB432C@fh2k093.fhmis.net> Poly Scientific (800-645-5825)makes a good Giemsa Stain stock solution. cat#s195-(16 oz) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda Hines Sent: Wednesday, July 20, 2005 10:19 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Rapid geimsa HI! All I am searching for the chemicals, and maker of the rapid geimsa staining for Helicobacter pylori. Anyone using this stain can let me know it is appreciated. Thanx and Have a Great Day!!! Linda Hines HT Histology Supervisor Specialty Diagnostics Phx. AZ 602-241-5134 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dusko.trajkovic <@t> pfizer.com Wed Jul 20 12:30:39 2005 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Confusing IHC results Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B2D89555@lajamrexm01.amer.pfizer.com> Well I have to tell you all that I am pleasantly surprised and elated that so many of you have responded regarding my IHC situation. Here is the additional info many of you have asked for: I'm staining rat liver. There are two protocols that have given me positive staining, with some non specific staining as well. I was trying to clean up the nonspecific staining, when I started doing the dilutions, and realized that it did not matter weather dilution is 1:100 or 1:2000. Slides were stained on Ventana Discovery XT, using DAB map kit, with heat. I also ran the same protocol without heat and slides were slightly cleaner, however, the signal was also weaker. Protease 2 treatment for 4 min. Avidin / Biotin blocking 20 min Serum free protein blocking from Dako I use Dako diluent for the primary and the secondary. I also tried adding 1% normal horse serum to the diluent. No change in the results. I played around with primary incubation time (32 min - 1 hour), did not alter the staining results. Longer (4 hours) incubation for the No heat protocol yielded similar results. Secondaries: Vector universal- Elite kit (PK-6200) 1:200 I also tried these: Vector- Rabbit anti Goat 1:200 Vector- Horse anti Goat 1:200 Santa Cruz - Bovine anti Goat. Tried 1:200, 1:400 and 1:600. Jackson Immuno- Donkey anti Goat 1:200 and 1:400. I also stained the same tissue on Dako autostainer, using Reveal, Borg and EDTA epitope retrieval, without any success. Here is another issue that surfaced during the writing of this email. That's why it took me 2 hours to write it. I had to go in the lab and get some specific information, as well as start some other IHC runs, when I came across information on my universal biotinylated secondary. I looked up the info in the Vector catalog for the ABC Elite kit PK-6200, and the information says to be used with mouse or rabbit Ab's. Nothing about goat Ab. It also says that this universal secondary should not be used on rat tissue. None of this information seems to be in the kit insert. I had a couple of people read it and we could not find it. What is interesting, on the slide I have two different sections of rat liver. One is normal and the other is necrotic. Only the necrotic section is staining as described in Toxicology and Applied Pharmacology publication (191 (2003) 211-226). The other section has just slight non specific staining. I hope I've given you enough info. I am now even more confused, but the original deal is still on the table (or the bar). Cheers, nazdrovije, ziveli, skol, ..... Dusko Trajkovic PS. To ms. Rae Ann S. who responded with a, "Hey fella, you better watch out what you are offering!!!! You can't just go around kissing anybody!!!! IT'S NOT ANYBODY. IT'S MY FELLOW HISTOTECHS. THE BEST PEOPLE IN THE WORLD. 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 JMacDonald <@t> mtsac.edu Wed Jul 20 13:31:01 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] PAP Staining Message-ID: If EA was not available, what would be a suitable replacement for EA in the PAP stain, eosin or light green? Thank you. Jennifer MacDonald From gkrasinski <@t> covx.com Mon Jul 18 18:40:37 2005 From: gkrasinski <@t> covx.com (Glenn Krasinski) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Truly human-specific cytokeratin... Message-ID: <6837C00A0CC6594197C2F0EEF256561805AA73@covxmail.covx.com> Help!! Does a truly human-specific (no murine cross-reactivity) antibody to most cytokeratins actually exist? With conjugates? Or am I just asking way too much. I want to distinguish human solid tumor cells of epithelial origin from mouse tissue in xenografts via immunohistochemistry. Glenn M. Krasinski Scientist, Biology CovX Research LLC 9381 Judicial Drive, Suite 200 San Diego, CA 92121 From szeev <@t> bgu.ac.il Tue Jul 19 03:31:30 2005 From: szeev <@t> bgu.ac.il (ze'ev silverman) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] transparent tissue Message-ID: We are trying to embed 300 micron thick, mouse olfactory bulb explants. The explants are about 3 mm diameter and become transparent following exposure to xylene, making it very difficult to orient them. Does anyone know how we might lightly stain or label the tissue, perhaps by some particulate material? Different stains and materials we have tried have not survived the ethanol or xylene steps. Thanks, ZS Ze'ev Silverman, Ph.D. Zlotowski Center for Neuroscience Dept. of Morphology Faculty of Health Sciences P.O. Box 653 Beer Sheva 84105 Israel 972 8 647-7307 (office) 8 647-7304 (lab) 8 647-7627 (fax) http://medic.bgu.ac.il/brain From yli <@t> mednet.swmed.edu Tue Jul 19 15:18:40 2005 From: yli <@t> mednet.swmed.edu (Yun Li) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] IHC with neomarker Rabbit anti Ki67 on paraffin brain sections? Message-ID: <42DD6020.7020104@mednet.swmed.edu> Greeting all, I am trying to look at adult neurogenesis in the brain and would like to do IHC with proliferation markers such as Ki67 and PCNA. I came across this new monoclonal rabbit anti-ki67 antibody and would like to know how well it works, particularly on paraffin embedded tissues. If anyone has used it and wouldn't mind sharing with me condition and result, please drop a line. I appreciate it!! yun From t3zprestegard <@t> mmm.com Wed Jul 20 14:15:56 2005 From: t3zprestegard <@t> mmm.com (t3zprestegard@mmm.com) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] immuno fluorescent stains on frozen rodent tissues Message-ID: I was hoping to find another researcher who is doing immuno fluorescent stains on frozen rat tissues. Especially, anyone who is using an Autostainer to stain these slides. I am attempting to stain with both CD3 and CD8a (biotinylated antibodies) on frozen spleen tissue. I can get both stain protocols to work but once I put the 2 of the protocols back-to-back on the Autostainer, the tissues want to either fold over or completely wash off the slides. I am using a good quality charged slides, cut 7 micron sections, allow to dry, fix with acetone, dry again and then store in the -80C freezer until the next day. I then remove the slides, allow to dry again, fix again with cold acetone, hydrate quickly with buffer, then begin my stain run, I have the Dako autostainer and have removing all forceful "wash buffer" steps with simply buffer dropped on as a reagent, and then blown off. The tissues have held on through the first primary antibody but can't make it to through the second primary protocol. Any suggestions would be very welcome. The only alternative will be to go back to manual stains and this group I'm working this up for has many, many tissues to stain. Warmest regards, terri Terri A. Prestegard 3M Center, 270-3S-05 3M Pharmaceuticals Pathology/Toxicology St. Paul, MN 55144-1000 Histology Archivist Senior Histotechnologist Tel: (651)-736-7519 Fax: (651)-737-5506 t3zprestegard@mmm.com From Jackie.O'Connor <@t> abbott.com Wed Jul 20 14:17:51 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Truly human-specific cytokeratin... Message-ID: I use a human mitochondria marker for exactly the same thing - works perfectly. Jackie O' "Glenn Krasinski" Sent by: histonet-bounces@lists.utsouthwestern.edu 07/18/2005 06:40 PM To: cc: Subject: [Histonet] Truly human-specific cytokeratin... Help!! Does a truly human-specific (no murine cross-reactivity) antibody to most cytokeratins actually exist? With conjugates? Or am I just asking way too much. I want to distinguish human solid tumor cells of epithelial origin from mouse tissue in xenografts via immunohistochemistry. Glenn M. Krasinski Scientist, Biology CovX Research LLC 9381 Judicial Drive, Suite 200 San Diego, CA 92121 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHosp.org Wed Jul 20 10:52:18 2005 From: Janet.Bonner <@t> FLHosp.org (Bonner, Janet) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Rapid Giemsa Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB432C@fh2k093.fhmis.net> Poly Scientific (800-645-5825)makes a good Giemsa Stain stock solution. cat#s195-(16 oz) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda Hines Sent: Wednesday, July 20, 2005 10:19 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Rapid geimsa HI! All I am searching for the chemicals, and maker of the rapid geimsa staining for Helicobacter pylori. Anyone using this stain can let me know it is appreciated. Thanx and Have a Great Day!!! Linda Hines HT Histology Supervisor Specialty Diagnostics Phx. AZ 602-241-5134 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Wed Jul 20 14:50:23 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] transparent tissue Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171757B@lsexch.lsmaster.lifespan.org> Two possibilities are: - A small amount of eosin added to the last absolute alcohol in the processing cycle. The specimens will pick up the stain and then move directly into xylene so that the eosin has no opportunity to wash out. I keep a small vial of very strong eosin (almost saturated) dissolved in absolute alcohol for this purpose. A few drops of this "superconcentrate" added to the final alcohol is sufficient to tint small specimens pink. - Mark the specimens with India Ink prior to processing. This is in fact a particulate material since it is a suspension of very fine carbon particles, not a true solution. I use this often, for example to distinguish between the proximal and distal ends of a vessel or nerve. Blot the specimen on paper towel, so that it is damp but not too wet. I use a wooden applicator stick to apply the ink. Dip the stick into the ink, blot it to remove excess ink, then touch it to the specimen. If there isn't a lot of liquid on the specimen and there isn't a lot of ink on the stick, the ink can be applied quite precisely. The ink doesn't penetrate into the tissue, so in the finished sections it appears as a black line along the marked surface only. The carbon is chemically inert, and is resistant to all solvents normally used in processing histological specimens. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of ze'ev > silverman > Sent: Tuesday, July 19, 2005 1:31 AM > To: Histonet@pathology.swmed.edu > Subject: [Histonet] transparent tissue > > We are trying to embed 300 micron thick, mouse olfactory bulb explants. > The explants are about 3 mm diameter and become transparent following > exposure to xylene, making it very difficult to orient them. Does > anyone know how we might lightly stain or label the tissue, perhaps by > some particulate material? Different stains and materials we have tried > have not survived the ethanol or xylene steps. > > Thanks, ZS > > > Ze'ev Silverman, Ph.D. > Zlotowski Center for Neuroscience > Dept. of Morphology > Faculty of Health Sciences > P.O. Box 653 > Beer Sheva 84105 > Israel > > 972 8 647-7307 (office) > 8 647-7304 (lab) > 8 647-7627 (fax) > http://medic.bgu.ac.il/brain > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From lcheung <@t> cvmed.Stanford.EDU Wed Jul 20 14:58:09 2005 From: lcheung <@t> cvmed.Stanford.EDU (Cheung, Lauren) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] unsubscribe Message-ID: <8022E9EE6D5ED511A39200A0C9DED0E902FF363B@cvexchange> ------------------------------ Lauren Cheung Rockson Lab Lab: 650-723-5354 Email: Lcheung@cvmed.stanford.edu From PMonfils <@t> Lifespan.org Wed Jul 20 14:59:54 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] One Step Trichrome. Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171757C@lsexch.lsmaster.lifespan.org> I use the Gomori technique as my standard trichrome stain. It's a "one-step procedure" in that the collagen stain (light green FCF in the original procedure, though I prefer to substitute aniline blue) and the plasma stain (chromotrope 2R) are combined in one solution with phosphotungstic acid and acetic acid, and the stain is progressive (no differentiation required). Staining time is 20 minutes, though good results can be obtained in half that time. The solution keeps well, at least six months at room temperature, at least a year refrigerated. If you need the protocol I can send it to you. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ian > Montgomery > Sent: Wednesday, July 20, 2005 9:11 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] One Step Trichrome. > > I'm considering using a one step trichrome for a zoology > undergraduate teaching class. As the material will come from invertebrates > > is there a one step technique which would offer the best and most > consistent result. > > Dr. Ian Montgomery, > Histotechnology, > Graham Kerr Building, > Institute of Biomedical & Life Sciences, > University of Glasgow, > Glasgow, > G12 8QQ. > Tel: 0141 339 8855 > Office: 4652 > Lab: 6644. > Pager: 07623 975451 > e-mail: ian.montgomery@bio.gla.ac.uk > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From godsgirlnow <@t> msn.com Wed Jul 20 15:26:21 2005 From: godsgirlnow <@t> msn.com (Roxanne Soto) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Rapid geimsa References: <20050720141858.36383.qmail@web33005.mail.mud.yahoo.com> Message-ID: Linda Hi, BBC Biochemical has a great dif quik for h pylori that takes less than a minute to do. Call BBC and ask for Adrian. Tell him that I told you about the dif quik, so he will know which one it is because they have a lot of stains. Roxanne Soto ----- Original Message ----- From: Linda Hines To: histonet@pathology.swmed.edu Sent: Wednesday, July 20, 2005 10:18 AM Subject: [Histonet] Rapid geimsa HI! All I am searching for the chemicals, and maker of the rapid geimsa staining for Helicobacter pylori. Anyone using this stain can let me know it is appreciated. Thanx and Have a Great Day!!! Linda Hines HT Histology Supervisor Specialty Diagnostics Phx. AZ 602-241-5134 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PIXLEYSK <@t> UCMAIL.UC.EDU Wed Jul 20 15:48:21 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] RE: fixing cultured cells on coverslips Message-ID: Dear LaCinda: For lightly attached cells grown on glass coverslips, like neurons, we have found the fixation protocol below will help (but not always!) keep them on the slip through IHC. Two things were important: 1) having the fixative at the same temperature (and pH) as the cells so that they didn't shrink from the temp (or pH) change and 2) not rinsing until after they are fixed. Delicately attached cells do not do well when exposed to air during a medium change. 1.Warm 4% paraformaldehyde (pH 7.4 in 0.1 M phosphate buffer) to 37 degrees C. 2. Add small volume of 4% PF directly into the medium that the cells are growing in. (In other words, don't remove the culture medium so that you don't expose the cells to an air-water interface.) Use approximately the same volume as the cells are growing in, so that you have roughly 2% para. Incubate 5 minutes. 3. Remove all the solution over the cells after the 5 minutes. Add fresh 4% para. Incubate 10 minutes. 4. Gently wash 3 times with room temperature PBS or PB. Keep being gentle during all rinses. Large bore plastic transfer pipets are what we currently use. I would also like to know about a list serve for culture people. Best of luck! Sarah Pixley Univ. Cincinnati College of Med. From jqb7 <@t> cdc.gov Wed Jul 20 15:53:08 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] One Step Trichrome. Message-ID: Newcomer Supply (www.newcomersupply.com) has a nice one. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ian Montgomery Sent: Wed 7/20/2005 12:11 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] One Step Trichrome. I'm considering using a one step trichrome for a zoology undergraduate teaching class. As the material will come from invertebrates is there a one step technique which would offer the best and most consistent result. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jbaez <@t> interscopepath.com Wed Jul 20 16:19:23 2005 From: jbaez <@t> interscopepath.com (Baez, Janet) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Rapid geimsa Message-ID: <9E956D8FEB06C2408B08AC16498325E9FE21@scopemx1.interscope.com> Fisher HealthCare has a good stain for H.Pylori, FSH Hema 3 Solution I and FSH Hema 3 Solution II. Janet -----Original Message----- From: Linda Hines [mailto:linhines@yahoo.com] Sent: Wednesday, July 20, 2005 7:19 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Rapid geimsa HI! All I am searching for the chemicals, and maker of the rapid geimsa staining for Helicobacter pylori. Anyone using this stain can let me know it is appreciated. Thanx and Have a Great Day!!! Linda Hines HT Histology Supervisor Specialty Diagnostics Phx. AZ 602-241-5134 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Wed Jul 20 16:55:09 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:21 2005 Subject: AW: [Histonet] IHC with neomarker Rabbit anti Ki67 on paraffin brainsections? In-Reply-To: <42DD6020.7020104@mednet.swmed.edu> Message-ID: Hi, we tested this antibody on mamma-tissue. The result was very good. We will use it in the future. Benchmark XT, iview DAB, 1:100, 16 min; retrieval with EDTA Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Yun Li Gesendet: Dienstag, 19. Juli 2005 22:19 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] IHC with neomarker Rabbit anti Ki67 on paraffin brainsections? Greeting all, I am trying to look at adult neurogenesis in the brain and would like to do IHC with proliferation markers such as Ki67 and PCNA. I came across this new monoclonal rabbit anti-ki67 antibody and would like to know how well it works, particularly on paraffin embedded tissues. If anyone has used it and wouldn't mind sharing with me condition and result, please drop a line. I appreciate it!! yun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Jul 20 17:20:26 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] IHC with neomarker Rabbit anti Ki67 on paraffinbrainsections? In-Reply-To: <200507202158.j6KLwXDJ072744@pro12.abac.com> Message-ID: <200507202220.j6KMKPpf010680@chip.viawest.net> I have used rab. Mono ki67 from Lab Vision on several species with really good results also with HIER with citrate ph6 buffer and rab. Labelled polymer detection system. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Wednesday, July 20, 2005 2:55 PM To: Histonetliste (Histonetliste) Subject: AW: [Histonet] IHC with neomarker Rabbit anti Ki67 on paraffinbrainsections? Hi, we tested this antibody on mamma-tissue. The result was very good. We will use it in the future. Benchmark XT, iview DAB, 1:100, 16 min; retrieval with EDTA Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Yun Li Gesendet: Dienstag, 19. Juli 2005 22:19 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] IHC with neomarker Rabbit anti Ki67 on paraffin brainsections? Greeting all, I am trying to look at adult neurogenesis in the brain and would like to do IHC with proliferation markers such as Ki67 and PCNA. I came across this new monoclonal rabbit anti-ki67 antibody and would like to know how well it works, particularly on paraffin embedded tissues. If anyone has used it and wouldn't mind sharing with me condition and result, please drop a line. I appreciate it!! yun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Jul 20 17:33:04 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Confusing IHC results In-Reply-To: <3AD0BD3142459B4E9B12CBEAFF2B89B2D89555@lajamrexm01.amer.pfizer.com> Message-ID: <200507202233.j6KMX4pf015016@chip.viawest.net> In my experience the endogenous biotin in liver cannot be completely removed even with the most aggressive AB blocking, this is why I do not use an avidin/biotin detection system, I use labeled polymer instead. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Trajkovic, Dusko Sent: Wednesday, July 20, 2005 10:31 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Confusing IHC results Well I have to tell you all that I am pleasantly surprised and elated that so many of you have responded regarding my IHC situation. Here is the additional info many of you have asked for: I'm staining rat liver. There are two protocols that have given me positive staining, with some non specific staining as well. I was trying to clean up the nonspecific staining, when I started doing the dilutions, and realized that it did not matter weather dilution is 1:100 or 1:2000. Slides were stained on Ventana Discovery XT, using DAB map kit, with heat. I also ran the same protocol without heat and slides were slightly cleaner, however, the signal was also weaker. Protease 2 treatment for 4 min. Avidin / Biotin blocking 20 min Serum free protein blocking from Dako I use Dako diluent for the primary and the secondary. I also tried adding 1% normal horse serum to the diluent. No change in the results. I played around with primary incubation time (32 min - 1 hour), did not alter the staining results. Longer (4 hours) incubation for the No heat protocol yielded similar results. Secondaries: Vector universal- Elite kit (PK-6200) 1:200 I also tried these: Vector- Rabbit anti Goat 1:200 Vector- Horse anti Goat 1:200 Santa Cruz - Bovine anti Goat. Tried 1:200, 1:400 and 1:600. Jackson Immuno- Donkey anti Goat 1:200 and 1:400. I also stained the same tissue on Dako autostainer, using Reveal, Borg and EDTA epitope retrieval, without any success. Here is another issue that surfaced during the writing of this email. That's why it took me 2 hours to write it. I had to go in the lab and get some specific information, as well as start some other IHC runs, when I came across information on my universal biotinylated secondary. I looked up the info in the Vector catalog for the ABC Elite kit PK-6200, and the information says to be used with mouse or rabbit Ab's. Nothing about goat Ab. It also says that this universal secondary should not be used on rat tissue. None of this information seems to be in the kit insert. I had a couple of people read it and we could not find it. What is interesting, on the slide I have two different sections of rat liver. One is normal and the other is necrotic. Only the necrotic section is staining as described in Toxicology and Applied Pharmacology publication (191 (2003) 211-226). The other section has just slight non specific staining. I hope I've given you enough info. I am now even more confused, but the original deal is still on the table (or the bar). Cheers, nazdrovije, ziveli, skol, ..... Dusko Trajkovic PS. To ms. Rae Ann S. who responded with a, "Hey fella, you better watch out what you are offering!!!! You can't just go around kissing anybody!!!! IT'S NOT ANYBODY. IT'S MY FELLOW HISTOTECHS. THE BEST PEOPLE IN THE WORLD. 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 Linke_Noelle <@t> Allergan.com Wed Jul 20 17:57:45 2005 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] pharmaceutical industry Message-ID: Hi everyone, I'm looking to get in touch with folks working in the pharmaceutical industry (particularly those involved in safety evaluation/tox) off-line. I have a few questions I would like to ask if you wouldn't mind! Thank you, Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 From jim.manavis <@t> imvs.sa.gov.au Wed Jul 20 18:56:45 2005 From: jim.manavis <@t> imvs.sa.gov.au (Jim Manavis) Date: Fri Sep 16 15:25:21 2005 Subject: FW: [Histonet] muscle histochemistry Message-ID: <001701c58d86$af246820$636c140a@itp36533> Dr. Girish, my name is Bernice Gutschmidt and I work in the Muscle and Nerve Laboratory in Adelaide, Australia. We routinely do enzyme histochemistry on diagnostic human and occasionally on mouse muscle for research projects. It is critical to freeze the muscle in precooled isopentane to preserve the archetecture and enzyme activity of the muscle and and reduce any ice crystal artefact that you may be seeing in your muscles at present. The atpase method we use is a variation of the method on page 32 in DUBOWITZ AND BROOKE - Muscle biopsy, a Modern approach. If you are interested I could fax a copy of our method to you. Re the incubation medium. I find a ph of about 9.5 is ideal as the ph can drop when the sections are rinsed in distilled water pre incubating, and the reaction is depleted. I quickly looked at the bancroft and stevens method and it seems a very round about and tedious way to do it. Our method is proven and is a lot more straight forward (and it works!) Good luck and let me know if you require our method Regards, Bernice Gutschmidt Kathy Cash Senior Technician Muscle & Nerve Lab Hanson Institute Centre for Neurological Diseases Frome Rd, Adelaide, South Australia ph 61-08-82223588 fax 61-08-82223392 email: kathy.cash@imvs.sa.gov.au > -----Original Message----- > From: Jim Manavis [mailto:jim.manavis@imvs.sa.gov.au] > Sent: Thursday, 21 July 2005 08:05 AM > To: Kathy Cash > Subject: FW: [Histonet] muscle histochemistry > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of bcgirish > Sent: Wednesday, 20 July 2005 11:25 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] muscle histochemistry > > > ? > Hi all > I am Dr Girish,toxicological pathologist working for a pharmaceutical > company in India. We are recently standardizing myosin atpse > histochemistry > to study the effect of ppar compounds on skeletal muscle. > We are using liquid nitrogen without isopentane to collect the muscle > gastrocnemius from rats.The reaction is givinig nonspecific reaction and > artifacts. Is using of isopentane must? Is there a better method than the > one described by Bancroft? > > How to preserve the enzyme activity?How long we have to incubate in > incubating solution if we are using Bancroft method?we are using > atp at the > concentration of 5mg/5ml solution b of bancroft method.How long this > solution can be kept and at what temperature? > > Is the pH maintained to be 9.4 or 10.4 (alkaline)? We are getting > a sort of > cavity in the muscle fibers, is it because of ice crystal artifacts? > > I am waiting for quick suggestions.Do share ur experience . > .......................... > > > > REGARDS > DR.GIRISH B.CHANDRASHEKAR > DEPT OF PRECLINICAL SAFETY EVALUATION > DR REDDYS LAB > HYDERABAD-49 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From IKirbis <@t> onko-i.si Thu Jul 21 00:01:15 2005 From: IKirbis <@t> onko-i.si (=?iso-8859-2?Q?Kirbi=B9_Srebotnik_Irena?=) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] RE: fixing cultured cells on coverslips Message-ID: we obtained good results (morphology and immunocytochemistry - GFAP) by fixing cells growing on the slides in METHANOL at 4 degrees Irena Srebotnik Kirbi?, MSc Institute of Oncology Dept. of Cytopathology Zalo?ka 2 1000 Ljubljana Slovenia phone +386 1 522 3826 fax +38615879400 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Sent: Wednesday, July 20, 2005 10:48 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: fixing cultured cells on coverslips Dear LaCinda: For lightly attached cells grown on glass coverslips, like neurons, we have found the fixation protocol below will help (but not always!) keep them on the slip through IHC. Two things were important: 1) having the fixative at the same temperature (and pH) as the cells so that they didn't shrink from the temp (or pH) change and 2) not rinsing until after they are fixed. Delicately attached cells do not do well when exposed to air during a medium change. 1.Warm 4% paraformaldehyde (pH 7.4 in 0.1 M phosphate buffer) to 37 degrees C. 2. Add small volume of 4% PF directly into the medium that the cells are growing in. (In other words, don't remove the culture medium so that you don't expose the cells to an air-water interface.) Use approximately the same volume as the cells are growing in, so that you have roughly 2% para. Incubate 5 minutes. 3. Remove all the solution over the cells after the 5 minutes. Add fresh 4% para. Incubate 10 minutes. 4. Gently wash 3 times with room temperature PBS or PB. Keep being gentle during all rinses. Large bore plastic transfer pipets are what we currently use. I would also like to know about a list serve for culture people. Best of luck! Sarah Pixley Univ. Cincinnati College of Med. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Thu Jul 21 03:03:11 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] RE: Rapid stain for H pylori Message-ID: It's called Hemacolor 3 (blue) from Merck, code: 1,11957.2500 in 2.5L bottle. Agitate slide for 10 seconds. Works well too! Jacqui Malam Lancaster DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From Kemlo.Rogerson <@t> elht.nhs.uk Thu Jul 21 03:04:38 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] PAP Staining Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4A9@elht-exch1.xelht.nhs.uk> Not with your question Jennifer; EA is a mix of eosin, light green and Bismarck Green. If you run out of EA you can make it up from the constituent dyes and chemicals. However the mix of dyes and chemicals depend on the type of EA you require. EA65, EA50 whatever depends on altering the amounts of dye and mordents. Classically EA50 is for Gynae whilst EA65 is for Non-Gynae; the latter is 'pinker'. If you want to make it up then tell me what you want it for or what your staining preference is and I'll send you the recipe. TTFN -----Original Message----- From: Jennifer MacDonald [mailto:JMacDonald@mtsac.edu] Sent: 20 July 2005 19:31 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PAP Staining If EA was not available, what would be a suitable replacement for EA in the PAP stain, eosin or light green? Thank you. Jennifer MacDonald _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Thu Jul 21 08:41:40 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] postmortem tissue collection In-Reply-To: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4A5@elht-exch1.xelht.nhs.uk> References: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4A5@elht-exch1.xelht.nhs.uk> Message-ID: <42DFA614.8060105@umdnj.edu> Many years ago (I hate beginning my responses like that but it is true) several colleagues were able to get viable human retinal pigment epithelial cells more than 48 hours after death. Also, I know of viable olfactory (sensory) epithelial cells being harvested 24 hours after death. Of course, ambient temperature is a likely variable and not all cells die at the same time. I would think that the adipose tissue in the hypodermis would have a relatively slow metabolic rate and might be viable for 24 hours? You have little to lose by trying. Geoff Rogerson Kemlo (ELHT) Pathology wrote: >Interestingly I was reading about tissue death after death (so as to >speak) on a Web Site dealing with Death (I'm strange like that); I was >interested in rigor. I hadn't realised that some tissue deep within the >cadaver can survive for many hours after death. I suppose it's those >tissues that require little or no oxygen to survive; brain dies very >rapidly. If I could remember the Site then I'd tell you but a search in >Google under rigor may help. Wonder which bit of you dies last? Same in >men and women? > >-----Original Message----- >From: Y. Wang [mailto:ynwang@u.washington.edu] >Sent: 20 July 2005 00:20 >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] postmortem tissue collection > >Dear histonetters, > >I have a question regarding collection of human tissue. A colleague >would >like to collect human tissue (skin and underlying adipose tissue) for >mechanical testing, histological analysis (cellular and structural >evaluation), IHC and protein analysis (extracellular matrix structure >and >concentrations). They asked what would be a fair cut off time for tissue > >collection so that the effects of decay would not be a factor. Currently > >they have given the tissue bank a time of 12 hours postmortem (I'm not >very sure how the body is stored during this time or how this is >calculated). > >I've read that human decomposition starts approx. 4 min after death and >autolysis is quicker in tissues with high enzyme and water content. >However, in terms of the skin and underlying adipose tissue I didn't >know >if there was an accepted 'cut off time' postmortem after which tissue >is considered far from representative of 'live' tissue and not worth >analysis. > >Can anyone give some insight? Any information or references would be >greatly appreciated. > >Thank you >Yak-Nam > >Senior Fellow >Department of Bioengineering >University of Washington >Box 357962 >Seattle, WA 98195 > >Tel.: (206)-221-5873 >Fax.: (206)-221-5874 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >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 mcauliff <@t> umdnj.edu Thu Jul 21 09:07:42 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Deformol In-Reply-To: <002301c58077$9ead7ac0$112b5c9f@Carlos> References: <002301c58077$9ead7ac0$112b5c9f@Carlos> Message-ID: <42DFAC2E.9070806@umdnj.edu> You might want to look at " A comparison of deformalinizing techniques" by Lhotka and Ferreira, Stain Technology 25(1): 27-32, 1950. Geoff Carl Hobbs wrote: > High , > would be very grateful for any info on "deformol". Only ref I found > was by Elvers in 1943....can't get the paper, tho. Would like to know > what's in it. Said to be able to remove formalin from tissue! > Thanks > Carl > > _______________________________________________ > 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 mcauliff <@t> umdnj.edu Thu Jul 21 09:11:37 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] One Step Trichrome. In-Reply-To: References: Message-ID: <42DFAD19.4000304@umdnj.edu> Ian: Have a look at Horobin and Flemming, Histochemical Journal 20:29-34, 1988. "One bath trichrome staining: investigation of a general mechanism based on structure-staining correlation analysis". Geoff > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ian Montgomery > Sent: Wed 7/20/2005 12:11 PM > To: histonet@lists.utsouthwestern.edu > Cc: > Subject: [Histonet] One Step Trichrome. > > > > I'm considering using a one step trichrome for a zoology > undergraduate teaching class. As the material will come from invertebrates > is there a one step technique which would offer the best and most > consistent result. > > Dr. Ian Montgomery, > Histotechnology, > Graham Kerr Building, > Institute of Biomedical & Life Sciences, > University of Glasgow, > Glasgow, > G12 8QQ. > Tel: 0141 339 8855 > Office: 4652 > Lab: 6644. > Pager: 07623 975451 > e-mail: ian.montgomery@bio.gla.ac.uk > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >------------------------------------------------------------------------ > >_______________________________________________ >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 c.m.vanderloos <@t> amc.uva.nl Thu Jul 21 09:29:08 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] RE: Confusing IHC results Message-ID: <1275aef1270882.12708821275aef@amc.uva.nl> Dusco, Patty is completely right about this! I have observed the same problem with FFPE kidney sections that were pronase pretreated. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Wed, 20 Jul 2005 16:33:04 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] Confusing IHC results To: "'Trajkovic, Dusko'" , In my experience the endogenous biotin in liver cannot be completely removed even with the most aggressive AB blocking, this is why I do not use an avidin/biotin detection system, I use labeled polymer instead. Patsy From jcarpenter764 <@t> aol.com Thu Jul 21 09:42:57 2005 From: jcarpenter764 <@t> aol.com (jcarpenter764@aol.com) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Gram staining Message-ID: <8C75C199E7DA460-E70-20212@FWM-D35.sysops.aol.com> Can anyone give me some tips on how to get Gram stain controls or how to substitute for the control. Thanks Jennell From beingmary53 <@t> sbcglobal.net Thu Jul 21 09:49:02 2005 From: beingmary53 <@t> sbcglobal.net (MARY JOHNSON) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] tranpan tissue Message-ID: <20050721144903.62499.qmail@web81608.mail.yahoo.com> Hi Silverman, If you would make up a 1% phloxine B solution and put it in your first Abs. alcohol., about 10 ml, I think this will work. Good luck Mary From MICHAEL.OWEN <@t> fda.gov Thu Jul 21 09:51:53 2005 From: MICHAEL.OWEN <@t> fda.gov (Owen, Michael P) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Re: Gram Stain Controls Message-ID: Dear Jennell, ATCC is good source for Gram stain controls. I have routinely seen Staphylococcus aureus ATCC 25923 used as a Gram-positive control and Escherichia coli 25922 used as a Gram-negative control. In my laboratory we grow cultures of these strains on Plate Count Agar or Tryptic Soy Agar with Yeast Extract for use with Gram staining performed the next day. Is this the type of information you wanted or did I misread your question? Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From juan.gutierrez <@t> christushealth.org Thu Jul 21 09:57:56 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Gram staining Message-ID: I had some pieces of placenta put into growth media and inoculated by our micro department. Let them grow a couple of days and process. Make sure your tissue is fresh and not fixed in formalin. Good luck. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of jcarpenter764@aol.com Sent: Thursday, July 21, 2005 9:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Gram staining Can anyone give me some tips on how to get Gram stain controls or how to substitute for the control. Thanks Jennell _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Jul 21 10:29:38 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] RE: more questions on IF In-Reply-To: Message-ID: <200507211529.j6LFTapf024354@chip.viawest.net> Terri, I think 1hr.h202 block of frozen is way too harsh, even with 1% compared to 3%, I have found with the T cell markers the endogenous peroxidase block is very critical, in fact although for most of my antibodies I block after the primary antibody, for T cell markers I have to do the peroxidase block before the antibody. What enzyme are you using for your second cd8? There is a great product from DAKO now called Dual Block which blocks both for peroxidase and alk. Phos. In the same solution. It seems to be gentle enough for frozen sections. I use it on frozen rat brains when I do cd3, cd4, and cd8. I block for 10 min. with this BEFORE the antibody and then use a labeled polymer detection system to avoid endogenous biotin issues. My antibody is rat anti-mouse so I then use rab. Anti-rat secondary and then the anti-rab. Labelled polymer with great results. I use a serum free protein block before the primary antibody for 5 min. and then I use that again before the secondary and before the labelled polymer, if makes for really clean results. Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: t3zprestegard@mmm.com [mailto:t3zprestegard@mmm.com] Sent: Thursday, July 21, 2005 5:59 AM To: pruegg@ihctech.net Subject: more questions on IF Hi Patsy, Thank you for answering my note to histonet. I have a fewe more detailed questions if you don't mind. Do you stain with an autostainer or by hand? Do you think it's necessary to block for an entire hour between the 2 abs. The protocol I was given has the entire CD3 stain protocol followed by 1 hour incubation in 1% H2O2 then the second protocol for staining with CD8a. My tissues seem to loosen starting with the hour long peroxide step. What type of slides do you mount these tissues on? What are your thoughts? Regards, terri Terri A. Prestegard 3M Center, 270-3S-05 3M Pharmaceuticals Pathology/Toxicology St. Paul, MN 55144-1000 Histology Archivist Senior Histotechnologist Tel: (651)-736-7519 Fax: (651)-737-5506 t3zprestegard@mmm.com From PMonfils <@t> Lifespan.org Thu Jul 21 11:14:38 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Gram staining Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171757E@lsexch.lsmaster.lifespan.org> I have been making my own gram controls for years. I purchase cultures of non-pathogenic gram positive and gram negative organisms grown in liquid suspension (rather than on an agar plate) from a biological supply company. These are inexpensive, less than $10.00 per culture. When I receive the tubes I spin them down to concentrate the organisms, remove most of the supernatant, resuspend the culture in a small volume of its original culture medium, then inject it into a piece of unfixed lung through a fine gauge needle. I use lung because there are many small natural spaces to retain the culture. I push the needle most of the way through the tissue, then slowly withdraw it as I inject, to distribute the culture through the tissue. Then inject again perpendicular to the first injection. Drop the tissue into formalin, fix overnight, process and embed as usual. I embed a gram positive piece and a gram negative piece side by side in the same block. Many different organisms can be used, but I am currently using a mix of Bacillus subtilis and Micrococcus roseus for the gram positive control; and a mix of Escherichia coli and Serratia liquefaciens for the gram negative. I purchase the cultures here: http://www.ctvalleybio.com Click on "living organisms", then "bacteria". Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > jcarpenter764@aol.com > Sent: Thursday, July 21, 2005 7:42 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Gram staining > > Can anyone give me some tips on how to get Gram stain controls or how to > substitute for the control. Thanks Jennell > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From m-degutes <@t> northwestern.edu Thu Jul 21 11:23:06 2005 From: m-degutes <@t> northwestern.edu (Mathew DeGutes) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] CD25 staining on frozen tissue Message-ID: <200507211623.j6LGNFNO015860@casbah.it.northwestern.edu> Hi Everyone, I am trying to get CD25 staining to work in order to identify T reg cells in mouse tissue sections. So far I really haven't had much luck. I have both a 7D4 biotinylated anti-CD25 from Pharmingen and a PC61 Fitc-conjugated anti-CD25 from eBioscience and I am using Spleen as a positive control. I am using fresh frozen sections that have not been fixed at all. I have tried various fixations with Acetone and PFA. I would think PFA might block staining as many people have had trouble getting staining on parafin sections. I have played with many different antibody concentrations and we have secondaries and amplification protocols that have worked for other biotinylated and fitc-conjugated antibodies quite well using strep-hrp and anti-fitc-hrp along with a TSA cy3 or fluorescein kit. Does anyone have a protocol or some tips on making these antibodies stain for immunofluorescence? I know this is a common stain, but I simply have had no luck with it. Any help is appreciated thanks. From cfavara <@t> niaid.nih.gov Thu Jul 21 11:56:20 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] FDC-M1 Message-ID: Does anyone have any experience with FDC[follicular dendritic cell] -M1 antibody clone 4C11 in mouse tissue? I do have some references that I am tracking down, but am having difficulty with the source Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From zyu <@t> mail.med.upenn.edu Thu Jul 21 12:02:49 2005 From: zyu <@t> mail.med.upenn.edu (Zhenming Yu) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] tissue falling off slides Message-ID: <1121965369.42dfd539b88cb@webmail.pobox.upenn.edu> Thanks to so many of you for the very helpful responses to my last question regarding chatter in fly head paraffin section. It turned out that we probably had over-processed the fly heads in alcohol and xylene. We are modifying our protocols and keeping our fingers crossed that the problem will be solved soon. Also I got another question that I hope you guys can help with---we sometimes ran across the problem of patches of tissue coming off slides. Could this also be attributed to over exposure of the heads to alcohol and xylene during processing? We have been using Superfrost Plus slides from Fisher---is any further slide treatment necessary? We usually lay ribbons (5 or 7 micron thick) of paraffin sections on the water surface in a 38?C water bath for less than one minute (too short?) before loading to a slide. Then the slide was kept in the vertical position for about 30 minutes to dry and incubated in a 37 incubator overnight for further drying. Any thoughts would be gratefully received. Thanks. **************************** Zhenming Yu Univ. of Penn Philadelphia, PA 19104 **************************** From ksverlow <@t> cahfs.ucdavis.edu Thu Jul 21 12:20:46 2005 From: ksverlow <@t> cahfs.ucdavis.edu (Sverlow, Karen) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] RE: Chlamydia antibody and Aspergillus IHC Message-ID: Hi Everyone, Has anyone found a replacement antibody for the Virostat Chlamydia monoclonal product #1631 (no longer available)? Specifically, we are looking for a monoclonal that will detect Chlamydia in FFPE mammalian and avian tissues. On another note, I would like to talk with those of you doing Aspergillus IHC. Please email me. Thank you, Karen Sverlow Staff Research Associate California Animal Health and Food Safety Laboratory University of California, Davis ksverlow@cahfs.ucdavis.edu From MICHAEL.OWEN <@t> fda.gov Thu Jul 21 12:23:58 2005 From: MICHAEL.OWEN <@t> fda.gov (Owen, Michael P) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Reagent Search: PDTS Message-ID: Dear Histonet Members, A colleague of mine is looking for a reagent called "Phenolphthalein Disulfate, Tripotassium Salt" for a staining project. The Sigma-Aldrich product number is P0251. Sigma-Aldrich has stated the chemical is no longer available. Does anyone on this list know of an alternative source? Thanks in advance. Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From gcallis <@t> montana.edu Thu Jul 21 12:27:05 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] FDC-M1 antibody Message-ID: <6.0.0.22.1.20050721112507.01b18438@gemini.msu.montana.edu> Cynthia, Yes, we stain for this using IFA, but it also works IHC. We use frozens sections only, and it was an easy protocol. The antibody came from BD Pharmingen. You wrote: Does anyone have any experience with FDC[follicular dendritic cell] -M1 antibody clone 4C11 in mouse tissue?I do have some references that I am tracking down, but am having difficulty with the source Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Thu Jul 21 12:33:08 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] CD25 on murine tissue woes Message-ID: <6.0.0.22.1.20050721112807.01b13c90@gemini.msu.montana.edu> Matthew, You wrote: I am trying to get CD25 staining to work in order to identify T reg cells in mouse tissue sections. So far I really haven't had much luck. I have both a 7D4 biotinylated anti-CD25 from Pharmingen and a PC61 Fitc-conjugated anti-CD25 from eBioscience and I am using Spleen as a positive control. I am using fresh frozen sections that have not been fixed at all. I have tried various fixations with Acetone and PFA. I would think PFA might block staining as many people have had trouble getting staining on parafin sections. I have played with many different antibody concentrations and we have secondaries and amplification protocols that have worked for other biotinylated and fitc-conjugated antibodies quite well using strep-hrp and anti-fitc-hrp along with a TSA cy3 or fluorescein kit. Does anyone have a protocol or some tips on making these antibodies stain for immunofluorescence? I know this is a common stain, but I simply have had no luck with it. Any help is appreciated thanks. Could you provide more details on HOW you are doing the staining, both as IHC and or immunofluorescence. If you are trying to do the primary antibody directly conjugated to FITC on a tissue section, you may not get positive results - this is not uncommon with some of this type conjugate as a direct IFA stain. I can't make it work with CD4-FITC or CD8-FITC. As for biotinylated antibodies (primaries) one can do either IHC and IFA - we do this on many other rat antimouse primaries, CD markers - so provide concentrations, times, etc, etc - all those good things. Thanks Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Thu Jul 21 12:40:16 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Re: Reagent Search: PDTS In-Reply-To: References: Message-ID: <6.0.0.22.1.20050721113905.01b6cf68@gemini.msu.montana.edu> Try some other chemical companies - ACROS vis Fisher, and ICN are possibilities. At 11:23 AM 7/21/2005, you wrote: >Dear Histonet Members, > >A colleague of mine is looking for a reagent called "Phenolphthalein >Disulfate, Tripotassium Salt" for a staining project. The Sigma-Aldrich >product number is P0251. > >Sigma-Aldrich has stated the chemical is no longer available. Does anyone on >this list know of an alternative source? 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 PMonfils <@t> Lifespan.org Thu Jul 21 12:51:08 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Reagent Search: PDTS Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171757F@lsexch.lsmaster.lifespan.org> Lab Depot carries this compound. www.labdepotinc.com Also, you can search for suppliers of chemicals at www.chemexper.com Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Owen, > Michael P > Sent: Thursday, July 21, 2005 10:23 AM > To: 'Histonet' > Subject: [Histonet] Reagent Search: PDTS > > > > Dear Histonet Members, > > A colleague of mine is looking for a reagent called "Phenolphthalein > Disulfate, Tripotassium Salt" for a staining project. The Sigma-Aldrich > product number is P0251. > > Sigma-Aldrich has stated the chemical is no longer available. Does anyone > on > this list know of an alternative source? > > Thanks in advance. > > > Michael P. Owen, Regulatory Microbiologist > U.S. FDA Pacific Regional Lab Northwest > 22201 23rd Drive SE Bothell, WA 98021-4421 > Phone: 425-483-4865 E-Mail: michael.owen@fda.gov > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gcallis <@t> montana.edu Thu Jul 21 12:54:25 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] type of mouse for spleen control Re: CD25 on murine tissue woes In-Reply-To: References: Message-ID: <6.0.0.22.1.20050721115143.01b4be20@gemini.msu.montana.edu> Mouse was C57 black. At 11:44 AM 7/21/2005, you wrote: >Just curious, Gayle - what type of mouse is your spleen from? > >Jacqueline M. O'Connor HT(ASCP) QIHC >Assistant Scientist >Discovery Cancer R4N2 AP3 >847-938-4919 >Jackie.O'Connor@abbott.com Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From LewisS <@t> ccri.net Thu Jul 21 12:55:54 2005 From: LewisS <@t> ccri.net (Lewis, Sarah) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Gram staining Message-ID: <714B9F12B4E18C4C843B66E8E190F2AD44755C@res2k3ms01.CRII.ORG> rumor has it that *slim Jim* beef jerky works for both gram positive and gram negative.>>>?? -----Original Message----- From: jcarpenter764@aol.com [mailto:jcarpenter764@aol.com] Sent: Thursday, July 21, 2005 10:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Gram staining Can anyone give me some tips on how to get Gram stain controls or how to substitute for the control. Thanks Jennell From TillRenee <@t> uams.edu Thu Jul 21 14:19:32 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] MGMT Message-ID: Hello. Does anyone know of a good MGMT antibody for rat tissues? I have found several that say weakly reactive with rat tissue, and many that don't specify. Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 From shive003 <@t> umn.edu Thu Jul 21 14:33:34 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] RE: Chlamydia antibody and Aspergillus IHC References: Message-ID: <00c801c58e2b$15c5ce30$41065486@auxs.umn.edu> I don't use a monoclonal for Chlamydia, but I do use a rabbit polyclonal from Biodesign (B65256R). It's made against Chlamydia trachomatis (EB's all antigens), but cross-reacts with Chlamydia psittacii and pneumoniae. It requires no tissue pretreatment. Jan Shivers Univ. of Minnesota Vet Diag Lab ----- Original Message ----- From: "Sverlow, Karen" To: Sent: Thursday, July 21, 2005 12:20 PM Subject: [Histonet] RE: Chlamydia antibody and Aspergillus IHC Hi Everyone, Has anyone found a replacement antibody for the Virostat Chlamydia monoclonal product #1631 (no longer available)? Specifically, we are looking for a monoclonal that will detect Chlamydia in FFPE mammalian and avian tissues. On another note, I would like to talk with those of you doing Aspergillus IHC. Please email me. Thank you, Karen Sverlow Staff Research Associate California Animal Health and Food Safety Laboratory University of California, Davis ksverlow@cahfs.ucdavis.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From m-degutes <@t> northwestern.edu Thu Jul 21 15:00:54 2005 From: m-degutes <@t> northwestern.edu (Mathew DeGutes) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] RE: CD25 on murine tissue woes Message-ID: <200507212001.j6LK14Mv008154@casbah.it.northwestern.edu> If you want all the nitty gritty details. General protocol is this 1. Cut sections. Let dry and store at -80c until use. 2. Let sections return to room temperature and then fix. (Have tried Acetone 10 min -20c, 2%PFA 5 min room temp) 3. Wash 3x5 PBS 4. Wash 5 min in 3% H2O2 (have tried without this step) 5. Wash 3x5 PBS 6. Encircle and Avidin Block 10 min. (avidin biotin blocks are dependent on whether a biotinylated antibody is used). 7. Wash off Avidin in PBS 8. Biotin block 10 min 10. Aspirate Biotin block and add Block (have tried various serum at 10% and TNB, either with CD16/32 added at 1:50) for 30 min minimum. 11. Aspirate block and add Primary ab (have tried both 7D4 biotinylated anti-CD25 from Pharmingen and a PC61 Fitc-conjugated anti-CD25 from eBioscience at concentrations from 1:25 to 1:500) diluted in TNB or a 2% serum + .2% Triton in PBS buffer for 1 hour at room temp (have also tried overnight at 4c, but generally do the 1hour room temp). 12. Wash 3x5 PBS 13. Add 2ndary ab (Either Streptavidin-HRP or Anti-Fitc HRP) 1:100 in staining buffer from step 11 and inc for 30 min. 14. Wash 3x5 PBS 15. Use TSA Cy3 or Fluorescein kit. Dilute 1:100 in diluent and develop 5 minutes. 16. Wash 3x5 PBS 17. Mount with Vectashield with DAPI and coverslip Now we have used these 2ndaries and TSA kits extensively and have found them to work very well on any number of antigens. As I said before I haven't attempted to make it work with a direct. I come back and amplify with anti-fitc-hrp and the TSA kit. Also we are generally working in SJL mouse strains. I would really like to get this to work to go alongside FoxP3 staining for regs. I realize that regs are supposedly lower in SJL strains, but I have attempted this on Naive BalbC spleens as well and also have attempted on spleens from mice that were given injections of CD25+ CD4+ Tcells and so should have a far greater number of cells that are CD25+ (whether these cells will actually show up in anything other than flow I am not sure). You Wrote: Could you provide more details on HOW you are doing the staining, both as IHC and or immunofluorescence. If you are trying to do the primary antibody directly conjugated to FITC on a tissue section, you may not get positive results - this is not uncommon with some of this type conjugate as a direct IFA stain. I can't make it work with CD4-FITC or CD8-FITC. As for biotinylated antibodies (primaries) one can do either IHC and IFA - we do this on many other rat antimouse primaries, CD markers - so provide concentrations, times, etc, etc - all those good things. From pruegg <@t> ihctech.net Thu Jul 21 15:23:42 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] BIOGENEX Message-ID: <200507212023.j6LKNepf006377@chip.viawest.net> Can someone from BioGenex please contact me about a MW of theirs I am demoing. Patsy Ruegg Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From emerald_lake77 <@t> yahoo.com Thu Jul 21 17:30:22 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Murine Blood Smear / Smears IHC Message-ID: <20050721223022.30674.qmail@web31710.mail.mud.yahoo.com> Hello everyone, I've searched but my answers so far have come up too vague. I need to know if it is possible to do IHC on mouse blood smears???? If so, how would one go about preparing and fixing the sample? Is there anything I should watch out for? I just want to run normal IHC (probably to mark certain white blood cells (for example, monocytes to start). Any assistance is valued and greatly appreciated. Thanks! Gustave Hebert Scientist II Wyeth Research Cambridge MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From ynwang <@t> u.washington.edu Thu Jul 21 17:47:19 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] postmortem tissue collection In-Reply-To: <42DFA614.8060105@umdnj.edu> References: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4A5@elht-exch1.xelht.nhs.uk> <42DFA614.8060105@umdnj.edu> Message-ID: Thank you all for your thoughts on this question. I haven't found anything more enlightening on the web searches I've done so far. However, I will continue searching and will let you know if and when I find some thing exciting (aside from the strange but interesting nuggets that one always happens across in these searches on tissue death after death). Thanks again Yak-Nam > Many years ago (I hate beginning my responses like that but it is true) > several colleagues were able to get viable human retinal pigment epithelial > cells more than 48 hours after death. Also, I know of viable olfactory > (sensory) epithelial cells being harvested 24 hours after death. Of course, > ambient temperature is a likely variable and not all cells die at the same > time. I would think that the adipose tissue in the hypodermis would have a > relatively slow metabolic rate and might be viable for 24 hours? You have > little to lose by trying. > > Geoff > > Rogerson Kemlo (ELHT) Pathology wrote: > >> Interestingly I was reading about tissue death after death (so as to >> speak) on a Web Site dealing with Death (I'm strange like that); I was >> interested in rigor. I hadn't realised that some tissue deep within the >> cadaver can survive for many hours after death. I suppose it's those >> tissues that require little or no oxygen to survive; brain dies very >> rapidly. If I could remember the Site then I'd tell you but a search in >> Google under rigor may help. Wonder which bit of you dies last? Same in >> men and women? >> >> -----Original Message----- >> From: Y. Wang [mailto:ynwang@u.washington.edu] Sent: 20 July 2005 00:20 >> To: Histonet@lists.utsouthwestern.edu >> Subject: [Histonet] postmortem tissue collection >> >> Dear histonetters, >> >> I have a question regarding collection of human tissue. A colleague >> would like to collect human tissue (skin and underlying adipose tissue) for >> mechanical testing, histological analysis (cellular and structural >> evaluation), IHC and protein analysis (extracellular matrix structure >> and concentrations). They asked what would be a fair cut off time for >> tissue >> >> collection so that the effects of decay would not be a factor. Currently >> >> they have given the tissue bank a time of 12 hours postmortem (I'm not very >> sure how the body is stored during this time or how this is calculated). >> >> I've read that human decomposition starts approx. 4 min after death and >> autolysis is quicker in tissues with high enzyme and water content. >> However, in terms of the skin and underlying adipose tissue I didn't >> know if there was an accepted 'cut off time' postmortem after which tissue >> is considered far from representative of 'live' tissue and not worth >> analysis. >> >> Can anyone give some insight? Any information or references would be >> greatly appreciated. >> >> Thank you >> Yak-Nam >> >> Senior Fellow >> Department of Bioengineering >> University of Washington >> Box 357962 >> Seattle, WA 98195 >> >> Tel.: (206)-221-5873 >> Fax.: (206)-221-5874 >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> _______________________________________________ >> 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 kccatunda <@t> terra.com.br Thu Jul 21 18:22:32 2005 From: kccatunda <@t> terra.com.br (Katia Cristina Catunda) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Workloads vs Hiso staffing References: Message-ID: <015b01c58e4b$12750f00$a279fea9@privatexx> We do have a great interest on that question either. My supervisor sent me an FAQ he got on CAP telling that the ideal workload would be 50 H/E slides per histotechnician http://www.cap.org/apps/docs/cap_today/q_and_a/qa_1102.html - answered by Richard W. Brown, MD In our lab we do have 12 histotechnicians with a routine that is about 470 blocks (for 1/3 of them we generate two slides, one for HE and one for H. pilory) and 600 citological slides per day. They alternate their function every week: - embedding - 3 of them - microtomy 4 of them - staining/mounting and organizing slides - 2 of them - specific staining -the same of embedding - archiving blocks - the same of staining - citology routine - 2 of them - one is allways on vacation - immunohistochemistry - 2 of them are moved to other room twice a week I was wondering which is the best way to determinate their workload in order to analyse if we have too many or too less employees. My personal opinion is that 50 H/E slides per employee would be very hard-working and out of our range. Does anybody has any idea? But all of these informations are international since we are from Brazil... Katia ----- Original Message ----- From: "Maray Weirauch" To: Sent: Monday, July 18, 2005 3:44 PM Subject: [Histonet] Workloads vs Hiso staffing > ** PRIVATE ** > > Our hospital is conducting an analysis of laboratory workload, workflow > and staffing. This will include Histology as well, and we are being > asked for input with any national benchmarks available for surgical > histology tasks (i.e. average number cases accessioned, blocks embedded, > slides cut../some type of time unit) Does anyone know of some realistic > published values or ranges? Thanks! > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From cfavara <@t> niaid.nih.gov Thu Jul 21 19:15:57 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Murine Blood Smear / Smears IHC Message-ID: Questions - why are you dong this? What is the ultimate goal? If you just want a differential then do a differential. If you are looking for various populations of cells then FACS might be more reasonable and far simpler. Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: GT Hebert [mailto:emerald_lake77@yahoo.com] Sent: Thursday, July 21, 2005 3:30 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Murine Blood Smear / Smears IHC Hello everyone, I've searched but my answers so far have come up too vague. I need to know if it is possible to do IHC on mouse blood smears???? If so, how would one go about preparing and fixing the sample? Is there anything I should watch out for? I just want to run normal IHC (probably to mark certain white blood cells (for example, monocytes to start). Any assistance is valued and greatly appreciated. Thanks! Gustave Hebert Scientist II Wyeth Research Cambridge MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From steven.p.postl <@t> abbott.com Thu Jul 21 19:40:26 2005 From: steven.p.postl <@t> abbott.com (Steven P Postl) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. From opiecurt <@t> yahoo.com Thu Jul 21 20:05:31 2005 From: opiecurt <@t> yahoo.com (curt tague) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] anyone know who sells these bottles? Message-ID: <20050722010531.85013.qmail@web80101.mail.yahoo.com> there are 2 picutres under histo net images. i'd love to find a vendor for these bottles. others are a tad too small for my labels. thanks. From kbroomal <@t> NEMOURS.ORG Fri Jul 22 07:24:34 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824394@wlmmsx02.nemours.org> I would think it would be difficult to find something to write on a black cassette & be visible. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Steven P Postl Sent: Thursday, July 21, 2005 8:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Black paraffin and cassettes A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bcgpath <@t> rediffmail.com Fri Jul 22 08:21:05 2005 From: bcgpath <@t> rediffmail.com (bcgirish) Date: Fri Sep 16 15:25:21 2005 Subject: [Histonet] muscle histochemistry- Message-ID: <20050722132105.13765.qmail@webmail29.rediffmail.com> Hi Bernice Gut Schmidt I am very much interested in the method you are using for muscle atpse histochemistry(Dubowitz and Brooke).If you can send me the same it will be very much useful to us. My Fax ------91-40-23045438 Phone ----914023045439 REGARDS Dr.Girish B.Chandrashekar Dept of Preclinical Safety Evaluation Dr Reddys Lab Hyderabad-49 From PMonfils <@t> Lifespan.org Fri Jul 22 09:10:58 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717580@lsexch.lsmaster.lifespan.org> I believe the dye Sudan Black B is soluble in paraffin. Black cassettes? Nope. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Steven P Postl > Sent: Thursday, July 21, 2005 5:40 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Black paraffin and cassettes > > A researcher is wondering (now so am I [I never heard this request > before]) if any company sells black paraffin and black embedding > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > > be possible to add something to paraffin to make it stay a black color and > > remain strong as our commercial paraffin products? Thanks. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From pmarcum <@t> vet.upenn.edu Fri Jul 22 09:16:24 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE01717580@lsexch.lsmaster.l ifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE01717580@lsexch.lsmaster.lifespan.org> Message-ID: <6.1.1.1.2.20050722101403.019bfb80@mail.vet.upenn.edu> At 10:10 AM 7/22/2005, Monfils, Paul wrote: >I believe the dye Sudan Black B is soluble in paraffin. > >Black cassettes? Nope. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > > Steven P Postl > > Sent: Thursday, July 21, 2005 5:40 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] Black paraffin and cassettes > > > > A researcher is wondering (now so am I [I never heard this request > > before]) if any company sells black paraffin and black embedding > > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > > > > be possible to add something to paraffin to make it stay a black color and > > > > remain strong as our commercial paraffin products? 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 It is soluble in paraffin however it will be removed by reagents used in deparaffinization and staining. Due to the additional materials added to raw paraffin it will be spotty. It will also leave some precipitate in the tissue. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From JWEEMS <@t> sjha.org Fri Jul 22 09:16:54 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA461D8@sjhaexc02.sjha.org> Perhaps clear cassettes would work? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Monfils, Paul Sent: Friday, July 22, 2005 10:11 AM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Black paraffin and cassettes I believe the dye Sudan Black B is soluble in paraffin. Black cassettes? Nope. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Steven P Postl > Sent: Thursday, July 21, 2005 5:40 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Black paraffin and cassettes > > A researcher is wondering (now so am I [I never heard this request > before]) if any company sells black paraffin and black embedding > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > > be possible to add something to paraffin to make it stay a black color and > > remain strong as our commercial paraffin products? 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From ree3 <@t> leicester.ac.uk Fri Jul 22 09:17:34 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] AMACR Message-ID: Looking for an antibody to alpha-methylacyl co-enzyme A racemose(AMACR) which works on paraffin sections of mouse tissues. Many thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER.U.K..... From sjchtascp <@t> yahoo.com Fri Jul 22 09:22:10 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] glycerol Message-ID: <20050722142210.84803.qmail@web90203.mail.scd.yahoo.com> I need to have someone refresh my memory why some special stains call for glycerol? Steve --------------------------------- Do you Yahoo!? Yahoo! Mail - You care about security. So do we. From cfavara <@t> niaid.nih.gov Fri Jul 22 09:28:03 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: Osmium will make everything black! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Steven P Postl [mailto:steven.p.postl@abbott.com] Sent: Thursday, July 21, 2005 5:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Black paraffin and cassettes A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From steven.p.postl <@t> abbott.com Fri Jul 22 09:31:55 2005 From: steven.p.postl <@t> abbott.com (Steven P Postl) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: Let me add a few points after touching base with the researcher. There will be no tissue processed, nor labeling needed. Just a plain paraffin block made. This block will be used like a "cork board" but with paraffin instead to pin a tissue flat to the surface. Odd request, but I have to ask. Thanks again for those who have answered, and thanks to all of you who are scratching your head, drinking your coffee and thinking about this. TGIF ----- Forwarded by Steven P Postl/LAKE/PPRD/ABBOTT on 07/22/2005 09:24 AM ----- Steven P Postl 07/21/2005 07:40 PM To: histonet@lists.utsouthwestern.edu cc: Subject: Black paraffin and cassettes A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. From Kemlo.Rogerson <@t> elht.nhs.uk Fri Jul 22 09:34:59 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698DF6@elht-exch1.xelht.nhs.uk> That's a bit like black bed sheets; a trifle strange! -----Original Message----- From: Monfils, Paul [mailto:PMonfils@Lifespan.org] Sent: 22 July 2005 15:11 To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Black paraffin and cassettes I believe the dye Sudan Black B is soluble in paraffin. Black cassettes? Nope. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Steven P Postl > Sent: Thursday, July 21, 2005 5:40 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Black paraffin and cassettes > > A researcher is wondering (now so am I [I never heard this request > before]) if any company sells black paraffin and black embedding > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > > be possible to add something to paraffin to make it stay a black color and > > remain strong as our commercial paraffin products? 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 Kemlo.Rogerson <@t> elht.nhs.uk Fri Jul 22 09:36:44 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] glycerol Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698DF7@elht-exch1.xelht.nhs.uk> Doesn't it prevent over oxidation? -----Original Message----- From: Steven Coakley [mailto:sjchtascp@yahoo.com] Sent: 22 July 2005 15:22 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] glycerol I need to have someone refresh my memory why some special stains call for glycerol? Steve --------------------------------- Do you Yahoo!? Yahoo! Mail - You care about security. So do we. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kbroomal <@t> NEMOURS.ORG Fri Jul 22 09:36:33 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824395@wlmmsx02.nemours.org> Take a cassette & color it with a Sharpie or slide pen. ;) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Steven P Postl Sent: Friday, July 22, 2005 10:32 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Black paraffin and cassettes Let me add a few points after touching base with the researcher. There will be no tissue processed, nor labeling needed. Just a plain paraffin block made. This block will be used like a "cork board" but with paraffin instead to pin a tissue flat to the surface. Odd request, but I have to ask. Thanks again for those who have answered, and thanks to all of you who are scratching your head, drinking your coffee and thinking about this. TGIF ----- Forwarded by Steven P Postl/LAKE/PPRD/ABBOTT on 07/22/2005 09:24 AM ----- Steven P Postl 07/21/2005 07:40 PM To: histonet@lists.utsouthwestern.edu cc: Subject: Black paraffin and cassettes A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri Jul 22 09:45:17 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: Steve - You can get black WAX for this purpose from an art supply store, where they have candlemaking supplies. I'm pretty sure someone (maybe surgipath) makes grey cassettes. Jackie O' "Steven P Postl" Sent by: histonet-bounces@lists.utsouthwestern.edu 07/22/2005 09:31 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Black paraffin and cassettes Let me add a few points after touching base with the researcher. There will be no tissue processed, nor labeling needed. Just a plain paraffin block made. This block will be used like a "cork board" but with paraffin instead to pin a tissue flat to the surface. Odd request, but I have to ask. Thanks again for those who have answered, and thanks to all of you who are scratching your head, drinking your coffee and thinking about this. TGIF ----- Forwarded by Steven P Postl/LAKE/PPRD/ABBOTT on 07/22/2005 09:24 AM ----- Steven P Postl 07/21/2005 07:40 PM To: histonet@lists.utsouthwestern.edu cc: Subject: Black paraffin and cassettes A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Fri Jul 22 09:46:07 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717582@lsexch.lsmaster.lifespan.org> Steven, If the block doesn't have to be processed and sectioned, the paraffin could easily be colored by stirring in powdered carbon, which is essentially the same colorant found in black drawing ink. And the cassettes could be colored with one of the new spray paints designed specifically for painting plastics. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Steven P Postl > Sent: Friday, July 22, 2005 7:31 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Black paraffin and cassettes > > Let me add a few points after touching base with the researcher. There > will be no tissue processed, nor labeling needed. Just a plain paraffin > block made. This block will be used like a "cork board" but with paraffin > > instead to pin a tissue flat to the surface. Odd request, but I have to > ask. Thanks again for those who have answered, and thanks to all of you > who are scratching your head, drinking your coffee and thinking about > this. TGIF > > > ----- Forwarded by Steven P Postl/LAKE/PPRD/ABBOTT on 07/22/2005 09:24 AM > ----- > > > Steven P Postl > 07/21/2005 07:40 PM > > > To: histonet@lists.utsouthwestern.edu > cc: > Subject: Black paraffin and cassettes > > A researcher is wondering (now so am I [I never heard this request > before]) if any company sells black paraffin and black embedding > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > > be possible to add something to paraffin to make it stay a black color and > > remain strong as our commercial paraffin products? Thanks. > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gcallis <@t> montana.edu Fri Jul 22 09:50:31 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] using glycerin aka glycerol Message-ID: <6.0.0.22.1.20050722084938.01b6ab20@gemini.msu.montana.edu> Prevent evaporation? 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 PMonfils <@t> Lifespan.org Fri Jul 22 09:52:33 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717583@lsexch.lsmaster.lifespan.org> Biological supply companies sell dissecting pans with a layer of BLACK wax in them. Unlike embedding wax, this wax is specifically designed to be easily penetrated by pins. And you don't have to buy the whole pan. They also sell the wax itself. 5 pounds for $12.50! You can find it here: www.ctvalleybio.com > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Steven P Postl > Sent: Friday, July 22, 2005 7:31 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Black paraffin and cassettes > > Let me add a few points after touching base with the researcher. There > will be no tissue processed, nor labeling needed. Just a plain paraffin > block made. This block will be used like a "cork board" but with paraffin > > instead to pin a tissue flat to the surface. Odd request, but I have to > ask. Thanks again for those who have answered, and thanks to all of you > who are scratching your head, drinking your coffee and thinking about > this. TGIF > > > ----- Forwarded by Steven P Postl/LAKE/PPRD/ABBOTT on 07/22/2005 09:24 AM > ----- > > > Steven P Postl > 07/21/2005 07:40 PM > > > To: histonet@lists.utsouthwestern.edu > cc: > Subject: Black paraffin and cassettes > > A researcher is wondering (now so am I [I never heard this request > before]) if any company sells black paraffin and black embedding > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > > be possible to add something to paraffin to make it stay a black color and > > remain strong as our commercial paraffin products? Thanks. > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From AFeatherstone <@t> KaleidaHealth.Org Fri Jul 22 10:01:02 2005 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Ventana ES for sale Message-ID: <139141F8BAF4A642A945ECC528511AF0012A77F4@kalmb02.kaleidahealth.org> Is anyone out there interested in purchasing one or two working Ventana ES stainers? Annette Featherstone HT/MLT Supervisor Anatomic Pathology Kaleida Health, Buffalo General Hospital 100 High Street Buffalo NY 14203 716-859-2625 FAX: 716-859-1853 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Friday, July 22, 2005 10:37 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 29 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. tissue falling off slides (Zhenming Yu) 2. RE: Chlamydia antibody and Aspergillus IHC (Sverlow, Karen) 3. Reagent Search: PDTS (Owen, Michael P) 4. FDC-M1 antibody (Gayle Callis) 5. CD25 on murine tissue woes (Gayle Callis) 6. Re: Reagent Search: PDTS (Gayle Callis) 7. RE: Reagent Search: PDTS (Monfils, Paul) 8. type of mouse for spleen control Re: CD25 on murine tissue woes (Gayle Callis) 9. RE: Gram staining (Lewis, Sarah) 10. MGMT (Till, Renee) 11. Re: RE: Chlamydia antibody and Aspergillus IHC (Jan Shivers) 12. RE: CD25 on murine tissue woes (Mathew DeGutes) 13. BIOGENEX (Patsy Ruegg) 14. Murine Blood Smear / Smears IHC (GT Hebert) 15. Re: postmortem tissue collection (Y. Wang) 16. Re: Workloads vs Hiso staffing (Katia Cristina Catunda) 17. RE: Murine Blood Smear / Smears IHC (Favara, Cynthia (NIH/NIAID)) 18. Black paraffin and cassettes (Steven P Postl) 19. anyone know who sells these bottles? (curt tague) 20. RE: Black paraffin and cassettes (Kristen Broomall) 21. muscle histochemistry- (bcgirish) 22. RE: Black paraffin and cassettes (Monfils, Paul) 23. RE: Black paraffin and cassettes (Pamela Marcum) 24. RE: Black paraffin and cassettes (Weems, Joyce) 25. AMACR (Edwards, R.E.) 26. glycerol (Steven Coakley) 27. RE: Black paraffin and cassettes (Favara, Cynthia (NIH/NIAID)) 28. Black paraffin and cassettes (Steven P Postl) 29. RE: Black paraffin and cassettes (Rogerson Kemlo (ELHT) Pathology) ---------------------------------------------------------------------- Message: 1 Date: Thu, 21 Jul 2005 13:02:49 -0400 From: Zhenming Yu Subject: [Histonet] tissue falling off slides To: histonet@lists.utsouthwestern.edu Message-ID: <1121965369.42dfd539b88cb@webmail.pobox.upenn.edu> Content-Type: text/plain; charset=ISO-8859-1 Thanks to so many of you for the very helpful responses to my last question regarding chatter in fly head paraffin section. It turned out that we probably had over-processed the fly heads in alcohol and xylene. We are modifying our protocols and keeping our fingers crossed that the problem will be solved soon. Also I got another question that I hope you guys can help with---we sometimes ran across the problem of patches of tissue coming off slides. Could this also be attributed to over exposure of the heads to alcohol and xylene during processing? We have been using Superfrost Plus slides from Fisher---is any further slide treatment necessary? We usually lay ribbons (5 or 7 micron thick) of paraffin sections on the water surface in a 38?C water bath for less than one minute (too short?) before loading to a slide. Then the slide was kept in the vertical position for about 30 minutes to dry and incubated in a 37 incubator overnight for further drying. Any thoughts would be gratefully received. Thanks. **************************** Zhenming Yu Univ. of Penn Philadelphia, PA 19104 **************************** ------------------------------ Message: 2 Date: Thu, 21 Jul 2005 10:20:46 -0700 From: "Sverlow, Karen" Subject: [Histonet] RE: Chlamydia antibody and Aspergillus IHC To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi Everyone, Has anyone found a replacement antibody for the Virostat Chlamydia monoclonal product #1631 (no longer available)? Specifically, we are looking for a monoclonal that will detect Chlamydia in FFPE mammalian and avian tissues. On another note, I would like to talk with those of you doing Aspergillus IHC. Please email me. Thank you, Karen Sverlow Staff Research Associate California Animal Health and Food Safety Laboratory University of California, Davis ksverlow@cahfs.ucdavis.edu ------------------------------ Message: 3 Date: Thu, 21 Jul 2005 10:23:58 -0700 From: "Owen, Michael P" Subject: [Histonet] Reagent Search: PDTS To: 'Histonet' Message-ID: Content-Type: text/plain Dear Histonet Members, A colleague of mine is looking for a reagent called "Phenolphthalein Disulfate, Tripotassium Salt" for a staining project. The Sigma-Aldrich product number is P0251. Sigma-Aldrich has stated the chemical is no longer available. Does anyone on this list know of an alternative source? Thanks in advance. Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov ------------------------------ Message: 4 Date: Thu, 21 Jul 2005 11:27:05 -0600 From: Gayle Callis Subject: [Histonet] FDC-M1 antibody To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050721112507.01b18438@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Cynthia, Yes, we stain for this using IFA, but it also works IHC. We use frozens sections only, and it was an easy protocol. The antibody came from BD Pharmingen. You wrote: Does anyone have any experience with FDC[follicular dendritic cell] -M1 antibody clone 4C11 in mouse tissue?I do have some references that I am tracking down, but am having difficulty with the source 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: 5 Date: Thu, 21 Jul 2005 11:33:08 -0600 From: Gayle Callis Subject: [Histonet] CD25 on murine tissue woes To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050721112807.01b13c90@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Matthew, You wrote: I am trying to get CD25 staining to work in order to identify T reg cells in mouse tissue sections. So far I really haven't had much luck. I have both a 7D4 biotinylated anti-CD25 from Pharmingen and a PC61 Fitc-conjugated anti-CD25 from eBioscience and I am using Spleen as a positive control. I am using fresh frozen sections that have not been fixed at all. I have tried various fixations with Acetone and PFA. I would think PFA might block staining as many people have had trouble getting staining on parafin sections. I have played with many different antibody concentrations and we have secondaries and amplification protocols that have worked for other biotinylated and fitc-conjugated antibodies quite well using strep-hrp and anti-fitc-hrp along with a TSA cy3 or fluorescein kit. Does anyone have a protocol or some tips on making these antibodies stain for immunofluorescence? I know this is a common stain, but I simply have had no luck with it. Any help is appreciated thanks. Could you provide more details on HOW you are doing the staining, both as IHC and or immunofluorescence. If you are trying to do the primary antibody directly conjugated to FITC on a tissue section, you may not get positive results - this is not uncommon with some of this type conjugate as a direct IFA stain. I can't make it work with CD4-FITC or CD8-FITC. As for biotinylated antibodies (primaries) one can do either IHC and IFA - we do this on many other rat antimouse primaries, CD markers - so provide concentrations, times, etc, etc - all those good things. Thanks Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 6 Date: Thu, 21 Jul 2005 11:40:16 -0600 From: Gayle Callis Subject: [Histonet] Re: Reagent Search: PDTS To: "Owen, Michael P" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050721113905.01b6cf68@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Try some other chemical companies - ACROS vis Fisher, and ICN are possibilities. At 11:23 AM 7/21/2005, you wrote: >Dear Histonet Members, > >A colleague of mine is looking for a reagent called "Phenolphthalein >Disulfate, Tripotassium Salt" for a staining project. The Sigma-Aldrich >product number is P0251. > >Sigma-Aldrich has stated the chemical is no longer available. Does anyone on >this list know of an alternative source? 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: 7 Date: Thu, 21 Jul 2005 13:51:08 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] Reagent Search: PDTS To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171757F@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="ISO-8859-1" Lab Depot carries this compound. www.labdepotinc.com Also, you can search for suppliers of chemicals at www.chemexper.com Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Owen, > Michael P > Sent: Thursday, July 21, 2005 10:23 AM > To: 'Histonet' > Subject: [Histonet] Reagent Search: PDTS > > > > Dear Histonet Members, > > A colleague of mine is looking for a reagent called "Phenolphthalein > Disulfate, Tripotassium Salt" for a staining project. The Sigma-Aldrich > product number is P0251. > > Sigma-Aldrich has stated the chemical is no longer available. Does anyone > on > this list know of an alternative source? > > Thanks in advance. > > > Michael P. Owen, Regulatory Microbiologist > U.S. FDA Pacific Regional Lab Northwest > 22201 23rd Drive SE Bothell, WA 98021-4421 > Phone: 425-483-4865 E-Mail: michael.owen@fda.gov > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 8 Date: Thu, 21 Jul 2005 11:54:25 -0600 From: Gayle Callis Subject: [Histonet] type of mouse for spleen control Re: CD25 on murine tissue woes To: "Jackie M O'Connor" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050721115143.01b4be20@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Mouse was C57 black. At 11:44 AM 7/21/2005, you wrote: >Just curious, Gayle - what type of mouse is your spleen from? > >Jacqueline M. O'Connor HT(ASCP) QIHC >Assistant Scientist >Discovery Cancer R4N2 AP3 >847-938-4919 >Jackie.O'Connor@abbott.com Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 9 Date: Thu, 21 Jul 2005 13:55:54 -0400 From: "Lewis, Sarah" Subject: RE: [Histonet] Gram staining To: , Message-ID: <714B9F12B4E18C4C843B66E8E190F2AD44755C@res2k3ms01.CRII.ORG> Content-Type: text/plain; charset="iso-8859-1" rumor has it that *slim Jim* beef jerky works for both gram positive and gram negative.>>>?? -----Original Message----- From: jcarpenter764@aol.com [mailto:jcarpenter764@aol.com] Sent: Thursday, July 21, 2005 10:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Gram staining Can anyone give me some tips on how to get Gram stain controls or how to substitute for the control. Thanks Jennell ------------------------------ Message: 10 Date: Thu, 21 Jul 2005 14:19:32 -0500 From: "Till, Renee" Subject: [Histonet] MGMT To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Hello. Does anyone know of a good MGMT antibody for rat tissues? I have found several that say weakly reactive with rat tissue, and many that don't specify. Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 ------------------------------ Message: 11 Date: Thu, 21 Jul 2005 14:33:34 -0500 From: "Jan Shivers" Subject: Re: [Histonet] RE: Chlamydia antibody and Aspergillus IHC To: "Sverlow, Karen" Cc: histonet Message-ID: <00c801c58e2b$15c5ce30$41065486@auxs.umn.edu> Content-Type: text/plain; charset="iso-8859-1" I don't use a monoclonal for Chlamydia, but I do use a rabbit polyclonal from Biodesign (B65256R). It's made against Chlamydia trachomatis (EB's all antigens), but cross-reacts with Chlamydia psittacii and pneumoniae. It requires no tissue pretreatment. Jan Shivers Univ. of Minnesota Vet Diag Lab ----- Original Message ----- From: "Sverlow, Karen" To: Sent: Thursday, July 21, 2005 12:20 PM Subject: [Histonet] RE: Chlamydia antibody and Aspergillus IHC Hi Everyone, Has anyone found a replacement antibody for the Virostat Chlamydia monoclonal product #1631 (no longer available)? Specifically, we are looking for a monoclonal that will detect Chlamydia in FFPE mammalian and avian tissues. On another note, I would like to talk with those of you doing Aspergillus IHC. Please email me. Thank you, Karen Sverlow Staff Research Associate California Animal Health and Food Safety Laboratory University of California, Davis ksverlow@cahfs.ucdavis.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Thu, 21 Jul 2005 15:00:54 -0500 From: Mathew DeGutes Subject: [Histonet] RE: CD25 on murine tissue woes To: Histonet@lists.utsouthwestern.edu Message-ID: <200507212001.j6LK14Mv008154@casbah.it.northwestern.edu> Content-Type: text/plain If you want all the nitty gritty details. General protocol is this 1. Cut sections. Let dry and store at -80c until use. 2. Let sections return to room temperature and then fix. (Have tried Acetone 10 min -20c, 2%PFA 5 min room temp) 3. Wash 3x5 PBS 4. Wash 5 min in 3% H2O2 (have tried without this step) 5. Wash 3x5 PBS 6. Encircle and Avidin Block 10 min. (avidin biotin blocks are dependent on whether a biotinylated antibody is used). 7. Wash off Avidin in PBS 8. Biotin block 10 min 10. Aspirate Biotin block and add Block (have tried various serum at 10% and TNB, either with CD16/32 added at 1:50) for 30 min minimum. 11. Aspirate block and add Primary ab (have tried both 7D4 biotinylated anti-CD25 from Pharmingen and a PC61 Fitc-conjugated anti-CD25 from eBioscience at concentrations from 1:25 to 1:500) diluted in TNB or a 2% serum + .2% Triton in PBS buffer for 1 hour at room temp (have also tried overnight at 4c, but generally do the 1hour room temp). 12. Wash 3x5 PBS 13. Add 2ndary ab (Either Streptavidin-HRP or Anti-Fitc HRP) 1:100 in staining buffer from step 11 and inc for 30 min. 14. Wash 3x5 PBS 15. Use TSA Cy3 or Fluorescein kit. Dilute 1:100 in diluent and develop 5 minutes. 16. Wash 3x5 PBS 17. Mount with Vectashield with DAPI and coverslip Now we have used these 2ndaries and TSA kits extensively and have found them to work very well on any number of antigens. As I said before I haven't attempted to make it work with a direct. I come back and amplify with anti-fitc-hrp and the TSA kit. Also we are generally working in SJL mouse strains. I would really like to get this to work to go alongside FoxP3 staining for regs. I realize that regs are supposedly lower in SJL strains, but I have attempted this on Naive BalbC spleens as well and also have attempted on spleens from mice that were given injections of CD25+ CD4+ Tcells and so should have a far greater number of cells that are CD25+ (whether these cells will actually show up in anything other than flow I am not sure). You Wrote: Could you provide more details on HOW you are doing the staining, both as IHC and or immunofluorescence. If you are trying to do the primary antibody directly conjugated to FITC on a tissue section, you may not get positive results - this is not uncommon with some of this type conjugate as a direct IFA stain. I can't make it work with CD4-FITC or CD8-FITC. As for biotinylated antibodies (primaries) one can do either IHC and IFA - we do this on many other rat antimouse primaries, CD markers - so provide concentrations, times, etc, etc - all those good things. ------------------------------ Message: 13 Date: Thu, 21 Jul 2005 14:23:42 -0600 From: "Patsy Ruegg" Subject: [Histonet] BIOGENEX To: "'Histonet'" Message-ID: <200507212023.j6LKNepf006377@chip.viawest.net> Content-Type: text/plain; charset="US-ASCII" Can someone from BioGenex please contact me about a MW of theirs I am demoing. Patsy Ruegg Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. ------------------------------ Message: 14 Date: Thu, 21 Jul 2005 15:30:22 -0700 (PDT) From: GT Hebert Subject: [Histonet] Murine Blood Smear / Smears IHC To: Histonet@lists.utsouthwestern.edu Message-ID: <20050721223022.30674.qmail@web31710.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello everyone, I've searched but my answers so far have come up too vague. I need to know if it is possible to do IHC on mouse blood smears???? If so, how would one go about preparing and fixing the sample? Is there anything I should watch out for? I just want to run normal IHC (probably to mark certain white blood cells (for example, monocytes to start). Any assistance is valued and greatly appreciated. Thanks! Gustave Hebert Scientist II Wyeth Research Cambridge MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 15 Date: Thu, 21 Jul 2005 15:47:19 -0700 (PDT) From: "Y. Wang" Subject: Re: [Histonet] postmortem tissue collection To: Geoff McAuliffe Cc: Histonet@lists.utsouthwestern.edu, "Rogerson Kemlo \(ELHT\) Pathology" Message-ID: Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed Thank you all for your thoughts on this question. I haven't found anything more enlightening on the web searches I've done so far. However, I will continue searching and will let you know if and when I find some thing exciting (aside from the strange but interesting nuggets that one always happens across in these searches on tissue death after death). Thanks again Yak-Nam > Many years ago (I hate beginning my responses like that but it is true) > several colleagues were able to get viable human retinal pigment epithelial > cells more than 48 hours after death. Also, I know of viable olfactory > (sensory) epithelial cells being harvested 24 hours after death. Of course, > ambient temperature is a likely variable and not all cells die at the same > time. I would think that the adipose tissue in the hypodermis would have a > relatively slow metabolic rate and might be viable for 24 hours? You have > little to lose by trying. > > Geoff > > Rogerson Kemlo (ELHT) Pathology wrote: > >> Interestingly I was reading about tissue death after death (so as to >> speak) on a Web Site dealing with Death (I'm strange like that); I was >> interested in rigor. I hadn't realised that some tissue deep within the >> cadaver can survive for many hours after death. I suppose it's those >> tissues that require little or no oxygen to survive; brain dies very >> rapidly. If I could remember the Site then I'd tell you but a search in >> Google under rigor may help. Wonder which bit of you dies last? Same in >> men and women? >> >> -----Original Message----- >> From: Y. Wang [mailto:ynwang@u.washington.edu] Sent: 20 July 2005 00:20 >> To: Histonet@lists.utsouthwestern.edu >> Subject: [Histonet] postmortem tissue collection >> >> Dear histonetters, >> >> I have a question regarding collection of human tissue. A colleague >> would like to collect human tissue (skin and underlying adipose tissue) for >> mechanical testing, histological analysis (cellular and structural >> evaluation), IHC and protein analysis (extracellular matrix structure >> and concentrations). They asked what would be a fair cut off time for >> tissue >> >> collection so that the effects of decay would not be a factor. Currently >> >> they have given the tissue bank a time of 12 hours postmortem (I'm not very >> sure how the body is stored during this time or how this is calculated). >> >> I've read that human decomposition starts approx. 4 min after death and >> autolysis is quicker in tissues with high enzyme and water content. >> However, in terms of the skin and underlying adipose tissue I didn't >> know if there was an accepted 'cut off time' postmortem after which tissue >> is considered far from representative of 'live' tissue and not worth >> analysis. >> >> Can anyone give some insight? Any information or references would be >> greatly appreciated. >> >> Thank you >> Yak-Nam >> >> Senior Fellow >> Department of Bioengineering >> University of Washington >> Box 357962 >> Seattle, WA 98195 >> >> Tel.: (206)-221-5873 >> Fax.: (206)-221-5874 >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > > -- > -- > ********************************************** > Geoff McAuliffe, Ph.D. > Neuroscience and Cell Biology > Robert Wood Johnson Medical School > 675 Hoes Lane, Piscataway, NJ 08854 > voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu > ********************************************** > > > ------------------------------ Message: 16 Date: Thu, 21 Jul 2005 20:22:32 -0300 From: "Katia Cristina Catunda" Subject: Re: [Histonet] Workloads vs Hiso staffing To: Message-ID: <015b01c58e4b$12750f00$a279fea9@privatexx> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original We do have a great interest on that question either. My supervisor sent me an FAQ he got on CAP telling that the ideal workload would be 50 H/E slides per histotechnician http://www.cap.org/apps/docs/cap_today/q_and_a/qa_1102.html - answered by Richard W. Brown, MD In our lab we do have 12 histotechnicians with a routine that is about 470 blocks (for 1/3 of them we generate two slides, one for HE and one for H. pilory) and 600 citological slides per day. They alternate their function every week: - embedding - 3 of them - microtomy 4 of them - staining/mounting and organizing slides - 2 of them - specific staining -the same of embedding - archiving blocks - the same of staining - citology routine - 2 of them - one is allways on vacation - immunohistochemistry - 2 of them are moved to other room twice a week I was wondering which is the best way to determinate their workload in order to analyse if we have too many or too less employees. My personal opinion is that 50 H/E slides per employee would be very hard-working and out of our range. Does anybody has any idea? But all of these informations are international since we are from Brazil... Katia ----- Original Message ----- From: "Maray Weirauch" To: Sent: Monday, July 18, 2005 3:44 PM Subject: [Histonet] Workloads vs Hiso staffing > ** PRIVATE ** > > Our hospital is conducting an analysis of laboratory workload, workflow > and staffing. This will include Histology as well, and we are being > asked for input with any national benchmarks available for surgical > histology tasks (i.e. average number cases accessioned, blocks embedded, > slides cut../some type of time unit) Does anyone know of some realistic > published values or ranges? Thanks! > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 17 Date: Thu, 21 Jul 2005 20:15:57 -0400 From: "Favara, Cynthia (NIH/NIAID)" Subject: RE: [Histonet] Murine Blood Smear / Smears IHC To: 'GT Hebert' , Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Questions - why are you dong this? What is the ultimate goal? If you just want a differential then do a differential. If you are looking for various populations of cells then FACS might be more reasonable and far simpler. Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: GT Hebert [mailto:emerald_lake77@yahoo.com] Sent: Thursday, July 21, 2005 3:30 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Murine Blood Smear / Smears IHC Hello everyone, I've searched but my answers so far have come up too vague. I need to know if it is possible to do IHC on mouse blood smears???? If so, how would one go about preparing and fixing the sample? Is there anything I should watch out for? I just want to run normal IHC (probably to mark certain white blood cells (for example, monocytes to start). Any assistance is valued and greatly appreciated. Thanks! Gustave Hebert Scientist II Wyeth Research Cambridge MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Thu, 21 Jul 2005 19:40:26 -0500 From: "Steven P Postl" Subject: [Histonet] Black paraffin and cassettes To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. ------------------------------ Message: 19 Date: Thu, 21 Jul 2005 18:05:31 -0700 (PDT) From: curt tague Subject: [Histonet] anyone know who sells these bottles? To: histonet@lists.utsouthwestern.edu Message-ID: <20050722010531.85013.qmail@web80101.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 there are 2 picutres under histo net images. i'd love to find a vendor for these bottles. others are a tad too small for my labels. thanks. ------------------------------ Message: 20 Date: Fri, 22 Jul 2005 08:24:34 -0400 From: "Kristen Broomall" Subject: RE: [Histonet] Black paraffin and cassettes To: "'Steven P Postl'" , histonet@lists.utsouthwestern.edu Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824394@wlmmsx02.nemours.org> Content-Type: text/plain; charset=iso-8859-1 I would think it would be difficult to find something to write on a black cassette & be visible. Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Steven P Postl Sent: Thursday, July 21, 2005 8:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Black paraffin and cassettes A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: 22 Jul 2005 13:21:05 -0000 From: "bcgirish" Subject: [Histonet] muscle histochemistry- To: histonet@lists.utsouthwestern.edu Message-ID: <20050722132105.13765.qmail@webmail29.rediffmail.com> Content-Type: text/plain; charset=iso-8859-1 Hi Bernice Gut Schmidt I am very much interested in the method you are using for muscle atpse histochemistry(Dubowitz and Brooke).If you can send me the same it will be very much useful to us. My Fax ------91-40-23045438 Phone ----914023045439 REGARDS Dr.Girish B.Chandrashekar Dept of Preclinical Safety Evaluation Dr Reddys Lab Hyderabad-49 ------------------------------ Message: 22 Date: Fri, 22 Jul 2005 10:10:58 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] Black paraffin and cassettes To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717580@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="ISO-8859-1" I believe the dye Sudan Black B is soluble in paraffin. Black cassettes? Nope. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Steven P Postl > Sent: Thursday, July 21, 2005 5:40 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Black paraffin and cassettes > > A researcher is wondering (now so am I [I never heard this request > before]) if any company sells black paraffin and black embedding > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > > be possible to add something to paraffin to make it stay a black color and > > remain strong as our commercial paraffin products? Thanks. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 23 Date: Fri, 22 Jul 2005 10:16:24 -0400 From: Pamela Marcum Subject: RE: [Histonet] Black paraffin and cassettes To: "Monfils, Paul" , "'histonet@lists.utsouthwestern.edu'" Message-ID: <6.1.1.1.2.20050722101403.019bfb80@mail.vet.upenn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed At 10:10 AM 7/22/2005, Monfils, Paul wrote: >I believe the dye Sudan Black B is soluble in paraffin. > >Black cassettes? Nope. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > > Steven P Postl > > Sent: Thursday, July 21, 2005 5:40 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] Black paraffin and cassettes > > > > A researcher is wondering (now so am I [I never heard this request > > before]) if any company sells black paraffin and black embedding > > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > > > > be possible to add something to paraffin to make it stay a black color and > > > > remain strong as our commercial paraffin products? 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 It is soluble in paraffin however it will be removed by reagents used in deparaffinization and staining. Due to the additional materials added to raw paraffin it will be spotty. It will also leave some precipitate in the tissue. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu ------------------------------ Message: 24 Date: Fri, 22 Jul 2005 10:16:54 -0400 From: "Weems, Joyce" Subject: RE: [Histonet] Black paraffin and cassettes To: Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA461D8@sjhaexc02.sjha.org> Content-Type: text/plain; charset="iso-8859-1" Perhaps clear cassettes would work? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Monfils, Paul Sent: Friday, July 22, 2005 10:11 AM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Black paraffin and cassettes I believe the dye Sudan Black B is soluble in paraffin. Black cassettes? Nope. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Steven P Postl > Sent: Thursday, July 21, 2005 5:40 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Black paraffin and cassettes > > A researcher is wondering (now so am I [I never heard this request > before]) if any company sells black paraffin and black embedding > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > > be possible to add something to paraffin to make it stay a black color and > > remain strong as our commercial paraffin products? 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. ------------------------------ Message: 25 Date: Fri, 22 Jul 2005 15:17:34 +0100 From: "Edwards, R.E." Subject: [Histonet] AMACR To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Looking for an antibody to alpha-methylacyl co-enzyme A racemose(AMACR) which works on paraffin sections of mouse tissues. Many thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER.U.K..... ------------------------------ Message: 26 Date: Fri, 22 Jul 2005 07:22:10 -0700 (PDT) From: Steven Coakley Subject: [Histonet] glycerol To: Histonet@lists.utsouthwestern.edu Message-ID: <20050722142210.84803.qmail@web90203.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I need to have someone refresh my memory why some special stains call for glycerol? Steve --------------------------------- Do you Yahoo!? Yahoo! Mail - You care about security. So do we. ------------------------------ Message: 27 Date: Fri, 22 Jul 2005 10:28:03 -0400 From: "Favara, Cynthia (NIH/NIAID)" Subject: RE: [Histonet] Black paraffin and cassettes To: 'Steven P Postl' , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Osmium will make everything black! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Steven P Postl [mailto:steven.p.postl@abbott.com] Sent: Thursday, July 21, 2005 5:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Black paraffin and cassettes A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 28 Date: Fri, 22 Jul 2005 09:31:55 -0500 From: "Steven P Postl" Subject: [Histonet] Black paraffin and cassettes To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Let me add a few points after touching base with the researcher. There will be no tissue processed, nor labeling needed. Just a plain paraffin block made. This block will be used like a "cork board" but with paraffin instead to pin a tissue flat to the surface. Odd request, but I have to ask. Thanks again for those who have answered, and thanks to all of you who are scratching your head, drinking your coffee and thinking about this. TGIF ----- Forwarded by Steven P Postl/LAKE/PPRD/ABBOTT on 07/22/2005 09:24 AM ----- Steven P Postl 07/21/2005 07:40 PM To: histonet@lists.utsouthwestern.edu cc: Subject: Black paraffin and cassettes A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. ------------------------------ Message: 29 Date: Fri, 22 Jul 2005 15:34:59 +0100 From: "Rogerson Kemlo (ELHT) Pathology" Subject: RE: [Histonet] Black paraffin and cassettes To: "Monfils, Paul" , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698DF6@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" That's a bit like black bed sheets; a trifle strange! -----Original Message----- From: Monfils, Paul [mailto:PMonfils@Lifespan.org] Sent: 22 July 2005 15:11 To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Black paraffin and cassettes I believe the dye Sudan Black B is soluble in paraffin. Black cassettes? Nope. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Steven P Postl > Sent: Thursday, July 21, 2005 5:40 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Black paraffin and cassettes > > A researcher is wondering (now so am I [I never heard this request > before]) if any company sells black paraffin and black embedding > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > > be possible to add something to paraffin to make it stay a black color and > > remain strong as our commercial paraffin products? 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 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 29 **************************************** CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From funderwood <@t> mcohio.org Fri Jul 22 09:56:30 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes Message-ID: Maybe mixing some India ink into the melted paraffin will work. >>> "Steven P Postl" 07/22/05 10:31AM >>> Let me add a few points after touching base with the researcher. There will be no tissue processed, nor labeling needed. Just a plain paraffin block made. This block will be used like a "cork board" but with paraffin instead to pin a tissue flat to the surface. Odd request, but I have to ask. Thanks again for those who have answered, and thanks to all of you who are scratching your head, drinking your coffee and thinking about this. TGIF ----- Forwarded by Steven P Postl/LAKE/PPRD/ABBOTT on 07/22/2005 09:24 AM ----- Steven P Postl 07/21/2005 07:40 PM To: histonet@lists.utsouthwestern.edu cc: Subject: Black paraffin and cassettes A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From psanquin <@t> lugo.usc.es Fri Jul 22 10:11:28 2005 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Endogenous peroxidase blocking Message-ID: <3.0.6.32.20050722171128.0081b100@pop.lugo.usc.es> Dear Histonetters, I ask for your help. I am doing an immunohistochemical procedure in really bloddy specimens. The endogenous peroxidase blocking solution is 3% v/v hydrogen peroxyde in distilled water for 15 minutes. I am afraid that this blocking is not enough. How can I be sure that I have totally removed the endogenous activity? Are there stronger treatments than mine? Should the endogenous peroxidase staining appear only in red cells and granulocites or could it also produce a background staining in the whole tissue? Best regards Pablo Sanchez From stancelb <@t> msn.com Fri Jul 22 10:30:11 2005 From: stancelb <@t> msn.com (Barbara Stancel) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] RE: Black Paraffin Message-ID: Dear Steven, If you just want to make a paraffin board, try black or brown crayons. Or go to a candle store and get some candle/wax coloring made for coloring wax. Good Luck Histologically yours, Barbara Barbara Stancel Eastern Laboratory, Pathology Athens, Georgia 30604 Original Message: From: Steven P Postl [mailto:] Sent: Thursday, July 21, 2005 5:40 PM A researcher is wondering (now so am I [I never heard this request before]) if any company sells black paraffin and black embedding cassettes? No, this isn't a Gothic thing, a legitimate request. Would it be possible to add something to paraffin to make it stay a black color and remain strong as our commercial paraffin products? Thanks. From jfish <@t> gladstone.ucsf.edu Sat Jul 23 10:38:04 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Black paraffin and cassettes In-Reply-To: References: Message-ID: Dear Steven, Here is what I have learned to do for aorta pinouts: Melt about half a pound of pink dental wax along with about 1/4 piece of black candle dye. I'm sorry, there is no weight on the package, so it is actually a piece about the size of my thumbnail. Just mix and stir until it is a color you would like. I use a small cardboard box, with no seams, to mold the "plate" of wax. It comes out easily after chilling (I use one of my cold plates to chill them). The cassettes are too small for what we are pinning out. I hope this helps, and good luck! Jo Dee At 9:31 AM -0500 7/22/05, Steven P Postl wrote: >Let me add a few points after touching base with the researcher. There >will be no tissue processed, nor labeling needed. Just a plain paraffin >block made. This block will be used like a "cork board" but with paraffin >instead to pin a tissue flat to the surface. Odd request, but I have to >ask. Thanks again for those who have answered, and thanks to all of you >who are scratching your head, drinking your coffee and thinking about >this. TGIF > > >----- Forwarded by Steven P Postl/LAKE/PPRD/ABBOTT on 07/22/2005 09:24 AM >----- > > >Steven P Postl >07/21/2005 07:40 PM > > > To: histonet@lists.utsouthwestern.edu > cc: > Subject: Black paraffin and cassettes > >A researcher is wondering (now so am I [I never heard this request >before]) if any company sells black paraffin and black embedding >cassettes? No, this isn't a Gothic thing, a legitimate request. Would it >be possible to add something to paraffin to make it stay a black color and >remain strong as our commercial paraffin products? Thanks. > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** ********** Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 734-5766 Fax: (415) 355-0824 E-mail: jfish@gladstone.ucsf.edu Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158 From jseaton <@t> wlgore.com Fri Jul 22 10:58:15 2005 From: jseaton <@t> wlgore.com (Janella Seaton) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] specimen database Message-ID: Hello! We want to implement a database for specimens we receive in our lab. We would use this database starting with logging in specimens and ending with archiving of specimens. It would be used to track when various steps (gross photos, processing, staining, etc) have occurred and by whom. We do want a system that is user friendly. Any suggestions or advice? Thanks for the help..... j.s. From renafail <@t> bellsouth.net Fri Jul 22 11:17:11 2005 From: renafail <@t> bellsouth.net (renafail@bellsouth.net) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] AMACR In-Reply-To: Message-ID: <000601c58ed8$d13308b0$0301a8c0@RENAD4YK9B8ABE> Try Biocare -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Edwards, R.E. Sent: Friday, July 22, 2005 10:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] AMACR Looking for an antibody to alpha-methylacyl co-enzyme A racemose(AMACR) which works on paraffin sections of mouse tissues. Many thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER.U.K..... _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Jul 22 11:18:08 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:22 2005 Subject: Complete Endog peroxidase blocking Re: [Histonet] Endogenous peroxidase blocking In-Reply-To: <3.0.6.32.20050722171128.0081b100@pop.lugo.usc.es> References: <3.0.6.32.20050722171128.0081b100@pop.lugo.usc.es> Message-ID: <6.0.0.22.1.20050722101641.01b02cf0@gemini.msu.montana.edu> Pablo, I will be sending via private email - a method that may help. It is the glucose oxidase blocking method, and used nicely for frozen sections but also paraffin sections. At 09:11 AM 7/22/2005, you wrote: >Dear Histonetters, > >I ask for your help. I am doing an immunohistochemical procedure in really >bloddy specimens. The endogenous peroxidase blocking solution is 3% v/v >hydrogen peroxyde in distilled water for 15 minutes. > >I am afraid that this blocking is not enough. How can I be sure that I have >totally removed the endogenous activity? Are there stronger treatments than >mine? > >Should the endogenous peroxidase staining appear only in red cells and >granulocites or could it also produce a background staining in the whole >tissue? > >Best regards > >Pablo Sanchez > > > > >_______________________________________________ >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 PMonfils <@t> Lifespan.org Fri Jul 22 11:46:54 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Endogenous peroxidase blocking Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717584@lsexch.lsmaster.lifespan.org> We have repeatedly observed that aqueous peroxide solutions do not quench thoroughly when large amounts of endogenous peroxidase are present. We use the following solution, which works much better for us: Methanol ................ 300 ml Hydrogen Peroxide ... 5.0 ml HCl ........................ 0.6 ml Usual time is 20 minutes. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Pablo > S?nchez Quinteiro > Sent: Friday, July 22, 2005 8:11 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Endogenous peroxidase blocking > > Dear Histonetters, > > I ask for your help. I am doing an immunohistochemical procedure in really > bloddy specimens. The endogenous peroxidase blocking solution is 3% v/v > hydrogen peroxyde in distilled water for 15 minutes. > > I am afraid that this blocking is not enough. How can I be sure that I > have > totally removed the endogenous activity? Are there stronger treatments > than > mine? > > Should the endogenous peroxidase staining appear only in red cells and > granulocites or could it also produce a background staining in the whole > tissue? > > Best regards > > Pablo Sanchez > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gsennello <@t> osip.com Fri Jul 22 11:46:53 2005 From: gsennello <@t> osip.com (Sennello, Gina) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] specimen database Message-ID: I would be interested in this information also. Please address replies to everyone. Thanks Gina OSIP Boulder, CO -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Janella Seaton Sent: Friday, July 22, 2005 9:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] specimen database Hello! We want to implement a database for specimens we receive in our lab. We would use this database starting with logging in specimens and ending with archiving of specimens. It would be used to track when various steps (gross photos, processing, staining, etc) have occurred and by whom. We do want a system that is user friendly. Any suggestions or advice? Thanks for the help..... j.s. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Fri Jul 22 12:00:16 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] 2"x3" slides Message-ID: Does anyone out there have a good way to stain 2"x3" glass slides (whole-mount prostates) using a Sakura stainer? I have a 2000 and 601 model and have tried adapting the rack so I can stain 4 slides at a time, but it's so inefficient. Any ideas? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From contact <@t> excaliburpathology.com Fri Jul 22 12:07:53 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Specimen Database Message-ID: <20050722170753.97109.qmail@web50305.mail.yahoo.com> I created my entire patient database myself in Microsoft Access. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From Janet.Bonner <@t> FLHOSP.ORG Fri Jul 22 12:18:13 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] specimen database Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4332@fh2k093.fhmis.net> We're using Copath with alot of success - it is very user-friendly. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 7/22/2005 11:58 AM Subject: [Histonet] specimen database Hello! We want to implement a database for specimens we receive in our lab. We would use this database starting with logging in specimens and ending with archiving of specimens. It would be used to track when various steps (gross photos, processing, staining, etc) have occurred and by whom. We do want a system that is user friendly. Any suggestions or advice? Thanks for the help..... j.s. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Jul 22 12:53:51 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] 2"x3" slides In-Reply-To: References: Message-ID: <6.0.0.22.1.20050722115129.01b58ce8@gemini.msu.montana.edu> Years ago, I think the Shandon stainer had a rack to accomodate the larger slides, maybe Sakura needs to do this too. Otherwise, and we still do it now - hand staining, and can do either 20 or 30 at a time - kinda old fashioned but very efficient. At 11:00 AM 7/22/2005, you wrote: >Does anyone out there have a good way to stain 2"x3" glass slides >(whole-mount prostates) using a Sakura stainer? I have a 2000 and 601 >model and have tried adapting the rack so I can stain 4 slides at a >time, but it's so inefficient. Any ideas? > >Angela Bitting, HT(ASCP) >Technical Specialist, Histology >Geisinger Medical Center >100 N Academy Ave. MC 23-00 >Danville, PA 17822 >phone 570-214-9634 >fax 570-271-5916 > > >IMPORTANT WARNING: The information in this message (and the documents >attached to it, if any) is confidential and may be legally privileged. It >is intended solely for the addressee. Access to this message by anyone >else is unauthorized. If you are not the intended recipient, any >disclosure, copying, distribution or any action taken, or omitted to be >taken, in reliance on it is prohibited and may be unlawful. If you have >received this message in error, please delete all electronic copies of >this message (and the documents attached to it, if any), destroy any hard >copies you may have created and notify me immediately by replying to this >email. Thank you. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 psanquin <@t> lugo.usc.es Fri Jul 22 13:14:53 2005 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Endogenous peroxidase blocking In-Reply-To: <3.0.6.32.20050722171128.0081b100@pop.lugo.usc.es> Message-ID: <3.0.6.32.20050722201453.007b35e0@pop.lugo.usc.es> Dear histonetters, Thanks a lot for all your tips! One further question. When doing a control immunohistochemical procedure omitting the primary antibody I guess that if I have not blocked the endogenous peroxidase activity properly I should get staining. Isnt'it? If in case I do not get any stain may I understand that the endogenous activity is not a real problem? I am working with free floating sections, polyclonal antibody, ABC, DAB. Thanks once and again Pablo From JosefaNava <@t> texashealth.org Fri Jul 22 13:22:49 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Feedback on Fc Blocker and Background buster for Ventana IHC Machines Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944D5@phdex03.txhealth.org> Hello Everyone, Can someone give me the protocol of the FC Blocker and Background Buster reagents if you are using them for your Ventana IHC Machines. Please give me your feedback about these 2 reagents on your Ventana Machines. Thank you. Josie Nava Presbyterian Hospital of Dallas The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From kbowden <@t> ucsd.edu Fri Jul 22 13:28:48 2005 From: kbowden <@t> ucsd.edu (K.Bowden) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] specimen database In-Reply-To: <07AB60D5D7B9754EBF56F360F98D083DEB4332@fh2k093.fhmis.net> References: <07AB60D5D7B9754EBF56F360F98D083DEB4332@fh2k093.fhmis.net> Message-ID: <42E13AE0.4040304@ucsd.edu> I made a database using FileMaker Pro. It was fairly easy to create and it is totally custom. -- Karen Bowden Staff Research Associate II University of CA, San Diego Department of Orthopedics 9500 Gilman Dr. 0630 La Jolla, CA 92093-0630 858-534-4655 CONFIDENTIALITY NOTICE: THE INFORMATION TRANSMITTED IN THIS E-MAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVED THIS E-MAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER. >Hello! >We want to implement a database for specimens we receive in our lab. >We would use this database starting with logging in specimens and ending >with archiving of specimens. It would be used to track when various >steps >(gross photos, processing, staining, etc) have occurred and by whom. >We do want a system that is user friendly. >Any suggestions or advice? >Thanks for the help..... >j.s. > > >_______________________________________________ >Histonet mailing list >Histonet@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 gu.lang <@t> gmx.at Fri Jul 22 13:34:30 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:22 2005 Subject: AW: [Histonet] 2"x3" slides In-Reply-To: Message-ID: Sakura sells special forms to put in their usual racks for the bigger slides. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Angela Bitting Gesendet: Freitag, 22. Juli 2005 19:00 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] 2"x3" slides Does anyone out there have a good way to stain 2"x3" glass slides (whole-mount prostates) using a Sakura stainer? I have a 2000 and 601 model and have tried adapting the rack so I can stain 4 slides at a time, but it's so inefficient. Any ideas? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JosefaNava <@t> texashealth.org Fri Jul 22 13:35:08 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] background on the negative controls Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944D6@phdex03.txhealth.org> Hello, Has anyone had a problem on their negative controls having heavy brown staining that looks like real DAB staining on negative mouse and Rabbit controls that were retirieved using heat but the negative control that used only protease treatment was very clean. I did a lot of IHCs on adrenal mass tissue and my Rabbit and Mouse negatives look like they have positive staining. H2O2 treatment did not correct the problem, I also used the Avidin Biotin block. Anymore suggestions? Thank you. Josie The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From LINDA.MARGRAF <@t> childrens.com Fri Jul 22 13:41:54 2005 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Free Histology Learning Resources Message-ID: Dear Histonetters: I got an email from the marketing director for Visual Histology and he wanted to know if it was ok to put a message out on Histonet about it. I thought it looked like something interesting so here are his comments..... I want to make you aware of a relatively new website that provides some valuable free resources for histology teachers and students. The website is www.VisualHistology.com and anyone who registers with the site can download the Visual Histology Atlas (a 285 page comprehensive histology textbook from Moran and Rowley) and also get free access to a 30-minute video tutorial called "The Cell." The site also sells access to the complete Visual Histology video tutorial series (26 titles) through a premium membership. To learn more, visit: www.VisualHistology.com If you have questions please inquire at the website as I haven't had any personal experience with these products. Thanks Linda M Histonet administrator From mprice26 <@t> juno.com Fri Jul 22 13:55:50 2005 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] RE: Limit on amount of formalin you can pour down sink? Message-ID: <20050722.115646.4572.95444@webmail28.nyc.untd.com> Hi Histonetters, Does anyone know of any regulations as to the amount of formalin one can pour down the sink? I am in Texas. Thank you. Marsha Price From Jackie.O'Connor <@t> abbott.com Fri Jul 22 14:08:06 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] RE: Limit on amount of formalin you can pour down sink? Message-ID: Yeah - nothing. It's a hazardous chemical. I don't know of any state where local regulations allow formalin in the wastewater. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist Discovery Cancer R4N2 AP3 847-938-4919 Jackie.O'Connor@abbott.com "mprice26@juno.com" bellsouth.net Fri Jul 22 14:15:03 2005 From: kallred <@t> bellsouth.net (Kim Allred) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] CD 31 Message-ID: <20050722190950.UPKM23081.ibm59aec.bellsouth.net@D6GV9G51> I am having a problem getting my CD31 to work. The protocol was for an overnight incubation and pepsin pretreatment. I couldn't get it to work this way so I altered it a bit by using trilogy for AR and a two hour incubation for the primary ab. Does anyone have any ideas? This seemed to work well for a while but once again, I'm having problems with it. Any input would be appreciated. Thanks in advance! Kim Allred Dermatopathology Associates Jackson, MS From renafail <@t> bellsouth.net Fri Jul 22 14:18:24 2005 From: renafail <@t> bellsouth.net (renafail@bellsouth.net) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] CD 31 In-Reply-To: <20050722190950.UPKM23081.ibm59aec.bellsouth.net@D6GV9G51> Message-ID: <000701c58ef2$21f48530$0301a8c0@RENAD4YK9B8ABE> Hi Kim, We use CD31 at 1:50, pretreatment HIER using Citrate buffer Rena Fail -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Allred Sent: Friday, July 22, 2005 3:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD 31 I am having a problem getting my CD31 to work. The protocol was for an overnight incubation and pepsin pretreatment. I couldn't get it to work this way so I altered it a bit by using trilogy for AR and a two hour incubation for the primary ab. Does anyone have any ideas? This seemed to work well for a while but once again, I'm having problems with it. Any input would be appreciated. Thanks in advance! Kim Allred Dermatopathology Associates Jackson, MS _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Fri Jul 22 14:27:02 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] cryosectioning liver/sucrose Message-ID: <20050722192702.5732.qmail@web90205.mail.scd.yahoo.com> Something a "tad" new for me here. I've never sectioned 4%Para liver infiltrated with 30% sucrose. They sectioned well but the slide appear to be loaded with what appears to be folds throughout. Not what I routinely see on regular unfixed FS. Is this normal for fixed sucrose infiltrated sections. Are there any special techniques when dealing with this special tissue. Steve --------------------------------- Start your day with Yahoo! - make it your home page From marktarango <@t> earthlink.net Fri Jul 22 14:27:41 2005 From: marktarango <@t> earthlink.net (Mark Tarango) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Re: Black paraffin and cassettes References: <200507221035.1dVYD03N53Nl3490@mx-nebolish.atl.sa.earthlink.net> Message-ID: <001801c3d161$eebf6760$6401a8c0@TARANGO> I'm wondering how anyone could embed in a black tar kind of paraffin. I'm guessing that this researcher doesn't have much experience in histology. How would this help them? Mark Tarango > Date: Thu, 21 Jul 2005 19:40:26 -0500 > From: "Steven P Postl" > Subject: [Histonet] Black paraffin and cassettes > To: histonet@lists.utsouthwestern.edu > Message-ID: > > > Content-Type: text/plain; charset="us-ascii" > > A researcher is wondering (now so am I [I never heard this request > before]) if any company sells black paraffin and black embedding > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > be possible to add something to paraffin to make it stay a black color and > remain strong as our commercial paraffin products? Thanks. From Janet.Bonner <@t> FLHOSP.ORG Fri Jul 22 14:36:27 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] RE: Limit on amount of formalin you can pour down sink? Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4339@fh2k093.fhmis.net> Orlando Florida allows it. A few of the large cities are equipped to handle formalin in the system. You have to contact the municipal water system people to see if they are set up to handle the formalin and receive written confirmation for your records (and the inspectors) before you start dumping ANY of it in the sink. We just started recycling ours with a Creative Waste Solutions,Inc Formalin recycler (rex@cwsincorp.com) and have been very pleased with the results. How and why it works I leave to him to explain, but it does. Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of mprice26@juno.com Sent: Friday, July 22, 2005 2:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Limit on amount of formalin you can pour down sink? Hi Histonetters, Does anyone know of any regulations as to the amount of formalin one can pour down the sink? I am in Texas. Thank you. Marsha Price _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Fri Jul 22 15:17:52 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] cryosectioning liver/sucrose Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717588@lsexch.lsmaster.lifespan.org> At any given temperature, tissues infiltrated with sucrose tend to be softer and less completely frozen than similar tissues without sucrose (and 30% is a very high level of sucrose), and therefore they compress and wrinkle more. Try cutting them at a colder temperature than you would normally use for liver. That will probably help produce flatter sections with less wrinkling. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Steven Coakley > Sent: Friday, July 22, 2005 12:27 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] cryosectioning liver/sucrose > > Something a "tad" new for me here. I've never sectioned 4%Para liver > infiltrated with 30% sucrose. They sectioned well but the slide appear to > be loaded with what appears to be folds throughout. Not what I routinely > see on regular unfixed FS. Is this normal for fixed sucrose infiltrated > sections. Are there any special techniques when dealing with this special > tissue. > > Steve > > > > --------------------------------- > Start your day with Yahoo! - make it your home page > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From emry <@t> u.washington.edu Fri Jul 22 16:44:04 2005 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] under decal Message-ID: Is there anything that can be done for under decalcified bone after it has been processed? Trisha Seattle From Janet.Bonner <@t> FLHOSP.ORG Fri Jul 22 17:13:21 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] under decal Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB433B@fh2k093.fhmis.net> OPTION 1: "redecal" Melt the paraffin block, put the tissue in a cassette as you would a fresh piece of tissue to process. Drop it in xylene for 15 minutes x2, then 100% abs. alcohol x2, 95% abs. alcohol. Wash in water and drop back into the decal. Process when soft. OPTION 2: "surface decal" Pour some decal solution into a small container or Petri dish and put the faced block face-down in the solution. Let soak for 15 min (time depends on hardness of the tissue) put on ice and try to section. Only the surface will be decaled so try to take a section off the surface. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Trisha Emry Sent: Friday, July 22, 2005 5:44 PM To: histo Subject: [Histonet] under decal Is there anything that can be done for under decalcified bone after it has been processed? Trisha Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Fri Jul 22 17:23:51 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] under decal Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717589@lsexch.lsmaster.lifespan.org> Sometimes you can get a decent section by pressing gauze soaked with decal fluid on the face of the block for a couple of minutes (wear gloves), then taking the first section off the face. Or you may get a few successive sections by soaking the faced block in decal fluid for 15 to 30 minutes, then chilling the block on the cold tray and cutting right off the face. However, if you are going to be doing any substantial amount of work with that block, probably the best approach is to reverse process it, decal it properly, and reprocess it. Remove the paraffin by soaking in xylene or whatever clearing agent you use, remove the xylene with absolute alcohol, bring the specimen down to water, then decal it, wash it and process it as usual. If you want you can place it in formalin overnight once it reaches the water stage, to ensure the best possible fixation before decalcifying it. Since most tissue processors won't run the stations in reverse order, you'll probably have to do the reverse processing by hand, in a beaker or flask. If the tissue was well fixed initially, it should withstand this treatment with little if any noticeable harm. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Trisha Emry > Sent: Friday, July 22, 2005 2:44 PM > To: histo > Subject: [Histonet] under decal > > Is there anything that can be done for under decalcified bone after it has > been processed? > > Trisha > Seattle > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From lrichey <@t> u.washington.edu Fri Jul 22 17:48:27 2005 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] CD 31 In-Reply-To: <20050722190950.UPKM23081.ibm59aec.bellsouth.net@D6GV9G51> References: <20050722190950.UPKM23081.ibm59aec.bellsouth.net@D6GV9G51> Message-ID: <42E177BB.2050905@u.washington.edu> We use DAKO CD31 1:25 , 18 minutes microwave, in citrate buffer. Kim Allred wrote: >I am having a problem getting my CD31 to work. The protocol was for an >overnight incubation and pepsin pretreatment. I couldn't get it to work >this way so I altered it a bit by using trilogy for AR and a two hour >incubation for the primary ab. Does anyone have any ideas? This seemed to >work well for a while but once again, I'm having problems with it. Any >input would be appreciated. Thanks in advance! > > > > > > > >Kim Allred > >Dermatopathology Associates > >Jackson, MS > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From IamNinaOwen <@t> aol.com Sat Jul 23 03:19:18 2005 From: IamNinaOwen <@t> aol.com (IamNinaOwen@aol.com) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Staining for Helicobacter pylori Message-ID: Hi everyone! I'm doing a project on different staining methods for H. pylori in gastric biopsies, and just out of interest really wondered which special stains different labs routinely use for this purpose. My lab uses the Modified Giemsa. Thanks Nina Owen From 41dmb41 <@t> gmail.com Sat Jul 23 10:39:27 2005 From: 41dmb41 <@t> gmail.com (Drew Meyer) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Staining for Helicobacter pylori In-Reply-To: References: Message-ID: We use the Wharton Stary method.... you can also use an Alcian Yellow Stain... there's another called the Toluidine Method I believe... Hope this helps... Drew Meyer On 7/23/05, IamNinaOwen@aol.com wrote: > Hi everyone! > > I'm doing a project on different staining methods for H. pylori in gastric > biopsies, and just out of interest really wondered which special stains > different labs routinely use for this purpose. My lab uses the Modified Giemsa. > > Thanks > > Nina Owen > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From beingmary53 <@t> sbcglobal.net Sat Jul 23 13:00:09 2005 From: beingmary53 <@t> sbcglobal.net (MARY JOHNSON) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] decal bone Message-ID: <20050723180009.98249.qmail@web81609.mail.yahoo.com> Hi Trisha, you can face off you blocks and put them in 1/2 HCL and 1/2 decal solution for about an hour. It works for me. Hope this work for you. Mary From lfidgen <@t> vt.edu Sat Jul 23 14:01:30 2005 From: lfidgen <@t> vt.edu (Laura Fidgen) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Rat brains Message-ID: <6.0.0.22.0.20050723145647.0267aa98@pop.vt.edu> Please help... A research project for our neuropathologist includes rat brains. These are extremely difficult to section as he profuses the whole rat with glutaraldehyde then takes brain samples. I have tried adding alcohol to my water bath but these sections still have many, many wrinkles. Can anyone offer suggestions? I would be so grateful for any and all advice. Thank You!! Laura L. Fidgen, MScF, BSc, HT(ASCP) Laboratory and Research Practioner VMRCVM, Histopatholgy Blacksburg, VA 24060-0443 From ohenry <@t> dfw.net Sat Jul 23 15:42:29 2005 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] RE: Limit on amount of formalin you can pour down the sink? Message-ID: <001501c58fc7$0f4bd8e0$13dd3040@your4f1261a8e5> I'm in Texas too. You will have to check with your local water / waste utility. It will vary from city to city and from lab to lab within the same city. Allot will depend on your use, and just how much you want to pour down the sink and how often,i.e.: daily,weekly,etc. Same is true with alcohol. Susan Subject: [Histonet] RE: Limit on amount of formalin you can pour down sink? Hi Histonetters, Does anyone know of any regulations as to the amount of formalin one can pour down the sink? I am in Texas. Thank you. Marsha Price From kweidenh <@t> montefiore.org Sun Jul 24 07:57:58 2005 From: kweidenh <@t> montefiore.org (Karen Weidenheim) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] muscle histochemistry Message-ID: Here is a recipe for ATPase at 9.4 that we use in our lab: ADENOSINE TRIPHOSPHATASE (ATP) at pH 9.4 Principle: The histochemical reaction for adenosine triphosphatase is the primary method for determining distribution, size, and proportion of muscle fiber types 1 and 2. Calcium ions in the incubating medium precipitate the enzymatic reaction product, orthophosphate, thereby localizing ATPase activity. Treatment with cobaltous chloride produces cobalt phosphate, which is visualized, when brought into contact with dilute ammonium sulfide, as black/brown cobalt sulfide deposits. Specimen Preparation: Freshly cut frozen sections cut at 8 micra on coverslips. Solutions 0.1 M Sodium barbital buffer 2.0629 g sodium barbital (sodium diethyl barbiturate, powder) 100 ml distilled water Mix well. Label with date of preparation and expiration date of 1 year. Store at 4o C. Replace if precipitate forms. 0.18 M calcium chloride 1.9989 g calcium chloride 100 ml distilled water Mix. Label with date of preparation and expiration date of 1 year. Store at 4o C. Replace if precipitate forms. 2% calcium chloride 2 g calcium chloride 100 ml distilled water Mix. Label with date of preparation and expiration date of 1 year. Store at 4o C. Replace if precipitate forms. Cobaltous chloride solution 2 gm cobaltous chloride 100 ml distilled water Mix well. Label with date of preparation and expiration date of 1 year. Store at 4o C. Replace if precipitate forms. Ammonium sulfide solution (make fresh in fume hood each time) 1 ml ammonium sulfide 23 ml distilled water Working solution (must be freshly made each time) 4 ml 0.lM Sodium barbital solution 2 ml 0.18M Calcium chloride solution 4 ml distilled water 30 mg ATP (disodium salt, crystalline, equine muscle, 99-100% pure, Sigma) Mix all. Adjust pH to 9.4 Formol-calcium fixative 100 ml 40% formaldehyde Excess of calcium carbonate 100 ml 10% calcium chloride 300 ml distilled water Mix. Store in a glass bottle (not a plastic bottle) at 4oC. Method. 1. Fix tissue in cold formol-calcium for 5 minutes. 2. Rinse in distilled water 1 minute. 3. Incubate in working solution at 37o C for 30 minutes. 4. Rinse in 2% calcium chloride for 2 minutes. Repeat 3 times. 5. Rinse in 2% cobaltous chloride 5 minutes. 6. Rinse in tap water for 5-10 minutes until water is clear. 7. Develop in ammonium sulfide for 3-5 minutes. 8. Dehydrate through alcohols to xylene and mount in Permount. Results: Type 1 fibers are pale, Type 2 fibers are dark. Blood vessels are dark. Intermediate fibers are not usually seen in normal muscle with this stain, but they do appear in denervation and reinnervation. Quality Control: Blood vessels in the specimen are black, providing a built-in control. If they are not black, stain is repeated. Reference: Khan MA, Papadimitriou JM, Holt PG, Kakulas BA. A modified histochemical technique for sarcoplasmic reticular ATPase. Histochemie 1972;30:329-333. Khan MA, Papadimitriou JM, Kakulas BA. On the specificity of the histochemical technique for sarcoplasmic reticular adenosine triphosphatase: a light and electron microscopic study. Histochemistry 1975;43:101-111. Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> "bcgirish" 07/20/05 9:54 AM >>> Hi all I am Dr Girish,toxicological pathologist working for a pharmaceutical company in India. We are recently standardizing myosin atpse histochemistry to study the effect of ppar compounds on skeletal muscle. We are using liquid nitrogen without isopentane to collect the muscle gastrocnemius from rats.The reaction is givinig nonspecific reaction and artifacts. Is using of isopentane must? Is there a better method than the one described by Bancroft? How to preserve the enzyme activity?How long we have to incubate in incubating solution if we are using Bancroft method?we are using atp at the concentration of 5mg/5ml solution b of bancroft method.How long this solution can be kept and at what temperature? Is the pH maintained to be 9.4 or 10.4 (alkaline)? We are getting a sort of cavity in the muscle fibers, is it because of ice crystal artifacts? I am waiting for quick suggestions.Do share ur experience . .......................... REGARDS DR.GIRISH B.CHANDRASHEKAR DEPT OF PRECLINICAL SAFETY EVALUATION DR REDDYS LAB HYDERABAD-49 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Sun Jul 24 15:12:47 2005 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] RE: Limit on amount of formalin you can pour down sink? In-Reply-To: Message-ID: <200507242012.j6OKCp94071629@pro12.abac.com> There are new regs on formaldehyde, it is now a KNOWN human carcinogen according to our H&S consultant, the exposure limits are 2 parts per million to humans, I would think that pouring it down a drain would expose the pourer and anyone else in the area to limits higher than that unless your drain is in a fume hood. We need some haz. Chemicals experts to chime in here. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M O'Connor Sent: Friday, July 22, 2005 1:08 PM To: mprice26@juno.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Limit on amount of formalin you can pour down sink? Yeah - nothing. It's a hazardous chemical. I don't know of any state where local regulations allow formalin in the wastewater. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist Discovery Cancer R4N2 AP3 847-938-4919 Jackie.O'Connor@abbott.com "mprice26@juno.com" ihctech.net Sun Jul 24 15:17:39 2005 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:25:22 2005 Subject: Complete Endog peroxidase blocking Re: [Histonet] Endogenousperoxidase blocking In-Reply-To: <6.0.0.22.1.20050722101641.01b02cf0@gemini.msu.montana.edu> Message-ID: <200507242017.j6OKHhJk073165@pro12.abac.com> I had a particularly difficult time quenching endogenous peroxidase in sputum cytopreps full of granulocytes and it was suggested that I try the new DAKO Dual Block for HRP and Alk.Phos. and it worked a charm, don't know what's in it (proprietary you know), but perhaps it is similar to Gayles reagents for glucose oxidase blocking. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, July 22, 2005 10:18 AM To: Pablo S?nchez Quinteiro; Histonet@lists.utsouthwestern.edu Subject: Complete Endog peroxidase blocking Re: [Histonet] Endogenousperoxidase blocking Pablo, I will be sending via private email - a method that may help. It is the glucose oxidase blocking method, and used nicely for frozen sections but also paraffin sections. At 09:11 AM 7/22/2005, you wrote: >Dear Histonetters, > >I ask for your help. I am doing an immunohistochemical procedure in really >bloddy specimens. The endogenous peroxidase blocking solution is 3% v/v >hydrogen peroxyde in distilled water for 15 minutes. > >I am afraid that this blocking is not enough. How can I be sure that I have >totally removed the endogenous activity? Are there stronger treatments than >mine? > >Should the endogenous peroxidase staining appear only in red cells and >granulocites or could it also produce a background staining in the whole >tissue? > >Best regards > >Pablo Sanchez > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Sun Jul 24 15:20:19 2005 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] alk. phos. enzyme for osteoblasts Message-ID: <200507242020.j6OKKNxA073974@pro12.abac.com> Anybody doing alkaline phosphotase enzyme histochemistry on undecalcified bone for osteoblasts? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net From nienhuis <@t> ucla.edu Sun Jul 24 18:02:09 2005 From: nienhuis <@t> ucla.edu (NIENHUIS,ROBERT ) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] RE: Limit on amount of formalin you can pour down sink? In-Reply-To: <200507242012.j6OKCp94071629@pro12.abac.com> References: <200507242012.j6OKCp94071629@pro12.abac.com> Message-ID: <1122246129.42e41df17a98d@mail.ucla.edu> We use Formulex to treat formaldehyde before dumping down the drain. Bob Nienhuis UCLA / VA Medical Center Los Angeles, CA Quoting pruegg@ihctech.net: > There are new regs on formaldehyde, it is now a KNOWN human carcinogen > according to our H&S consultant, the exposure limits are 2 parts per > million > to humans, I would think that pouring it down a drain would expose the > pourer and anyone else in the area to limits higher than that unless > your > drain is in a fume hood. We need some haz. Chemicals experts to chime > in > here. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie > M > O'Connor > Sent: Friday, July 22, 2005 1:08 PM > To: mprice26@juno.com > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] RE: Limit on amount of formalin you can pour > down > sink? > > Yeah - nothing. It's a hazardous chemical. I don't know of any state > where local regulations allow formalin in the wastewater. > > > Jacqueline M. O'Connor HT(ASCP) QIHC > Assistant Scientist > Discovery Cancer R4N2 AP3 > 847-938-4919 > Jackie.O'Connor@abbott.com > > > > > > "mprice26@juno.com" Sent by: histonet-bounces@lists.utsouthwestern.edu > 07/22/2005 01:55 PM > > > To: histonet@lists.utsouthwestern.edu > cc: > Subject: [Histonet] RE: Limit on amount of formalin you > can > pour down sink? > > > > Hi Histonetters, > Does anyone know of any regulations as to the amount of formalin one > can > pour down the sink? I am in Texas. > > Thank you. > > Marsha Price > > _______________________________________________ > Histonet mailing list > Histonet@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 miffordclark <@t> optonline.net Sun Jul 24 19:48:07 2005 From: miffordclark <@t> optonline.net (clifford berger) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] RE: Limit on amount of formalin you can pour down sink? References: <200507242012.j6OKCp94071629@pro12.abac.com> Message-ID: <004801c590b2$85d144f0$0200a8c0@dellovo0ll7kuk> In order to facilitate this discussion, can we please just dintinguish between 10% Neutral Buffered Formalin and Formaldehyde. 10% NBF is what it is and Formaldehyde is the ingredient that is mixed with water and buffers to create 10% NBF. Again, just for this discussion. So, are the new regs which you refer to for 10% NBF or for Formaldehyde which is actually a corrosive, flammable and carinogen and is actually found in realtively small quanities in most pathology labs? Or are you talking about 10% NBF which is widely used in pathology labs throughout the world. And where can we find these regulations? ----- Original Message ----- From: To: "'Jackie M O'Connor'" ; Cc: Sent: Sunday, July 24, 2005 4:12 PM Subject: RE: [Histonet] RE: Limit on amount of formalin you can pour down sink? > There are new regs on formaldehyde, it is now a KNOWN human carcinogen > according to our H&S consultant, the exposure limits are 2 parts per > million > to humans, I would think that pouring it down a drain would expose the > pourer and anyone else in the area to limits higher than that unless your > drain is in a fume hood. We need some haz. Chemicals experts to chime in > here. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M > O'Connor > Sent: Friday, July 22, 2005 1:08 PM > To: mprice26@juno.com > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] RE: Limit on amount of formalin you can pour down > sink? > > Yeah - nothing. It's a hazardous chemical. I don't know of any state > where local regulations allow formalin in the wastewater. > > > Jacqueline M. O'Connor HT(ASCP) QIHC > Assistant Scientist > Discovery Cancer R4N2 AP3 > 847-938-4919 > Jackie.O'Connor@abbott.com > > > > > > "mprice26@juno.com" Sent by: histonet-bounces@lists.utsouthwestern.edu > 07/22/2005 01:55 PM > > > To: histonet@lists.utsouthwestern.edu > cc: > Subject: [Histonet] RE: Limit on amount of formalin you can > pour down sink? > > > > Hi Histonetters, > Does anyone know of any regulations as to the amount of formalin one can > pour down the sink? I am in Texas. > > Thank you. > > Marsha Price > > _______________________________________________ > Histonet mailing list > Histonet@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 adel2377 <@t> yahoo.com.sg Sun Jul 24 21:46:32 2005 From: adel2377 <@t> yahoo.com.sg (Adeline Chow) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] PGP 9.5 Message-ID: <20050725024632.91026.qmail@web60719.mail.yahoo.com> have a few questions.. 1) Do expired BDH polyLlysine slides have an effect on IHC staining besides the fact that they dun stick tissues on as well? 2) Does 4% paraformaldehyde as a fixative need to be prepared freshly before each use? It should be fine if its stored at 4C up to two months ? 3) Have been doing pgp 9.5 IHC on skin specimens (Hypothenar 3mm punches) for a while and sometimes looking for the fibres in the epidermis can be difficult. Sometimes its positive and sometimes its not. Staining protocol is , fixation in 4 % paraform. overnight followed by sucrose o/n and cut in a cryostat at 30um . Sections stained o/n 4C followed by use of DAKO envision kit . Anything wrong with this protocol? Pls help. Greatly appreciated. --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - 1GB free storage! From kappeler <@t> patho.unibe.ch Mon Jul 25 01:33:43 2005 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Pancreatic Stone Protein Message-ID: <00c701c590e2$df79d780$27955c82@patho.unibe.ch> Hi all does anybody know of a source for an antibody against (human) pancreatic stone protein (PSP)? Many thanks! Andi Kappeler Institute of Pathology, University of Bern, Switzerland From Jacqueline.Malam <@t> rli.mbht.nhs.uk Mon Jul 25 02:39:50 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Re: endogenous peroxidase blocking Message-ID: If you are performing IHC on really bloody specimens, you will be better off using an alkaline phosphatase method rather than HRP-based - it will give better results and be a lot less trouble. You may find you will affect the staining of your antigen if you increase the H2O2. Cheers Jacqui Malam Lancaster DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From holling <@t> med-in.uni-saarland.de Mon Jul 25 02:45:32 2005 From: holling <@t> med-in.uni-saarland.de (holling@med-in.uni-saarland.de) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] carstairs stain Message-ID: <2882.134.96.44.146.1122277532.squirrel@134.96.44.146> Hi, I would be grateful if somebody could give me the carstair stain procedure. TC Nicole Hollinger From angela.mcnabola.b <@t> bayer.com Mon Jul 25 05:40:13 2005 From: angela.mcnabola.b <@t> bayer.com (Angela McNabola) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] CD 31 Message-ID: Which antibody are you using? We have had great success with Santa Cruz (goat polyclonal) until I ordered a new lot this past month. When I called them, they indicated to me that they were having problems with their goat, literally. I guess it had stopped producing antibodies to CD31 that work in IHC (interestingly enough, they still work in Westerns). They hope to have the problem resolved within the next month, with a new goat. Angela McNabola, MS, HT(ASCP)SLS, QIHC Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 "Kim Allred" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] CD 31 western.edu 07/22/2005 03:15 PM I am having a problem getting my CD31 to work. The protocol was for an overnight incubation and pepsin pretreatment. I couldn't get it to work this way so I altered it a bit by using trilogy for AR and a two hour incubation for the primary ab. Does anyone have any ideas? This seemed to work well for a while but once again, I'm having problems with it. Any input would be appreciated. Thanks in advance! Kim Allred Dermatopathology Associates Jackson, MS _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From settembr <@t> umdnj.edu Mon Jul 25 06:38:08 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] CD 31 Message-ID: Kim, I use Dako's CD31 at an even more potent dilution than Rena does with Target Retieval Solution from Dako with a uterus as a positive control. Trilogy is good. You should stick with it and make your dilution more potent. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> 7/22/2005 3:18:24 PM >>> Hi Kim, We use CD31 at 1:50, pretreatment HIER using Citrate buffer Rena Fail -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Allred Sent: Friday, July 22, 2005 3:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD 31 I am having a problem getting my CD31 to work. The protocol was for an overnight incubation and pepsin pretreatment. I couldn't get it to work this way so I altered it a bit by using trilogy for AR and a two hour incubation for the primary ab. Does anyone have any ideas? This seemed to work well for a while but once again, I'm having problems with it. Any input would be appreciated. Thanks in advance! Kim Allred Dermatopathology Associates Jackson, MS _______________________________________________ Histonet mailing list Histonet@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 sbreeden <@t> nmda.nmsu.edu Mon Jul 25 07:15:08 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Under-Decal'd tissue Message-ID: Trisha - I swear by RDO, manufactured by Apex Engineering Products Corporation, Aurora, IL. The active ingredient is hydrochloric acid. If a block still contains calcification after either preliminary decalcification or if discovered after processing/facing, I drop the block into a small beaker with RDO and leave it for from 15-30 minutes (depending upon bone density). I've routinely used RDO for years on normal bony blocks, trephines, bone bx's, etc. and it works amazingly well. It is NOT, however, for LONG-TERM decalcification (i.e., overnight) because it works so well that it may render the bone devoid of any detail if left too long. The RDO must be washed off the block with a good dose of water before cutting, as the acid will discolor your knife holder. Their number is 800-451-6291 or you can contact them at www.rdo-apex.com. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Saturday, July 23, 2005 11:03 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 31 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. 2"x3" slides (Angela Bitting) 2. Specimen Database (Paula Pierce) 3. RE: specimen database (Bonner, Janet) 4. Re: 2"x3" slides (Gayle Callis) 5. Re: Endogenous peroxidase blocking (Pablo S?nchez Quinteiro) 6. Feedback on Fc Blocker and Background buster for Ventana IHC Machines (Nava, Josefa) 7. Re: specimen database (K.Bowden) 8. AW: [Histonet] 2"x3" slides (Gudrun Lang) 9. background on the negative controls (Nava, Josefa) 10. Free Histology Learning Resources (LINDA MARGRAF) 11. RE: Limit on amount of formalin you can pour down sink? (mprice26@juno.com) 12. Re: RE: Limit on amount of formalin you can pour down sink? (Jackie M O'Connor) 13. CD 31 (Kim Allred) 14. RE: CD 31 (renafail@bellsouth.net) 15. cryosectioning liver/sucrose (Steven Coakley) 16. Re: Black paraffin and cassettes (Mark Tarango) 17. RE: RE: Limit on amount of formalin you can pour down sink? (Bonner, Janet) 18. RE: cryosectioning liver/sucrose (Monfils, Paul) 19. under decal (Trisha Emry) 20. RE: under decal (Bonner, Janet) 21. RE: under decal (Monfils, Paul) 22. Re: CD 31 (Lori Richey) 23. Staining for Helicobacter pylori (IamNinaOwen@aol.com) 24. Re: Staining for Helicobacter pylori (Drew Meyer) ---------------------------------------------------------------------- Message: 1 Date: Fri, 22 Jul 2005 13:00:16 -0400 From: "Angela Bitting" Subject: [Histonet] 2"x3" slides To: Message-ID: Content-Type: text/plain; charset=US-ASCII Does anyone out there have a good way to stain 2"x3" glass slides (whole-mount prostates) using a Sakura stainer? I have a 2000 and 601 model and have tried adapting the rack so I can stain 4 slides at a time, but it's so inefficient. Any ideas? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. ------------------------------ Message: 2 Date: Fri, 22 Jul 2005 10:07:53 -0700 (PDT) From: Paula Pierce Subject: [Histonet] Specimen Database To: histonet@lists.utsouthwestern.edu Message-ID: <20050722170753.97109.qmail@web50305.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I created my entire patient database myself in Microsoft Access. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com ------------------------------ Message: 3 Date: Fri, 22 Jul 2005 13:18:13 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] specimen database To: "'Janella Seaton '" , "'histonet-bounces@lists.utsouthwestern.edu '" , "'histonet@lists.utsouthwestern.edu '" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4332@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 We're using Copath with alot of success - it is very user-friendly. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 7/22/2005 11:58 AM Subject: [Histonet] specimen database Hello! We want to implement a database for specimens we receive in our lab. We would use this database starting with logging in specimens and ending with archiving of specimens. It would be used to track when various steps (gross photos, processing, staining, etc) have occurred and by whom. We do want a system that is user friendly. Any suggestions or advice? Thanks for the help..... j.s. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Fri, 22 Jul 2005 11:53:51 -0600 From: Gayle Callis Subject: Re: [Histonet] 2"x3" slides To: "Angela Bitting" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050722115129.01b58ce8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Years ago, I think the Shandon stainer had a rack to accomodate the larger slides, maybe Sakura needs to do this too. Otherwise, and we still do it now - hand staining, and can do either 20 or 30 at a time - kinda old fashioned but very efficient. At 11:00 AM 7/22/2005, you wrote: >Does anyone out there have a good way to stain 2"x3" glass slides >(whole-mount prostates) using a Sakura stainer? I have a 2000 and 601 >model and have tried adapting the rack so I can stain 4 slides at a >time, but it's so inefficient. Any ideas? > >Angela Bitting, HT(ASCP) >Technical Specialist, Histology >Geisinger Medical Center >100 N Academy Ave. MC 23-00 >Danville, PA 17822 >phone 570-214-9634 >fax 570-271-5916 > > >IMPORTANT WARNING: The information in this message (and the documents >attached to it, if any) is confidential and may be legally privileged. It >is intended solely for the addressee. Access to this message by anyone >else is unauthorized. If you are not the intended recipient, any >disclosure, copying, distribution or any action taken, or omitted to be >taken, in reliance on it is prohibited and may be unlawful. If you have >received this message in error, please delete all electronic copies of >this message (and the documents attached to it, if any), destroy any hard >copies you may have created and notify me immediately by replying to this >email. Thank you. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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: 5 Date: Fri, 22 Jul 2005 20:14:53 +0200 From: Pablo S?nchez Quinteiro Subject: Re: [Histonet] Endogenous peroxidase blocking To: histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20050722201453.007b35e0@pop.lugo.usc.es> Content-Type: text/plain; charset="us-ascii" Dear histonetters, Thanks a lot for all your tips! One further question. When doing a control immunohistochemical procedure omitting the primary antibody I guess that if I have not blocked the endogenous peroxidase activity properly I should get staining. Isnt'it? If in case I do not get any stain may I understand that the endogenous activity is not a real problem? I am working with free floating sections, polyclonal antibody, ABC, DAB. Thanks once and again Pablo ------------------------------ Message: 6 Date: Fri, 22 Jul 2005 13:22:49 -0500 From: "Nava, Josefa" Subject: [Histonet] Feedback on Fc Blocker and Background buster for Ventana IHC Machines To: Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944D5@phdex03.txhealth.org> Content-Type: text/plain; charset="us-ascii" Hello Everyone, Can someone give me the protocol of the FC Blocker and Background Buster reagents if you are using them for your Ventana IHC Machines. Please give me your feedback about these 2 reagents on your Ventana Machines. Thank you. Josie Nava Presbyterian Hospital of Dallas The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. ------------------------------ Message: 7 Date: Fri, 22 Jul 2005 11:28:48 -0700 From: "K.Bowden" Subject: Re: [Histonet] specimen database To: "Bonner, Janet" Cc: "'histonet@lists.utsouthwestern.edu '" , "'histonet-bounces@lists.utsouthwestern.edu '" , 'Janella Seaton ' Message-ID: <42E13AE0.4040304@ucsd.edu> Content-Type: text/plain; charset=us-ascii; format=flowed I made a database using FileMaker Pro. It was fairly easy to create and it is totally custom. -- Karen Bowden Staff Research Associate II University of CA, San Diego Department of Orthopedics 9500 Gilman Dr. 0630 La Jolla, CA 92093-0630 858-534-4655 CONFIDENTIALITY NOTICE: THE INFORMATION TRANSMITTED IN THIS E-MAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVED THIS E-MAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER. >Hello! >We want to implement a database for specimens we receive in our lab. >We would use this database starting with logging in specimens and ending >with archiving of specimens. It would be used to track when various >steps >(gross photos, processing, staining, etc) have occurred and by whom. >We do want a system that is user friendly. >Any suggestions or advice? >Thanks for the help..... >j.s. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ------------------------------ Message: 8 Date: Fri, 22 Jul 2005 20:34:30 +0200 From: "Gudrun Lang" Subject: AW: [Histonet] 2"x3" slides To: "Histonetliste \(Histonetliste\)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Sakura sells special forms to put in their usual racks for the bigger slides. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Angela Bitting Gesendet: Freitag, 22. Juli 2005 19:00 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] 2"x3" slides Does anyone out there have a good way to stain 2"x3" glass slides (whole-mount prostates) using a Sakura stainer? I have a 2000 and 601 model and have tried adapting the rack so I can stain 4 slides at a time, but it's so inefficient. Any ideas? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Fri, 22 Jul 2005 13:35:08 -0500 From: "Nava, Josefa" Subject: [Histonet] background on the negative controls To: Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944D6@phdex03.txhealth.org> Content-Type: text/plain; charset="us-ascii" Hello, Has anyone had a problem on their negative controls having heavy brown staining that looks like real DAB staining on negative mouse and Rabbit controls that were retirieved using heat but the negative control that used only protease treatment was very clean. I did a lot of IHCs on adrenal mass tissue and my Rabbit and Mouse negatives look like they have positive staining. H2O2 treatment did not correct the problem, I also used the Avidin Biotin block. Anymore suggestions? Thank you. Josie The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. ------------------------------ Message: 10 Date: Fri, 22 Jul 2005 13:41:54 -0500 From: "LINDA MARGRAF" Subject: [Histonet] Free Histology Learning Resources To: Cc: doug@visualhistology.com Message-ID: Content-Type: text/plain; charset=US-ASCII Dear Histonetters: I got an email from the marketing director for Visual Histology and he wanted to know if it was ok to put a message out on Histonet about it. I thought it looked like something interesting so here are his comments..... I want to make you aware of a relatively new website that provides some valuable free resources for histology teachers and students. The website is www.VisualHistology.com and anyone who registers with the site can download the Visual Histology Atlas (a 285 page comprehensive histology textbook from Moran and Rowley) and also get free access to a 30-minute video tutorial called "The Cell." The site also sells access to the complete Visual Histology video tutorial series (26 titles) through a premium membership. To learn more, visit: www.VisualHistology.com If you have questions please inquire at the website as I haven't had any personal experience with these products. Thanks Linda M Histonet administrator ------------------------------ Message: 11 Date: Fri, 22 Jul 2005 18:55:50 GMT From: "mprice26@juno.com" Subject: [Histonet] RE: Limit on amount of formalin you can pour down sink? To: histonet@lists.utsouthwestern.edu Message-ID: <20050722.115646.4572.95444@webmail28.nyc.untd.com> Content-Type: text/plain Hi Histonetters, Does anyone know of any regulations as to the amount of formalin one can pour down the sink? I am in Texas. Thank you. Marsha Price ------------------------------ Message: 12 Date: Fri, 22 Jul 2005 14:08:06 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] RE: Limit on amount of formalin you can pour down sink? To: "mprice26@juno.com" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Yeah - nothing. It's a hazardous chemical. I don't know of any state where local regulations allow formalin in the wastewater. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist Discovery Cancer R4N2 AP3 847-938-4919 Jackie.O'Connor@abbott.com "mprice26@juno.com" Subject: [Histonet] CD 31 To: Message-ID: <20050722190950.UPKM23081.ibm59aec.bellsouth.net@D6GV9G51> Content-Type: text/plain; charset="us-ascii" I am having a problem getting my CD31 to work. The protocol was for an overnight incubation and pepsin pretreatment. I couldn't get it to work this way so I altered it a bit by using trilogy for AR and a two hour incubation for the primary ab. Does anyone have any ideas? This seemed to work well for a while but once again, I'm having problems with it. Any input would be appreciated. Thanks in advance! Kim Allred Dermatopathology Associates Jackson, MS ------------------------------ Message: 14 Date: Fri, 22 Jul 2005 15:18:24 -0400 From: Subject: RE: [Histonet] CD 31 To: "'Kim Allred'" , Message-ID: <000701c58ef2$21f48530$0301a8c0@RENAD4YK9B8ABE> Content-Type: text/plain; charset="us-ascii" Hi Kim, We use CD31 at 1:50, pretreatment HIER using Citrate buffer Rena Fail -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Allred Sent: Friday, July 22, 2005 3:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD 31 I am having a problem getting my CD31 to work. The protocol was for an overnight incubation and pepsin pretreatment. I couldn't get it to work this way so I altered it a bit by using trilogy for AR and a two hour incubation for the primary ab. Does anyone have any ideas? This seemed to work well for a while but once again, I'm having problems with it. Any input would be appreciated. Thanks in advance! Kim Allred Dermatopathology Associates Jackson, MS _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Fri, 22 Jul 2005 12:27:02 -0700 (PDT) From: Steven Coakley Subject: [Histonet] cryosectioning liver/sucrose To: Histonet@lists.utsouthwestern.edu Message-ID: <20050722192702.5732.qmail@web90205.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Something a "tad" new for me here. I've never sectioned 4%Para liver infiltrated with 30% sucrose. They sectioned well but the slide appear to be loaded with what appears to be folds throughout. Not what I routinely see on regular unfixed FS. Is this normal for fixed sucrose infiltrated sections. Are there any special techniques when dealing with this special tissue. Steve --------------------------------- Start your day with Yahoo! - make it your home page ------------------------------ Message: 16 Date: Fri, 2 Jan 2004 09:55:02 -0900 From: "Mark Tarango" Subject: [Histonet] Re: Black paraffin and cassettes To: Message-ID: <001801c3d161$eebf6760$6401a8c0@TARANGO> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original I'm wondering how anyone could embed in a black tar kind of paraffin. I'm guessing that this researcher doesn't have much experience in histology. How would this help them? Mark Tarango > Date: Thu, 21 Jul 2005 19:40:26 -0500 > From: "Steven P Postl" > Subject: [Histonet] Black paraffin and cassettes > To: histonet@lists.utsouthwestern.edu > Message-ID: > > > Content-Type: text/plain; charset="us-ascii" > > A researcher is wondering (now so am I [I never heard this request > before]) if any company sells black paraffin and black embedding > cassettes? No, this isn't a Gothic thing, a legitimate request. Would it > be possible to add something to paraffin to make it stay a black color and > remain strong as our commercial paraffin products? Thanks. ------------------------------ Message: 17 Date: Fri, 22 Jul 2005 15:36:27 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] RE: Limit on amount of formalin you can pour down sink? To: "'mprice26@juno.com'" , histonet@lists.utsouthwestern.edu Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4339@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 Orlando Florida allows it. A few of the large cities are equipped to handle formalin in the system. You have to contact the municipal water system people to see if they are set up to handle the formalin and receive written confirmation for your records (and the inspectors) before you start dumping ANY of it in the sink. We just started recycling ours with a Creative Waste Solutions,Inc Formalin recycler (rex@cwsincorp.com) and have been very pleased with the results. How and why it works I leave to him to explain, but it does. Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of mprice26@juno.com Sent: Friday, July 22, 2005 2:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Limit on amount of formalin you can pour down sink? Hi Histonetters, Does anyone know of any regulations as to the amount of formalin one can pour down the sink? I am in Texas. Thank you. Marsha Price _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Fri, 22 Jul 2005 16:17:52 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] cryosectioning liver/sucrose To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717588@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="ISO-8859-1" At any given temperature, tissues infiltrated with sucrose tend to be softer and less completely frozen than similar tissues without sucrose (and 30% is a very high level of sucrose), and therefore they compress and wrinkle more. Try cutting them at a colder temperature than you would normally use for liver. That will probably help produce flatter sections with less wrinkling. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Steven Coakley > Sent: Friday, July 22, 2005 12:27 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] cryosectioning liver/sucrose > > Something a "tad" new for me here. I've never sectioned 4%Para liver > infiltrated with 30% sucrose. They sectioned well but the slide appear to > be loaded with what appears to be folds throughout. Not what I routinely > see on regular unfixed FS. Is this normal for fixed sucrose infiltrated > sections. Are there any special techniques when dealing with this special > tissue. > > Steve > > > > --------------------------------- > Start your day with Yahoo! - make it your home page > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 19 Date: Fri, 22 Jul 2005 14:44:04 -0700 From: "Trisha Emry" Subject: [Histonet] under decal To: "histo" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Is there anything that can be done for under decalcified bone after it has been processed? Trisha Seattle ------------------------------ Message: 20 Date: Fri, 22 Jul 2005 18:13:21 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] under decal To: "'Trisha Emry'" , "histo" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB433B@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 OPTION 1: "redecal" Melt the paraffin block, put the tissue in a cassette as you would a fresh piece of tissue to process. Drop it in xylene for 15 minutes x2, then 100% abs. alcohol x2, 95% abs. alcohol. Wash in water and drop back into the decal. Process when soft. OPTION 2: "surface decal" Pour some decal solution into a small container or Petri dish and put the faced block face-down in the solution. Let soak for 15 min (time depends on hardness of the tissue) put on ice and try to section. Only the surface will be decaled so try to take a section off the surface. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Trisha Emry Sent: Friday, July 22, 2005 5:44 PM To: histo Subject: [Histonet] under decal Is there anything that can be done for under decalcified bone after it has been processed? Trisha Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Fri, 22 Jul 2005 18:23:51 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] under decal To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717589@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="ISO-8859-1" Sometimes you can get a decent section by pressing gauze soaked with decal fluid on the face of the block for a couple of minutes (wear gloves), then taking the first section off the face. Or you may get a few successive sections by soaking the faced block in decal fluid for 15 to 30 minutes, then chilling the block on the cold tray and cutting right off the face. However, if you are going to be doing any substantial amount of work with that block, probably the best approach is to reverse process it, decal it properly, and reprocess it. Remove the paraffin by soaking in xylene or whatever clearing agent you use, remove the xylene with absolute alcohol, bring the specimen down to water, then decal it, wash it and process it as usual. If you want you can place it in formalin overnight once it reaches the water stage, to ensure the best possible fixation before decalcifying it. Since most tissue processors won't run the stations in reverse order, you'll probably have to do the reverse processing by hand, in a beaker or flask. If the tissue was well fixed initially, it should withstand this treatment with little if any noticeable harm. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Trisha Emry > Sent: Friday, July 22, 2005 2:44 PM > To: histo > Subject: [Histonet] under decal > > Is there anything that can be done for under decalcified bone after it has > been processed? > > Trisha > Seattle > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 22 Date: Fri, 22 Jul 2005 15:48:27 -0700 From: Lori Richey Subject: Re: [Histonet] CD 31 To: Kim Allred Cc: histonet@lists.utsouthwestern.edu Message-ID: <42E177BB.2050905@u.washington.edu> Content-Type: text/plain; charset=us-ascii; format=flowed We use DAKO CD31 1:25 , 18 minutes microwave, in citrate buffer. Kim Allred wrote: >I am having a problem getting my CD31 to work. The protocol was for an >overnight incubation and pepsin pretreatment. I couldn't get it to work >this way so I altered it a bit by using trilogy for AR and a two hour >incubation for the primary ab. Does anyone have any ideas? This seemed to >work well for a while but once again, I'm having problems with it. Any >input would be appreciated. Thanks in advance! > > > > > > > >Kim Allred > >Dermatopathology Associates > >Jackson, MS > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 23 Date: Sat, 23 Jul 2005 04:19:18 EDT From: IamNinaOwen@aol.com Subject: [Histonet] Staining for Helicobacter pylori To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hi everyone! I'm doing a project on different staining methods for H. pylori in gastric biopsies, and just out of interest really wondered which special stains different labs routinely use for this purpose. My lab uses the Modified Giemsa. Thanks Nina Owen ------------------------------ Message: 24 Date: Sat, 23 Jul 2005 11:39:27 -0400 From: Drew Meyer <41dmb41@gmail.com> Subject: Re: [Histonet] Staining for Helicobacter pylori To: "IamNinaOwen@aol.com" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 We use the Wharton Stary method.... you can also use an Alcian Yellow Stain... there's another called the Toluidine Method I believe... Hope this helps... Drew Meyer On 7/23/05, IamNinaOwen@aol.com wrote: > Hi everyone! > > I'm doing a project on different staining methods for H. pylori in gastric > biopsies, and just out of interest really wondered which special stains > different labs routinely use for this purpose. My lab uses the Modified Giemsa. > > Thanks > > Nina Owen > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 31 **************************************** From bcgpath <@t> rediffmail.com Mon Jul 25 08:06:25 2005 From: bcgpath <@t> rediffmail.com (bcgirish) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] (no subject) Message-ID: <20050725130625.3008.qmail@webmail18.rediffmail.com> ? Dear friends, Does anyone do bronchoalveolar lavage in mice? we anaesthetize the animals with pentobarbitone.With an incision at the lower cervical region,trachea will be exposed,fluid will be collected after canulation and injecting 2ml normal saline.This method is yielding lot of RBCs which interferes with the interpretation.We tried with even exsanguination of the animals with renal artery severing but the result was not that much encouraging.can anyone help me in this regard by a better method? REGARDS Dr.Girish B.Chandrashekar Dept of Preclinical Safety Evaluation Dr Reddys Lab Hyderabad-49 From TMcNemar <@t> lmhealth.org Mon Jul 25 08:05:13 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Staining for Helicobacter pylori Message-ID: <6CD94D97ED7D924BA5C2B588FA9528213967E4@mail.lmhealth.org> We have been doing them by IHC for the last few years. Overkill, I know. The paths never like the Giemsa and we had so much trouble with the Warthin-Starry (consistency, repeats, etc) that I just switched to IHC. Works every time. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Drew Meyer [mailto:41dmb41@gmail.com] Sent: Saturday, July 23, 2005 11:39 AM To: IamNinaOwen@aol.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Staining for Helicobacter pylori We use the Wharton Stary method.... you can also use an Alcian Yellow Stain... there's another called the Toluidine Method I believe... Hope this helps... Drew Meyer On 7/23/05, IamNinaOwen@aol.com wrote: > Hi everyone! > > I'm doing a project on different staining methods for H. pylori in gastric > biopsies, and just out of interest really wondered which special stains > different labs routinely use for this purpose. My lab uses the Modified Giemsa. > > Thanks > > Nina Owen > _______________________________________________ > Histonet mailing list > Histonet@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 sa.drew <@t> hosp.wisc.edu Mon Jul 25 08:18:53 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Staining for Helicobacter pylori Message-ID: Our histology lab does Alcian Yellow, but we also just brought on H.pylori by IHC. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of IamNinaOwen@aol.com Sent: Saturday, July 23, 2005 3:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Staining for Helicobacter pylori Hi everyone! I'm doing a project on different staining methods for H. pylori in gastric biopsies, and just out of interest really wondered which special stains different labs routinely use for this purpose. My lab uses the Modified Giemsa. Thanks Nina Owen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Jul 25 09:27:33 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Re:cryosectioning liver/sucrose In-Reply-To: <20050722192702.5732.qmail@web90205.mail.scd.yahoo.com> References: <20050722192702.5732.qmail@web90205.mail.scd.yahoo.com> Message-ID: <6.0.0.22.1.20050725082212.01b69e98@gemini.msu.montana.edu> Steve, You did not say what temperature you were cutting at? However, with sucrose cryoprotected tissue, we prefer -26C (colder) or the sucrose begins to ooze out of the block like Karo syrup. That is a bit colder than what we would cut unfixed, fresh liver, at -17C for this very homogenous tissue. So play with the temperatures a bit. We also blot excess sucrose from the tissue a bit before snap freezing just to have a good interface with the OCT. With the cryoprotected tissue, cutting too arm may help cause the folding. We do use brush technic, and the sections should come off perfectly flat - sharp blade (new edge) is a good idea too. Good luck At 01:27 PM 7/22/2005, you wrote: >Something a "tad" new for me here. I've never sectioned 4%Para liver >infiltrated with 30% sucrose. They sectioned well but the slide appear to >be loaded with what appears to be folds throughout. Not what I routinely >see on regular unfixed FS. Is this normal for fixed sucrose infiltrated >sections. Are there any special techniques when dealing with this special >tissue. > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From POWELL_SA <@t> Mercer.edu Mon Jul 25 09:36:20 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] 2"x3" slides In-Reply-To: Message-ID: <01LR1F9BXXPI8X07SM@Macon2.Mercer.edu> I am not sure if the rack for the stainer can be modified in the same way, but I had the maintenance department take off one inch from one side of a regular metal slide rack that held 30 slides we used for manual staining. I was able to stain thirty 2x3" at one time, but it was by hand. If you want a picture of the rack I can email it to you. Shirley Powell -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Friday, July 22, 2005 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] 2"x3" slides Does anyone out there have a good way to stain 2"x3" glass slides (whole-mount prostates) using a Sakura stainer? I have a 2000 and 601 model and have tried adapting the rack so I can stain 4 slides at a time, but it's so inefficient. Any ideas? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Jul 25 09:40:02 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Re: Staining for Helicobacter pylori In-Reply-To: References: Message-ID: <6.0.0.22.1.20050725083514.01b7c948@gemini.msu.montana.edu> Besides the Warthin Starry method, you can use Steiner and Steiner (our preferred) but go to the Sakura Finetek website, and Histologic, there was a superb review of staining for Helicobacter a few years back, gave many methods. Labs are also doing an immunohistochemical stain for the organism - very specific and in a lot of ways, nicer than using silver staining methods. At 09:39 AM 7/23/2005, you wrote: >We use the Wharton Stary method.... you can also use an Alcian Yellow >Stain... there's another called the Toluidine Method I believe... > >Hope this helps... > >Drew Meyer 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 Jul 25 09:46:14 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Sorry, not the same RE: Complete Endog peroxidase blocking In-Reply-To: <200507242017.j6OKHhJk073165@pro12.abac.com> References: <6.0.0.22.1.20050722101641.01b02cf0@gemini.msu.montana.edu> <200507242017.j6OKHhJk073165@pro12.abac.com> Message-ID: <6.0.0.22.1.20050725084107.01b62460@gemini.msu.montana.edu> The DAKO Dual Block for HRP and Alk phos is NOT the same as the Glucose oxidase method. Glucose oxidase method is based on glucose oxidase + glucose = a slow steady release of hydrogen peroxide that interacts with the endogenous peroxidases and pseudoperoxidases in the tissues to knock them out. This is a mixture of these two components in a buffer containing a slight amount of sodium azide and the sections are incubated in this for 1 hour in a water bath at 37C. This block does NOT affect or kill endogenous alkaline phosphatase. The DAKO Dual blocker is probably the same as KPL Universal Block, a lovely ready to use reagent. I think it is one of the ways to block according to Jules Elias's list in his book. At 02:17 PM 7/24/2005, you wrote: >I had a particularly difficult time quenching endogenous peroxidase in >sputum cytopreps full of granulocytes and it was suggested that I try the >new DAKO Dual Block for HRP and Alk.Phos. and it worked a charm, don't know >what's in it (proprietary you know), but perhaps it is similar to Gayles >reagents for glucose oxidase blocking. >Patsy 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 PMonfils <@t> Lifespan.org Mon Jul 25 09:46:30 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] carstairs stain Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171758A@lsexch.lsmaster.lifespan.org> CARSTAIR'S METHOD FOR FIBRIN AND PLATELETS FIXATION: Formol-saline for at least 48 hours or longer. (Regular buffered formalin works fine, but fixation time is important - see results) Cut paraffin sections at 5 microns. SOLUTIONS: 5% aqueous ferric alum Mayer's Hematoxylin Picric Acid-Orange G Solution: Saturated aqueous picric acid ................ 20 ml Saturated picric acid in isopropanol ........ 80 ml Orange G ............................................. 0.2 gram Ponceau-fuchsin Solution: Distilled Water ...................................... 100 ml Acetic Acid .............................................. 1 ml Acid Fuchsin ..........................................0.5 gram Ponceau 2R ...........................................0.5 gram Aniline Blue Solution: 1% Acetic Acid ..................................... 100 ml Aniline Blue ............................................ 1.0 gram 1% aqueous phosphotungstic acid solution TECHNIQUE: 1. Deparaffibnize and hydrate sections 2. Mordant in 5% ferric alum, 5 minutes. Rinse in running trap water. 3. Stain with Mayer's Hematoxylin, 5 minutes. Rinse in running tap water. 4. Stain in Picric Acid-Orange G solution, 30 minutes to 1 hour. Rinse once in distilled water. 5. Stain in Ponceau-Fuchsin solution, 2 to 4 minutes. 6. Differentiate in 1% phosphotungstic acid solution until muscle is red and background is pale pink. Rinse in distilled water. 7. Stain in Aniline Blue solution, 1 hour. Rinse in several changes of distilled water. 8. Dehydrate, clear, coverslip using a synthetic mounting medium. RESULTS: Fibrin: (fixation 48 hours or more) - bright red; (less than 48 hours) - orange to red-orange Platelets: (fixation 48 hours or more) - gray-blue to navy blue; (less than 48 hours) - light gray Collagen - bright blue Muscle - red Red Blood Cells: (fixation 48 hours or more) - clear yellow; (less than 48 hours) - variable - red, green, or yellow. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > holling@med-in.uni-saarland.de > Sent: Monday, July 25, 2005 12:45 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] carstairs stain > > Hi, > > I would be grateful if somebody could give me the carstair stain > procedure. > > TC Nicole Hollinger > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From nancy.troiano <@t> yale.edu Mon Jul 25 10:22:04 2005 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] alkaline phosphatase enzyme for osteoblasts Message-ID: <5.2.1.1.2.20050725112036.00c0ab08@email.med.yale.edu> Hi Patsy - we routinely stain for alk phos on undecalcified MMA embedded bones. Do you need a protocol? If so, I can give you one. Nancy Troiano Yale Core Center for Musculoskeletal Disorders nancy.troiano@yale.edu From biswas.16 <@t> osu.edu Mon Jul 25 11:02:45 2005 From: biswas.16 <@t> osu.edu (SABYASACHI BISWAS) Date: Fri Sep 16 15:25:22 2005 Subject: [Histonet] Preferred brand of steamer Message-ID: <19a72da19a4e87.19a4e8719a72da@osu.edu> Hi Everyone Thanks to all who answered my previous query about pig skin sections. We were able to troubleshoot most problems regarding section adhesion except with heat induced epitope retrieval using a microwave. So we are planning to use the steamer method for HIER and were wondering if there is any particular brand that is suitable or preferred for this purpose. Thanks Sabya Biswas 524DHLRI The Ohio State University From dmarsha3 <@t> utmem.edu Mon Jul 25 11:10:26 2005 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] (no subject) References: <20050725130625.3008.qmail@webmail18.rediffmail.com> Message-ID: <001b01c59133$5e7d0b60$f623c084@DanaM> hi, we generally use less than that volume for rinse, approx 0.8 ml with gentle 2-3 time up and down. remove that 0.8 ml and repeat the process two more times, each time with 0.8ml.. the yield is good but it must be done very gently. we then did a quick rbc lysis step. hope this helps. dana marshall ----- Original Message ----- From: "bcgirish" To: Sent: Monday, July 25, 2005 8:06 AM Subject: [Histonet] (no subject) Dear friends, Does anyone do bronchoalveolar lavage in mice? we anaesthetize the animals with pentobarbitone.With an incision at the lower cervical region,trachea will be exposed,fluid will be collected after canulation and injecting 2ml normal saline.This method is yielding lot of RBCs which interferes with the interpretation.We tried with even exsanguination of the animals with renal artery severing but the result was not that much encouraging.can anyone help me in this regard by a better method? REGARDS Dr.Girish B.Chandrashekar Dept of Preclinical Safety Evaluation Dr Reddys Lab Hyderabad-49 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Mon Jul 25 11:42:49 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Staining for Helicobacter pylori Message-ID: I think immunohistochemistry is best because most positive cases can be identified within seconds and it will identify "intracellular" H. pylori which can be missed on conventional histochemical stains. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> 07/23/05 04:19AM >>> Hi everyone! I'm doing a project on different staining methods for H. pylori in gastric biopsies, and just out of interest really wondered which special stains different labs routinely use for this purpose. My lab uses the Modified Giemsa. Thanks Nina Owen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From godsgirlnow <@t> msn.com Mon Jul 25 11:49:49 2005 From: godsgirlnow <@t> msn.com (Roxanne Soto) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Staining for Helicobacter pylori In-Reply-To: Message-ID: I agree with you 100%, however most Pathologists that I know do not want to wait for 2-3 hours for something they feel is just as good because they can get it in 15 minutes. Roxanne Soto >From: "Richard Cartun" >To: , >Subject: Re: [Histonet] Staining for Helicobacter pylori >Date: Mon, 25 Jul 2005 12:42:49 -0400 > >I think immunohistochemistry is best because most positive cases can be >identified within seconds and it will identify "intracellular" H. pylori >which can be missed on conventional histochemical stains. > >Richard > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 >(860) 545-1596 >(860) 545-0174 Fax > > >>> 07/23/05 04:19AM >>> >Hi everyone! > >I'm doing a project on different staining methods for H. pylori in gastric >biopsies, and just out of interest really wondered which special stains >different labs routinely use for this purpose. My lab uses the Modified >Giemsa. > >Thanks > >Nina Owen >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >----------------------------------------------- >Confidentiality Notice > >This e-mail message, including any attachments, is for the sole use of the >intended recipient(s) and may contain confidential or proprietary >information which is legally privileged. Any unauthorized review, use, >disclosure, or distribution is prohibited. If you are not the intended >recipient, please promptly contact the sender by reply e-mail and destroy >all copies of the original message. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Jul 25 12:21:28 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:23 2005 Subject: gluteraldehyde fixation with Re: [Histonet] Rat brains In-Reply-To: <6.0.0.22.0.20050723145647.0267aa98@pop.vt.edu> References: <6.0.0.22.0.20050723145647.0267aa98@pop.vt.edu> Message-ID: <6.0.0.22.1.20050725111447.01b7ada0@gemini.msu.montana.edu> Question: is he planning to do EM on these tissues? If he had used paraformaldehyde, your sectioning would be much easier - but if he is also doing EM - you can try some things to counteract the dry/brittle brain. In general, gluteraldehyde creates some friable, brittle tissues when these are processed and embedded in paraffin. Check to see if you are processing too long as long dehydrations, etc and heat from paraffin contributes to the problem. A good long soak in ice water or solid ice block with water on top after trimming might help but be sure to not retrim the soak away, you have to get the first sections off the knife. A warm water or room temperature soak the going onto ice water for a time may help too, brain sometimes needs a bit of help. At 01:01 PM 7/23/2005, you wrote: >Please help... >A research project for our neuropathologist includes rat brains. These >are extremely difficult to section as he profuses the whole rat with >glutaraldehyde then takes brain samples. I have tried adding alcohol to >my water bath but these sections still have many, many wrinkles. Can >anyone offer suggestions? I would be so grateful for any and all advice. 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 Jul 25 12:25:52 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] IHC for RE:Staining for Helicobacter pylori In-Reply-To: <6CD94D97ED7D924BA5C2B588FA9528213967E4@mail.lmhealth.org> References: <6CD94D97ED7D924BA5C2B588FA9528213967E4@mail.lmhealth.org> Message-ID: <6.0.0.22.1.20050725112230.01b7b5c8@gemini.msu.montana.edu> Dear Tom, If I had to do Helicobacter, I would vote for IHC too!!! Very specific for identification of Helicobacter in human tissues, I only wished I had it for murine animal model work but no antibody was available - boo hoo!!! There is a nice method published in Histologic, Sakura Finetek website on doing this IHC, and it was wonderful. At 07:05 AM 7/25/2005, you wrote: >We have been doing them by IHC for the last few years. Overkill, I know. >The paths never like the Giemsa and we had so much trouble with the >Warthin-Starry (consistency, repeats, etc) that I just switched to IHC. >Works every time. >Tom McNemar, HT(ASCP) >Histology Co-ordinator >Licking Memorial Hospital >Newark, Ohio 43055 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 IamNinaOwen <@t> aol.com Mon Jul 25 12:30:46 2005 From: IamNinaOwen <@t> aol.com (IamNinaOwen@aol.com) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Re: Staining for Helicobacter pylori Message-ID: <65.49f13cd4.30167bc6@aol.com> Hi all Thanks for all your views on this matter, keep the replies rolling in! Nina Owen From nienhuis <@t> ucla.edu Mon Jul 25 13:55:40 2005 From: nienhuis <@t> ucla.edu (NIENHUIS,ROBERT ) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] TMB staining for WGA-HRP Message-ID: <1122317740.42e535ac936d3@mail.ucla.edu> We are about to cut a brain that was injected with WGA-HRP for tract tracing. We haven't done this for quite a while (decades?) and don't remember if it is possible to delay processing after cutting. If it is possible, how long can we delay, and what solution should the cut tissue be stored in? We plan to stain using the Olucha molybybdate modification to the Mesulam TMB procedure. Bob Nienhuis UCLA / VA Medical Center Los Angeles, CA From katherine-walters <@t> uiowa.edu Mon Jul 25 15:07:50 2005 From: katherine-walters <@t> uiowa.edu (Walters, Katherine S) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] historesin Message-ID: I have a colleague looking to buy some historesin. Apparently it is made by Leica (in Germany), and they will no longer ship it to the US. Is there a company here that can sell it, or a similar product? Thanks, Kathy From gcallis <@t> montana.edu Mon Jul 25 16:11:15 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:23 2005 Subject: equivalent to Re: [Histonet] historesin In-Reply-To: References: Message-ID: <6.0.0.22.1.20050725145504.01b275e0@gemini.msu.montana.edu> I don't recall plastic type of Historesin since it has been out of circulation for a while in USA, but if this is GMA, buy Technovits 7100 If it is the methylmethacrylate, MMA then buy Technovits 9100. Electron Microscopy Sciences sells both kits. These are excellent products. At 02:07 PM 7/25/2005, you wrote: >I have a colleague looking to buy some historesin. Apparently it is >made by Leica (in Germany), and they will no longer ship it to the US. >Is there a company here that can sell it, or a similar product? > >Thanks, >Kathy > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From tpmorken <@t> labvision.com Mon Jul 25 17:47:26 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Janella Seaton...please contact me Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CF55@usca0082k08.labvision.apogent.com> Janella, please contact me about the database you want. From: Janella Seaton ---------------------------------------------------------------------------- ---- Hello! We want to implement a database for specimens we receive in our lab. We would use this database starting with logging in specimens and ending with archiving of specimens. It would be used to track when various steps (gross photos, processing, staining, etc) have occurred and by whom. We do want a system that is user friendly. Any suggestions or advice? Thanks for the help..... j.s. Tim Morken From cmalc <@t> unimelb.edu.au Tue Jul 26 03:44:09 2005 From: cmalc <@t> unimelb.edu.au (Cathy Malcontenti-Wilson) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Background staining: mouse on mouse Broad spectrum kit Message-ID: <6.2.1.2.2.20050726182055.02ecbce8@mail.staff.unimelb.edu.au> Dear All, I really need some advice regarding an ongoing intense background problem. We use FFP sections of mouse liver. We are using an immuno kit called Zymed Histomouse Max Kit (It is a HRP- DAB one). Our primary ab is mouse anti human VEGF (1:1000). The kit is called a BROAD SPECTRUM kit because it is meant to be able to be used for mouse, rabbit, rat and guinea pig primaries. The secondary antibody is not completely explained in the kit but I think it is of goat host species. Question: If it is broad spectrum does this mean that the secondary antibody is a mixture of different antibodies? The kit contains its own non specific block, with no details of what it actually is. Question: If the secondary ab is raised in goat, is this non specific block normal goat serum? If not, what could it be? We know that when I incubate the tissue with DAB only, we get endogenous peroxidase staining which is pretty much abolished with H2O2 block. We know that when we omit the primary antibody and keep all the other layers, we get intense background staining of the sinusoidal (endothelial) lining of practically all the sinusoids, and we get brown staining in plasma cells, macrophages, connective tissue and other cells within vessels. Question: Omission of non specific blocking reagent showed increased background staining. Does this mean that the blocking agent is actually blocking some of the background but not all? Why not? We know that when we omit both the primary antibody and the non specific block then we get background staining. Question: What does this mean? We know if we omit the secondary antibody only, we get no staining. Question: What does this mean? I would greatly appreciate any input into this problem which has been plaguing us for a while now. I hope someone can help, I cant beleive what a great wealth of knowledge there is on this histonet list. Cathy Malcontenti-Wilson From Don.Birgerson <@t> leica-microsystems.com Tue Jul 26 07:32:06 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] historesin In-Reply-To: Message-ID: Hi Kathy, While we quit importing Historesin, there is product called "Technovit " that is sold in the US by "Energy Beam Science" at 413-786-9322. If you have further questions, please call me. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Walters, Katherine S" Sent by: histonet-bounces@lists.utsouthwestern.edu 07/25/2005 03:07 PM To cc Subject [Histonet] historesin I have a colleague looking to buy some historesin. Apparently it is made by Leica (in Germany), and they will no longer ship it to the US. Is there a company here that can sell it, or a similar product? Thanks, Kathy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com From Jackie.O'Connor <@t> abbott.com Tue Jul 26 07:43:00 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] CD 31 Message-ID: Santa Cruz gave me the same story - it's amazing that they depend on one goat for a product line. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist Discovery Cancer R4N2 AP3 847-938-4919 Jackie.O'Connor@abbott.com Angela McNabola Sent by: histonet-bounces@lists.utsouthwestern.edu 07/25/2005 05:40 AM To: histonet@lists.utsouthwestern.edu cc: Subject: Re: [Histonet] CD 31 Which antibody are you using? We have had great success with Santa Cruz (goat polyclonal) until I ordered a new lot this past month. When I called them, they indicated to me that they were having problems with their goat, literally. I guess it had stopped producing antibodies to CD31 that work in IHC (interestingly enough, they still work in Westerns). They hope to have the problem resolved within the next month, with a new goat. Angela McNabola, MS, HT(ASCP)SLS, QIHC Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 "Kim Allred" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] CD 31 western.edu 07/22/2005 03:15 PM I am having a problem getting my CD31 to work. The protocol was for an overnight incubation and pepsin pretreatment. I couldn't get it to work this way so I altered it a bit by using trilogy for AR and a two hour incubation for the primary ab. Does anyone have any ideas? This seemed to work well for a while but once again, I'm having problems with it. Any input would be appreciated. Thanks in advance! Kim Allred Dermatopathology Associates Jackson, MS _______________________________________________ Histonet mailing list Histonet@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 MajorFocus <@t> aol.com Tue Jul 26 08:18:10 2005 From: MajorFocus <@t> aol.com (MajorFocus@aol.com) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] North Carolina Histologist Position Available Message-ID: Frye Regional Medical Center in Hickory, North Carolina is currently seeking a Histology Technician or Technologist for full time employment. Qualified applicants must be certified by the ASCP or equivalent agency. Please apply on-line at _www.fryemedctr.com_ (http://www.fryemedctr.com/) or call toll-free (866) 494-4747. Frye Regional Medical Center Human Resources 420 North Center Street Hickory, NC 28601. From pmarcum <@t> vet.upenn.edu Tue Jul 26 08:25:08 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:23 2005 Subject: equivalent to Re: [Histonet] historesin In-Reply-To: <6.0.0.22.1.20050725145504.01b275e0@gemini.msu.montana.edu> References: <6.0.0.22.1.20050725145504.01b275e0@gemini.msu.montana.edu> Message-ID: <6.1.1.1.2.20050726092213.019799b0@mail.vet.upenn.edu> Hi, The Technovit 8100 is the same as JB-4 Plus which has a second polymer to make the GMA remain clearer and somewhat less brittle. As Don was kind enough to give you the number for EBS I shall not repeat and you can reach Polysciences at 800-523-2575. Pam Marcum At 05:11 PM 7/25/2005, Gayle Callis wrote: >I don't recall plastic type of Historesin since it has been out of >circulation for a while in USA, but if this is GMA, buy Technovits 7100 > >If it is the methylmethacrylate, MMA then buy Technovits 9100. > >Electron Microscopy Sciences sells both kits. These are excellent products. > >At 02:07 PM 7/25/2005, you wrote: > > >>I have a colleague looking to buy some historesin. Apparently it is >>made by Leica (in Germany), and they will no longer ship it to the US. >>Is there a company here that can sell it, or a similar product? >> >>Thanks, >>Kathy >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Luis.Chiriboga <@t> med.nyu.edu Tue Jul 26 08:49:42 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Looking 4 Antibodies Message-ID: HI everyone I'm looking for antibodies against MLH-1, MSH-2, BRCA-1 and BRCA-2 for use in human FF-PE tissue. recommendations as well as comments are welcome. feel free to respond directly to my address. Thanks in advance Luis From pathrm35 <@t> adelphia.net Tue Jul 26 09:55:00 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] DIF procedures Message-ID: <10787860.1122389700521.JavaMail.root@web2.mail.adelphia.net> Fellow Techs, Does anyone have Direct Immunofluorescence procedures they are willing to share? We are interested in CD3, fibrinogen, IgA, IgM and IgG. Thanks in advance. Ron Martin, BS HT (ASCP) HTL fax 561-721-1249 From pathrm35 <@t> adelphia.net Tue Jul 26 10:01:06 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] teaching heads for Nikon microscope Message-ID: <28528193.1122390066380.JavaMail.root@web2.mail.adelphia.net> Fellow Techs, Does anyone know of or where I can find binocular teaching heads for a (older model) multiheaded Nikon Optiphot microscope? Any leads or info is greatly appreciated. Thanks in advance. Ron Martin, BS HT (ASCP) HTL fax 561-721-1249 From sluhisto <@t> yahoo.com Tue Jul 26 10:35:52 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Integrase Interactor 1 (INI-1/hsnf5) antibody needed Message-ID: <20050726153552.54597.qmail@web51005.mail.yahoo.com> Hello All: I have a doc looking for the above antibody for use in FFPE tissue sections of rhabdoid tumor in children. Any help would be greatly appreciated. Thanks, in advance. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From pedro.louro <@t> spcorp.com Tue Jul 26 10:58:13 2005 From: pedro.louro <@t> spcorp.com (Louro, Pedro) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] looking for iNOS antibody Message-ID: <4508920F80C0D411B90200508BF9A9F4062B4A40@LAFMSG30.us.schp.com> Hi all, I'm starting a new experiment with rat mesentery artery looking for inducible nitric oxide synthase (iNOS, eNOS, and nNOS). What companies would carry this antibody that would work on FFPE rat tissue. Also, what would be a good positive control? Thanks in advance for your assistance Pedro ********************************************************************* 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 settembr <@t> umdnj.edu Tue Jul 26 11:13:31 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Integrase Interactor 1 (INI-1/hsnf5) antibody needed Message-ID: I purchased it from BD Biosciences from San Diego, CA cat# 612110 I pretreat with DakoCytomation's Target Retrieval Solution. and use it at 1:50 incubating for 15 minutes room temp. Works nicely but there's not much in the vial and I am looking to Santa Cruz, who sells 1ml of it. Santa Cruz Cat. sc-13055 Good Luck. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> Histology SLU 7/26/2005 11:35:52 AM >>> Hello All: I have a doc looking for the above antibody for use in FFPE tissue sections of rhabdoid tumor in children. Any help would be greatly appreciated. Thanks, in advance. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Tue Jul 26 11:48:10 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] IgG2a and IgG1 for guinea pig Message-ID: <000101c59201$ce33a1c0$a7d48a80@AMY> Hello everyone I have been looking for some reagents for guinea pig for both ELISA and IHC for IgG2a and IgG1. I did find a source for the IgG2a through Nordic Immuno Reagents. Has anyone had any dealings with them? They have several antibodies against guinea pig IgG2a all derived from different species (sheep, goat and rabbit). I was guessing that the rabbit antibody would be nice since I could use a polymer detection system if necessary. Any suggestions of a different vendor or anything else would be appreciated. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From IamNinaOwen <@t> aol.com Tue Jul 26 13:29:31 2005 From: IamNinaOwen <@t> aol.com (IamNinaOwen@aol.com) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Genta stain for Helicobacter pylori Message-ID: <20.4990d7aa.3017db0b@aol.com> Hello Histonetters! Has anyone ever used the Genta stain for Helicobacter pylori? I'm trying to find a protocol for this method, so any help will be very much appreciated! Thanks Nina Owen From sjchtascp <@t> yahoo.com Tue Jul 26 13:30:39 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] stain for pre-collagen and mature collagen Message-ID: <20050726183039.46878.qmail@web90204.mail.scd.yahoo.com> I've been working on Herovica's Method for differentiating pre-collagen(young) verses mature collagen using varies combinations of methy blue and van gieson's. Luna has a modified method on pg 453 of his "Histopathologic Methods...." The regular Herovica stain calls for tissue fixed in formol-acetic-alcohol. 10% NBF leaves the muscle a blue to blue-green and very little red (mature collagen). I tried post fixation in the FAA for 1 hr w/o allot of difference. Bouins post-fixation seems to work best. Certainly not getting the results as outlined. Is there anyway to check the saturated picric acid? Any ideas or suggestion would be appreciated, especially from those familiar with this stain. Steve --------------------------------- Start your day with Yahoo! - make it your home page From Kristopher.Kalleberg <@t> unilever.com Tue Jul 26 13:29:43 2005 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] re: iNOS Message-ID: If I remember right Neomarkers/Lab Vision Corp. has iNOS antibodies. I used them maybe 1 1/2 years ago but never got great results from the stain. I ran the test numerous times and was unable to solve the problem. If you decide to use Neomarkers and are able to achieve good results please send me your secret. Good luck. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull, CT 06611 203 381 5765 -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, July 26, 2005 1:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 35 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. gluteraldehyde fixation with Re: [Histonet] Rat brains (Gayle Callis) 2. IHC for RE:Staining for Helicobacter pylori (Gayle Callis) 3. Re: Staining for Helicobacter pylori (IamNinaOwen@aol.com) 4. TMB staining for WGA-HRP (NIENHUIS,ROBERT ) 5. historesin (Walters, Katherine S) 6. equivalent to Re: [Histonet] historesin (Gayle Callis) 7. Janella Seaton...please contact me (Morken, Tim - Labvision) 8. Background staining: mouse on mouse Broad spectrum kit (Cathy Malcontenti-Wilson) 9. Re: historesin (Don.Birgerson@leica-microsystems.com) 10. Re: CD 31 (Jackie M O'Connor) 11. North Carolina Histologist Position Available (MajorFocus@aol.com) 12. Re: equivalent to Re: [Histonet] historesin (Pamela Marcum) 13. Looking 4 Antibodies (Luis Chiriboga) 14. DIF procedures (pathrm35@adelphia.net) 15. teaching heads for Nikon microscope (pathrm35@adelphia.net) 16. Integrase Interactor 1 (INI-1/hsnf5) antibody needed (Histology SLU) 17. looking for iNOS antibody (Louro, Pedro) 18. Re: Integrase Interactor 1 (INI-1/hsnf5) antibody needed (Dana Settembre) 19. IgG2a and IgG1 for guinea pig (Elizabeth Chlipala) ---------------------------------------------------------------------- Message: 1 Date: Mon, 25 Jul 2005 11:21:28 -0600 From: Gayle Callis Subject: gluteraldehyde fixation with Re: [Histonet] Rat brains To: Laura Fidgen , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050725111447.01b7ada0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Question: is he planning to do EM on these tissues? If he had used paraformaldehyde, your sectioning would be much easier - but if he is also doing EM - you can try some things to counteract the dry/brittle brain. In general, gluteraldehyde creates some friable, brittle tissues when these are processed and embedded in paraffin. Check to see if you are processing too long as long dehydrations, etc and heat from paraffin contributes to the problem. A good long soak in ice water or solid ice block with water on top after trimming might help but be sure to not retrim the soak away, you have to get the first sections off the knife. A warm water or room temperature soak the going onto ice water for a time may help too, brain sometimes needs a bit of help. At 01:01 PM 7/23/2005, you wrote: >Please help... >A research project for our neuropathologist includes rat brains. These >are extremely difficult to section as he profuses the whole rat with >glutaraldehyde then takes brain samples. I have tried adding alcohol to >my water bath but these sections still have many, many wrinkles. Can >anyone offer suggestions? I would be so grateful for any and all advice. 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: 2 Date: Mon, 25 Jul 2005 11:25:52 -0600 From: Gayle Callis Subject: [Histonet] IHC for RE:Staining for Helicobacter pylori To: Tom McNemar , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050725112230.01b7b5c8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Dear Tom, If I had to do Helicobacter, I would vote for IHC too!!! Very specific for identification of Helicobacter in human tissues, I only wished I had it for murine animal model work but no antibody was available - boo hoo!!! There is a nice method published in Histologic, Sakura Finetek website on doing this IHC, and it was wonderful. At 07:05 AM 7/25/2005, you wrote: >We have been doing them by IHC for the last few years. Overkill, I know. >The paths never like the Giemsa and we had so much trouble with the >Warthin-Starry (consistency, repeats, etc) that I just switched to IHC. >Works every time. >Tom McNemar, HT(ASCP) >Histology Co-ordinator >Licking Memorial Hospital >Newark, Ohio 43055 Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 3 Date: Mon, 25 Jul 2005 13:30:46 EDT From: IamNinaOwen@aol.com Subject: [Histonet] Re: Staining for Helicobacter pylori To: histonet@lists.utsouthwestern.edu Message-ID: <65.49f13cd4.30167bc6@aol.com> Content-Type: text/plain; charset="US-ASCII" Hi all Thanks for all your views on this matter, keep the replies rolling in! Nina Owen ------------------------------ Message: 4 Date: Mon, 25 Jul 2005 11:55:40 -0700 From: "NIENHUIS,ROBERT " Subject: [Histonet] TMB staining for WGA-HRP To: Histonet@lists.utsouthwestern.edu Message-ID: <1122317740.42e535ac936d3@mail.ucla.edu> Content-Type: text/plain; charset=ISO-8859-1 We are about to cut a brain that was injected with WGA-HRP for tract tracing. We haven't done this for quite a while (decades?) and don't remember if it is possible to delay processing after cutting. If it is possible, how long can we delay, and what solution should the cut tissue be stored in? We plan to stain using the Olucha molybybdate modification to the Mesulam TMB procedure. Bob Nienhuis UCLA / VA Medical Center Los Angeles, CA ------------------------------ Message: 5 Date: Mon, 25 Jul 2005 15:07:50 -0500 From: "Walters, Katherine S" Subject: [Histonet] historesin To: Message-ID: Content-Type: text/plain; charset="us-ascii" I have a colleague looking to buy some historesin. Apparently it is made by Leica (in Germany), and they will no longer ship it to the US. Is there a company here that can sell it, or a similar product? Thanks, Kathy ------------------------------ Message: 6 Date: Mon, 25 Jul 2005 15:11:15 -0600 From: Gayle Callis Subject: equivalent to Re: [Histonet] historesin To: "Walters, Katherine S" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050725145504.01b275e0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I don't recall plastic type of Historesin since it has been out of circulation for a while in USA, but if this is GMA, buy Technovits 7100 If it is the methylmethacrylate, MMA then buy Technovits 9100. Electron Microscopy Sciences sells both kits. These are excellent products. At 02:07 PM 7/25/2005, you wrote: >I have a colleague looking to buy some historesin. Apparently it is >made by Leica (in Germany), and they will no longer ship it to the US. >Is there a company here that can sell it, or a similar product? > >Thanks, >Kathy > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 7 Date: Mon, 25 Jul 2005 18:47:26 -0400 From: "Morken, Tim - Labvision" Subject: [Histonet] Janella Seaton...please contact me To: "'Histonet@lists.utsouthwestern.edu'" Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CF55@usca0082k08.labvision.apogent.com> Content-Type: text/plain Janella, please contact me about the database you want. From: Janella Seaton ---------------------------------------------------------------------------- ---- Hello! We want to implement a database for specimens we receive in our lab. We would use this database starting with logging in specimens and ending with archiving of specimens. It would be used to track when various steps (gross photos, processing, staining, etc) have occurred and by whom. We do want a system that is user friendly. Any suggestions or advice? Thanks for the help..... j.s. Tim Morken ------------------------------ Message: 8 Date: Tue, 26 Jul 2005 18:44:09 +1000 From: Cathy Malcontenti-Wilson Subject: [Histonet] Background staining: mouse on mouse Broad spectrum kit To: histonet@lists.utsouthwestern.edu Message-ID: <6.2.1.2.2.20050726182055.02ecbce8@mail.staff.unimelb.edu.au> Content-Type: text/plain; charset="us-ascii"; format=flowed Dear All, I really need some advice regarding an ongoing intense background problem. We use FFP sections of mouse liver. We are using an immuno kit called Zymed Histomouse Max Kit (It is a HRP- DAB one). Our primary ab is mouse anti human VEGF (1:1000). The kit is called a BROAD SPECTRUM kit because it is meant to be able to be used for mouse, rabbit, rat and guinea pig primaries. The secondary antibody is not completely explained in the kit but I think it is of goat host species. Question: If it is broad spectrum does this mean that the secondary antibody is a mixture of different antibodies? The kit contains its own non specific block, with no details of what it actually is. Question: If the secondary ab is raised in goat, is this non specific block normal goat serum? If not, what could it be? We know that when I incubate the tissue with DAB only, we get endogenous peroxidase staining which is pretty much abolished with H2O2 block. We know that when we omit the primary antibody and keep all the other layers, we get intense background staining of the sinusoidal (endothelial) lining of practically all the sinusoids, and we get brown staining in plasma cells, macrophages, connective tissue and other cells within vessels. Question: Omission of non specific blocking reagent showed increased background staining. Does this mean that the blocking agent is actually blocking some of the background but not all? Why not? We know that when we omit both the primary antibody and the non specific block then we get background staining. Question: What does this mean? We know if we omit the secondary antibody only, we get no staining. Question: What does this mean? I would greatly appreciate any input into this problem which has been plaguing us for a while now. I hope someone can help, I cant beleive what a great wealth of knowledge there is on this histonet list. Cathy Malcontenti-Wilson ------------------------------ Message: 9 Date: Tue, 26 Jul 2005 07:32:06 -0500 From: Don.Birgerson@leica-microsystems.com Subject: Re: [Histonet] historesin To: "Walters, Katherine S" Cc: histonet-bounces@lists.utsouthwestern.edu, histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hi Kathy, While we quit importing Historesin, there is product called "Technovit " that is sold in the US by "Energy Beam Science" at 413-786-9322. If you have further questions, please call me. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Walters, Katherine S" Sent by: histonet-bounces@lists.utsouthwestern.edu 07/25/2005 03:07 PM To cc Subject [Histonet] historesin I have a colleague looking to buy some historesin. Apparently it is made by Leica (in Germany), and they will no longer ship it to the US. Is there a company here that can sell it, or a similar product? Thanks, Kathy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com ------------------------------ Message: 10 Date: Tue, 26 Jul 2005 07:43:00 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] CD 31 To: Angela McNabola Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Santa Cruz gave me the same story - it's amazing that they depend on one goat for a product line. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist Discovery Cancer R4N2 AP3 847-938-4919 Jackie.O'Connor@abbott.com Angela McNabola Sent by: histonet-bounces@lists.utsouthwestern.edu 07/25/2005 05:40 AM To: histonet@lists.utsouthwestern.edu cc: Subject: Re: [Histonet] CD 31 Which antibody are you using? We have had great success with Santa Cruz (goat polyclonal) until I ordered a new lot this past month. When I called them, they indicated to me that they were having problems with their goat, literally. I guess it had stopped producing antibodies to CD31 that work in IHC (interestingly enough, they still work in Westerns). They hope to have the problem resolved within the next month, with a new goat. Angela McNabola, MS, HT(ASCP)SLS, QIHC Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 "Kim Allred" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] CD 31 western.edu 07/22/2005 03:15 PM I am having a problem getting my CD31 to work. The protocol was for an overnight incubation and pepsin pretreatment. I couldn't get it to work this way so I altered it a bit by using trilogy for AR and a two hour incubation for the primary ab. Does anyone have any ideas? This seemed to work well for a while but once again, I'm having problems with it. Any input would be appreciated. Thanks in advance! Kim Allred Dermatopathology Associates Jackson, MS _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Tue, 26 Jul 2005 09:18:10 EDT From: MajorFocus@aol.com Subject: [Histonet] North Carolina Histologist Position Available To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Frye Regional Medical Center in Hickory, North Carolina is currently seeking a Histology Technician or Technologist for full time employment. Qualified applicants must be certified by the ASCP or equivalent agency. Please apply on-line at _www.fryemedctr.com_ (http://www.fryemedctr.com/) or call toll-free (866) 494-4747. Frye Regional Medical Center Human Resources 420 North Center Street Hickory, NC 28601. ------------------------------ Message: 12 Date: Tue, 26 Jul 2005 09:25:08 -0400 From: Pamela Marcum Subject: Re: equivalent to Re: [Histonet] historesin To: Gayle Callis , "Walters, Katherine S" , Histonet@lists.utsouthwestern.edu Message-ID: <6.1.1.1.2.20050726092213.019799b0@mail.vet.upenn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Hi, The Technovit 8100 is the same as JB-4 Plus which has a second polymer to make the GMA remain clearer and somewhat less brittle. As Don was kind enough to give you the number for EBS I shall not repeat and you can reach Polysciences at 800-523-2575. Pam Marcum At 05:11 PM 7/25/2005, Gayle Callis wrote: >I don't recall plastic type of Historesin since it has been out of >circulation for a while in USA, but if this is GMA, buy Technovits 7100 > >If it is the methylmethacrylate, MMA then buy Technovits 9100. > >Electron Microscopy Sciences sells both kits. These are excellent products. > >At 02:07 PM 7/25/2005, you wrote: > > >>I have a colleague looking to buy some historesin. Apparently it is >>made by Leica (in Germany), and they will no longer ship it to the US. >>Is there a company here that can sell it, or a similar product? >> >>Thanks, >>Kathy >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Tue, 26 Jul 2005 09:49:42 -0400 From: Luis Chiriboga Subject: [Histonet] Looking 4 Antibodies To: Histonet Message-ID: Content-Type: text/plain; charset="iso-8859-1" HI everyone I'm looking for antibodies against MLH-1, MSH-2, BRCA-1 and BRCA-2 for use in human FF-PE tissue. recommendations as well as comments are welcome. feel free to respond directly to my address. Thanks in advance Luis ------------------------------ Message: 14 Date: Tue, 26 Jul 2005 10:55:00 -0400 From: Subject: [Histonet] DIF procedures To: histonet@lists.utsouthwestern.edu Message-ID: <10787860.1122389700521.JavaMail.root@web2.mail.adelphia.net> Content-Type: text/plain; charset=utf-8 Fellow Techs, Does anyone have Direct Immunofluorescence procedures they are willing to share? We are interested in CD3, fibrinogen, IgA, IgM and IgG. Thanks in advance. Ron Martin, BS HT (ASCP) HTL fax 561-721-1249 ------------------------------ Message: 15 Date: Tue, 26 Jul 2005 11:01:06 -0400 From: Subject: [Histonet] teaching heads for Nikon microscope To: histonet@lists.utsouthwestern.edu Message-ID: <28528193.1122390066380.JavaMail.root@web2.mail.adelphia.net> Content-Type: text/plain; charset=utf-8 Fellow Techs, Does anyone know of or where I can find binocular teaching heads for a (older model) multiheaded Nikon Optiphot microscope? Any leads or info is greatly appreciated. Thanks in advance. Ron Martin, BS HT (ASCP) HTL fax 561-721-1249 ------------------------------ Message: 16 Date: Tue, 26 Jul 2005 08:35:52 -0700 (PDT) From: Histology SLU Subject: [Histonet] Integrase Interactor 1 (INI-1/hsnf5) antibody needed To: histonet@lists.utsouthwestern.edu Message-ID: <20050726153552.54597.qmail@web51005.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello All: I have a doc looking for the above antibody for use in FFPE tissue sections of rhabdoid tumor in children. Any help would be greatly appreciated. Thanks, in advance. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 17 Date: Tue, 26 Jul 2005 11:58:13 -0400 From: "Louro, Pedro" Subject: [Histonet] looking for iNOS antibody To: histonet@lists.utsouthwestern.edu Message-ID: <4508920F80C0D411B90200508BF9A9F4062B4A40@LAFMSG30.us.schp.com> Content-Type: text/plain Hi all, I'm starting a new experiment with rat mesentery artery looking for inducible nitric oxide synthase (iNOS, eNOS, and nNOS). What companies would carry this antibody that would work on FFPE rat tissue. Also, what would be a good positive control? Thanks in advance for your assistance Pedro ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 18 Date: Tue, 26 Jul 2005 12:13:31 -0400 From: Dana Settembre Subject: Re: [Histonet] Integrase Interactor 1 (INI-1/hsnf5) antibody needed To: histonet@lists.utsouthwestern.edu, sluhisto@yahoo.com Message-ID: Content-Type: text/plain; charset=US-ASCII I purchased it from BD Biosciences from San Diego, CA cat# 612110 I pretreat with DakoCytomation's Target Retrieval Solution. and use it at 1:50 incubating for 15 minutes room temp. Works nicely but there's not much in the vial and I am looking to Santa Cruz, who sells 1ml of it. Santa Cruz Cat. sc-13055 Good Luck. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> Histology SLU 7/26/2005 11:35:52 AM >>> Hello All: I have a doc looking for the above antibody for use in FFPE tissue sections of rhabdoid tumor in children. Any help would be greatly appreciated. Thanks, in advance. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 26 Jul 2005 10:48:10 -0600 From: "Elizabeth Chlipala" Subject: [Histonet] IgG2a and IgG1 for guinea pig To: "'Histonet'" Message-ID: <000101c59201$ce33a1c0$a7d48a80@AMY> Content-Type: text/plain; charset="US-ASCII" Hello everyone I have been looking for some reagents for guinea pig for both ELISA and IHC for IgG2a and IgG1. I did find a source for the IgG2a through Nordic Immuno Reagents. Has anyone had any dealings with them? They have several antibodies against guinea pig IgG2a all derived from different species (sheep, goat and rabbit). I was guessing that the rabbit antibody would be nice since I could use a polymer detection system if necessary. Any suggestions of a different vendor or anything else would be appreciated. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 35 **************************************** From PMonfils <@t> Lifespan.org Tue Jul 26 13:41:25 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Genta stain for Helicobacter pylori Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171758F@lsexch.lsmaster.lifespan.org> It's in the archives, here: http://www.histosearch.com/histonet/Nov99/RE.GENTASTAINA.html I read a paper which stated that lead nitrate can be substituted for uranium nitrate in this procedure, thereby avoiding the problems of handling and disposing of radioactive material. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > IamNinaOwen@aol.com > Sent: Tuesday, July 26, 2005 11:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Genta stain for Helicobacter pylori > > Hello Histonetters! > > Has anyone ever used the Genta stain for Helicobacter pylori? I'm trying > to > find a protocol for this method, so any help will be very much > appreciated! > > Thanks > > Nina Owen > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From dmccaig <@t> ckha.on.ca Tue Jul 26 13:43:45 2005 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] granular debris in paraffin blocks Message-ID: <3E5A3F039F0BD8118B4700C00D00202404361E@CKHA9> Does anyone have any suggestions to obtain sections from an autopsy case of a drowning victim. The lung tissue has granular debris from a muddy ditch. The pathologist wants this demonstrated but it is virtually impossible to obtain anything due to fragmentation of the blocks from this. Thanks Diana From Kristopher.Kalleberg <@t> unilever.com Tue Jul 26 13:48:38 2005 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] re: iNOS Message-ID: If I remember right Neomarkers/Lab Vision Corp. has iNOS antibodies. I used them maybe 1 1/2 years ago but never got great results from the stain. I ran the test numerous times and was unable to solve the problem. If you decide to use Neomarkers and are able to achieve good results please send me your secret. Good luck. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull, CT 06611 203 381 5765 From MAUGER <@t> email.chop.edu Tue Jul 26 13:51:00 2005 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Integrase Interactor 1 (INI-1/hsnf5) antibody needed Message-ID: Susan, You can get INI-1(BAF47) from BD Transduction labs- catalog # 612110 for 50 ug, or 61211 for 150 ug. Jo >>> Histology SLU 07/26/05 11:35 AM >>> Hello All: I have a doc looking for the above antibody for use in FFPE tissue sections of rhabdoid tumor in children. Any help would be greatly appreciated. Thanks, in advance. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Tue Jul 26 13:53:05 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Applied IHC Journal Message-ID: Does anyone have the Volume 3, Number 1 issue of Applied Immunohistochemistry from 1995? Thank you! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From PMonfils <@t> Lifespan.org Tue Jul 26 14:12:19 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] granular debris in paraffin blocks Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717590@lsexch.lsmaster.lifespan.org> Now there's a unique problem! No, sand really doesn't section very well. How about cutting a thick section? A 20 or 30 micron section may not be suitable for cellular detail, but it might contain enough of the material to demonstrate it adequately; and while the section might be full of striations, hopefully the striations might not be deep enough to shread the ribbon. Also, try to get the earliest section you can get, while knife damage is minimal. Use a fresh section of knife edge, and start rotating the microtome before the block touches the knife, so you can get the first couple of sections off the block face. If that doesn't work, your pathologist will probably have to demonstrate the debris by having the lung x-rayed. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Diana > McCaig > Sent: Tuesday, July 26, 2005 11:43 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] granular debris in paraffin blocks > > Does anyone have any suggestions to obtain sections from an autopsy case > of > a drowning victim. The lung tissue has granular debris from a muddy > ditch. > The pathologist wants this demonstrated but it is virtually impossible to > obtain anything due to fragmentation of the blocks from this. > Thanks > Diana > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From emerald_lake77 <@t> yahoo.com Tue Jul 26 14:19:51 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Serotec Rat Anti-mouse F4/80 : Biotin -- anyone know of good concentration? Message-ID: <20050726191951.92792.qmail@web31708.mail.mud.yahoo.com> Hello once again, I just received my Serotec Rat Anti-mouse F4/80 antibody and was wondering if anyone can share with me the concentration they use -- closest to optimized would be great! Thanks Gustave Hebert Scientist II CVMD Wyeth - Cambridge __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From emerald_lake77 <@t> yahoo.com Tue Jul 26 14:23:10 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Biotin conjugated Antibodies -- can we skip biotin?? Message-ID: <20050726192310.49951.qmail@web31711.mail.mud.yahoo.com> Hello, Just a general question to help increase my knowledge: If I had an antibody that was conjugated to biotin -- say for example, a goat anti-mouse P-selectin (biotin) -- can I just take a rabbit anti-goat conjugated to AP and run IHC that way -- hopefully avoiding the ABC step and background OR does the biotin from the primary Ab block secondaries from coming in? Thanks. --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour From gkrasinski <@t> covx.com Tue Jul 26 14:45:44 2005 From: gkrasinski <@t> covx.com (Glenn Krasinski) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] anti-mouse cleaved PARP for IHC... Message-ID: <6837C00A0CC6594197C2F0EEF256561805AADA@covxmail.covx.com> I am looking for a mouse-specific anti-cleaved PARP antibody that works in IHC. Cell Signaling has a rabbit polyclonal that work in IC and WB, but not in IHC. Many thanks. Glenn M. Krasinski Scientist, Biology CovX Research LLC 9381 Judicial Drive, Suite 200 San Diego, CA 92121 Ph: (858) 964-2053 Fax: (858) 964-2090 E-dress: gkrasinski@covx.com From ajohnson <@t> aipathology.com Tue Jul 26 14:45:22 2005 From: ajohnson <@t> aipathology.com (Amy Johnson) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] CK7 control/ CK20 contol Message-ID: <016A64931E1ED511B12C0002B302608052334F@SERV001> What do most of you out there use for a CK7 control?......CK20 control?.... We currently are using an adenocarcinoma of the lung for our CK7 and an adenocarcinoma of the colon for the CK20. We have been having problems and need some help. Thanks Amy Johnson From la.sebree <@t> hosp.wisc.edu Tue Jul 26 14:56:03 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] CK7 control/ CK20 contol Message-ID: We mainly use liver for CK7 (stains the bile ducts) and colon adenocarcinoma for CK20. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Johnson Sent: Tuesday, July 26, 2005 2:45 PM To: Histonet (E-mail) Subject: [Histonet] CK7 control/ CK20 contol What do most of you out there use for a CK7 control?......CK20 control?.... We currently are using an adenocarcinoma of the lung for our CK7 and an adenocarcinoma of the colon for the CK20. We have been having problems and need some help. Thanks Amy Johnson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Jul 26 15:13:26 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Re: Serotec Rat Anti-mouse F4/80 : Biotin -- anyone know of good concentration? In-Reply-To: <20050726191951.92792.qmail@web31708.mail.mud.yahoo.com> References: <20050726191951.92792.qmail@web31708.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20050726141256.01b7ea60@gemini.msu.montana.edu> Fro paraffin (FFPE) or frozen sections? At 01:19 PM 7/26/2005, you wrote: >Hello once again, > >I just received my Serotec Rat Anti-mouse F4/80 antibody and was wondering >if anyone can share with me the concentration they use -- closest to >optimized would be great! > >Thanks > >Gustave Hebert >Scientist II >CVMD >Wyeth - Cambridge > > >__________________________________________________ >Do You Yahoo!? >Tired of spam? Yahoo! Mail has the best spam protection around >http://mail.yahoo.com >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 emerald_lake77 <@t> yahoo.com Tue Jul 26 15:15:46 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Re: Serotec Rat Anti-mouse F4/80 : Biotin -- anyone know of good concentration? In-Reply-To: <6.0.0.22.1.20050726141256.01b7ea60@gemini.msu.montana.edu> Message-ID: <20050726201546.63399.qmail@web31711.mail.mud.yahoo.com> Sorry Gayle, This would be for FFPE. Thanks. Gustave Gayle Callis wrote: Fro paraffin (FFPE) or frozen sections? At 01:19 PM 7/26/2005, you wrote: >Hello once again, > >I just received my Serotec Rat Anti-mouse F4/80 antibody and was wondering >if anyone can share with me the concentration they use -- closest to >optimized would be great! > >Thanks > >Gustave Hebert >Scientist II >CVMD >Wyeth - Cambridge > > >__________________________________________________ >Do You Yahoo!? >Tired of spam? Yahoo! Mail has the best spam protection around >http://mail.yahoo.com >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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) --------------------------------- Start your day with Yahoo! - make it your home page From gcallis <@t> montana.edu Tue Jul 26 15:22:05 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Re: Biotin conjugated Antibodies -- can we skip biotin?? In-Reply-To: <20050726192310.49951.qmail@web31711.mail.mud.yahoo.com> References: <20050726192310.49951.qmail@web31711.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20050726141359.01b7ee00@gemini.msu.montana.edu> Yes, you do not need more biotin in the system. When you work with biotinylated primaries, all you need to do is use Strepavidin-HRP or Strepavidin-AP, followed by the appropriate chromogen. You eliminate the secondary conjugated to biotin completely, and also the ABC complex, and but be sure to use a good reliable Strepavidin-HRP or AP. Southern Biotechnology has these as does Biosource/TAGO. The primary conjugated to biotin provides all the biotin you will need. One of the perks of working with biotinylated primaries - it shortens the protocol! IF you use Vectors ABC complex, DON"T or you will experience loss of sensitivity and have poor staining. You will still need to do the appropriate blocking, i.e. avidin/biotion (OR Strepavidin/biotin block from Vector) normal serum, proper diluents with serums, etc. to avoid background problems. We do biotinylated primaries almost exclusively and with great success for murine CD markers IHC and IFA (using Molecular Probes Strepavidin-Alexa fluorophore conjugates.) If you want to discuss this more, contact me privately. At 01:23 PM 7/26/2005, you wrote: >Hello, > >Just a general question to help increase my knowledge: > >If I had an antibody that was conjugated to biotin -- say for example, a >goat anti-mouse P-selectin (biotin) -- can I just take a rabbit anti-goat >conjugated to AP and run IHC that way -- hopefully avoiding the ABC step >and background OR does the biotin from the primary Ab block secondaries >from coming in? > >Thanks. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From denise.woodward <@t> uconn.edu Tue Jul 26 15:25:10 2005 From: denise.woodward <@t> uconn.edu (Woodward, Denise) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Artifact pictures and cell block prep Message-ID: Hi all, I'm looking for pictures of sectioning, staining and processing artifacts. Anyone know where I might be able to find some on the web? Also, looking for a reference for making cell blocks from body fluids. Anyone know of a good source I can provide my histo students? Many thanks to you all, Denise Denise Long Woodward, MS, HT(ASCP), HTL, QIHC Dept. of Pathobiology and Veterinary Sciences University of Connecticut From Dndsomi <@t> aol.com Tue Jul 26 15:31:09 2005 From: Dndsomi <@t> aol.com (Dndsomi@aol.com) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Productivity Message-ID: <82.2cd8fb46.3017f78d@aol.com> We've just been asked to start turning in our histology procedures to be included in our labs daily productivity. How should we report our workload; # of cassettes, blocks, slides, special stains ? Any information will surely be appreciated. Thanks, DDietz, HT From cfranci <@t> rigel.com Tue Jul 26 15:34:36 2005 From: cfranci <@t> rigel.com (Christian Franci) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] bunny Ab's Message-ID: <5f251aaa45b280ca9b47daed7b9455c6@rigel.com> Hello Kids! Was just wondering if any of you have a good supplier of rabbit anti rat Ab's that are mouse absorbed (for real). Supposedly I purchased one but, sans primary antibody present, it makes my mouse tissue light up like a xmas tree.... talk about false positive! thanks in advance. Chris From mcauliff <@t> umdnj.edu Tue Jul 26 15:34:48 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Artifact pictures and cell block prep In-Reply-To: References: Message-ID: <42E69E68.1030302@umdnj.edu> The AFIP manual (3rd edition, 1968, I don't know about other editions) has photos of artifacts after the last numbered chapter. For electron microscopy, there is a book called "An Atlas of Artifacts", I don't remember the author off the top of my head. I can't offer any suggestions for your body fluid question. Geoff Woodward, Denise wrote: >Hi all, > >I'm looking for pictures of sectioning, staining and processing >artifacts. Anyone know where I might be able to find some on the web? > > >Also, looking for a reference for making cell blocks from body fluids. >Anyone know of a good source I can provide my histo students? > >Many thanks to you all, >Denise > >Denise Long Woodward, MS, HT(ASCP), HTL, QIHC >Dept. of Pathobiology and Veterinary Sciences >University of Connecticut > >_______________________________________________ >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 PMonfils <@t> Lifespan.org Tue Jul 26 15:40:58 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Artifact pictures and cell block prep Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717591@lsexch.lsmaster.lifespan.org> You might be interested in one of the following, if you can find a copy: Atlas of Microscopic Artifacts and Foreign Materials by Yeh, Brooks & Pietra An Atlas of Artifacts Encountered in the Preparation of Microscopic Tissue Sections by Thompson & Luna You might find them through www.bookfinder.com We do a fair amount of cell block work in our lab, and have several protocols for different kinds of fluids and cell suspensions. Is there anything in particular you are looking for? Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Woodward, Denise > Sent: Tuesday, July 26, 2005 1:25 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Artifact pictures and cell block prep > > Hi all, > > I'm looking for pictures of sectioning, staining and processing > artifacts. Anyone know where I might be able to find some on the web? > > > Also, looking for a reference for making cell blocks from body fluids. > Anyone know of a good source I can provide my histo students? > > Many thanks to you all, > Denise > > Denise Long Woodward, MS, HT(ASCP), HTL, QIHC > Dept. of Pathobiology and Veterinary Sciences > University of Connecticut > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From denise.woodward <@t> uconn.edu Tue Jul 26 15:52:16 2005 From: denise.woodward <@t> uconn.edu (Woodward, Denise) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Artifact pictures and cell block prep Message-ID: Sorry, I didn't specify, but I'm looking for artifact pictures ON-LINE, so my students can access them from home. Thanks, DLW Denise Long Woodward, MS, HT(ASCP), HTL, QIHC Dept. of Pathobiology and Veterinary Sciences University of Connecticut -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Tuesday, July 26, 2005 4:35 PM To: Woodward, Denise Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artifact pictures and cell block prep The AFIP manual (3rd edition, 1968, I don't know about other editions) has photos of artifacts after the last numbered chapter. For electron microscopy, there is a book called "An Atlas of Artifacts", I don't remember the author off the top of my head. I can't offer any suggestions for your body fluid question. Geoff Woodward, Denise wrote: >Hi all, > >I'm looking for pictures of sectioning, staining and processing >artifacts. Anyone know where I might be able to find some on the web? > > >Also, looking for a reference for making cell blocks from body fluids. >Anyone know of a good source I can provide my histo students? > >Many thanks to you all, >Denise > >Denise Long Woodward, MS, HT(ASCP), HTL, QIHC >Dept. of Pathobiology and Veterinary Sciences >University of Connecticut > >_______________________________________________ >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 Linke_Noelle <@t> Allergan.com Tue Jul 26 16:05:17 2005 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] bunny Ab's Message-ID: Hi Chris, Try Jackson, they usually have everything in the barnyard! Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Christian Franci Sent: Tuesday, July 26, 2005 1:35 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bunny Ab's Hello Kids! Was just wondering if any of you have a good supplier of rabbit anti rat Ab's that are mouse absorbed (for real). Supposedly I purchased one but, sans primary antibody present, it makes my mouse tissue light up like a xmas tree.... talk about false positive! thanks in advance. Chris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charlene.Henry <@t> STJUDE.ORG Tue Jul 26 16:46:40 2005 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] un-subscribe Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2099A199A@SJMEMXMB02.stjude.sjcrh.local> Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 From gkrasinski <@t> covx.com Tue Jul 26 17:27:25 2005 From: gkrasinski <@t> covx.com (Glenn Krasinski) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Anti-mouse specific CD178 (Fas ligand) for IHC (Fr.).... Message-ID: <6837C00A0CC6594197C2F0EEF256561805AAE0@covxmail.covx.com> I am looking for an anti-mouse specific CD178 (fas ligand) clone that will work in IHC(Fr.). My goal is to look for increased levels in endothelial cells treated with compound in Matrigel plug assays. Many thanks for any assistance. Glenn M. Krasinski Scientist, Biology CovX Research LLC 9381 Judicial Drive, Suite 200 San Diego, CA 92121 Ph: (858) 964-2053 FAX: (858) 964-2090 E-dress: gkrasinski@covx.com From TJasper <@t> smdc.org Tue Jul 26 17:59:32 2005 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Productivity Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E451A8@harrier> Dear D, We have a fairly detailed productivity tracking system in place. This is in large part the aftermath of a consultant group that basically tried to cut the department to the bone. Fortunately for us we were operating in an efficient manner and came through pretty much unscathed. I do however have a good deal more paperwork (electronic) to do as a result and my mantra to the staff about this is that it proves we do what we say we do. In a nutshell on a daily basis I track slide production, this includes histology (H+E), cytology (gyn and non-gyn), special stains, and IHC. We are also picking up the bone marrow cores and particles as we do H+E's and Fe stains on them. I also track hours worked for the histotechs, cytotechs, cyto assistants, transcriptionists and autopsy tech. I receive minutes transcribed as a production value for typed dictation on path reports. I also track quality and service indicators, for histology it is, 1st out/last out time, for slide trays. For cytology it is case TAT and for transcription it is reports pending. I have just begun (this month) with a quality report for IHC which is given at our management meeting (along with the other quality indicators). All of this data goes onto spreadsheets (daily) for 2 week intervals coinciding with pay periods to reconcile payroll. Weekly and at the end of each pay period certain figures are carried over onto other spreadsheets which are then accessible to and interpreted by senior administration. Having said all that, I do track blocks daily as a production value on my own. This was not mandated to me by the consultants. I do it because I believe it is a valid production indicator. This is because every block is handled at least once at the grossing bench, once at the embedding center, will generate at least one slide, is block checked against a slide at least once, and is filed at least once. Block counts are solid basic indicators of the minimum work involved as they are handled. No one can ever view them as inflated or exaggerated figures. Our pathologists also like to use the block count numbers (daily) as a gauge for how big a work day to expect. I believe they also base their own division of labor on this number. Just as a note I do spend roughly 50% of my time on the bench, the consultants target for me is 15%. That's just not real world at this time. Please feel free to contact me with any questions, I'll do my best to answer them. tj Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Dndsomi@aol.com [mailto:Dndsomi@aol.com] Sent: Tuesday, July 26, 2005 3:31 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Productivity We've just been asked to start turning in our histology procedures to be included in our labs daily productivity. How should we report our workload; # of cassettes, blocks, slides, special stains ? Any information will surely be appreciated. Thanks, DDietz, HT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Jul 26 17:59:58 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Re: Anti-mouse specific CD178 (Fas ligand) for IHC (Fr.).... In-Reply-To: <6837C00A0CC6594197C2F0EEF256561805AAE0@covxmail.covx.com> References: <6837C00A0CC6594197C2F0EEF256561805AAE0@covxmail.covx.com> Message-ID: <6.0.0.22.1.20050726165659.01b3a4f0@gemini.msu.montana.edu> BD Pharmingen has a monoclonal that works, Serotec may have one also. At 04:27 PM 7/26/2005, you wrote: >I am looking for an anti-mouse specific CD178 (fas ligand) clone that >will work in IHC(Fr.). My goal is to look for increased levels in >endothelial cells treated with compound in Matrigel plug assays. >Many thanks for any assistance. >Glenn M. Krasinski 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 Krat18 <@t> aol.com Tue Jul 26 18:49:15 2005 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] CK7 control/ CK20 contol Message-ID: <1dc.4156ac04.301825fb@aol.com> We use the same as you, but once we had problems because we were not using a lung primary adenoca but one with mets. After we corrected that, we have not had any difficulty. Karen Raterman _krat18@aol.com_ (mailto:krat18@aol.com) From katri <@t> cogeco.ca Tue Jul 26 20:09:52 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] DIF procedures References: <10787860.1122389700521.JavaMail.root@web2.mail.adelphia.net> Message-ID: <003101c59247$e46ab960$6a9a9618@Katri> Ron, we have used a simple direct IF method for years to do IgA, IgG, IgM, complement3c, fibrinogen, albumin, kappa and lambda on fresh frozen human renal biopsies. We collect the needle biopsy at the ultrasound department, when the biopsy is being taken. One core is put in 10% NBF, second one snap frozen and a small piece is fixed in 2% glutaraldehyde for EM. When IF is requested by the pathologist, we cut 5mu sections from the frozen piece and let slides ( use poly-l-lysine or + slides) dry at room temperature for minimum 30 minutes. H&E gets done on one slide to make sure there are glomeruli in the biopsy. We then do the direct immunofluorescence on unfixed sections: 1. Wash in PBS for 15 minutes 2. Apply fluorescein conjugated antibody on the sections for 30 minutes: Dako FITC/IgA at 1:20 in PBS Dako FITC/IgG at 1:30 Dako FITC/IgM at 1:20 Dako FITC/C3c at 1:50 Dako FITC/fibrinogen at 1:10 Dako FITC/kappa at 1:20 Dako FITC/lambda at 1:20 Dako FITC/albumin at 1:30 (keep slides away from light, cover with a towel) 3. Wash well in PBS x2 for 15 minutes each. 4. Coverslip with aqueous non-fluorescent mounting medium, store in fridge until ready to read. Photographic record is made of results, since these preparations are not considered permanent. Hope this helps, Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: To: Sent: Tuesday, July 26, 2005 10:55 AM Subject: [Histonet] DIF procedures > Fellow Techs, > Does anyone have Direct Immunofluorescence procedures they are willing to > share? We are interested in CD3, fibrinogen, IgA, IgM and IgG. > Thanks in advance. > Ron Martin, BS HT (ASCP) HTL > fax 561-721-1249 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From louise.renton <@t> gmail.com Wed Jul 27 01:38:15 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] granular debris in paraffin blocks In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE01717590@lsexch.lsmaster.lifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE01717590@lsexch.lsmaster.lifespan.org> Message-ID: Off the top of my head.... What about doing something like microincineration (same as is done for asbestos/asbestosis in lung)? That would show the inorganic, but not the organic component or perhaps xray to show the silica particles? Just a few lateral thoughts On 7/26/05, Monfils, Paul wrote: > Now there's a unique problem! No, sand really doesn't section very well. How > about cutting a thick section? A 20 or 30 micron section may not be > suitable for cellular detail, but it might contain enough of the material to > demonstrate it adequately; and while the section might be full of > striations, hopefully the striations might not be deep enough to shread the > ribbon. Also, try to get the earliest section you can get, while knife > damage is minimal. Use a fresh section of knife edge, and start rotating the > microtome before the block touches the knife, so you can get the first > couple of sections off the block face. If that doesn't work, your > pathologist will probably have to demonstrate the debris by having the lung > x-rayed. > > Paul M. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Diana > > McCaig > > Sent: Tuesday, July 26, 2005 11:43 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] granular debris in paraffin blocks > > > > Does anyone have any suggestions to obtain sections from an autopsy case > > of > > a drowning victim. The lung tissue has granular debris from a muddy > > ditch. > > The pathologist wants this demonstrated but it is virtually impossible to > > obtain anything due to fragmentation of the blocks from this. > > Thanks > > Diana > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From c.m.vanderloos <@t> amc.uva.nl Wed Jul 27 02:54:33 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] RE: Background staining: mouse on mouse Broad spectrum Message-ID: <1a566e61a53c78.1a53c781a566e6@amc.uva.nl> Dear Cathy, From your mail I understand this Zymed Histomouse Max Kit isn't working for you. I heard from a colleague here in the hospital this kit wasn't working background-free for her on rat tissue either. I realized the Dakocytomation ARKit (based on streptavidin-biotin) isn't appropriate here as well since you are dealing with liver full of non-blockable endogenous biotin. The best alternative for you is perhaps a Zenon kit ([1]http://probes.invitrogen.com/products/zenon). Like with the ARKit the Zenon kit works with an in vitro labeling of the primary antibody with a labeled Fab-fragment. Kits are available with many fluorochromes as label, including FITC. This allows further enzymatic visualization via Rabbit anti-FITC and goat anti-rabbit/HRP or whatever. This doesn't answer your questions, but might solve the background staining problem! Hope this works for you. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Tue, 26 Jul 2005 18:44:09 +1000 From: Cathy Malcontenti-Wilson Subject: [Histonet] Background staining: mouse on mouse Broad spectrum kit To: histonet@lists.utsouthwestern.edu Dear All, I really need some advice regarding an ongoing intense background problem. We use FFP sections of mouse liver. We are using an immuno kit called Zymed Histomouse Max Kit (It is a HRP- DAB one). Our primary ab is mouse anti human VEGF (1:1000). The kit is called a BROAD SPECTRUM kit because it is meant to be able to be used for mouse, rabbit, rat and guinea pig primaries. The secondary antibody is not completely explained in the kit but I think it is of goat host species. Question: If it is broad spectrum does this mean that the secondary antibody is a mixture of different antibodies? The kit contains its own non specific block, with no details of what it actually is. Question: If the second! ary ab is References 1. http://probes.invitrogen.com/products/zenon From lilliangarrett <@t> gmail.com Wed Jul 27 05:39:35 2005 From: lilliangarrett <@t> gmail.com (Lillian Garrett) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Optimising the vasopressin V1b receptor stain Message-ID: <81b03aa3050727033966889ed6@mail.gmail.com> Hi everybody, I'm just wondering whether anyone has had success using the vasopressin V1b receptor antibody from Alpha Diagnostics International in brain tissue... I am having trouble optimising this stain and get faint to no signal...... Could anyone suggest possible ways of getting the most from this staining..... The brain tissue is fixed in 4% paraformaldehyde.... I want to stain cryosectioned material using DAB as the cromogen.... So far I have increased the primary antibody incubation time to 48 hours and have heated the sections to no avail..... Any help with this problem would be greatly appreciated... Thanks, Lillian From jluis.palazon <@t> icman.csic.es Wed Jul 27 06:08:21 2005 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] ANTI-MYOSIN AND ANTIRENIN Message-ID: <20050727110821.0A9DD88D8B2@perceval.uca.es> Dear List-members I am finishing my doctoral thesis and would like to demonstrate renin in an aglomerular teleost kidney, smooth muscle in the iris, and muscle in the bulbus arteriosus. I know that this can be done inmunohistologically but my lab does not work in inmunohistochemistry so I do not have the antibodies. I would like to know if there is someone of our list here in Spain (hospital, University) that can do this kind of inmunos and the posibility of doing my samples with them or maybe sending the samples and paying for the inmunos. thanks in advance Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Jul 27 06:53:15 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] RE: CK7 and CK 20 positive control tissue Message-ID: We use liver for the CK 7 and colon+appendix for the CK 20. We make a multiblock with colon+appx+tonsil which does for quite a few antisera anchored saves the time it takes in keep putting in and taking out a single control block for each antisera. Just cut a ribbon and pick your sections off as you want them. Cheers Jacqui Malam Lancaster DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From c.m.vanderloos <@t> amc.uva.nl Wed Jul 27 07:14:22 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] RE: Biotin conjugated Antibodies -- can we skip biotin?? Message-ID: <1abd9271abb50e.1abb50e1abd927@amc.uva.nl> Hello, You may apply successfully any appropriate anti-goat reagent, ignoring the biotin-label of the primary antibody. Biotin groups at the primary are far too small to block anti-goat reagents. As long as there are no biotin-binding structures in the section, which is only very rarely the case, the biotin-label of the primary isn't influencing the final staining result. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Tue, 26 Jul 2005 12:23:10 -0700 (PDT) From: GT Hebert Subject: [Histonet] Biotin conjugated Antibodies -- can we skip biotin?? To: histonet@lists.utsouthwestern.edu Hello, Just a general question to help increase my knowledge: If I had an antibody that was conjugated to biotin -- say for example, a goat anti-mouse P-selectin (biotin) -- can I just take a rabbit anti-goat conjugated to AP and run IHC that way -- hopefully avoiding the ABC step and background OR does the biotin from the primary Ab block secondaries from coming in? Thanks. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Jul 27 08:05:19 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] RE: artefacts Message-ID: You might look in our EQA web site (http://www.cpteqa.pwp.blueyonder.co.uk/Crit or look in Woods and Ellis' "Laboratory Histopathology: A complete Reference". Churchill Livingstone 1994. Vol 1; 5.3. Cheers Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From Wanda.Smith <@t> HCAhealthcare.com Wed Jul 27 08:33:03 2005 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] subscribe Message-ID: Wanda G. Smith, HTL/HT(ASCP) Pathology Supervisor Trident Medical Center 9330 Medical Plaza Drive North Charleston, SC 29406 843-797-4586 fax 843-797-4296 wanda.smith@hcahealthcare.com From Terry.Marshall <@t> rothgen.nhs.uk Wed Jul 27 08:42:00 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] RE: artefacts Message-ID: All very well, but we get a 403 message - "You don't have permission to access those file(s)." 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: Malam Jacqueline [mailto:Jacqueline.Malam@rli.mbht.nhs.uk] Sent: 27 July 2005 14:05 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: artefacts You might look in our EQA web site (http://www.cpteqa.pwp.blueyonder.co.uk/Crit or look in Woods and Ellis' "Laboratory Histopathology: A complete Reference". Churchill Livingstone 1994. Vol 1; 5.3. Cheers Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jluis.palazon <@t> icman.csic.es Wed Jul 27 11:01:58 2005 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] retina neurons Message-ID: <20050727160158.10EC588F23A@perceval.uca.es> Dear list fellows is there any simple stain that I can use to differentiate between amacrine, horizontal and bipolar cells in the retina? thanks in advance jose luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From Eric <@t> ategra.com Wed Jul 27 12:34:42 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] HistoTech Job Openings Message-ID: Histonetters - My name is Eric Dye and I am the Histo Tech recruiter here at Ategra. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job openings are in Denver, Colorado who is seeking a Histotech Manager and Northern Oregon also seeking a Histotech Manager. I also have bench histo tech jobs available in: California,Michigan, Pennsylvania, Illinois, New Hampshire, Massachusetts, West Virginia, Georgia, Colorado and Florida. I have a opening for a pathologist's Assistant in Illinois. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------- From failm <@t> musc.edu Wed Jul 27 12:35:24 2005 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] ? For IHC labs with 1200-1500 slides per month Message-ID: Regarding controls. Do you place the patient tissue on the slide with the control tissue? If so who cuts the pt. tissue, who cuts the controls? If the pt tissue is cut in one lab and the controls in another, could you describe how the controls are labeled, stored, distributed ? We are going to be putting the pt tissue on the control slides, which will decrease the amt of slides placed on th e autostainer, increase the number of control slides needing to be cut, and the amt of reagent required for each slide. We have some ideas on how to distribute the controls, but I would be very interested in how some of you handle the controls for IHC. Thank you, Rena Fail From PMonfils <@t> Lifespan.org Wed Jul 27 13:07:04 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] non-aqueous, non-xylene mounting media Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717595@lsexch.lsmaster.lifespan.org> I am using VectaMount from Vector Laboratories to coverslip stained paraffin sections for special projects where the specimens/sections cannot be exposed to xylene or toluene, and the stains cannot be mounted with aqueous media. This mounting medium is non-aqueous, and xylene/toluene free. Are there any other commercially available mounting media that meet these criteria? And has anyone done any comparisons between brands? If so, which one do you prefer, and why? From kaleid11 <@t> yahoo.com Wed Jul 27 13:19:11 2005 From: kaleid11 <@t> yahoo.com (Adam Perry) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] ligand binding Message-ID: <20050727181912.60021.qmail@web31610.mail.mud.yahoo.com> hello all, I am trying to work up a protocol for detecting oxytocin receptors in rodent brain sections using a biotinylated ligand and subsequent avidin-biotin-HRP steps for visualization. Our lab currently uses autoradiography, but I'd like to use the biotinylated method for more discrete localization. Does anyone have experience with ligand binding studies or better yet non-isotopic binding? If so, my specific questions are: 1) How does fixation of sections alter binding? Currently we use fresh frozen brains, cut sections on cryostat and then lightly fix the sections in formaldehyde prior to binding incubation. Is it possible to perform ligand binding on tissue perfused with 4% formaldehyde and then sectioned- I would rather work with tissue fixed before cutting- of course I'm sure different ligands have different requirements... 2) Is it necessary to fix the sections after the binding step and washes to "trap" the ligand-receptor complex for A/B development? I've seen protocols that do both (fix or no fix after binding). 3) Has anyone done tyramine signal amplification in conjunction with biotinylated-ligand binding? Does it help, or not really required? Any other comments on your experiences would be most appreciated. Thanks in advance, Adam Perry Department of Physiology and Biophysics University of Illinois at Chicago --------------------------------- Start your day with Yahoo! - make it your home page From mclarke <@t> allsaintshealthcare.org Wed Jul 27 13:21:50 2005 From: mclarke <@t> allsaintshealthcare.org (Clarke, Mary) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] ck7 and ck20controls Message-ID: <6FE2A453DA437F4D96FAAF363F20ABB20FC868@WFEXBE04.wfsi.priv> We use a bladder carcinoma for the ck7 and a mucicarmine control for the ck20. Terri Clarke Histology Supervisor All Saints Laboratory 3801 Spring Street Racine, Wi 53405 mclarke@allsaintshealthcare.org 262-687-6609 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, July 26, 2005 4:56 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 36 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Genta stain for Helicobacter pylori (IamNinaOwen@aol.com) 2. stain for pre-collagen and mature collagen (Steven Coakley) 3. re: iNOS (Kristopher Kalleberg) 4. RE: Genta stain for Helicobacter pylori (Monfils, Paul) 5. granular debris in paraffin blocks (Diana McCaig) 6. re: iNOS (Kristopher Kalleberg) 7. Re: Integrase Interactor 1 (INI-1/hsnf5) antibody needed (Joanne Mauger) 8. Applied IHC Journal (Richard Cartun) 9. RE: granular debris in paraffin blocks (Monfils, Paul) 10. Serotec Rat Anti-mouse F4/80 : Biotin -- anyone know of good concentration? (GT Hebert) 11. Biotin conjugated Antibodies -- can we skip biotin?? (GT Hebert) 12. anti-mouse cleaved PARP for IHC... (Glenn Krasinski) 13. CK7 control/ CK20 contol (Amy Johnson) 14. RE: CK7 control/ CK20 contol (Sebree Linda A.) 15. Re: Serotec Rat Anti-mouse F4/80 : Biotin -- anyone know of good concentration? (Gayle Callis) 16. Re: Serotec Rat Anti-mouse F4/80 : Biotin -- anyone know of good concentration? (GT Hebert) 17. Re: Biotin conjugated Antibodies -- can we skip biotin?? (Gayle Callis) 18. Artifact pictures and cell block prep (Woodward, Denise) 19. Productivity (Dndsomi@aol.com) 20. bunny Ab's (Christian Franci) 21. Re: Artifact pictures and cell block prep (Geoff McAuliffe) 22. RE: Artifact pictures and cell block prep (Monfils, Paul) 23. RE: Artifact pictures and cell block prep (Woodward, Denise) 24. RE: bunny Ab's (Linke_Noelle) 25. un-subscribe (Henry, Charlene) ---------------------------------------------------------------------- Message: 1 Date: Tue, 26 Jul 2005 14:29:31 EDT From: IamNinaOwen@aol.com Subject: [Histonet] Genta stain for Helicobacter pylori To: histonet@lists.utsouthwestern.edu Message-ID: <20.4990d7aa.3017db0b@aol.com> Content-Type: text/plain; charset="US-ASCII" Hello Histonetters! Has anyone ever used the Genta stain for Helicobacter pylori? I'm trying to find a protocol for this method, so any help will be very much appreciated! Thanks Nina Owen ------------------------------ Message: 2 Date: Tue, 26 Jul 2005 11:30:39 -0700 (PDT) From: Steven Coakley Subject: [Histonet] stain for pre-collagen and mature collagen To: Histonet@lists.utsouthwestern.edu Message-ID: <20050726183039.46878.qmail@web90204.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I've been working on Herovica's Method for differentiating pre-collagen(young) verses mature collagen using varies combinations of methy blue and van gieson's. Luna has a modified method on pg 453 of his "Histopathologic Methods...." The regular Herovica stain calls for tissue fixed in formol-acetic-alcohol. 10% NBF leaves the muscle a blue to blue-green and very little red (mature collagen). I tried post fixation in the FAA for 1 hr w/o allot of difference. Bouins post-fixation seems to work best. Certainly not getting the results as outlined. Is there anyway to check the saturated picric acid? Any ideas or suggestion would be appreciated, especially from those familiar with this stain. Steve --------------------------------- Start your day with Yahoo! - make it your home page ------------------------------ Message: 3 Date: Tue, 26 Jul 2005 14:29:43 -0400 From: "Kristopher Kalleberg" Subject: [Histonet] re: iNOS To: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: TEXT/PLAIN; CHARSET=US-ASCII If I remember right Neomarkers/Lab Vision Corp. has iNOS antibodies. I used them maybe 1 1/2 years ago but never got great results from the stain. I ran the test numerous times and was unable to solve the problem. If you decide to use Neomarkers and are able to achieve good results please send me your secret. Good luck. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull, CT 06611 203 381 5765 -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, July 26, 2005 1:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 20, Issue 35 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. gluteraldehyde fixation with Re: [Histonet] Rat brains (Gayle Callis) 2. IHC for RE:Staining for Helicobacter pylori (Gayle Callis) 3. Re: Staining for Helicobacter pylori (IamNinaOwen@aol.com) 4. TMB staining for WGA-HRP (NIENHUIS,ROBERT ) 5. historesin (Walters, Katherine S) 6. equivalent to Re: [Histonet] historesin (Gayle Callis) 7. Janella Seaton...please contact me (Morken, Tim - Labvision) 8. Background staining: mouse on mouse Broad spectrum kit (Cathy Malcontenti-Wilson) 9. Re: historesin (Don.Birgerson@leica-microsystems.com) 10. Re: CD 31 (Jackie M O'Connor) 11. North Carolina Histologist Position Available (MajorFocus@aol.com) 12. Re: equivalent to Re: [Histonet] historesin (Pamela Marcum) 13. Looking 4 Antibodies (Luis Chiriboga) 14. DIF procedures (pathrm35@adelphia.net) 15. teaching heads for Nikon microscope (pathrm35@adelphia.net) 16. Integrase Interactor 1 (INI-1/hsnf5) antibody needed (Histology SLU) 17. looking for iNOS antibody (Louro, Pedro) 18. Re: Integrase Interactor 1 (INI-1/hsnf5) antibody needed (Dana Settembre) 19. IgG2a and IgG1 for guinea pig (Elizabeth Chlipala) ---------------------------------------------------------------------- Message: 1 Date: Mon, 25 Jul 2005 11:21:28 -0600 From: Gayle Callis Subject: gluteraldehyde fixation with Re: [Histonet] Rat brains To: Laura Fidgen , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050725111447.01b7ada0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Question: is he planning to do EM on these tissues? If he had used paraformaldehyde, your sectioning would be much easier - but if he is also doing EM - you can try some things to counteract the dry/brittle brain. In general, gluteraldehyde creates some friable, brittle tissues when these are processed and embedded in paraffin. Check to see if you are processing too long as long dehydrations, etc and heat from paraffin contributes to the problem. A good long soak in ice water or solid ice block with water on top after trimming might help but be sure to not retrim the soak away, you have to get the first sections off the knife. A warm water or room temperature soak the going onto ice water for a time may help too, brain sometimes needs a bit of help. At 01:01 PM 7/23/2005, you wrote: >Please help... >A research project for our neuropathologist includes rat brains. These >are extremely difficult to section as he profuses the whole rat with >glutaraldehyde then takes brain samples. I have tried adding alcohol to >my water bath but these sections still have many, many wrinkles. Can >anyone offer suggestions? I would be so grateful for any and all advice. 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: 2 Date: Mon, 25 Jul 2005 11:25:52 -0600 From: Gayle Callis Subject: [Histonet] IHC for RE:Staining for Helicobacter pylori To: Tom McNemar , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050725112230.01b7b5c8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Dear Tom, If I had to do Helicobacter, I would vote for IHC too!!! Very specific for identification of Helicobacter in human tissues, I only wished I had it for murine animal model work but no antibody was available - boo hoo!!! There is a nice method published in Histologic, Sakura Finetek website on doing this IHC, and it was wonderful. At 07:05 AM 7/25/2005, you wrote: >We have been doing them by IHC for the last few years. Overkill, I know. >The paths never like the Giemsa and we had so much trouble with the >Warthin-Starry (consistency, repeats, etc) that I just switched to IHC. >Works every time. >Tom McNemar, HT(ASCP) >Histology Co-ordinator >Licking Memorial Hospital >Newark, Ohio 43055 Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 3 Date: Mon, 25 Jul 2005 13:30:46 EDT From: IamNinaOwen@aol.com Subject: [Histonet] Re: Staining for Helicobacter pylori To: histonet@lists.utsouthwestern.edu Message-ID: <65.49f13cd4.30167bc6@aol.com> Content-Type: text/plain; charset="US-ASCII" Hi all Thanks for all your views on this matter, keep the replies rolling in! Nina Owen ------------------------------ Message: 4 Date: Mon, 25 Jul 2005 11:55:40 -0700 From: "NIENHUIS,ROBERT " Subject: [Histonet] TMB staining for WGA-HRP To: Histonet@lists.utsouthwestern.edu Message-ID: <1122317740.42e535ac936d3@mail.ucla.edu> Content-Type: text/plain; charset=ISO-8859-1 We are about to cut a brain that was injected with WGA-HRP for tract tracing. We haven't done this for quite a while (decades?) and don't remember if it is possible to delay processing after cutting. If it is possible, how long can we delay, and what solution should the cut tissue be stored in? We plan to stain using the Olucha molybybdate modification to the Mesulam TMB procedure. Bob Nienhuis UCLA / VA Medical Center Los Angeles, CA ------------------------------ Message: 5 Date: Mon, 25 Jul 2005 15:07:50 -0500 From: "Walters, Katherine S" Subject: [Histonet] historesin To: Message-ID: Content-Type: text/plain; charset="us-ascii" I have a colleague looking to buy some historesin. Apparently it is made by Leica (in Germany), and they will no longer ship it to the US. Is there a company here that can sell it, or a similar product? Thanks, Kathy ------------------------------ Message: 6 Date: Mon, 25 Jul 2005 15:11:15 -0600 From: Gayle Callis Subject: equivalent to Re: [Histonet] historesin To: "Walters, Katherine S" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050725145504.01b275e0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I don't recall plastic type of Historesin since it has been out of circulation for a while in USA, but if this is GMA, buy Technovits 7100 If it is the methylmethacrylate, MMA then buy Technovits 9100. Electron Microscopy Sciences sells both kits. These are excellent products. At 02:07 PM 7/25/2005, you wrote: >I have a colleague looking to buy some historesin. Apparently it is >made by Leica (in Germany), and they will no longer ship it to the US. >Is there a company here that can sell it, or a similar product? > >Thanks, >Kathy > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 7 Date: Mon, 25 Jul 2005 18:47:26 -0400 From: "Morken, Tim - Labvision" Subject: [Histonet] Janella Seaton...please contact me To: "'Histonet@lists.utsouthwestern.edu'" Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CF55@usca0082k08.labvision.apogent. com> Content-Type: text/plain Janella, please contact me about the database you want. From: Janella Seaton ------------------------------------------------------------------------ ---- ---- Hello! We want to implement a database for specimens we receive in our lab. We would use this database starting with logging in specimens and ending with archiving of specimens. It would be used to track when various steps (gross photos, processing, staining, etc) have occurred and by whom. We do want a system that is user friendly. Any suggestions or advice? Thanks for the help..... j.s. Tim Morken ------------------------------ Message: 8 Date: Tue, 26 Jul 2005 18:44:09 +1000 From: Cathy Malcontenti-Wilson Subject: [Histonet] Background staining: mouse on mouse Broad spectrum kit To: histonet@lists.utsouthwestern.edu Message-ID: <6.2.1.2.2.20050726182055.02ecbce8@mail.staff.unimelb.edu.au> Content-Type: text/plain; charset="us-ascii"; format=flowed Dear All, I really need some advice regarding an ongoing intense background problem. We use FFP sections of mouse liver. We are using an immuno kit called Zymed Histomouse Max Kit (It is a HRP- DAB one). Our primary ab is mouse anti human VEGF (1:1000). The kit is called a BROAD SPECTRUM kit because it is meant to be able to be used for mouse, rabbit, rat and guinea pig primaries. The secondary antibody is not completely explained in the kit but I think it is of goat host species. Question: If it is broad spectrum does this mean that the secondary antibody is a mixture of different antibodies? The kit contains its own non specific block, with no details of what it actually is. Question: If the secondary ab is raised in goat, is this non specific block normal goat serum? If not, what could it be? We know that when I incubate the tissue with DAB only, we get endogenous peroxidase staining which is pretty much abolished with H2O2 block. We know that when we omit the primary antibody and keep all the other layers, we get intense background staining of the sinusoidal (endothelial) lining of practically all the sinusoids, and we get brown staining in plasma cells, macrophages, connective tissue and other cells within vessels. Question: Omission of non specific blocking reagent showed increased background staining. Does this mean that the blocking agent is actually blocking some of the background but not all? Why not? We know that when we omit both the primary antibody and the non specific block then we get background staining. Question: What does this mean? We know if we omit the secondary antibody only, we get no staining. Question: What does this mean? I would greatly appreciate any input into this problem which has been plaguing us for a while now. I hope someone can help, I cant beleive what a great wealth of knowledge there is on this histonet list. Cathy Malcontenti-Wilson ------------------------------ Message: 9 Date: Tue, 26 Jul 2005 07:32:06 -0500 From: Don.Birgerson@leica-microsystems.com Subject: Re: [Histonet] historesin To: "Walters, Katherine S" Cc: histonet-bounces@lists.utsouthwestern.edu, histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hi Kathy, While we quit importing Historesin, there is product called "Technovit " that is sold in the US by "Energy Beam Science" at 413-786-9322. If you have further questions, please call me. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Walters, Katherine S" Sent by: histonet-bounces@lists.utsouthwestern.edu 07/25/2005 03:07 PM To cc Subject [Histonet] historesin I have a colleague looking to buy some historesin. Apparently it is made by Leica (in Germany), and they will no longer ship it to the US. Is there a company here that can sell it, or a similar product? Thanks, Kathy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com ------------------------------ Message: 10 Date: Tue, 26 Jul 2005 07:43:00 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] CD 31 To: Angela McNabola Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Santa Cruz gave me the same story - it's amazing that they depend on one goat for a product line. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist Discovery Cancer R4N2 AP3 847-938-4919 Jackie.O'Connor@abbott.com Angela McNabola Sent by: histonet-bounces@lists.utsouthwestern.edu 07/25/2005 05:40 AM To: histonet@lists.utsouthwestern.edu cc: Subject: Re: [Histonet] CD 31 Which antibody are you using? We have had great success with Santa Cruz (goat polyclonal) until I ordered a new lot this past month. When I called them, they indicated to me that they were having problems with their goat, literally. I guess it had stopped producing antibodies to CD31 that work in IHC (interestingly enough, they still work in Westerns). They hope to have the problem resolved within the next month, with a new goat. Angela McNabola, MS, HT(ASCP)SLS, QIHC Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 "Kim Allred" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] CD 31 western.edu 07/22/2005 03:15 PM I am having a problem getting my CD31 to work. The protocol was for an overnight incubation and pepsin pretreatment. I couldn't get it to work this way so I altered it a bit by using trilogy for AR and a two hour incubation for the primary ab. Does anyone have any ideas? This seemed to work well for a while but once again, I'm having problems with it. Any input would be appreciated. Thanks in advance! Kim Allred Dermatopathology Associates Jackson, MS _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Tue, 26 Jul 2005 09:18:10 EDT From: MajorFocus@aol.com Subject: [Histonet] North Carolina Histologist Position Available To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Frye Regional Medical Center in Hickory, North Carolina is currently seeking a Histology Technician or Technologist for full time employment. Qualified applicants must be certified by the ASCP or equivalent agency. Please apply on-line at _www.fryemedctr.com_ (http://www.fryemedctr.com/) or call toll-free (866) 494-4747. Frye Regional Medical Center Human Resources 420 North Center Street Hickory, NC 28601. ------------------------------ Message: 12 Date: Tue, 26 Jul 2005 09:25:08 -0400 From: Pamela Marcum Subject: Re: equivalent to Re: [Histonet] historesin To: Gayle Callis , "Walters, Katherine S" , Histonet@lists.utsouthwestern.edu Message-ID: <6.1.1.1.2.20050726092213.019799b0@mail.vet.upenn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Hi, The Technovit 8100 is the same as JB-4 Plus which has a second polymer to make the GMA remain clearer and somewhat less brittle. As Don was kind enough to give you the number for EBS I shall not repeat and you can reach Polysciences at 800-523-2575. Pam Marcum At 05:11 PM 7/25/2005, Gayle Callis wrote: >I don't recall plastic type of Historesin since it has been out of >circulation for a while in USA, but if this is GMA, buy Technovits 7100 > >If it is the methylmethacrylate, MMA then buy Technovits 9100. > >Electron Microscopy Sciences sells both kits. These are excellent products. > >At 02:07 PM 7/25/2005, you wrote: > > >>I have a colleague looking to buy some historesin. Apparently it is >>made by Leica (in Germany), and they will no longer ship it to the US. >>Is there a company here that can sell it, or a similar product? >> >>Thanks, >>Kathy >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Tue, 26 Jul 2005 09:49:42 -0400 From: Luis Chiriboga Subject: [Histonet] Looking 4 Antibodies To: Histonet Message-ID: Content-Type: text/plain; charset="iso-8859-1" HI everyone I'm looking for antibodies against MLH-1, MSH-2, BRCA-1 and BRCA-2 for use in human FF-PE tissue. recommendations as well as comments are welcome. feel free to respond directly to my address. Thanks in advance Luis ------------------------------ Message: 14 Date: Tue, 26 Jul 2005 10:55:00 -0400 From: Subject: [Histonet] DIF procedures To: histonet@lists.utsouthwestern.edu Message-ID: <10787860.1122389700521.JavaMail.root@web2.mail.adelphia.net> Content-Type: text/plain; charset=utf-8 Fellow Techs, Does anyone have Direct Immunofluorescence procedures they are willing to share? We are interested in CD3, fibrinogen, IgA, IgM and IgG. Thanks in advance. Ron Martin, BS HT (ASCP) HTL fax 561-721-1249 ------------------------------ Message: 15 Date: Tue, 26 Jul 2005 11:01:06 -0400 From: Subject: [Histonet] teaching heads for Nikon microscope To: histonet@lists.utsouthwestern.edu Message-ID: <28528193.1122390066380.JavaMail.root@web2.mail.adelphia.net> Content-Type: text/plain; charset=utf-8 Fellow Techs, Does anyone know of or where I can find binocular teaching heads for a (older model) multiheaded Nikon Optiphot microscope? Any leads or info is greatly appreciated. Thanks in advance. Ron Martin, BS HT (ASCP) HTL fax 561-721-1249 ------------------------------ Message: 16 Date: Tue, 26 Jul 2005 08:35:52 -0700 (PDT) From: Histology SLU Subject: [Histonet] Integrase Interactor 1 (INI-1/hsnf5) antibody needed To: histonet@lists.utsouthwestern.edu Message-ID: <20050726153552.54597.qmail@web51005.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello All: I have a doc looking for the above antibody for use in FFPE tissue sections of rhabdoid tumor in children. Any help would be greatly appreciated. Thanks, in advance. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 17 Date: Tue, 26 Jul 2005 11:58:13 -0400 From: "Louro, Pedro" Subject: [Histonet] looking for iNOS antibody To: histonet@lists.utsouthwestern.edu Message-ID: <4508920F80C0D411B90200508BF9A9F4062B4A40@LAFMSG30.us.schp.com> Content-Type: text/plain Hi all, I'm starting a new experiment with rat mesentery artery looking for inducible nitric oxide synthase (iNOS, eNOS, and nNOS). What companies would carry this antibody that would work on FFPE rat tissue. Also, what would be a good positive control? Thanks in advance for your assistance Pedro ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 18 Date: Tue, 26 Jul 2005 12:13:31 -0400 From: Dana Settembre Subject: Re: [Histonet] Integrase Interactor 1 (INI-1/hsnf5) antibody needed To: histonet@lists.utsouthwestern.edu, sluhisto@yahoo.com Message-ID: Content-Type: text/plain; charset=US-ASCII I purchased it from BD Biosciences from San Diego, CA cat# 612110 I pretreat with DakoCytomation's Target Retrieval Solution. and use it at 1:50 incubating for 15 minutes room temp. Works nicely but there's not much in the vial and I am looking to Santa Cruz, who sells 1ml of it. Santa Cruz Cat. sc-13055 Good Luck. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> Histology SLU 7/26/2005 11:35:52 AM >>> Hello All: I have a doc looking for the above antibody for use in FFPE tissue sections of rhabdoid tumor in children. Any help would be greatly appreciated. Thanks, in advance. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 26 Jul 2005 10:48:10 -0600 From: "Elizabeth Chlipala" Subject: [Histonet] IgG2a and IgG1 for guinea pig To: "'Histonet'" Message-ID: <000101c59201$ce33a1c0$a7d48a80@AMY> Content-Type: text/plain; charset="US-ASCII" Hello everyone I have been looking for some reagents for guinea pig for both ELISA and IHC for IgG2a and IgG1. I did find a source for the IgG2a through Nordic Immuno Reagents. Has anyone had any dealings with them? They have several antibodies against guinea pig IgG2a all derived from different species (sheep, goat and rabbit). I was guessing that the rabbit antibody would be nice since I could use a polymer detection system if necessary. Any suggestions of a different vendor or anything else would be appreciated. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 35 **************************************** ------------------------------ Message: 4 Date: Tue, 26 Jul 2005 14:41:25 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] Genta stain for Helicobacter pylori To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171758F@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="ISO-8859-1" It's in the archives, here: http://www.histosearch.com/histonet/Nov99/RE.GENTASTAINA.html I read a paper which stated that lead nitrate can be substituted for uranium nitrate in this procedure, thereby avoiding the problems of handling and disposing of radioactive material. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > IamNinaOwen@aol.com > Sent: Tuesday, July 26, 2005 11:29 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Genta stain for Helicobacter pylori > > Hello Histonetters! > > Has anyone ever used the Genta stain for Helicobacter pylori? I'm trying > to > find a protocol for this method, so any help will be very much > appreciated! > > Thanks > > Nina Owen > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 5 Date: Tue, 26 Jul 2005 14:43:45 -0400 From: Diana McCaig Subject: [Histonet] granular debris in paraffin blocks To: histonet@lists.utsouthwestern.edu Message-ID: <3E5A3F039F0BD8118B4700C00D00202404361E@CKHA9> Content-Type: text/plain Does anyone have any suggestions to obtain sections from an autopsy case of a drowning victim. The lung tissue has granular debris from a muddy ditch. The pathologist wants this demonstrated but it is virtually impossible to obtain anything due to fragmentation of the blocks from this. Thanks Diana ------------------------------ Message: 6 Date: Tue, 26 Jul 2005 14:48:38 -0400 From: "Kristopher Kalleberg" Subject: [Histonet] re: iNOS To: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: TEXT/PLAIN; CHARSET=US-ASCII If I remember right Neomarkers/Lab Vision Corp. has iNOS antibodies. I used them maybe 1 1/2 years ago but never got great results from the stain. I ran the test numerous times and was unable to solve the problem. If you decide to use Neomarkers and are able to achieve good results please send me your secret. Good luck. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull, CT 06611 203 381 5765 ------------------------------ Message: 7 Date: Tue, 26 Jul 2005 14:51:00 -0400 From: "Joanne Mauger" Subject: Re: [Histonet] Integrase Interactor 1 (INI-1/hsnf5) antibody needed To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Susan, You can get INI-1(BAF47) from BD Transduction labs- catalog # 612110 for 50 ug, or 61211 for 150 ug. Jo >>> Histology SLU 07/26/05 11:35 AM >>> Hello All: I have a doc looking for the above antibody for use in FFPE tissue sections of rhabdoid tumor in children. Any help would be greatly appreciated. Thanks, in advance. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Tue, 26 Jul 2005 14:53:05 -0400 From: "Richard Cartun" Subject: [Histonet] Applied IHC Journal To: Message-ID: Content-Type: text/plain; charset=US-ASCII Does anyone have the Volume 3, Number 1 issue of Applied Immunohistochemistry from 1995? Thank you! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 9 Date: Tue, 26 Jul 2005 15:12:19 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] granular debris in paraffin blocks To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717590@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="ISO-8859-1" Now there's a unique problem! No, sand really doesn't section very well. How about cutting a thick section? A 20 or 30 micron section may not be suitable for cellular detail, but it might contain enough of the material to demonstrate it adequately; and while the section might be full of striations, hopefully the striations might not be deep enough to shread the ribbon. Also, try to get the earliest section you can get, while knife damage is minimal. Use a fresh section of knife edge, and start rotating the microtome before the block touches the knife, so you can get the first couple of sections off the block face. If that doesn't work, your pathologist will probably have to demonstrate the debris by having the lung x-rayed. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Diana > McCaig > Sent: Tuesday, July 26, 2005 11:43 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] granular debris in paraffin blocks > > Does anyone have any suggestions to obtain sections from an autopsy case > of > a drowning victim. The lung tissue has granular debris from a muddy > ditch. > The pathologist wants this demonstrated but it is virtually impossible to > obtain anything due to fragmentation of the blocks from this. > Thanks > Diana > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 10 Date: Tue, 26 Jul 2005 12:19:51 -0700 (PDT) From: GT Hebert Subject: [Histonet] Serotec Rat Anti-mouse F4/80 : Biotin -- anyone know of good concentration? To: histonet@lists.utsouthwestern.edu Message-ID: <20050726191951.92792.qmail@web31708.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello once again, I just received my Serotec Rat Anti-mouse F4/80 antibody and was wondering if anyone can share with me the concentration they use -- closest to optimized would be great! Thanks Gustave Hebert Scientist II CVMD Wyeth - Cambridge __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 11 Date: Tue, 26 Jul 2005 12:23:10 -0700 (PDT) From: GT Hebert Subject: [Histonet] Biotin conjugated Antibodies -- can we skip biotin?? To: histonet@lists.utsouthwestern.edu Message-ID: <20050726192310.49951.qmail@web31711.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello, Just a general question to help increase my knowledge: If I had an antibody that was conjugated to biotin -- say for example, a goat anti-mouse P-selectin (biotin) -- can I just take a rabbit anti-goat conjugated to AP and run IHC that way -- hopefully avoiding the ABC step and background OR does the biotin from the primary Ab block secondaries from coming in? Thanks. --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour ------------------------------ Message: 12 Date: Tue, 26 Jul 2005 12:45:44 -0700 From: "Glenn Krasinski" Subject: [Histonet] anti-mouse cleaved PARP for IHC... To: Message-ID: <6837C00A0CC6594197C2F0EEF256561805AADA@covxmail.covx.com> Content-Type: text/plain; charset="us-ascii" I am looking for a mouse-specific anti-cleaved PARP antibody that works in IHC. Cell Signaling has a rabbit polyclonal that work in IC and WB, but not in IHC. Many thanks. Glenn M. Krasinski Scientist, Biology CovX Research LLC 9381 Judicial Drive, Suite 200 San Diego, CA 92121 Ph: (858) 964-2053 Fax: (858) 964-2090 E-dress: gkrasinski@covx.com ------------------------------ Message: 13 Date: Tue, 26 Jul 2005 14:45:22 -0500 From: Amy Johnson Subject: [Histonet] CK7 control/ CK20 contol To: "Histonet (E-mail)" Message-ID: <016A64931E1ED511B12C0002B302608052334F@SERV001> Content-Type: text/plain; charset="iso-8859-1" What do most of you out there use for a CK7 control?......CK20 control?.... We currently are using an adenocarcinoma of the lung for our CK7 and an adenocarcinoma of the colon for the CK20. We have been having problems and need some help. Thanks Amy Johnson ------------------------------ Message: 14 Date: Tue, 26 Jul 2005 14:56:03 -0500 From: "Sebree Linda A." Subject: RE: [Histonet] CK7 control/ CK20 contol To: "Amy Johnson" , "Histonet \(E-mail\)" Message-ID: Content-Type: text/plain; charset="us-ascii" We mainly use liver for CK7 (stains the bile ducts) and colon adenocarcinoma for CK20. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Johnson Sent: Tuesday, July 26, 2005 2:45 PM To: Histonet (E-mail) Subject: [Histonet] CK7 control/ CK20 contol What do most of you out there use for a CK7 control?......CK20 control?.... We currently are using an adenocarcinoma of the lung for our CK7 and an adenocarcinoma of the colon for the CK20. We have been having problems and need some help. Thanks Amy Johnson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Tue, 26 Jul 2005 14:13:26 -0600 From: Gayle Callis Subject: [Histonet] Re: Serotec Rat Anti-mouse F4/80 : Biotin -- anyone know of good concentration? To: GT Hebert , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050726141256.01b7ea60@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Fro paraffin (FFPE) or frozen sections? At 01:19 PM 7/26/2005, you wrote: >Hello once again, > >I just received my Serotec Rat Anti-mouse F4/80 antibody and was wondering >if anyone can share with me the concentration they use -- closest to >optimized would be great! > >Thanks > >Gustave Hebert >Scientist II >CVMD >Wyeth - Cambridge > > >__________________________________________________ >Do You Yahoo!? >Tired of spam? Yahoo! Mail has the best spam protection around >http://mail.yahoo.com >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 16 Date: Tue, 26 Jul 2005 13:15:46 -0700 (PDT) From: GT Hebert Subject: [Histonet] Re: Serotec Rat Anti-mouse F4/80 : Biotin -- anyone know of good concentration? To: Gayle Callis Cc: Histonet@lists.utsouthwestern.edu Message-ID: <20050726201546.63399.qmail@web31711.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Sorry Gayle, This would be for FFPE. Thanks. Gustave Gayle Callis wrote: Fro paraffin (FFPE) or frozen sections? At 01:19 PM 7/26/2005, you wrote: >Hello once again, > >I just received my Serotec Rat Anti-mouse F4/80 antibody and was wondering >if anyone can share with me the concentration they use -- closest to >optimized would be great! > >Thanks > >Gustave Hebert >Scientist II >CVMD >Wyeth - Cambridge > > >__________________________________________________ >Do You Yahoo!? >Tired of spam? Yahoo! Mail has the best spam protection around >http://mail.yahoo.com >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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) --------------------------------- Start your day with Yahoo! - make it your home page ------------------------------ Message: 17 Date: Tue, 26 Jul 2005 14:22:05 -0600 From: Gayle Callis Subject: [Histonet] Re: Biotin conjugated Antibodies -- can we skip biotin?? To: GT Hebert , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050726141359.01b7ee00@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Yes, you do not need more biotin in the system. When you work with biotinylated primaries, all you need to do is use Strepavidin-HRP or Strepavidin-AP, followed by the appropriate chromogen. You eliminate the secondary conjugated to biotin completely, and also the ABC complex, and but be sure to use a good reliable Strepavidin-HRP or AP. Southern Biotechnology has these as does Biosource/TAGO. The primary conjugated to biotin provides all the biotin you will need. One of the perks of working with biotinylated primaries - it shortens the protocol! IF you use Vectors ABC complex, DON"T or you will experience loss of sensitivity and have poor staining. You will still need to do the appropriate blocking, i.e. avidin/biotion (OR Strepavidin/biotin block from Vector) normal serum, proper diluents with serums, etc. to avoid background problems. We do biotinylated primaries almost exclusively and with great success for murine CD markers IHC and IFA (using Molecular Probes Strepavidin-Alexa fluorophore conjugates.) If you want to discuss this more, contact me privately. At 01:23 PM 7/26/2005, you wrote: >Hello, > >Just a general question to help increase my knowledge: > >If I had an antibody that was conjugated to biotin -- say for example, a >goat anti-mouse P-selectin (biotin) -- can I just take a rabbit anti-goat >conjugated to AP and run IHC that way -- hopefully avoiding the ABC step >and background OR does the biotin from the primary Ab block secondaries >from coming in? > >Thanks. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 18 Date: Tue, 26 Jul 2005 16:25:10 -0400 From: "Woodward, Denise" Subject: [Histonet] Artifact pictures and cell block prep To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Hi all, I'm looking for pictures of sectioning, staining and processing artifacts. Anyone know where I might be able to find some on the web? Also, looking for a reference for making cell blocks from body fluids. Anyone know of a good source I can provide my histo students? Many thanks to you all, Denise Denise Long Woodward, MS, HT(ASCP), HTL, QIHC Dept. of Pathobiology and Veterinary Sciences University of Connecticut ------------------------------ Message: 19 Date: Tue, 26 Jul 2005 16:31:09 EDT From: Dndsomi@aol.com Subject: [Histonet] Productivity To: histonet@pathology.swmed.edu Message-ID: <82.2cd8fb46.3017f78d@aol.com> Content-Type: text/plain; charset="US-ASCII" We've just been asked to start turning in our histology procedures to be included in our labs daily productivity. How should we report our workload; # of cassettes, blocks, slides, special stains ? Any information will surely be appreciated. Thanks, DDietz, HT ------------------------------ Message: 20 Date: Tue, 26 Jul 2005 13:34:36 -0700 From: Christian Franci Subject: [Histonet] bunny Ab's To: histonet@lists.utsouthwestern.edu Message-ID: <5f251aaa45b280ca9b47daed7b9455c6@rigel.com> Content-Type: text/plain; charset=US-ASCII; format=flowed Hello Kids! Was just wondering if any of you have a good supplier of rabbit anti rat Ab's that are mouse absorbed (for real). Supposedly I purchased one but, sans primary antibody present, it makes my mouse tissue light up like a xmas tree.... talk about false positive! thanks in advance. Chris ------------------------------ Message: 21 Date: Tue, 26 Jul 2005 16:34:48 -0400 From: Geoff McAuliffe Subject: Re: [Histonet] Artifact pictures and cell block prep To: "Woodward, Denise" Cc: histonet@lists.utsouthwestern.edu Message-ID: <42E69E68.1030302@umdnj.edu> Content-Type: text/plain; format=flowed; charset=us-ascii The AFIP manual (3rd edition, 1968, I don't know about other editions) has photos of artifacts after the last numbered chapter. For electron microscopy, there is a book called "An Atlas of Artifacts", I don't remember the author off the top of my head. I can't offer any suggestions for your body fluid question. Geoff Woodward, Denise wrote: >Hi all, > >I'm looking for pictures of sectioning, staining and processing >artifacts. Anyone know where I might be able to find some on the web? > > >Also, looking for a reference for making cell blocks from body fluids. >Anyone know of a good source I can provide my histo students? > >Many thanks to you all, >Denise > >Denise Long Woodward, MS, HT(ASCP), HTL, QIHC >Dept. of Pathobiology and Veterinary Sciences >University of Connecticut > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** ------------------------------ Message: 22 Date: Tue, 26 Jul 2005 16:40:58 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] Artifact pictures and cell block prep To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717591@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="ISO-8859-1" You might be interested in one of the following, if you can find a copy: Atlas of Microscopic Artifacts and Foreign Materials by Yeh, Brooks & Pietra An Atlas of Artifacts Encountered in the Preparation of Microscopic Tissue Sections by Thompson & Luna You might find them through www.bookfinder.com We do a fair amount of cell block work in our lab, and have several protocols for different kinds of fluids and cell suspensions. Is there anything in particular you are looking for? Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Woodward, Denise > Sent: Tuesday, July 26, 2005 1:25 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Artifact pictures and cell block prep > > Hi all, > > I'm looking for pictures of sectioning, staining and processing > artifacts. Anyone know where I might be able to find some on the web? > > > Also, looking for a reference for making cell blocks from body fluids. > Anyone know of a good source I can provide my histo students? > > Many thanks to you all, > Denise > > Denise Long Woodward, MS, HT(ASCP), HTL, QIHC > Dept. of Pathobiology and Veterinary Sciences > University of Connecticut > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 23 Date: Tue, 26 Jul 2005 16:52:16 -0400 From: "Woodward, Denise" Subject: RE: [Histonet] Artifact pictures and cell block prep To: "Geoff McAuliffe" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Sorry, I didn't specify, but I'm looking for artifact pictures ON-LINE, so my students can access them from home. Thanks, DLW Denise Long Woodward, MS, HT(ASCP), HTL, QIHC Dept. of Pathobiology and Veterinary Sciences University of Connecticut -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Tuesday, July 26, 2005 4:35 PM To: Woodward, Denise Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artifact pictures and cell block prep The AFIP manual (3rd edition, 1968, I don't know about other editions) has photos of artifacts after the last numbered chapter. For electron microscopy, there is a book called "An Atlas of Artifacts", I don't remember the author off the top of my head. I can't offer any suggestions for your body fluid question. Geoff Woodward, Denise wrote: >Hi all, > >I'm looking for pictures of sectioning, staining and processing >artifacts. Anyone know where I might be able to find some on the web? > > >Also, looking for a reference for making cell blocks from body fluids. >Anyone know of a good source I can provide my histo students? > >Many thanks to you all, >Denise > >Denise Long Woodward, MS, HT(ASCP), HTL, QIHC >Dept. of Pathobiology and Veterinary Sciences >University of Connecticut > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** ------------------------------ Message: 24 Date: Tue, 26 Jul 2005 14:05:17 -0700 From: "Linke_Noelle" Subject: RE: [Histonet] bunny Ab's To: "Christian Franci" , Message-ID: Content-Type: text/plain; charset="us-ascii" Hi Chris, Try Jackson, they usually have everything in the barnyard! Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Christian Franci Sent: Tuesday, July 26, 2005 1:35 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bunny Ab's Hello Kids! Was just wondering if any of you have a good supplier of rabbit anti rat Ab's that are mouse absorbed (for real). Supposedly I purchased one but, sans primary antibody present, it makes my mouse tissue light up like a xmas tree.... talk about false positive! thanks in advance. Chris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 25 Date: Tue, 26 Jul 2005 16:46:40 -0500 From: "Henry, Charlene" Subject: [Histonet] un-subscribe To: Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2099A199A@SJMEMXMB02.stjude.sjcrh.local> Content-Type: text/plain; charset="US-ASCII" Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 20, Issue 36 **************************************** Privileged/Confidential information may be contained in this message. 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Opinions, conclusions and other information in this message that do not relate to the official business of the firm shall be understood as neither given nor endorsed by it. From gu.lang <@t> gmx.at Wed Jul 27 14:58:52 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] endogen biotin in prostata? Message-ID: Hi, Since a few weeks our 34betaE12 shows granular spots in the cytoplasma of some cells in the prostata-slide. They look really sprinkled, but not in all. Could that be endogen biotin? Or appears biotin more homogen? Please help an unknowing histotech. Gudrun Lang From Eric <@t> ategra.com Wed Jul 27 13:08:45 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] HistoTech Job Openings Message-ID: Histonetters - My name is Eric Dye and I am the Histo Tech recruiter here at Ategra. I am presently on a search for my best client companies Nationwide who are seeking to hire fulltime Histo Techs. Right now my hottest job openings are in Denver, Colorado who is seeking a Histotech Manager and Northern Oregon also seeking a Histotech Manager. I also have bench histo tech jobs available in: California,Michigan, Pennsylvania, Illinois, New Hampshire, Massachusetts, West Virginia, Georgia, Colorado and Florida. I have a opening for a pathologist's Assistant in Illinois. My openings are permanent fulltime positions with excellent benefits. If you are interested in any of these positions - please CALL ME at 800 466 9919 ext 223 Thank You !! Eric - 800 466 9919 ext 223 PS: If you are interested any other state, also please call me at 800 466 9919 ext 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------- From katri <@t> cogeco.ca Wed Jul 27 22:40:22 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] endogen biotin in prostata? References: <20050727200606.72F876A8@fep5.cogeco.net> Message-ID: <004601c59326$165a63b0$6a9a9618@Katri> Gudrun, Do you use HIER to antigen retrieve? I have seen this type of endogenous biotin in heat retrieved tissue. Do a biotin block and see, if it's gone. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Gudrun Lang" To: "Histonetliste (Histonetliste)" Sent: Wednesday, July 27, 2005 3:58 PM Subject: [Histonet] endogen biotin in prostata? > Hi, > > Since a few weeks our 34betaE12 shows granular spots in the cytoplasma of > some cells in the prostata-slide. They look really sprinkled, but not in > all. Could that be endogen biotin? > > Or appears biotin more homogen? Please help an unknowing histotech. > > > > Gudrun Lang > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Wed Jul 27 23:16:05 2005 From: anh2006 <@t> med.cornell.edu (anh2006@med.cornell.edu) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] endogen biotin in prostata? In-Reply-To: <004601c59326$165a63b0$6a9a9618@Katri> References: <20050727200606.72F876A8@fep5.cogeco.net> <004601c59326$165a63b0$6a9a9618@Katri> Message-ID: <4509.66.108.127.19.1122524165.squirrel@webmail> You can just add your strep-HRP or ABC step to a section (without primary or secondary) and if you get signal it is surely from the biotin endogenous to the tissue. It is a good control to run in any given experiment. Biotin is tricky as it often appears in very "specific" patterns and is often misinterpreted as real staining. I also agree HIER can dramatically increase biotin signal. > ----- Original Message ----- > From: "Gudrun Lang" > To: "Histonetliste (Histonetliste)" > Sent: Wednesday, July 27, 2005 3:58 PM > Subject: [Histonet] endogen biotin in prostata? > > >> Hi, >> >> Since a few weeks our 34betaE12 shows granular spots in the cytoplasma >> of >> some cells in the prostata-slide. They look really sprinkled, but not in >> all. Could that be endogen biotin? >> >> Or appears biotin more homogen? Please help an unknowing histotech. >> >> >> >> Gudrun Lang >> From Julie.Sanders <@t> med.va.gov Thu Jul 28 06:46:21 2005 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] "waviness" Message-ID: <457381D92B01BD44B21CF37CC02EBDFD029276DD@vhacinexc5.v10.med.va.gov> One of our pathologists is complaining about having to focus up and down too much while reading slides. I have tried many things to resolve the problem but nothing is making him happy. I have changed the way we pick up from water bath to be sure slides are perpendicular (straight up and down) when picking up sections, we use only plus slides, I've asked that slides not be placed on the warming platform after picking up. Does anyone have any ideas about how I can fix this problem? Julie Julie Sanders Supervisor, Anatomic Pathology Cincinnati VA From ian.montgomery <@t> bio.gla.ac.uk Thu Jul 28 07:21:57 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:25:23 2005 Subject: Fwd: [Histonet] "waviness" Message-ID: <6.2.3.4.2.20050728131737.03657150@udcf.gla.ac.uk> Julie, Fix the problem, easy, plug his dangly bits into the electricity supply and switch on. A microscopist who doesn't use the focus controls, I'm shocked and alarmed. Ian. >One of our pathologists is complaining about having to focus up and down too >much while reading slides. I have tried many things to resolve the problem >but nothing is making him happy. I have changed the way we pick up from >water bath to be sure slides are perpendicular (straight up and down) when >picking up sections, we use only plus slides, I've asked that slides not be >placed on the warming platform after picking up. Does anyone have any ideas >about how I can fix this problem? > >Julie > >Julie Sanders >Supervisor, Anatomic Pathology >Cincinnati VA > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From Terry.Marshall <@t> rothgen.nhs.uk Thu Jul 28 08:19:10 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] "waviness" Message-ID: Not as shocked as he would be were this strategy to be followed! 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: Ian Montgomery [mailto:ian.montgomery@bio.gla.ac.uk] Sent: 28 July 2005 13:22 To: histonet@lists.utsouthwestern.edu Subject: Fwd: [Histonet] "waviness" Julie, Fix the problem, easy, plug his dangly bits into the electricity supply and switch on. A microscopist who doesn't use the focus controls, I'm shocked and alarmed. Ian. >One of our pathologists is complaining about having to focus up and down too >much while reading slides. I have tried many things to resolve the problem >but nothing is making him happy. I have changed the way we pick up from >water bath to be sure slides are perpendicular (straight up and down) when >picking up sections, we use only plus slides, I've asked that slides not be >placed on the warming platform after picking up. Does anyone have any ideas >about how I can fix this problem? > >Julie > >Julie Sanders >Supervisor, Anatomic Pathology >Cincinnati VA > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From louise.renton <@t> gmail.com Thu Jul 28 08:19:48 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] "waviness" In-Reply-To: <457381D92B01BD44B21CF37CC02EBDFD029276DD@vhacinexc5.v10.med.va.gov> References: <457381D92B01BD44B21CF37CC02EBDFD029276DD@vhacinexc5.v10.med.va.gov> Message-ID: Are you using a tape coverslipping system? If you are, this might be the cause of all your woes. Try recoverslippig the offending slides with glass and see if the problem is solved Best regards On 7/28/05, Julie.Sanders@med.va.gov wrote: > > One of our pathologists is complaining about having to focus up and down too > much while reading slides. I have tried many things to resolve the problem > but nothing is making him happy. I have changed the way we pick up from > water bath to be sure slides are perpendicular (straight up and down) when > picking up sections, we use only plus slides, I've asked that slides not be > placed on the warming platform after picking up. Does anyone have any ideas > about how I can fix this problem? > > Julie > > Julie Sanders > Supervisor, Anatomic Pathology > Cincinnati VA > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From donald <@t> labcareer.com Thu Jul 28 08:51:30 2005 From: donald <@t> labcareer.com (Donald Knowles) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Histology Placement Professional Message-ID: <26A7728EF768DA478F45954252293C202E4A0F@matrix.HCCI.local> Dear Histology Professionals: I specialize in recruiting laboratory professionals throughout the United States and I come to you to ask for your assistance. I am currently on a search for Certified Medical Technologists throughout the country and I was hoping you might be able to recommend a professional that would be interested in hearing about some great opportunities. If you know someone and pass their information on to me and that person is placed in a position I will be more then happy to give you a referral bonus. We currently have positions in California, Florida, Georgia, Iowa, Illinois, Massachusetts, Maine, New Hampshire, Pennsylvania, Texas and Washington State. These positions are at all levels from entry level to management. I am also here to assist in career planning and would be more then happy to talk to anyone about their future employment plans. I have over ten years experience in placing individuals in healthcare settings that include Pharmacy's, Radiological and Research and Clinical Laboratory's. I also have links to many other recruiters and would be happy to refer people over to other hard-working honest Placements professionals. I look forward to answering any questions you might throw my way! I appreciate any help you may be able to provide and I look forward to speaking with you! Don Knowles Healthcare Connections Inc. 866-346-8522 ext 311 www.labcareer.com From Emily.Wiesner <@t> medecine.unige.ch Thu Jul 28 09:38:31 2005 From: Emily.Wiesner <@t> medecine.unige.ch (Emily Jane Wiesner-Camm) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] Golgi Staining Message-ID: <42E8EDE7.1030009@medecine.unige.ch> Hi All, I was wondering whether anyone could let me know of a reliable Golgi staining method. I would like to stain perfused rat brain sections that have been cut on the cryostat (10um sections). Thanks in advance. Emily -- Dr. Emily J. Camm Fondation pour Recherches M?dicales Avenue de la Roseraie 64 1205 Gen?ve Phone: +41 22 382 4569 Fax: + 41 22 347 5979 From cbass <@t> bidmc.harvard.edu Thu Jul 28 09:52:33 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Sep 16 15:25:23 2005 Subject: [Histonet] HER2 elisa/protein Message-ID: Hey guys, I am attempting to quantify the amount of HER2 in some cell lysates. I have gotten a lot of advice about HER2 on this forum so I thought someone might be able to help. Is there a cheap HER2 ELISA that someone could recommend? I have found a couple but $700 for this small experiment is too much. I was also thinking about doing a semi- quantitative western with known amounts of HER2, so if someone could recommend a source for HER2 protein that I could use for a standard that would be great. Any advice would be appreciated. I don't know if hospitals routinely do HER2 measurements. If it is I thought perhaps I could pay to have my samples quantified at our medical center. Thanks, Caroline Bass From cornettl <@t> hotmail.com Thu Jul 28 10:18:52 2005 From: cornettl <@t> hotmail.com (Lorraine Cornett) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] "waviness" In-Reply-To: <457381D92B01BD44B21CF37CC02EBDFD029276DD@vhacinexc5.v10.med.va.gov> Message-ID: Julie, we have had this problem in the past as well. One thing to look at, especially with using charged slides, is to make absolutely sure that the slides are DRY, no water at all, before staining. Are you using a auto stainer or hand staining? Somtimes we noticed that water was being trapped under the paraffin (had a bad lot of charged or plus slides) and would usually result in the pathologist complaining about waviness. Make sure you water bath temps are warm enough to flatten the sections before picking up as well. Keep me posted as to what resolves it for you. You never know, this monster may show back up in my lab as well. Lorraine Cornett, HT (ASCP) Highlands Pathology Blue Ridge Division, Kingsport, TN 423 224-5793 fax 423 224-5349 >From: Julie.Sanders@med.va.gov >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] "waviness" Date: Thu, 28 Jul 2005 06:46:21 -0500 > > >One of our pathologists is complaining about having to focus up and down >too >much while reading slides. I have tried many things to resolve the problem >but nothing is making him happy. I have changed the way we pick up from >water bath to be sure slides are perpendicular (straight up and down) when >picking up sections, we use only plus slides, I've asked that slides not be >placed on the warming platform after picking up. Does anyone have any >ideas >about how I can fix this problem? > >Julie > >Julie Sanders >Supervisor, Anatomic Pathology >Cincinnati VA > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From liz <@t> premierlab.com Thu Jul 28 10:21:28 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] anti-talin antibody Message-ID: <000501c59388$09446600$a7d48a80@AMY> Hello everyone I have been working on an Immunohistochemical protocol for talin on human skin samples. I currently have the chemicon antibody. I have tried it on both frozen and paraffin sections, but I'm not having much success. I get minimal staining with retrieval in the paraffin sections. When I tried the frozen sections I got no staining at all. Has anyone used this antibody before? Any help would be appreciated, different antibody source, etc. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From gcallis <@t> montana.edu Thu Jul 28 11:32:34 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Re: "waviness" In-Reply-To: <457381D92B01BD44B21CF37CC02EBDFD029276DD@vhacinexc5.v10.me d.va.gov> References: <457381D92B01BD44B21CF37CC02EBDFD029276DD@vhacinexc5.v10.med.va.gov> Message-ID: <6.0.0.22.1.20050728101353.01b84e00@gemini.msu.montana.edu> Julie, It may be more than how you pick up from the water bath. Make sure your disposable blades are sharp, of high quality and maybe change more often to a sharp edge. Make sure the knife angle is correct for the blade you are using. The sections should come off the block onto blade without compression. Check all levers, microtome knife holder adjustments have to be tight. Check you water bath temperature for the paraffin you use, the sections need to flatten out without exploding, and you are picking up nicely - do this quickly so section hits slide surface evenly - Marcia from Newcomer Supply gave excellent hints on getting sections to the surface so it STICKS onto plus charge. If your waterbath is too cool, the section will not be flat at times - a problem I have when I am sectioning too fast and don't take the time to wait for sections to flatten - I then up the temp a few degrees to accomodate speed coupled with impatience. Be careful doing the latter, you don't want to have sections come apart. Do not put any adhesives in the waterbath, it should be distilled water (if you use tap and water is hard, i.e. mineralized, that will afftect how sections sticks) adhesives coat the plus charge and negate the plus effect. This is all given on the package insert inside Plus Charge slide boxes. When you pick up a section onto Plus slides, drain the section vertically so the water drains OFF slide - down, away and out from under section. You are correct in not going to warming platform immediately after picking up section as water will pool under the section, creates uneven adherence. Another thing, additives in paraffins are heavy and can settle to bottom of dispenser when paraffin melts. Stir the paraffin before embedding to redistribute these additives. You want the paraffin well mixed to have benefits of the additives for good sectioning. Good luck At 05:46 AM 7/28/2005, you wrote: >One of our pathologists is complaining about having to focus up and down too >much while reading slides. I have tried many things to resolve the problem >but nothing is making him happy. I have changed the way we pick up from >water bath to be sure slides are perpendicular (straight up and down) when >picking up sections, we use only plus slides, I've asked that slides not be >placed on the warming platform after picking up. Does anyone have any ideas >about how I can fix this problem? > >Julie > >Julie Sanders >Supervisor, Anatomic Pathology >Cincinnati VA > >_______________________________________________ >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 Frederick.Fifield <@t> sunhealth.org Thu Jul 28 12:00:16 2005 From: Frederick.Fifield <@t> sunhealth.org (Frederick.Fifield@sunhealth.org) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Fred on APL. Message-ID: I will be out of the office starting 07/28/2005 and will not return until 08/04/2005. I will be out of the lab beginning on 07/28/05. I will respond to your message when I return on Thursday, 08/4/05. From JosefaNava <@t> texashealth.org Thu Jul 28 12:57:34 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] VEGF ANTIBODY Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944E1@phdex03.txhealth.org> For Ventana users can you please tell me what company carries a good VEGF antibody . Thank you so much. Josie The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From JosefaNava <@t> texashealth.org Thu Jul 28 13:23:27 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] EGFR Message-ID: <2C515C1049EAF5459EFD8C9B929078A41944E3@phdex03.txhealth.org> Hello Everyone, Can someone tell me what is the best clone for EGFR that works best on Ventana. Is there any other company aside from Dako that is FDA approved for EGFR ? I appreciate any information. Thanks.. Josie The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From JMacDonald <@t> mtsac.edu Thu Jul 28 14:49:03 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] H. Pylori Message-ID: Somebody asked for methods for staining H. pylori. I just came across a 2003 publication by Poly Scientific that has seven staining methods for H. pylori. Included are the procedures and references. Perhaps you could get this from Poly Sci. Jennifer MacDonald From sjchtascp <@t> yahoo.com Thu Jul 28 15:19:03 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Bone processing Message-ID: <20050728201904.59382.qmail@web90206.mail.scd.yahoo.com> New to research, just received the "Animal Processing Manual". I'm about to process mouse joints and I noticed a much longer time especially in the infiltration paraffins. Along these lines I also noticed processing for mouse/rat tissue calls for only RT processing and no vacuum/pressure.. I have a clinical background so some or allot of this is a "tad" new. Anyone up to explaining the big picture difference between human tissue verses animal tissue processing. Steve --------------------------------- Start your day with Yahoo! - make it your home page From liz <@t> premierlab.com Thu Jul 28 15:34:17 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Bone processing In-Reply-To: <20050728201904.59382.qmail@web90206.mail.scd.yahoo.com> Message-ID: <000001c593b3$ba1aba30$a7d48a80@AMY> Steve We process mouse joints all of time and use vacuum/pressure during the processing cycle. The cycle we use is long approximately 14 + hours. 50% alcohol - 60 minutes 70% alcohol - 60 minutes 80% alcohol - 60 minutes 95% alcohol - 60 minutes 100% alcohol - 60 minutes 100% alcohol - 60 minutes 100% alcohol - 60 minutes Xylene - 60 minutes Xylene - 60 minute Xylene - 60 minutes Paraffin - 60 minutes Paraffin - 60 minutes Paraffin - 60 minutes Paraffin - 60 minutes Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Thursday, July 28, 2005 1:19 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone processing New to research, just received the "Animal Processing Manual". I'm about to process mouse joints and I noticed a much longer time especially in the infiltration paraffins. Along these lines I also noticed processing for mouse/rat tissue calls for only RT processing and no vacuum/pressure.. I have a clinical background so some or allot of this is a "tad" new. Anyone up to explaining the big picture difference between human tissue verses animal tissue processing. Steve --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Jul 28 16:14:36 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Bone processing In-Reply-To: <20050728201904.59382.qmail@web90206.mail.scd.yahoo.com> References: <20050728201904.59382.qmail@web90206.mail.scd.yahoo.com> Message-ID: <6.0.0.22.1.20050728150001.01b1b5e0@gemini.msu.montana.edu> Steven, Those protocols are from people who did their animal tissues in a certain way, and not everyone does it just exactly like the protocol submitted for the Animal Processing Manual - the protocols are not written in stone, but are good guidelines for successful animal tissue work. As for our mouse bone work, we use a Sakura Finetek VIP, and extend processing for mouse joints (denser due to the collagen matrix of decalcified bone). We do ambient i.e RT temperatures for dehydration and clearing and use vacuum and pressure throughout. Adding temperatures to alcohols, clearing agents sometimes adds to hardness of tissue at sectioning - animal tissues, in particular the tiny mouse soft tissues. In general, animal tissues are much leaner, less fatty than human tissue and overdehydration tends to dry out rodent tissue more. This species is not the only one that suffers from lean i.e birds, reptiles, rabbit, and many others. We tend to do custom processing for rodent work, and have special schedules for mouse brain versus hamster brain versus rat brain versus lung and other soft tissues. Larger animals processing can be more like human tissue processing i.e. cow, horse, dog, cat, but sometimes these can be dry also. What you want to do is remove the free water from the tissue spaces and not the bound water found on the proteins (that is what makes mouse spleen hard little nuggets sometimes referred to as overprocessing). If your tissue seem dry at sectioning - you can soak a faced block on ice with water on top and reevaluate if you need to cut down on total time of processing, in small increments of time however. Good luck At 02:19 PM 7/28/2005, you wrote: >New to research, just received the "Animal Processing Manual". I'm about >to process mouse joints and I noticed a much longer time especially in the >infiltration paraffins. > >Along these lines I also noticed processing for mouse/rat tissue calls for >only RT processing and no vacuum/pressure.. > >I have a clinical background so some or allot of this is a "tad" >new. Anyone up to explaining the big picture difference between human >tissue verses animal tissue processing. > >Steve > > > >--------------------------------- > Start your day with Yahoo! - make it your home page >_______________________________________________ >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 JMacDonald <@t> mtsac.edu Thu Jul 28 18:51:17 2005 From: JMacDonald <@t> mtsac.edu (JMacDonald@mtsac.edu) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] autostainers Message-ID: I have a colleague that is looking for a autostainer (H&E) that is less than 3 feet in length. As usual in a histology lab, space is very limited. Vendors?? Jennifer MacDonald From HornHV <@t> archildrens.org Fri Jul 29 08:05:18 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] autostainers Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDDBB@EMAIL.archildrens.org> We have a Shandon Gemini that is less than 28 inches in wide. But it does have a computer that would take up another 16 inches. We like our stainer very much. Have her/him contact the ThermoShandon folks. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JMacDonald@mtsac.edu Sent: Thursday, July 28, 2005 6:51 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] autostainers I have a colleague that is looking for a autostainer (H&E) that is less than 3 feet in length. As usual in a histology lab, space is very limited. Vendors?? Jennifer MacDonald _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From wecare <@t> qualityhistology.com Fri Jul 29 09:00:02 2005 From: wecare <@t> qualityhistology.com (wecare@qualityhistology.com) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] autostainers References: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDDBB@EMAIL.archildrens.org> Message-ID: <017801c59445$dd0258e0$0300a8c0@internetconnect.net> Dear Ms. MacDonald, RUSHABH Instruments makes an H & E autostainer that is only 34" wide including display. It has 30 positions, 5 rinse stations, 2 ovens and provides multiple programs with multiple block. Please contact me if I can provide any additional information. Preyas Shah 215-491-0081 ----- Original Message ----- From: "Horn, Hazel V" To: ; Sent: Friday, July 29, 2005 9:05 AM Subject: RE: [Histonet] autostainers We have a Shandon Gemini that is less than 28 inches in wide. But it does have a computer that would take up another 16 inches. We like our stainer very much. Have her/him contact the ThermoShandon folks. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JMacDonald@mtsac.edu Sent: Thursday, July 28, 2005 6:51 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] autostainers I have a colleague that is looking for a autostainer (H&E) that is less than 3 feet in length. As usual in a histology lab, space is very limited. Vendors?? Jennifer MacDonald _______________________________________________ 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. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From relia1 <@t> earthlink.net Fri Jul 29 09:06:53 2005 From: relia1 <@t> earthlink.net (Pam Barker) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] RELIA Histology Opportunity Update 7/29/05 - From Pam Barker Message-ID: Hello Histonetters, I hope you are having a great summer. I just wanted to drop you a line and tell you about my current histology openings. All the positions I represent are permanent positions with great companies who offer excellent compensation, benefits and relocation/sign on bonuses. Wherever you want to go, wherever you want to stay if you are looking for a new opportunity I can help. I offer over 20 years of recruiting experience and a recruiting practice solely dedicated to permanent placement in the Histology profession. In addition to representing you to the positions you are interested in with my clients I offer to you FREE of Charge: ? Assistance with updating or creating your resume ? Tips on interviewing ? Encouragement and assistance during the course of your job search ? Complete Confidentiality ( I will only represent you to jobs you tell me you are interested in looking into) Here is a list of my current openings: HISTOLOGY MANAGEMENT: 1. Anatomic Pathology Manager ? South Carolina 2. Anatomic Pathology Manager ? Boston, MA 3. Histology Supervisor ? Boston, MA 4. Histology Supervisor ? Pennsylvania 5. Histology Supervisor ? South Florida HISTOTECHNICIAN/TECHNOLOGIST: 1. Histo Tech (clinical setting) ? Northern, CA 2. Histo Tech (research setting) ? Northern, CA 3. Histo Tech (clinical setting) ? Central Florida 4. Histo Tech (clinical setting) - South Florida 5. Histo Tech (clinical setting) ? Boston, MA 6. MOHS/Histo Tech ? South Florida 7. Histo Tech (clinical setting) - Minnesota 8. Histo Tech (clinical setting) ? Alaska 9. Histo Tech (Night shift) - Colorado If you or any of your friends would like more information on any of these positions. Or if you would like to discuss opportunities in other areas or future job searches please contact me. I would be more than happy to assist you. You can reach me at 866-607-3542 or relia1@earthlink.net I work with facilities nationwide and I will keep your resume confidential. I will only represent you to jobs you tell me you are interested in looking into. Remember It never hurts to keep an eye open even if you are happy in your present job. Thank you, Pam ? 866-607-3542 (866-60RELIA) Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net From vsailes <@t> nd.edu Fri Jul 29 09:54:30 2005 From: vsailes <@t> nd.edu (vsailes@nd.edu) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Looking for Antibody Message-ID: <1122648870.42ea4326a615e@webmail.nd.edu> Hi I'm looking to stain platelets in formalin fixed paraffin embedded mouse tissues. I wanted to know if anyone out there has used CD41 or CD9 antibodies? If so did you get good results and what company did you use? Any information would be most appreciated. Thanks, Valerie From juan.gutierrez <@t> christushealth.org Fri Jul 29 10:36:57 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Looking for Antibody Message-ID: We use Dakocytomation's CD41 at 1:50 for 1hr at room temp. Pre-treat with proteinase k for 10 min. We don't use CD9. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of vsailes@nd.edu Sent: Friday, July 29, 2005 9:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Looking for Antibody Hi I'm looking to stain platelets in formalin fixed paraffin embedded mouse tissues. I wanted to know if anyone out there has used CD41 or CD9 antibodies? If so did you get good results and what company did you use? Any information would be most appreciated. Thanks, Valerie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sbreeden <@t> nmda.nmsu.edu Fri Jul 29 11:26:55 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] New Lab Design Message-ID: A new histology lab (in a new building) is in the works (yipee!!). I have some definite ideas about what I'll need in my new digs, but I need more opinions! Within the next couple weeks, I will be asked by the architects for my input. This is a State veterinary lab where I do "surgicals", necropsies, standard special stains, with a rapidly expanding IHC workload. My total workload was roughly 10,000 blocks last year and I expect it to grow by 5% each year. I expect to have roughly 500 sq ft to work with in this new space. I work alone for three DVM pathologists. What would you consider absolutely essential in a new histology lab? What would you do differently if you'd had a chance? What kind of: bench space and type of surface, ventilation, lighting, hoods, haz mat storage, supply storage, windows? I only get once chance at this and I want to make it count...I have 8 more years until retirement and I'd rather not kick myself for missing something obvious. Please reply directly to me. Thank you in advance!! I rely on 'netters for great information. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From vd38 <@t> georgetown.edu Fri Jul 29 12:06:56 2005 From: vd38 <@t> georgetown.edu (Vernon Dailey) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Biotinylating Primary Antibodies Message-ID: <75ae06e90c.6e90c75ae0@imap.georgetown.edu> Hi Histonet, I am working on a project in which I need to use a VEGF monoclonal primary on FFPE mouse tissue. I plan on using sc-7269 from Santa Cruz. I am trying to avoid mouse on mouse issues and would like to biotinylate the primary. Are there and good commercial kits or protocols available for this procedure? Or, does anyone have another method they can recommend to get rid of the mouse on mouse artifacts. I have tried vector's mouse on mouse kit but would rather try something else. Thanks in advance. -Vernon Vernon Keith Dailey IHC Laboratory Manager Histopathology and Tissue Shared Resource Lombardi Comprehensive Cancer Center Georgetown University Phone: 202-687-0047 From altaytamer <@t> hotmail.com Fri Jul 29 13:44:04 2005 From: altaytamer <@t> hotmail.com (tamer altay) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] gfp-mice Message-ID: Hi, I have Tie2-GFP labeled mice. I try to image the cerebral vasculature through a glass cranial window mounted on the surface of the brain after I remove the bone. A week after the window installment, I wanted to image the animal under a fluorescein microscope, but I could not see anything. The vessels were supposed to be emitting the green fluorescein light. I am wondering if the light emitted from GFP source, under a glass cranial window where the vessels are exposed to daylight all the time, bleaches despite the continuos GFP production. I would appreciate any information or idea you guys come up with to help me solve this issue. Thanks a lot!! Tamer _________________________________________________________________ Her tür saldiri ve virüslerden korunmanin en güvenli sekli - MSN PC Koruma'da! [1]Burayi tiklayin! References 1. http://g.msn.com/8HMAENTR/2746??PS=47575 From sjchtascp <@t> yahoo.com Fri Jul 29 14:00:45 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Cryojane/gentle jane Message-ID: <20050729190045.45048.qmail@web90202.mail.scd.yahoo.com> One of the scientist I work with is looking into purchasing a cryojane. Does anyone have any experiance with this piece of equipment. Also the gentle jane for snap frozens. Steve --------------------------------- Start your day with Yahoo! - make it your home page From leahcox27 <@t> yahoo.com Fri Jul 29 14:06:05 2005 From: leahcox27 <@t> yahoo.com (Leah Cox) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Position in Seattle Message-ID: <20050729190605.62143.qmail@web50205.mail.yahoo.com> Medical Laboratory Associates in Seattle WA is currently looking for a histologist. We are a small stable company with great employee's and advancement opportunities. Hours are 6 AM to 2PM. ASCP certified or registry eligible. Relocation advances negotiable. Contact info: Ken Pierce 206-623-3814 kpierce@cancer-test.com __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From slappycraw <@t> yahoo.com Fri Jul 29 14:06:41 2005 From: slappycraw <@t> yahoo.com (Larry Woody) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] collagen precursors Message-ID: <20050729190641.62861.qmail@web52610.mail.yahoo.com> Hi all: Does anyone out there know of any special stains or immuno stains used for collagen precursors? Thanks Larry __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From rocan <@t> mac.com Fri Jul 29 14:20:20 2005 From: rocan <@t> mac.com (Rocan) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Cryojane/gentle jane In-Reply-To: <20050729190045.45048.qmail@web90202.mail.scd.yahoo.com> References: <20050729190045.45048.qmail@web90202.mail.scd.yahoo.com> Message-ID: <0DD63920-160E-49FD-9272-DAEEB8AFE68D@mac.com> Steven, We use both and have had a very good experience with the items. The freezing thing is excellent for our purposes and works well. Their UV curable mounting media is also quite nice in our hands. The Cryojane is something you need to develop a good technique since sometimes the tape can retain some of the section. It is a little tricky. In my opinion, the Cryojane is not for routine use but for specific applications. We do mouse skin wounds and it has been invaluable for us. However, when we cut other organs we do not need it or use it. The supplies are expensive ($1 per slide) and you are always dependent on them for the supplies. Also, very importantly, their customer service can be on the "defensive" side, so when you have a problem with their products do not expect them to think that the customer is always right. I hope this information helps you. ----- Dr.Rocio Sierra-Honigmann Director Engineered Wound Repair Laboratory Cedars Sinai Research Institute Davis 1091 310-423-1882 Honigmannr@cshs.org On Jul 29, 2005, at 12:00 PM, Steven Coakley wrote: > One of the scientist I work with is looking into purchasing a > cryojane. Does anyone have any experiance with this piece of > equipment. Also the gentle jane for snap frozens. > > Steve > > > --------------------------------- > Start your day with Yahoo! - make it your home page > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From la.sebree <@t> hosp.wisc.edu Fri Jul 29 14:23:44 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Cryojane/gentle jane Message-ID: I used it for sectioning frozen rabbit myocardium where it was important to get nearly identical adjacent sections. I worked very well for this purpose. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Friday, July 29, 2005 2:01 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Cryojane/gentle jane One of the scientist I work with is looking into purchasing a cryojane. Does anyone have any experiance with this piece of equipment. Also the gentle jane for snap frozens. Steve --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ynwang <@t> u.washington.edu Fri Jul 29 15:08:04 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] collagen precursors In-Reply-To: <20050729190641.62861.qmail@web52610.mail.yahoo.com> References: <20050729190641.62861.qmail@web52610.mail.yahoo.com> Message-ID: Larry, There are some abs against type I and type III procollagen available. There is a company called DiaSorin that has a radioimmunoassay kit for these too (human). They also may have the straight antibodies. I've never tied any of them but there are a few companies that have these abs (mainly for human). I believe Chemicon, mdbiosciences and novotec have them, to name a few. I hope this is of some help. Yak-Nam > Hi all: Does anyone out there know of any special stains or immuno > stains used for collagen precursors? Thanks Larry > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From TJJ <@t> Stowers-Institute.org Fri Jul 29 16:35:16 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Embedding mouse tibia + posterior 1/2 femur Message-ID: Has anybody done PMMA embedding/sectioning of mouse tibia and posterior/distal 1/2 of femur? If so, what mold/vessel did you use for embedment? Must one use glass for PMMA or are there other suitable molds out there that will work? Thanks as always! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From gcallis <@t> montana.edu Fri Jul 29 17:22:06 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Molds for PMMA Re: Embedding mouse tibia + posterior 1/2 femur In-Reply-To: <200507292144.j6TLivJU027411@mail4.msu.montana.edu> References: <200507292144.j6TLivJU027411@mail4.msu.montana.edu> Message-ID: <6.0.0.22.1.20050729155619.01b3a098@gemini.msu.montana.edu> Teri, To avoid breaking glass and having glass shards around, use a polypropylene container with screw on lid to avoid any evaporation of the monomer. If evaporation occurs, it changes the consistency of the plastic mixture and it ends up funky, rubbery, and hard to cut, Peel a way molds became a no no. Been there, done that - not good! Polypropylene scintillation (sp?) vials will work, although you can't see into them - but cheap. After polymerization and curing, simply cut off the end, or grind it away, and push the block out so it can be shaped to fit in block clamp for microtoming or thin 100 um slabs for thick sections. A prepolymerized layer helps as it raises the bone above an uneven flat bottom, and the layer does not interfer with polymerization in fact it interfaces perfectly with embedding mixture, creates a very clear block. These vials have a lip on them so you have to push away from the lip or cut it off too. A mini hobby band saw works well for this purpose. We used small plastic, almost clear specimen containers that have wonderful flat bottoms - always a plus when working with PMMA. Erie Scientific under the SAMCO Scientific Corp, 800-522-3359, name has O ring lids platic lids with these non sterile containers. They are available in 40 ml/48 mm diameter, 20 ml/35 mm diameter, along with larger containers. Contact Erie for samples and info. Other companies have these things, including Evergreen Scientific - remember M and M candies in the specimen container samples at NSH conventions - those are the little containers I am referring to, and there are some even smaller than the diameters given. If the bone was small enough - we made prepolymerized layers in the bottom of a 15 ml or 50 ml conical centrifuge tube, embedded on top of the layer, then polyermized with the cap on in a 37C waterbath, incubators are shunned do to uneven heating. Or let them polymerize at RT inside a hood, tightly capped. After block is cured, cut off conical end, shove block out, shape and microtome. Tubes must held down with a weight to prevent floating but perfect blocks resulted. At 03:35 PM 7/29/2005, you wrote: >Has anybody done PMMA embedding/sectioning of mouse tibia and >posterior/distal 1/2 of femur? If so, what mold/vessel did you use for >embedment? Must one use glass for PMMA or are there other suitable molds >out there that will work? Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From aptafreshi <@t> yahoo.com Sat Jul 30 03:30:29 2005 From: aptafreshi <@t> yahoo.com (azita parvaneh tafreshi) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Problems of processing brain for paraffin embedding Message-ID: <20050730083029.26833.qmail@web52102.mail.yahoo.com> Dear Members, I have problems with paraffin processing of mouse and hamster brains and spinal cords. I have perfused the animals with PFA 4% and I postfixed them in the same buffer for 7-8 days and then stored in Alc 70% (up to now). After sectioning, tissues wrinkle in a way that it is torn after flattenning (on a slide warmer), therefore the quality is not good at all. Can anyone give me hints to overcome the problem. A detailted protocol of paraffin processing would be great. Regard A.P. Tafreshi __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From jkiernan <@t> uwo.ca Sat Jul 30 23:26:01 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Problems of processing brain for paraffin embedding References: <20050730083029.26833.qmail@web52102.mail.yahoo.com> Message-ID: <42EC52D9.940503CB@uwo.ca> Dear A.P.Tafreshi, Have you discussed your problems with an experienced colleague? Someone in the histopathology department of a nearby hospital could probably advise you. Your statement, "I postfixed them in the same buffer" indicates that you mmy not know what the words "fix" and "buffer" mean. You need on-the-spot advice, and it will be available in abundance in your own city. Histotechnicians worldwide are generous in giving advice to colleagues. Visit a local lab, and meet the people who will give you advice that's based on a combination of education, training and practical experience. John Kiernan London, Canada. -------------------------------------------------------- azita parvaneh tafreshi wrote: > > Dear Members, > I have problems with paraffin processing of mouse and hamster brains and spinal cords. I have perfused the animals with PFA 4% and I postfixed them in the same buffer for 7-8 days and then stored in Alc 70% (up to now). After sectioning, tissues wrinkle in a way that it is torn after flattenning (on a slide warmer), therefore the quality is not good at all. > > Can anyone give me hints to overcome the problem. A detailted protocol of paraffin processing would be great. > > Regard > A.P. Tafreshi > From lachlan.smith <@t> imvs.sa.gov.au Sun Jul 31 05:18:09 2005 From: lachlan.smith <@t> imvs.sa.gov.au (lachlan.smith@imvs.sa.gov.au) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] (no subject) Message-ID: <1122805089.42eca561c7451@mail.imvs.org> Dear Subscribers I am staining cartilage using Millers elastic fibre stain, and while elastic fibres are staining well, there is a lot of blue background staining which makes thresholding for histoquantitation almost impossible. My sections are 30 micron paraffin embedded. I am also oxidising with 0.5% aqueous potassium permanaganate and bleaching with 2% oxalic acid. Any suggestings for reducing this background staining would be greatly appreciated. Kind regards, Lachlan Smith Institute of Medical and Veterinary Science Adelaide, Australia From cfavara <@t> niaid.nih.gov Sun Jul 31 06:52:21 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Problems of processing brain for paraffin embeddin g Message-ID: I do mouse and hamster and would be happy to help. If you will send me your processing schedule, reagents used, I will take a look and see if I can pick up on anything. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: azita parvaneh tafreshi [mailto:aptafreshi@yahoo.com] Sent: Saturday, July 30, 2005 1:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Problems of processing brain for paraffin embedding Dear Members, I have problems with paraffin processing of mouse and hamster brains and spinal cords. I have perfused the animals with PFA 4% and I postfixed them in the same buffer for 7-8 days and then stored in Alc 70% (up to now). After sectioning, tissues wrinkle in a way that it is torn after flattenning (on a slide warmer), therefore the quality is not good at all. Can anyone give me hints to overcome the problem. A detailted protocol of paraffin processing would be great. Regard A.P. Tafreshi __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Sun Jul 31 09:34:49 2005 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] RE: Biotin conjugated Antibodies -- can we skip biotin?? In-Reply-To: <1abd9271abb50e.1abb50e1abd927@amc.uva.nl> Message-ID: <200507311434.j6VEYrKM050646@pro12.abac.com> Chris Is correct here I use what ever secondary I have that is against the primary antibody species even if it is conjugated with biotin or fitc with a labeled polymer hrp detection system to avoid the problem of unblockable endogenous biotin in the tissue. I just got a labeled polymer detection system for primary antibodies made in goats from Invitrogen (I had trouble finding one for goat primary antibodies). Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of C.M. van der Loos Sent: Wednesday, July 27, 2005 6:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Biotin conjugated Antibodies -- can we skip biotin?? Hello, You may apply successfully any appropriate anti-goat reagent, ignoring the biotin-label of the primary antibody. Biotin groups at the primary are far too small to block anti-goat reagents. As long as there are no biotin-binding structures in the section, which is only very rarely the case, the biotin-label of the primary isn't influencing the final staining result. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Tue, 26 Jul 2005 12:23:10 -0700 (PDT) From: GT Hebert Subject: [Histonet] Biotin conjugated Antibodies -- can we skip biotin?? To: histonet@lists.utsouthwestern.edu Hello, Just a general question to help increase my knowledge: If I had an antibody that was conjugated to biotin -- say for example, a goat anti-mouse P-selectin (biotin) -- can I just take a rabbit anti-goat conjugated to AP and run IHC that way -- hopefully avoiding the ABC step and background OR does the biotin from the primary Ab block secondaries from coming in? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lachlan.smith <@t> imvs.sa.gov.au Sun Jul 31 19:03:17 2005 From: lachlan.smith <@t> imvs.sa.gov.au (Lachlan Smith) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Excessive background staining with Millers elastic stain Message-ID: <000001c5962c$6b43cca0$e46b140a@ITP36161> Dear Subscribers I am staining cartilage using Millers elastic fibre stain, and while elastic fibres are staining well, there is a lot of blue background staining which makes thresholding for histoquantitation almost impossible. My sections are 30 micron paraffin embedded. I am also oxidising with 0.5% aqueous potassium permanaganate and bleaching with 2% oxalic acid. Any suggestings for reducing this background staining would be greatly appreciated. Kind regards, Lachlan Smith Institute of Medical and Veterinary Science Adelaide, Australia From scoop <@t> mail.nih.gov Sun Jul 31 20:39:40 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] staining for iron in microglia Message-ID: Dear Histonetters, I have a semi-scientific question: macrophages in almost all tissues can be stained for iron with Perl's stain to give a blue color - is this also true for microglia? If not, is there any other way to see how much iron is in microglia (DAB enhanced Perl's or some other method)? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892